LaoABO Tony I1.00 -O Vol. 19, No. 5, p. US, INS

Viral Agents in Hepatitis

A Review

DAVID F. HERSEY, PH.D., AND EUGENE D. SHAW, PH.D.

Smit hsol~In stituIttI on, Washington, D.C. 20036, and Armed Forces Institute of Pathology, Washington, D.C. 20305

An annotated bibliography of investigations con- various investigators from a number of foreign coun-cerned with the isolation and characterization of tries has been presented to emphasize the interna-viral agents from human cases of infectious hepatitis tional and universal problem of ascertaining etiologichas been assembled. Documentation of the host sys- agents for this disease and to provide a means for cor-tems, which included both conventional and noncon- relation of the isolated agents. The classified viruses,ventional laboratory animals and tissue cultures, unclassified viral agents, and host systems employedutilized in the recovery of 23 classified viruses and have been tabulated. The bibliography includes onesix or more strains of unclassified viral agents has hundred and sixty-five references.been presented. In addition to those reports catego-rized under various host systems used in isolation, Additional key words: Infectious disease, Tissuesupplementary documentation of published reports by culture, Viral isolation.

This paper represents an attempt to bring together Much of the literature which dlescribedl early isolationan annotatedl bibliography of studies on tile isolation attemplts referred to the use of more than one testand characterization of etiologic agents of infectious systenm or species of animal. Such papers have beenhepatitis. Ini addition, certain thoughts which OCCUrredl included tinder one test systemn and are not repeated.to the authors duiring their review of the literature A cross-reference listing has beens prepared and is in-are p~resentedl to suggest ways in wihich tile present chided as 'Table 3. In many of the cases cited, secondarystate of the art might be viewed and possibly improved, passages were performned in test systems different fromThere have been nunmerotus reviews whsich have ap- those used for primary isolations. The studies on non-peare in ~th ie literatuire. 2

--." ,.-,6. 3.11011:1. 114. 142-. 164. 1641 hutman iprilllates have bteens separated andi have beenSeveral of these reviews (lid not inlclud~e many of the dealt with individually b~ecause of their special interest.recent tissue (ulittre sttudies or significant p~roportionsof tihe foreign literatture. Ilie p~resenlt report hlas at- ATTEMPTS AT PROPAGATION IN THEtempted to in(orlMorate the miore recent tissule culture C:HIC:K EMIBRYOisolationl studies andl to include niore of tile foreign There hlave been aI numbiler of attempts to isolaterefereuc es inl an extensive review of the literature. Al- tile etiologic agenlts of hlepatitis in enlt)ronated eggs.thloughl it is possible and evenl likely tilat references wsith almost all of tile inlvestigators reporting some dle-htave bleenl overlookedl, no references were inltentionally gree of success ill isolationl of an agent. InI mlost cases,omitted since tile purpose of thlis report was to provide hlowever, tlile subseqtuent investigations oil tile isolatedan extensive list of references frons aroundt tile world, agents htave not1 beenl extenlsive enloughl to evaltuate theand thuis pirovidie ain oplM~rttInity for a more detailed relationshlip of tile recovered agenlts to tlse clinicalstudyl of tile ifldisldllal papers. rathler tihanl as aI critical sylldromie. Amiong theC earlier reports whlichl have ap-evalulation. peitred i ll tile literatture are those of Siede and Med.

Two stinlila r% tabiles havse beef) prlesenlted ill whiich a ing,'-'-' Dresel. Nleditig. ;and Weineck.2 9 Dollnen,27

numnber of isolations hase beenl tabu~latedI. Known viral Pe~lll.I,13s Heille et 14*,13 and Drake ('I al.24agenlts whiidi were either initialls or later idlentified as Both Siede and Nletinlg",I anld Dresel et 0t.2" notedstu I and whticih were isolated in connect ion with hepatitis dleatht of etubr~onated eggs within .1 to .1 (lays whsenare listed in ']*able 1. Table 2 has been pr~eparled to char- the citorioallantoic sac was inocuilated with dutodlenalacteri/e mid brinig togethler. in a ibroad (lassification., tltcse jutice. No tonlsistenlt pathlologic flllditigs were observed;as %(et un~idlentiflied agetnts that hame been isolated ill -onl- however, tile lethlal factor was repolrtedi to have beenjtln( lion wig II ases oi hepatitis anld hlave received rathler tranlsmlitted for- two to eight passages. Urineit and bloodCxt('tlsile stttck. Thiis list was not inltendedh to be all-in- were seldom found to inlfect (htick embryos.clttsive. as tllere ale iti existence num~lerotus other cases Pell(

1 53 per(lortlled expi~einiellts witi 7-dIa%-old em-

most of whih Itinvols edi sinlgle jptibli( at ions with n1o cor- ibryollated eggs bN tusinig both filltred dtuodenal juticeroiboratile studies. frotil p~atients aud~ liver jutice of experilllettally in-

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Vol. lt, No.1 Pi t~a VIRAL. AGENT.S IN HIEPATIT~IS55

let te(I pigs. Both iti.ltrates caiised death of the eiiibrso followed Iiy passage into the aninijotic cavit% of thean lit 1(011 ipte growl Ili ofi tile allan toi ineibiiilra ie. -li ic k Ini the lolloiA-tip stud toHrlsfrst paper,Ihliese restilts were traianimittecl iii 17 serial passages in Drake v/ njY/. rep~orted on tlIr development of liepatic

eggs. Viriilent e was cletreaseti with scict eeding passages disease resulting from hinman volunteer ino tilations,Ih ie agenit was repoirted as filterabile. Dohvinienl*7 at- with heeagents. Inoculated suihjects were anictericlclipled -ti cioiisatioli of %irtis oni the clrorioiallantoi: [but showed other symiptomns suaggestive of naiciral in-iieiiihaiie iii emnbr~onrted eggs. and tile incwtalat ion of fectioii. Thiese investigators also lotitd that anl antigenI' iiiithi l rls~age ma telial in to mite prlticl cicelIlness. prep~ared fromi infected a nin jot ic flucid an tic flat ivated

Beginnminig in 19-15. He-letiI antd associates"'0 at teiimptedh witli tiltira itilet light gave at positive skin test iii alllie i stil ;tic ii of ci iologic agentis ofi in ec~t iciis liepat it is. coiia lescenit pat ien is, whether in fect ion was cleriI liee wvoikers reported thre isolation tof tw.%o viral agents iiini~al or nmattaral.front separate outbreaks of inlec-tiotis hecpatitis. Original Henle'12 later pubillishied at review of various attemptsintitxclation was I-i formied with tissue cuilttices ol rabbit to adapt thfe virtus for propagation iii the cluck embryo.lii ci tells in a-oiler tuibes anid oif minced chictk cinbinos. Serigoku14l" reported serial p)assage cof six strainis oif an

agent thircotghlt uck enihr)os andI then into cultoresoif Elirlic i's asdites (tinor cells, HeLa cells, cells from

TABLE 1 I E1N-ti ABLE ISOLATES .\SSOUiATEO Wi ni htnnan ailinictiti nmeimbraine, antI 1, tells. Cells front thleVIRAL tEIEPAriUis ORi IEPArIC INVOriVEMENI -- ician amnioictic membrane showed aI high stiso eptihil.

Comsackic- % trus it) to the 5-irtis. Segagni, Ansaldi. anic N igro"'' tmtilizedGroup A\ sera fromt 31) thildren at an earl% stage of jaunclice. The

Type- 4 \Morris ef al. useia were inoticalated on the lrorioalloncoi- iembraneTyp, 1 Etohil cI aI.io of emibrsoiratecl eggs antI in tissue c cilttire, ofii chik

Types 1It, Itt j~-ziquet and Steiner77 eminrso cells. Thmese workers relxirted tie development(:isackje 5 irus oif chorioa llaintciic and liepa tic lesicits. hotilt iiia(rco-

Group B aintd mni-rcicopic . Essen. Lenibke. and Sc ilec it:er re-IType 4 Kibrick and Benirschkel'Iitdta lati.iotltosrsle ntefrTyspe -) OXShautl-hncssey anod Bue-chner'3 areltatalnociictltcn eule niefr

'I vc .5 Siegel el mat ion of nectrcrtic areas. oir in death of the emnbryo;T yps-s 2,4 losicr and Newon',' hiowever, these manifestations were insuflic icott lot titra-Is p- Ip Suitrand Stoith 11r lion of inetii\ .ir\(i l lradjcskiewiuI26

Echos iruses repcirtecl pa~g of af virus cif inlet tiotis hepatitis, strainIs pr t I Kisleva' NlNIG. iii embrooonatetl eggs. Electrtmn miiicrigraphls

Typ, 4 Karzon 1 ifa. ' fromm infected anti dead enibrios. demonstrated spherical,Tyvpe 4 MIa Ihaerhr-. I tarw in, andt Smtich' ' irregul ar b~ot ies a pprox itia teli ISXO inp in clialiietr.Ts-pr 9i Sain,. KrumbtiegeI , atnd Wi~ant rid Similar particles were seeli in t isscae c tilttinres i ncx ciIa ted

h-Pc 1t L apittleni d ci recth Iwii thm Iihoth of sick i ntd ua ilaIs. I'le tissue c ca-'I yps 7, 12 Slrarlai t It. L" ttire lireparatiin uisecd was not clestriliecl Esseil and

.Sdeno% ris Ii rlegun.av ulihdie5r~c i= ~~~I ype 1 1 hitch anrd Stein:"i scm egtesh eit lilec Itn e fp

_rviipr. IDa'is" idrirt ilicat ion as air adrenosirus dlealin g withr their i solatittin at ellipts in eggs. antd morerited hvDi' n jthardt an Ho tlottes " and lcet d c iii 1 ii 963. onl tle cIlse ofii skiln test withI ma-Itlrtch ai ntit Sicir-r * c-ia I grown ill emhr cila ted eggs.1i At least( three oithier

I~ vp, 1. 5 .Srranwsrt f/ I lmprý-` 63, liase aIppeared in whithI skin test ainigensIs ypr 1, 4 Keriirrr./,deý! pllcl edrt in cnihbrysiiiaecl eggs hiaste beeii ieveltipecT% Pc I tm~ ti e I t l.", Htill is" andc ciii ll))ccl wit !m somiie re 1x~rrct c del, ee iii si i-cess.

N t" Kohleri t al. IThewt are li sc is~setl in greater cdet a iIc under tilie originalIRerts tnti ,e, isoilaIition a cieiiipt s ctmi nectet w sithI i lieimm.

ts.pt I Felsenfr.ItiI vpvr Strucstsmikar i "2 cited in Russian vdi- ATlEMNIPTS AT ISOL.A]IO*N IN\

toriril- EXPERIMENTAL. ANIMALS

JvcI NMcK rr" , Hurling~thmn and;rrr Mc tee- i Ant erse i i l and Ilfiliiitis". CIlill i\ci tilie oiraI atllia nis-Entrmrust-. S rtirilio let inlictilc- of bmile toi uniite andt p~igs anid oibtainedMIiscellaineous S arlai - serial trallsin issir n oii an ii ii c-ct icics agent. Pentfl."i~

\Nv-s\iruts us filt crc-c chnit lena I ciiice froim G ermanI scltdiers andWH1 Virus G iles. A,-ihhrtier Krugr iii.r and I ii i- ill~iii icc d ct it in[ I t igs. HIiice, andic em hr ciia ted eggs.

inc-rr-- fit nh e i it-t IaSiliwed rate( cIi weighl iit reaw i- illo cic-Liclhhr her. andt ;cssixi es"' WT h ie pcI igs. Ili ione o f thei pigs, sac ri ficed l aItcr 63~ cl;tys

1)5 virus Prris. halk . I chhabrte, rin(t Krugnirit t hle til scri cc I tIia i gc it) ill it'Iiser, ishi i I i msebAquceiithy1)\Vrs fshiiing. Ism-scirt anrd Mc( illrtin' cotuihl hit inii~chicl In sIia trniisi of thle liver

MN, (il pi~cr N i e e "psc ..... ;, al issctc aI tcr flthi c- passages. A irti lent e let reaseth withi3f, a~l "Ptrum(Y..Illy ad anocitc., fiassagte liii raieXriltiIe;il incHcilatiti~n cif Iliic witilI ~~Refer-er-rce nrumb ier, aned wotrker tor nriu p ofr workers filtecrdc I cticic enalIj it ic ind1th i -l li s-tr lesioits restiiil i ng

N 'INot yet c%-pabil. thoise ill hiTatt. lT-e inc tiilmailin hwricx inl mitc- was

mW HERSEY AND SHAW LAMORATOUY I NV~.srIGATION

TABLE 2. REPRESENTATIVE AGENTs, NOT YET IDENTIFIED AS KNOWN VIRUSES, ISOLATED F'ROM INI'LCTiOU4 lHPATri5 PATIENTS

Camumcm, mamme original description and confirming references Host Range Seology and immus..oey'

Motol visrus 1Originally isolated and described by Originally isolated in allantoic cav- Varies according to host system, butKubelka 4,t a!.t7 further studies by ity of 'I-day-old chick emtbryos; under various conditionsa has yielded +Kubeka and associates'4 . 1.; grown in mionkey kidney. Chang's CF, + neutralization, and + heinag-Chlap et a!.";, Schon et a!.147; liver, and KB tissue cultures; glutination; positive skin tests; heinag-Kowalczykowa rt al." "after passage in MK and Chang's glutinating fraction is antigtenically

liver cells, it is pathogenic for related to type 5) simian virus and DAInice. myxmovirus.

Laszlo agent Isolated by Laszlo et al. between Grows and produces cytopathic + CIF tests; + neutral iza tion.1957 and 1964; Laszlo and asso- effect in tissue cultures of einbry-ciates.10

15 '5 5

onic human liver and Detroit-6cells; induces hepatitis lesions inhamssters.

K3 v irus Isolated in I960 from feces of patient Grown i n human renal cell culture + CF tests; + neutralization iby patientwith infectious hepatitis. Ananiev and Detroit-6 cells; 50 passages in sera; Cross-reacts with Mitroiu's agentand associates" t ; Barinskiy and humian kidney cells; does not grow fromn Ruijiania.11associates." - 1 in mionkey kidney cells.

Mitroiu virus Viruses originally isolated from Grows equally well in KB, HeLa, Neutralized by patient sera; cross-reactschildren with epidemic hepatitis; HEp2, Detroit-6, and monkey with Moscow K3 strain (two of tiveMitroiu and associate3.1"''5 ' heart cells; nonpathogenic for strains from Rumiania showed cross-

suckling mice and rabbits. neutralization); virus was neutralizedby ECIl10 7 ant iserumi after 20) passages.plus later ones; types 19 and 20) echo-virus antiserum neutralized virus strains

BF at 20th passage. but not after 36th

one; original isolates did not reactwith erhovirus antiseruum.

Rightsel and Viral strains MR-I. AR-17, 'sW3,Limited to Detroit-6 (PD) cells; + S'-ruuuu neutralization (7 years post-McLean RA-214, and LG-217 recovered AR-17 produced clinical jaundice disease); - serum neutralization withagents from serum or plasma of patients in volunteer subjects, Boggs rt a!.14; viral recovery (3 moriths postdiseasel

with viral hepatitis hin Detroit-() heat-stable at 60' C~. for 30) min.; + hemagglutinatioi, of rhesus erythro-(PD) cells; McLean et a!.'" 34; filterable through 1)3 Selas filter, cytes only by patient sertim~ not byand Rightsel el al."'~ Rightsel.1l" viral agents, Rightsel.111

+,Positive; -, negative; CF, comnplemsent fixation test.

TABLE 3. REFERENCE LIST To HOST SYSTEMS USED FOR inistillastion andI isstraperitone~aI injectionu ot bile troils1sOLATION AND STUDY OF ETIOLOGIC; AGENIs patients with infectious hsepatitis into mitse. flaa~gess

ASSOCIATED WITH VIRAL HEPATITIS and( NWywiorski-Sclseele'" protlticetl a liver p);tlholog)

Host ym similar to that fotund in tlie paltients: eleusseitary bodlies- - -- -- -- __---- ____ were seens in liver atnt Itings of muie, as well as its the

Emryte Mc and or other Subhuman Timu ~ultu- huminai patients. 'I'se agent wass filterab~le andt (otill b~eF~mbeynsted nosprssate primaesImaitn a i neldi trough serial pas~sage.

6, 7, 8.29,5,646 5l34,26 42' 4,Ii IISIll I 2 Thue Niotol virus, described later in this ieview, hasb,27 28, 19, 3,64, 51-5, '2b, 42, 21., 8:1, 14, 28, 20, beets reported, by several inlvestigaItors,"' 'ý 97 1 as in-6,2,1 63, 79, 16S,90$, 104, 118s, 431 45, 41), 50, 57, (4 fectiotis for mice. A brief review ots thle earlN list- of

35), 10,7 12b, log',. 11t),- 156, h3, 67, 6H, 70. 74, rodents in isolation attempts was preparedl inl I 95i b%Ill, 132. IA'0. I122, 11I), I64" 77, 79, 812, PA, 118, 1K) NIi iit k.''" TIwo othler psaperss. byN lacad ; t ll "' ;in(l1449, 149, 15.1, I V2, 136~, ii, -4, 17, I I10 1017, Ntrr'

2 ave appeairetl these (over thle perioil tilt tolI #4, 1317, 1 49I, Il1 W 112, 1 1,_, 117, 1951I The relxlrt of Mac( alltgin'"' p)rovidetd ;I sit ns;iar

16' I t)"1.1 124. 121) 127, of sttidlies in atininals (c;Its, dlogs. slllt', ferrets, etc.)111, 1 14, 117, 1 4(1, tstially associatetl withi experimnent a I IaiNmra orN work,141, 141. 144 146, 148, wherea's thle suIN til b~v Morris''poie.terslso14'I 1 T2, 1l74, 18, . Pl~kdterettso

SI IA,41 tI, ,10 , studies inolving it wide range tof both tloiiestjt andlItA.' 16wil anim~als. im-Itising Wioti nmammials and birds. A~

It if (of 9.3 nmammalIiain and six avia n species were-Review articles. tested, all withi negative resutils. Dresel 0i aI.,'29 1cr,-

berg,'14 .0 and( Oni/sakat:I,.13"1 have reported onl theirvariable, with file allnimals thing alter Ilii hours to 29 attempshts toI propagate the viruis in canaries. Dreseldlays. Passage of infected pig mnaterial has alreaidy beens v! al.-'! testetd not only canaries buti crossbreedls of gold-mentioned in (onnel lion with Pendlls work on ino~tula- finches withs can;Iries. Filtered tirine was inot-tilated intotion into embryonated eggs. By mleasis of intranasal the b~reast intim(ics (If birds, Disease symptoms ap)pearedl

Vol. 111, No. 5, 1 %8 VIRAL AGENTS IN HEPATITIS !Ob1

withini I to 2 this alter ii iectioli andt coiisiste't (if also ill(lutled a statemlenit to the effec t that fxx)ICtl ser-a ofru ffled p~l umage. shiort ness of b rea.th li Iistlessness, andI tiornia I marmosets firtHI ttl similar pa tlIerns of liveractelerateif heart rates. Death usually ot cirretl on tile eli/vine and liistopiatftologit thanges similar to thoseftitii tli(Li. Crossbreeds were mlost resistant: eight notedt in animals infec ted with eithier the Bar ker strain or

own(MCIS Stirs i eti two or' three piassages anidt, inI one Witlit thelc own c linical spet i mens. A move deta iled i efiortaýsc. ninile passages. of these observations will tantotibtetlly be borithonhing.

Soici of tife carIN repo~rts onl the efforts of jaipanieseWorkers to isolate hlieftitis virtis in stinaff animials have TISSU E CU LTU REbeen revie'setl hi Arakawa 0i al."1 A series of reports I) b% issuie cultore stutdies ha~ve been performed exten-Hara atnd his (oworkers52,- are amtotg the miore extensive sively over the past 15 to 20 years in attempts to isolatestUlifie. Thiese investigators.` hnomiliated liver cells. of)- etiologit agents for inlet tiotis hepatitis. It should sufifce,tained l io011 a fatal case of hepatitis. intraperitoneally ;it th~is point, to cite several papers that have involved a

inomice. whlit h tdied IantI 10t 2 tdays I ter. Pas wihurit ftellles. v.g., the studies of Bang andsage was matde scrialhN at least 20 times, with stilise- Warwick,"'Hfenle."'l NcLean,' 2 Ananiev, Kaverin, Nat-

twit t dea thI at 5 da. s. H istol ogit lesions iii mice closely ski%, antI Barinskiv,:i ;nd C;ole, fDanks, and Campbell.--reseitblt-u those seen in humian patients. V'irtis was tieti- Iti the McLean sdy'2IS cell linies were tested, eightcrali/ed b\ humian consalescent sc-ia iti tdilutions till) t) of whit I gave at least somte positive restilts althotigh

10 I". Elec(tion microst opi tdemonstrated virus liarticlescil the results Withi all but one (PD391') have not beenIf t to 15 ntI ip. In Liter articles Hai-a anid his asstxia tesi.-:. d onlsistent or clear-tint. Amiong the cell lines showingrelitirt't the istolatiton of a similar agent froni other somle promiise were HeLa. human aminion and thepatitents, and tlescribetl the preparation oif a skin test Detroit-li (strain PD). Anianiev cl' al.:' studiedti de effectantiiigen frioni in fectetl mouse t issue, of K 3 virtus in a witde range tof primary tissue (tiltutres

A rat hetr impressive list of investigators who have andI cell linies, and refitired a cytopauliic effect inl bothalt tei pied i sillat ion of thle virus in stbi itiman pii 'niiites homiani k idnei and aoirta primary cultures as well asiiol Gi iIle i 11( otet in t Iiis review. The li tera ture p~ritor someic -%o ti effect in Detroit-ti. KV. SK humanto I 191I ha% fieeii reviewetd hi Evan. 42' Iii order not to kitlnei aind SO"B' tell lilies. Highest titers ofl tie agentdtipilit ate puiflit ations that have covered the stiliject were 'obtained in htiman kidney and aoirta primaryrathecr exteiisively. referente has been made onily to tissue ctilttmres, the highest reached being 10-" TClID.,.,seerAl of thle more retent reviews.2 .1- i69t ]56ie re- Alintng thle primarN tissue ctilttires trietd were those ofvieici% I% laras/ti", inticited a comnprehensive table of monke% , pig. gutina pig, calf, cat. rabbiit, mouse, sheep.at teimpts atit resitlis Wilit tin appiropriate bibliography, chick einbr~o, and (log. Cell linies inicltuted, other thanI-lte aiueinpis to (late, although promising. still provitde those showing imsitive cytopalthic effect were HeLa.,

io bireakthfroughi in termns of a tisefl" atlinIm hos"t for M~ 10, SPEX' and 11.0. Cole ei (11.22 emlployed primiarylife riout ine isola tion oif life agent for v-iral Iepa tit is, wilil)I tissu~e cttltutres of mnonkey kidney. humian aminion antIItit' lit isililt' except iti tof a recent r fx~rt by Dei iiIiardt It. iiig. atd ic Iuna i enihl)r n ic kidneuiy and intestine. Tile

Ifof intes. Caps f ;fis.a t Popper'-"; onl imarmtset s. '1 hese H EI2 anid HeLa cell lutes andl thle Fa irfielt inhes ofalihlt rs rejioreif thIiat i nocu Lit itis of litiimani serimi litiimani epith el iumi an iit noiikev embrvon it t issues asor I h silia. otita inetldi trinig thle a~t ite phase of viral iwellI as Det roi t-fli were tesied for stiste ptibiIi t% to t heirI ep a litis- t it I d ie(hIfeinhifa anrd nilorfilologit I iepa tit agent: otilv~ (lie l)Cet roi t_-i tissueC ctil ti ie sitfpt~ortetf growthIdiista s in ima rmosets iii to isoiut of fiv i-ex liermn miita of tlt is. aiget'i. Ga rtf andI Al in'" a ttemiptedl to ( til~t iate tilieseries. Thie lisease isas transmissible in series frotti agenti of1 infeitiouis liepa tit is in o~ ster t issue. btt thecianut iset to mia rimoset . andit resutiIieuf it] I litreas ing ietsii Is were doubtfuti I in Ilighi of tilie tIi flittilt i thlese

ri lc-it t. i .i i-r biopjsý sfxiitcimens examiiinted uindter t(ode a tit fitrs re poretl iii growing satisf act on tissue material.-xitil ii i-il somte of Ilife tha rat trist iis of hiumania viral A t tittparison of isola tion sttttesses in tilie i-aritits tellI

hepatitis anid were reatlii tdist tingui shiabfle fronti notn- lilties woult Ible of little signi lita iie bet;. use of tlie witdesf't ifit if a uges. Ntio it# 7,I no sira I istila t in has been'i i-ari at ion tof ma teriaI antdc its sotmre. \[;fit,, of thlet' iCii l.vd althIouighi biopfil sical mid ant Csnlogic dlai a~ttri- a~gents reported to (la te. as will be' notedl below. hai-e/aiin si i ldi's io f tite itilet tIis fllstiili frtonti nia rmosets been it ul-ii ifiet ias i ariotis p)t-ottit les of lfassi tied vi-i

art'- iii pro grtess. Fimil ap froof ifhiat lifei it hepaiit is obsered rtises fiat iim or inia n ot bear 'a tIi ret rela tionshiipin ifhest niiarm isets is fami etd by agentts of lIiiman viiiral I o inlt~io' t ts lit'pa tit is. Soiti of thle tel Ililties t hat htav-eI ep aliiiis is still In k ing an irn mist await fuirt her i iiiesti . f'eI c lii 1)11plo edl hotl Wiilthi aniit with loat success incI luetdga iii ii Parks. Voss. andt Mel liiik'' hiavie reetell if ret- hi-f tk em bro, I).fi'!.a. litiiman eii plie ia I K B. fiuitnitfjxmitd on si titit's if life BaII.rker st ra int. i sofa led b y t'iifrs ottit Iltong antit kidrims tiitkei andl rabfihit k itftes.I)ito fiatd tIi1ia.1 in ina i iii set s. as wt'll as twto iiriises aind Ol ft'Ie'troil-ti linte. Cell i lii's of if te Iaftter in vai-anattI% itsoil a ted in nalrmi st-Is. IlThe stmh o I. Parks tivI' (1.3 ol foritiS. VA~ aind f 'l) i". ti; ic alfso fieen tested. B othI

tfit- Bat ket sit a ii slitowedf tif agentito lime (ic ariattens- slat o l-r- andit ro-e iltriiiliem fitxflos limse feen C111f4lvi e

6iis iiof ike if ost' t ia ri 'rist its lotr Iitmati lit'alia iiis %i itis, witlli rt'fit rtet i sttim-ss. O rgan ii tufttres of livuer havtse alsoin Hifma ifit' agent Was %ens~itii e to ci her an titl iut tie (le't' been tist'olIor isoila tion attem fipts.',%iitime'd I Im italintg at 56" C . loii 34) miiiii ites. Nvidic Itt l- II ill is ;indi I hanmg 7

l ent pft ta itil ftr itt le (iit it tts ofmtalli -/-gh f nilll ni itt tonilaleset ii i na ritst't selti i aitttl liii i 1111a i andl ioti st' cut irso livier. ais wt'l ;Ias at lot tion-

ifec icli~iiis of ile ... %iriis il marmosets Thius velxmril" kt'i, orgain laltores oii fiuaniati ~ liver. antI tryf)-

vat agntsof nfetios hpattisallwitan tigten s atip tmpra trs f6C. Theorina isoat tsoe of fiv agnutes.

indirect fltiorescent antibody~ tests. Eight of thle speci. These agents were noninfectious for suckling miie.ineiss inocuilated had been demonstrated, in p~reviouis A report by Kibrick anti Benirschke14 involved twohuman tests, to (ofltaiil agents capable of p)rotdticing fatal cases of generalized infection in the neonatal periodtypical dijsease with jatindice. Tissue culture teduhnques with isolation of type 4. group B Coxsackie viruis. Iso-have provided ani additional mleans for stud) ing tile lation of tile virus front tile liver of one of thle infantsisolated agents by litiorestence Inicroscop)'. 'M and was reportetd. The report by O'Shauighinessey and Buieclo-other immunologic teSt%.:I. 4- 7,0 142 0 Comparisons have ner13 7 involved the isolation of a type 5. group Bbeen reported concerning the cytopatlsic effect of various Coxsackie viruis fromt a pregnant woman suffering froinisolates front different sotircesý-1 and hase suggested a clinit ally dletectab~le hepatitis. The paper by Siegel v'tsuiplleienetal methsod of itlentification. which might be al.154 tdestribetd the isolation of type 5. grotup B Cox-employed if the resuilt% totild ble consistently repeated sackie virtis fromt young children during a 1961 out-and tonffirmed by other investigators. break of pleorodynia and pneumonia; iat least 19 of

AGEN S IDNTIIED SCASSIIEDthle cases involved liejatomnegaly. The clinical patternA(NTSIDETIIRUED S ASFE in these cases mnakes a direct association with in-

VIRUSESfectious hepatitis appear unlikely. The cases cited bN

A large number of virus~es have been isolated andlt Hosier antit( Newton-,: involving thle isolation of sariotisidentified in association with cases of infectious hepm- typ~es of group B Coxsackie virus were cllaracteri/edl bytitis. Proof of at causative relationship to the disease severe focal hiepatic necrosis without other evidence ofhas been dlifficult, andt( thle stattis of these viruses, for infectiotas hepatitis. Sunl and Smith1"" reported at casethle most pat has remained unclear. Mlany of the which stiggestetl that infectious hepatitis mna' have oc--idlentifiedl agents have been isolated fromt patients with coirred along the more classic symiptomus of (oxsa kie vi-'. ntptomns not suggestive of infectious hepatitis, andi rtis infections. The incidence of hepatitis ass(N. iated withthe infectivity of the isolateid agents and( their ability (Coxsackie virus infection has been low, and tilie actualito inmittce hepatitis have not been tested in man or role of these virtises in infectious hepatitis has not beenexperi mentalI animtals. Those agents which have been established.inoculated into man andt( test animals requtiie further The role of tile echoviruses ill hiepatiti~s would apfx'arstittl antI confirmation, to be more obscure. There hame been ie1 Miits"I" 145-. 1.9

Kibrickss has recently. reviewed the literature onl thle noting milti liver dysfunction or lmepatomegalh assuxirole of thme (;oxsackie viruses antI eclmovirtises. li-e statedl atetl with echiovirtms infections. Liapinieimiil". Im re-that "'tile relationship between Coxsackie B viruis infec- ported tin thme isolation, in 1959. of tp 6wI eclioirumslion andt hepatitis in newborns is clear.- and thimi front Iwo severe cases during a miild oiuihreak of*.'there is suggestive evitlene that Coxsackie A and B hepatitis in Helsinki. Finlantd. Virus particles and anti-virus~es mna' ble relatetd to hepatitis in patients beyontd botl tti ecioviruis type 6 were demonstrated. The atilhortile newborn period.'' Althotigh the nuimber of refereiices suggested that it mnight only have been a liepatotropicited b'. K ibrick. both c ail-.* and reent. wi aild tent'. 'o St rait] bitl that its close assot ia tion with titlie tdise'ase mnat I

support hiis assumption that these agents hav, e Ix en it a potential eI ologic agent. 1l pci I an ml Ct loilt)' rmfretjtentl. isolated iii tonnection wvithi cases of hepatitis, were reportetd bm Kiseleva'0 to have bween isolattet hum

arev-iew' of tile (itetl papewrs itdentified onlN a few cases of infet tio;us hepatitis in Imir-kilitia (U .S.S. R.).age nts that were a~t a~l' reco'.eretl from (ases oif in- Neti Ira li/inig anti bothN agains thIle isolatmet I virises wsasfe( t otis h epa tit is. ()ie sIlt) Iiase is that of NI urns. present tin 63.3 per tent of thle sera examined i ttI vit IF'lisberg. Pond. and W~ebbl''2 who isolatetd a t) 1w 1, titers tof 1/10( to I 6il0--one tase hiatd a titer of I 84(1m).grouip A Coxsackie virni'. froni thle bltood of a t lid with siiI In a parallel studitl withI sera I romil 60 I a Iit'll" Is.t\IN.' I!ever, rash, antI a (Ii nica I pit totre of lit'patlitis. Both ethiovi rtis was iieuIra Ii ed in 12 cast's, andmt onl I'. isneutralizing anti coiipletieiit-fixing antibodlies deel ases neutralized anl adenovirtis (unspet ihied ti jx). Thieopeul against thle agent tdutring lie patient's ( onva les- st1ra ins (if eclioviri s istola ted woouId a ppear to ie 1w losel-'teitte. W~hethier the agent existetd as a m oncuirretit ill- associatetl with the descrihwtl (pitleisi of hmepatit is.fed mull or act a~l) prtxldieul thle IihepaItitis SNliIpIlliii Shla la i. Nlo~oieiiko. antI TahI.i k" lih'.e re~et tit ie-t annot ble astertaimied. Embil. %'al Ruoo.en. a~ni Nag. portetd the isolation of tdiverse tylws of agetits Ihftoitler:!ý reportedtl le i solat ion of a Iip 1t'Ill group A C ox- paltientIs stiffen ng front iilifed iou'. hetpatit is whmithI arc'sack ie viruis froni thle stools of Ii:ý uli a tIs in al (il nt - associa tet wsi th viruses (if tilie intlest inalI andtt resp iira torybreak of iimfectiouis hepatitis at a housing tdevelopmnent. grtoups. Thmese sttditits were perfuorame'd titi liiltrt'i withiFotur isolations wcre matde fromt tilie blood Io(f te li' oi- lion i~terit foriii ofi in let iotis hiepat it is. I xs7 a iit(1tat% I. JtVMttiel and Steinmer-, retentlN rein rtedthilie itlent i- 12 et ho i ruis wsert' inivolved~ i ii a noumbexr if i lit' (itedfl atIion of tileI original Bunitliner an tu Shireeve" isolate. (ases.

withI whidii i tilie'. have- worked ex teisi tel'." .- as 4 ross- Tylpes I antd 2 retiviro rums beas- en istola ted in asot i a-

Vol. V), No. 5, 1%18 VIRAL. AGENTS IN IIEPATl'TlS 563

lion wsilth inifect ions hie palit is. Thle Brown adli Buclietlnr agent s isola ted front sera of ho~t i huciman patlien ts anrdstrains of a ty-pc I-like reoiirtis wecre tlescriIbel In1 chimpanizees with s'. ipiftoiiis of inlettionis hlepatitis, anti.M(Kee " a nil b) Burlaingh ami a in I Mt K ce.'' andl were these agentis have been identified as simil ar to the typeisolated front the fet es of pat ientIs withi in fe~tiious hepa- 5 adenovirus.""1titis. Both virus strajins (o111( be prtopagatedt in 1 liik 1-ciman experimental infections with at least oneeinhr'os hi% either tile cliorioaflantojt cavity or yolk t)pe of adenovirus have failed to induce hepatitis.sat( rotite. and bo~th lprodltietf a lieinoi~ lii, which was Kauel, Loda. and Kniglht" utilized at type 1ii aileno-heat-stable atl 1210 C. for 21 minutes. Thie ienioNl'sjn virus, recently isolatedi fromt childtren, in an attempt towits not firmnl) Worildl to the %irion and could be infect voltinteer subIjects via the conjunctiva. Vii usseparated bN dlial~sis,. The liemolbsii (fill not requiire cotald be isolated fromt the inocuilated eyes, the throat,recivirtis~ porsie to hio/e rtlirm{~tel, buit anti recttlin, btit no antibodies were demtonstrable fol-

the ret cftor sites a pixared to he adjaimen to tile lowinzg in jellion. During the course of 7 mnont lis ofrein iruis rel eptor. -1h. role of these lirilses in relation ohservi7lion, iio jaunidite or signs of liver dvsitin tiou,to viral heptatitis hias nti) bteen t eans1 established. was noted. Hartwell, Love, and EidenbockV7 reportedStruitsoskaia.'' 2 ats replxrtelf in at Russian editorial,'1 the isolation of type 5 adenovirus fromt bloodI clots ofrectiveretl ti pe 2 reovirtis stisfpl tell as possiblfe Ctio- patients iii sporadic outbreaks of hepatitis in Arizonalogii tgent iii ant otitbreak iof enterii. infections with (UI. S. A.). all in the vicinity of the city, Phoenix. Tohepatitis iii children. joske ot al.-' repo~rtedI onl the iso- offset the possibility that adenovirus might be prevalentlattion of tIf! 2 remi-Irtis fturont a uii ti ner of patients, in tIile normal population. 70 1 b(xxl samples were ex-severalI of whi un h ad cliniiial lhepat it is.AIthlotigh it wats arniOnell front inorimalI patients of a si milfar age grotip,diffuiclt to ascribe a definite collneitioii between the with onii ont isolation of at t) PC 5 adent'virtis.reovi ruses isolatetd and thl e cblI reai s illness, *joske and Dinring a I 2-monit f eri Kiihlei. AaxxIaca, and;issoiciates felt that, at least ini tie oiie fatal case. Springer" examined 288 sertumi samplles fromt liniicafcl tofs'. fiindlings in te leIiier were tyvpica I of those oh- fiepaltit is patie-nts a nil miade 162 adenroviruis isolatlions.

served tii reovirus infections o~f infant mice. The 01- Dcirinig the samre pe~riodl. exaiminationl of serniil speci-a itsioni I x lrrence of enieplia lit is in in I t ious liepa- niens fromt I103 pat ien ts wist hotat evidence of hiepatitis

(itris intlii atetl thfat, atl least in slome tof theicir cases, a '.ield et on ly 10 isol atitons ofl adenovi rtis isolations wereic-n irtis etiofiig) cioild not be rnled outt made iii primary pig kidlney teli cutuitres. No (yto-

Felsenfeldl' has reported repeatedl isolations of type I fiat iic effecit was observetd: however, identificatioii wasret ivirlis front stools of at ps~ ciologv ted ilmi~ii ia w4ho mate I .. IIiistologic examiilna tiioi of thme (eil iiionolayers.hlandlfed a grouip of ),oumg cfminipalizees. and also fromt Repeatedl examiniiatiomis were perfornied ott 62 patients;the stoiols of the lijiies.Liver b~iopsies, froni both W1 per cenit of these cases gave refpeatedl positivesdlemionistratedl t ianges suiggestive of infc t ious hepatitis duiring the first 20 tlavs after onset of ictertms. None ofbalset onl t(lie Sllneta na ' `" dest ripft illi iA f sulilloglaiitl t- th li'f0 i sofaC. i ds emionst ra ted anN, similarity with the I1Lii iia iisIoria t ,on if f).irendi li iloa filer t-Iswats a f- kim iwi (%x-s cif at erioviri lws. Nen Ira Iiziog aintibhod iespari-ntif' illen lit aI ito ifit observe. ill c pitfcniii ot(f t ott d lit4t 1 be iin Inst ri ell w5ith hIoimologous a ntiserat.fhepatitis in D~elhii. India.",' aindt iii Almir, (.hiaiia.i At le'asi iwo tfiserse %erot~lfws have bweii itdentified

Thle niol of aifenocirlises tin assolaiol u-101 W d in fe~l ilis anrltaig the isolates,. ThIiis refxiri was a follow-iup toliepaltiiis hast peiliaps bweii ci ciinium c illielslin1g, A- ilie eallier report bn Ktileie antI Apoixlat iii 1%6.thouighi equtafll as midi ear. Das is,2i" in 196 I. isolated Aft fhongli adllelosiltises hail been recoseret front%ia I agents known ;Is ihe Sit)) Cirllos agen is In no if JI~ a ses of ili e~tlit ns liepaiiitis. tI e etioulog ic b asis for thle

stool is of Noiting (hldI reti fitspitIaIi /et I if iiig aim out- di seale was not i eintle-I esiabl)1ish eid for iit-ese agents.fbreak of hepatfalit is o~ii a inl Iniani rt'sen atiou . Rei imitI. When it IIa fest mion ~ i of iit-e atelenmiiuis-assotxiatet . delet -Deinihiardtl an ii 111)1ohes2 5 an Htin di ;ita (1 Siei~'m poreiteirtl Ii st pa iltites was IvXriinccl 6%' I ItgganI an hist ls asoi-ithulifitaltion oif thitse aIgents is (predouniiamith) ;cliiio- it Ics7' and tit 1~ 1 %(I Iiso)II. (';sto, ant I lanioin.7 Ifleir%munses. Mfilrti aiid Siemiu~t Ionfirmiecf ilie idellil% otft smiall size( stmlatei fa li Itirler i-Searth Ii iii a IoSsiileIlie Davis agents as t% )ws I and .3 ahenin-iruis. In Ci iologit role tlliift inftions lielmitiis. fIleal'nist-

addtIt iion, an i sillaie i(Ietflemii lit-ls I'. tw 13 i'dIios iias atieth v-iral urtidi es were tdItpexndenlt siil llt'fvIoli tilie%-as found i . Iflie t x' 3 at enini rtis ifill creif r iim thle f resemie toif illi at cm is i r s lori propfagatIion. 1 I icy weref~rotoi ~x' 3 acleiinirums In fit-uima~gglua lialiil~iti-ilifiiliiticin. afpifxii iniiiiae 2201 to '2M1 A iin tiaiiier t'ias obseis-ec inB%. it-e use oif tIile I emiiigglil tit i lit I in-in ibiiliit II ion lei. - tegaInise (onitsii c a le t-f'ron iiiitrtisltil , a nd c cimsistetl ofni(ii1 le, a reflat ion ship tf o Iti p If 1 at hrmiti ints wis i lidi- a uleox riblit iii( mi t tid ifcore. II awl k rn5 ' I tisitleretla incl. K-ni m-Zaiel-' uilif d izecf ( c~ ellIs fur I inmimma itheii anma figt ii isto If ac sia el liie f alri itIt' of tit lt itbacto

isilaiitin aniid ret os-ret 21 I ci ipa i ngen it agemi s I ruml imectum5is %ii is. II oggani. 11 liik iou, Raiij kit, antd RoweC72,

let es of 50 patiaentIs with1 iin~letioiiis fipt- idlii . Of th ese destili lwc ftlf in i r o lmiie scltiog it sit lt~peI of tlit atelemo-agent s, 23 were foimnri to he anlt igen iAIal' andt scro- is ircisassxi a ted f ia mit Ic..s A ci xii elotedl~Nm isi Mol cx ten-big it all% refaine'd to tuv pcI 1an tI- at ermiuos ts. 1fT'e site si tudil-s f1;%a not miilxeii sin essullo if) ithe i-st ablishllnt-strains intltiiet intra~ellular intuisitins in moonkes kid- iif ;in li ologic melal iolsliif flii c-ill h- the atlenosmuses ortit'%. F It' ma antit H fr)2 t(el l%..A sigivi n diai lioptifliiion l'sti %ubt~p, I f 1)i ntm si IlasiI ' . ic It' to in let-of pair-if sera tested shoitwedl an iii rcawt iii loili p~l- Iiiiis l' hepatitis.miii11 fixaltion ;iil it. ill is'1 61 ri-I onte oil ciisiralI Two aoitfa i i faid--S f rs, I i e ii iclla tilie XVIII

564 HERSEY AND SHAW LABORATORY INVESTIGATION

Scientific Session devoted to problems of infectious in volunteer studie-s and was knowns to contain tile virushepatitis held in Moswow (U.S.S.R.) during 1%65, have for serum htepatitis. Interestingly enouigh, serum speci.recently appearedi in the Ii terat ure.2 1 In one of these, mens from .30 voltunteers, re~ijienis of file original poolKorenblit andi Pancltenko'" reported that viruses. iso- of plasmna, revealed rising levels of antibody against thlelated from tile blood of jisfectiok hepatitis patients A-I agent, whereas sertans specimens froms patients with,n a Detroit cell ctaltur. were neutralized with infectious hepatitis failed, for the most part, to reveal any

canine serum to type I adenovirus; however, in contra- anti bodies against the A-I agent. Only recently, O'Mialley.distinction to the general properties of the adenovirus McGee, Barile, and Barker 133 reported the identificationgroup. the virtus catused death of newborn and adtult of the A-I agent as AMycoplasina galliseptir urn, whichmice tupon inoculation in the brain and abdominal would account for lack of !setotrali/ation by infectiouscavity. The viral titers in the brain of the sick inice hepatitis patient sera. Bolin, Alsever. Berger, andi jarsis'7

were 10-`:1 and 10-4 L1 ,D,,, :and the viruis retained the reported mnultip~le isolations of virtuses from thle bloodl ofcapacity to evoke typical cytopathic changes in tissue volunteers inoculated with the NIH plasmra posol 6.culture. In a~fdition. the viruis could be pas ed in Further studies".' 17 repoirted by this group will be10- to 12-daN chick embryos by inuculation into the discussed later in this fpaper. Somse of the original iso-allantoic and amniotic cavities. The properties men- lates were infectious for mice andl produced a tyto-tioned are similar to those of the known virus strain jpathic effect on a wide range of host cells.called NMotol, isolated by Kubelka. Slavik. and Sousek 9 7 If the list of references associated with knowsn agentsin Czechoslosaktia, and will be described later. The seems lengthy, that of unitdentified agents is even moresecond paper by Ananiev el al.'- was concernedl with a implfressiv'e although many of those originallN reportedstud%~ of the role of latent virtuses in infections hepatitis. as uinidlentifiatble have now beetn classified. These isola-Virologic studies were performed by tusing tisstue cul. tiois have been reported from around tfile world. Attures of Detroit-6 and trypsinized ctultures of human p~resent, there has been little or no opportunity for oneembr~onic kidney tissue, from which 89 strains of vi- laboratory to check and confirm the results of another.ruses were isolated its I1120) satmples from 1023 patients This interrelationship should increase as insvestigatorswith infec tiotas hepatitis. The majority of tlse strains are provided[ more opj)prttltity to conic togethier andiisolated were classified as belonging to the latent group dlisctuss their observations. A recent exampfle of thisof t~pes I and 5 adenovirus and to the enterovirus occturred in the IX International Congress for Micro-group according to their biologic. physicochiemical. and biology, which dlevotedl a focal topic session to infectiousserologic properties. hepatitis.) The problem of evalttatitng viral isolations in

Kruginan andi associates.`,. ~"~"over a long period infectious hepatitis has been compounded by the lackof time. sttudied cytopatlsic agents isolated from boths of good, reliable serologic tests, lack of aI suitable ani-inapparetnt antd apparent infections hepatitis andl (let- mial hiost, aind the dlifficulty involved in conductingonstratedlfthe time the agents appeared in the bloodl and sttudies in volttnteer subjects.urine. Recently, this group"17 , 1411 hias reported tile iden- In Canada. virologists have been active in this field.tification of the Willowbrook (WBI) virus (isolated fromn beginning with the report by) Franklin ansd Siniclair 4`patients. with inapparent andi apparent hiepatitis) as a who attempted the isolation of the agent of infectiotusmsr-xovlrus ;aidI a member of the parainfluenza group hepatitis by using thle Detroit-6 cell line. These resultsof' viruses. This adds another previously knowtn virus were negatilve or inconclusive. Buchmner and Sitreeve"las a fpotenlial etiologic agent in infectiotns hepatitis. A reportedfithe isolation of five agents from stooks ofrelationship of WB virus to SV-5 is suggested by a patients withi infectiotis hepatitis, two of which werereciprotal rise of SVs-5 antibody titers in W1B-inmtinized studied in (detail. The original sttudies indicatted itiatanimals althotuagh foniologous antibody titers were it thle agents were tiot identified as known elsueroviruises.least 8-fold higher than heterologotis titers. Thse ap- Intermediary reports by .1&6quael aind Steiner---" de-parent relationshtip to SV-5 raises thle possibility that scribed fuarthier attempts to idlentify and dflaracteriieWB virus might have been isolated as a contaminant these agents. Electron microscopy revealed viral par-frotm tissue d tlttires infected with aI simian virtus al- ticles with diameters of 2501 A. A recent report brvthotigh it seents ttnlikely in this stu dy. since (1) only j~i~qutel and Steiner 7 7 citedl the serologic dross-relation-WVf-38 tissue cuiltutres were usedl for primary isolations; ship (of one of thle original isolates wsilth two strains.(2) reisolations were performed in new stocks of XVI-38 of C;oxsackie virus. Enibil 0i a/.32 hav, already beentultuires: (3) negative splecimens remained negative on citedl for their recent repcorted isolatiots of a t~ pe 10.repeated blind tissue ctulture passage; (4I) isolatiotn was groutp B Coxsackie virus, in connection with] ats ott-made in at least one case in another l~aboratory directly break (of infectious htepatitis.from aI serutm specimen taken 33 (lays after inoctulation, InI Ciechoslovakia in 1957, Kaibelka rt al."', reportedbut not from the preinoculation specimen; andI[ (5) the isolation of an agent known as ' otol virus fromtcad I of the patient, from whtom virus was isolated poo~led sera (of two patients with hepatitis. Under cer-laitsdeveloped a I-fold ristc in serum-neutralizing antibody. conditions, the virus agglutinated erythrocy'tes and was

In 1961. O'Nalley. Meyer. and Smadel' 3' isolated an infettiotis for- mice. (;eorgiades,-tnd Zacek in personalagent dfesignatedf as A-I front a Nagional lInstittttes of commtinmucataions to Ktabelkal'- have sugge'sted that it isHealth (NI H) plasma psool. The pool was p~rev'ioutsly tused probably aI deoxytibonticleic acidl virtus, approximately

IMM Ai n diail metStilt 01The C5 sr~ios of t-Iilie Nlotl sn.l wereolitainvletd insolat ion anf hepatitis ofsinfwithithetid NiaI(,tiit,'an ote~ilt h ossbeln be a i ma ies to saea NIeaits a t iri s in ant igei b lrain iltraiong iteis bux r-a

Npotti ol uiiishas been 2iWe bA with hexaoa tll11 til sera fbaie d isl gromp febril 0( hepatiti poi-

Moll ills as ee) tudedflýotergroups in Po- formed bs Arakawaan higru-fndtled m tlaiittl." "I 21ý The inifeltiouls nlature of Niotol viruis ill nitice of filtratio'n at 580 A. and elettron microscopy shtowedhias fbeen amnply descti bed"' 9.~ 9-, and( t) fi a I signs of a (lia meter of 5MN to 6001 A. Ntiraoka2 1's. 21 stutdied thf-atulit hfepa~t it is ha se becn reported, withI viralI isola tion fbiotfhemtli(a I and bin1 if it ua I piopeities of III I Mit ahoit mo1(st oligains andI jatftologic (flanges Consistent with strain aind fouind piositise (ottiplemlent fixation tests inl

htepat itis. all of the patieints whose sera had been (ollected beforeRe~eit reprsi""- I'l fronm Fitnland liase Andicated the thle 20tht (la of their illness. lit all other sera. (-ollectell

po~ssilile role of t~ pe 6 elliovirtis ii aI hlepatitis out- after the 26th fas of illness. only 60 per tent werefbreaik in H elsinki. A futtrher (limutssioti onl this work ,xositisc for (oiitpleinent fixatiotn antibody with the Mitd

ait-rs earlier tinder tite possiblie role oIf the echo- ,traiin of %irtis. Average inettralizationt iiidi(es of 320.5viruses iii tie etiology of hiepatitis. atnd 207.6. resfie( tivels, were observed for these two

G.ermtan investigators have been active in hiepatitis groups of sera. Six cases of itifectiotts hlepatitis frontresea rh Iior a h long little. NIci tif ltheir earls Isolk wats the Medi( al IC ollege. Ok a~anita . Ja pain hladl negatIiv s itor-tles( ribet I iti er the sect ion intvolvsing egg, and ot her plernent fixfat ion tests bil ;itn a ver-age neutri a lia ion i nd exanimiial isolation at tenl) pts.,-'7 .20. 139 Part Iii Ia rlv protit - (if 21 .7. Sera I root five h eaIt h% indiv~idu~ltals wsere negativ etIke os-er a long spant of )cars has been the work front for liothi tests. CJomplemtent fixation tests were negativetile lab1or;irors of Esseni and ],is tjlleagoieS.34

-4

1 Thlese itt sera fronm five cases oif serumn hepatitis, b~itl onesitidies hias c iistiled isol at ion at temnpt s, electron in i- intdlis idua I had a ii eqjuiivoca I sertuno-vi rts neutira liztaIioniros( (f)ý. and1( sk in testing tof liepat it is pat ieiits wsi th t iter. MI tiraoka 1:11 also tfescri bedl histopath ofogi( 1 -fanges

the use of siruts material propagated in eiillrsonated in liver tissues (if mite that had receivedh itiultilationsIii( ken eggs. Re cot repot ts have appeared by' Ka. witht the fixed viruis. FtijiWara, 4

11-41 Kuratachi,!"' (gasa-fIalaiii alln, spies I'..,6 (oncerti ing the isolationt of %ital ita ra * 13L antil On ittka"' "';. il repor-ted extenlsivye in-

.igciiis asso( iatetf with] illfel tiotis lepatitis. The work of vestigatiolts of viral agenits rclovcredl front cises of in-Dr. Spies htas -etiteretf primtarily around the isolation fectious hlepatitis in the Oka~ania prefecture. The sir;aiitf i faracteri/at ion oif atitointerferin, virtises found strains we-ce isoilatedf front patient nitaterials bys suc-iii ;sso~imttion with ;15es of inifect ious htepatitis. His (essive transmiissioni through mice and~ criitfiroiiatedfCoist isolatiotn wsas found in associationl Withl thte Notol (hiwketi eggs. (mirlfitg to Ogasahiara 1:11. 13L2 ieý (thicksiruisorigirm~ll lisolated bN Kulelkaeai 0.'7discussed liiN a otr isepil ote virus. but the

linettf lessthan. Unfilluhte material or inaerldi leotn ie. Althoughtigta few of ifI( meiiit e dIied oser t ftis peth~xl

lind lssthi ' ý,di no sow I \toatic ff (St (l() ays) an tilhad nlaiked f ath11011ig Icha oges, most (ifin helina epthlil i~e tll o Chngolin tile mtOie develop ed a tl I-ottic infec t ioi wiifth (egeitera tion

sw)ittnfi ftd fioanittf kde iu Iltr h s of il tand te(is oif thle paWlelfl( mna ItellIs an 1(1 cl luiat- itifil-20-et\ hoanhrne ppard t cihn~cgrwt w tih a itaitn. hIt th mi. iouse litngs. thle fnofi feia lion (If loll iltIh lli.(o etatdla ti(la.Bif il~tr-(eCfo Iit(limia tlsaoidhernlii and bloodx

toi tlait fits have ilct bieer idfentifiedl moire sptecficaffy. tiotise to lie cfth k eniiblo wsas estabilishted, stiggestitiglIt ft a f. Segagnti ft ( it." hiave re jmirtet t heir isola- thiat the s irhlsIad ireta inietl iit thle mtittse olga its fot all

lionis inii fali tissue (utltitre anf emhrs-rona ted eggs of cx telnsi S ~c telitlf oif litile wSilth telpea ted Si iaInul ii 11lplica.-igetiIs ftn oi (1I thle sera (if 31) (hlIdreit with initfect iouts tion anttd (Ifi ll ll iiiionl. t IljiAwara1

4 11-4 d~escri bed usew of

lie fixitit is. Iliepat i( lesions were reliortetf in lie eggs 72 i-tintpleimetit fixati ion altid neot raiiiatjolt tests wh iach em-oot rs after i not it at i n. Ba lildhieri rt a I., repiortedf I;%- f io~-d aI mouIIse f i se hiomtogen ate as ani ant igen.

So sitett iore i solat ion at temipfts anid if te t se oif fitiores- iThrouigh aI imod1i fied Ab.4isai img u . 'es.i ff or cross-pro-(cmi jillbth- Io le ieilos in) an oll enda ou- to fentoit- tel- utioi lest Ibte lest ifis of t fu- ( Otl fih itct t fi xa tiont

st rai bo fNth Ii si-r and1( anti bod-. Theiir resutiis. however, and ilc t raIi. 't oit tests tIuild lie ti rrefalt e andt~ were

a re t t~otdoit iisc aIt hotughi re portet as encouiragintg Ii-by toid to ef itit aI (oiiiiin att tigetti~t is iitoig the iso-thlemi. fate t v i ral st raitns. fThe mtod iflied .4N4lsfgfii ung z~i.%c wfi i(ias

Japianese woirkers havec repojrtetl work oii the etiologic seroilogi( alf% sigtififuait wsith htighi titeref sera bitl notigenlt s )If hiepatitis lot seseal Nearis cesi thfotiglt intam of witlli sera of a fins titer against til ftsira I agenlts.

theiri re ic N ris hiav s- ot fieen reatdifl- asa ilable ill tftis ( li/tuk a :l testri lied studies of lIi ss-514 firet ibi fitv to(otttir. Ntim ft of tfie sta~ts (If filte earls- work htas bieen six sarijed straitis of viruises (Isllifara. Kaitaitmiit. Notfa.Iics fewetl in ;I afImipfel 1), Araikatwa 0f(t. IA 1hle wsork of lhiis ( gawa N , Itimotit t. Aomoi1 n) u lid f were isofa tetd from

jt.h 1MlRSE'N AND SIHAWV LsiHncA bsI svc INVItIAIiUN

patiells withil ill ettIiol heitpatitis in lil t- ( ka~ alila pire- Si Vhs prO(t ~c I tie l a issule ciiilitrye t iii ge ((Jil ar able tofemitre. Iii criIt-nia lor in i-c t in. iii cigi t swc ies of th oste dCS( Ii ibed i\ n dIicir I for G( clotp I. " Il I iciils ( Wi ie(

OIAod. were tile Sp~t illt pailIi iii g it t anlges ill file Civ ser i opal itguil it glol 9 of s iloses Were- cesi ii et as plit-anid buig. W~ithi fixationl of lilt S iiat strajils adltitscit ill (fillinig fiolphlr lolcgic c ianlge% Simii la to (Ile seccond gi ouijfi se 511cc es se tassagcs of lit-e S rat agenlts. pa t iologic cot Eiiuics. The 01. Seserowa 51ra in tpl xlticet a c I(Nij tIaiiiialIteratIions were jilt rca~ins gi ses c-i. The mlost mlarked elcii t I oilli Ii a tue to thle thii (\iiIlt~e of dicgeiclia 1(1tio de-effects Ivere oltserle ati ;ila I ir ildct i~if of ii ) ii ini stilit-il hN ii nbitiis, Thles (ont I itied that IS11(till elt Ihuhl.%eripUSilii (u iar, an lci i "1 illIn ut Ur lit/it diiiie I lii, polottgit cliielenli s 0cc ii iec ini tile Iin ilIro c 111111 C iot-with suc tessitil resI~oI el Ill(. ci Niirtus front lowl tol this t IC oli iil tililia Iicli to1 I~l0SittI a I isci iiul~ Splemientil

mite lin all c ases. Iii i Iis vc-ptiiiinieiis icf t- 11011it illicjt- to1 tilt ieliit ifica ticoll of newls isola ted s i tiscs. The- lreporttijol ill Jowls, ()ni/tika"' iliti ulaiti lilt- Ishluitara. b%~ \Ior/\tk;i adli Tai~lclI*2-_ l~iclltci outi ithat diiise u thitisKa ilamilsi i 51 and1( NtcIa strVaills cii sir ses intoIc 37 cania ries. wuieith timile to isolat e agents Is l Wool cait litcii u

1 ii sSie I)C i -

33 sisk iiis (Ch/i (is siniiiia min inni . & N~.). anid 201 itise- minsl ls iiepaltit is pat it-uts in tilie lDetiloit-lt straiallsoci lissti

binics. (h)ser a c titse of I 1(11 6 s liiniic iiile~t itlis (([littile esC-il thounghli sluls isoltioiinis had bieein deiiltii-were. jutdged 11 beiut preseto iN alterat-ion ls iii [i ic pa tiiol- st rated i il othter h ost %stsiells. 1The oirig Iinla isolta imnop~ oflilit- user antd IItiig as weti ;Ii in liet w like (tci were ilade it hiuimi ran eiriibrs n(Il it issiit c tlt ii ls. us tie-

((1(1111 ait io blood~t pitzzrc. A itew iethal at a ss wire iiii- sribVeh b\S \Ior/\(iSi ef (ii.2*scr~ec: i itwes cr. in dil apprent Ior ciirt mit iii ectio 1(111 tlt l Rtii uianiiian wo rkers hia se been st-r% a t1t-iii in epiatitisilsolatls tie csia Ill islic- will) illirape ioryit jiioctlaticoi olf rseard i iniil at asi two laboraiories. I as/ll":'

0 and hit iis

stis il sios prprd fotnitus t ist rs o il o- a ma te.s hia St Vel e t Iile i stota tion of %i rat tl aet s inilidteii eggs thiiat ha l m rtc 1i Id thle abiiis thiree sVa ills (it bthci iih1umani aiiltnd e broi (li dit i tckti t i er ti ssuie ciil-

\ironi se. Remiarkable lt et-ikt eicOia was iiisi-rsec in t i e lii r. 1lThese stuoi es jilt iticlet scrtlltg ii Ct i ig as %t It-.tri st ages of inii(o liiiitllsi ii a sevsere ilt rea se of as ciei tr rcii111crtlstcpit st ildies oli tilit ageiits isolatec.

.it~pitai li an Samciikl-itgt-ic-ra Ci itiikcit ~tts. Livt-r atl- At Io i these agent s mlea suiret ll0 tilt ranlgt- ol i00 ttoIteraIii Ils icilsi sttci of ronilld ct-il infllittrationil ill thle 7001 A. Miore rC(~tc enis as/Io vI~f hav SCIci l tct o ill

licri pcinaiitl ilcit st~ t issuec wiitt i a ricits itws~ of (cc-- a sttud\ ofi 21 strSt isostlat-ed be-twetein 1957 an iilI96-iIiila r ch-gt-otra ic m scic h a s inoii iit- an tiii lci rot it Al i ft hei ise wi-re i solatted in Det rci t -i (telis. Both i iit

cii aniges. w itt icic ali/(( ,c ct cruisi% sma ttered ill t lit-jiter- I Id i/aia on1 an iii clii enet fiixa t ionii tests werte Ciil-

miediatle Pa rts ol thlt amill i. Thei si skills wert- itit- most5 din ecl to est abliish seil oogic irelit ioniships a 111111g I liesi ic tpt ibi i ot til liteIsted lowss w-isjthi lilan11 lethiiat cases strlainis i soltated fromi tilie piat ietuis. Sjiltc t I9ii2 MIi tiiti

ill w i thi icrs si-c-rt- litc rtisi andi t- oL Velua inilt Iratjioni anlIit his iossorkersl- -I-"2 ita-c- iteporlted the ir studieiis cit

ot c l-rtec ill tilt-Iis-er. Thit c hanges ill thte It ings ap- ageiits o rigin;]]]ii\ i solated IfIriont ipat-ilnts with IitpidC kin iVec iLi i- late tani those iii thle Ii ser a tic

1 persistd fclcr liepat~tit is. Thesie ageints grews swell iii ii h iman epCatitheliat .

6i lilt0liltS. willt) ii-olri 11COI IIA it-iluiiL- inIi it ia tili andl K B. e La, HEI) 2 Dci rciit-. .an iil lciikt-s Ihltdrt (ellIIlltici ;tiscciiiIis w51ill] hii-llolilliagt a iiiiiii th li- iooiii - cii, ( tiri-s. Thei viruist-s c l titd not It- idcenitiflitel sCrl-aiscoi ii aliii bloid iiis tsseis. icigic al It wii thi an\s knoiwn iii-itelcis urs a it iscra a ilititigl

K iillma anit Hilt Ia' lepillted thut lid iasliissit 1tin of tieN wsie icuiiiii d 1)titt ,i tile scra f rom mit ilsttie lit

aI s-i ra a geint Vtiocr~ froCV~ IInitil p at iet" t d iirinig all epm- pa ltiets5. hit liiid Itc iilcmor reich t stuii s itSwas filit detei-

dciiiii of hetpati tis mticcirilig froni autoit oill 1i 915 onlstralio of11 ;Ii sercittgic iclatioinshiip lit-ieetn-i I lit %tratin

to spin ol it9i6 iii tit-, K~otc-Osakat-Koble district of isolted~c us Milrillt tiit .isscuiattS aiid ilie Htissiaii

Japani. Ilejldriniii-t bloodii taktii fromii paieitils ii itit siraili. K3. tiesribntct b\ Aiiaiiit- i- al..' as swell asfebtrilie siagi- oif iiiiitss. wa~s ili(tLIt~ u iitac ile hi- l~tr- t liltiigts in file isolateis of MNllitltt (.1 (1. ciil adaptllaition

jtc(6) l oftl a %sttsic-sion ofi lis-ir i issoc- fromi ;tui inic~tei-c Ruissian wotrkers hiaste benqit-ti t aiit-cli~te. Prutlah i Ilit-

Intilist was gist-il tol a lit-aillis acill mail. A~lmt 23 class. mlosIt tim live i-t- bieten AIIatliC lc I i and Ili% ].o iatt-s.2

. 21

a ( iiiitjijonl tit-Stttiiec iiilit silt \loiiittC sllbjic Issidic whotic has c rejlml-t-c till Botikiiis tdi- ase (a tiii Irt--

iiitIci liouis hiepatitis. Hasashii. Kawaso -. aidc Itaka- hieptitiis). liT-e %traini tril~ic iill\ ICssiati i utc ith thisii1511i rVtiicrtt-cl mort- re-tiiintll on it- isoliatiton iii siral dtisease is caticci K3. andc thie smiticis ofi .\ midiit anid.Igtills assoicdllu with Iletualilis ini J~taii. tIllt Situices hlis ccwtirkers liasc iuiscist-t temits ltitius oi lilt- s urat

ofl tiara ailici ilis scil-s wer s t-nt-te oilil-it- l m jrtojirlies ill iisstt c taittires. as well Is cili l~lt scroitigit

.ttlaji. (.orgimlecs, amidiItrj~hiicst reticirit-t tin relt~tionlshlip livelseei Illis siraii atiid lith R~umlaniani

A tillijaralist slluul tof itlir gntllljs cit srat agc-iis: isotait-s has atreaici ticiel initlcitti.i' Kisac iko ;iiit[(1) Mitot s l \ms. (2) OI Sec-ti-tiwa. (3) aceit-lti I iss. andt (Galkc'. hiast- rIixir(lt thie isolaltion (ill Dc-tloilh6 ciiiI I) I gilill) f i ' itihdiogt-ilit 5illise"Sstioicd ill ;ssoc ia- c ilittirts,) ill sinai lgc-ii fromi paltisi wtithi iiiltc litititiln with1 lhi-r c tarmcterislic (%tol;Ilalii elictIS ill ilie litpalilis. \it-si- hit~ lait tictil tarlitie iii stiiatKB line of lissiic t tiilrt. Bs itsilig flei d assiliataiulil pasttaiir itheir isoltioniii i1`011l bloodit SjIKc-( 0 i itIieatCi~

1 Tiicria ofi tidut%.:`1 I tte n-ilc thal~t (lilt- Niclolt o lif) C~l~(. Aul six strinils of sirmu appeadr tobl onhttig toi a

hortogecousgrop ad ar no ne tralife b \G bewen lfrsi mononucleosis and th clineiso

No~kovl"attmped ira iolaios front 2001 paItientis isolates hias becen mcade bhsY R igftsel, Keltsch , Tek cishan,with inf~tiois epattisbN ising prinmar% huminan acid NMcLearil 4 who reported, in 1956, the isolation

eiiibrs olic kidrie% antd Detroit-li (ell c nltutres. ifore thtan (if numerou~s agentts front sera anti stools of hepatitis10 slrailts of ci opiatlic- virlises were isolated, 20 of ptes.Several reports were publlishiedl by this group)c-inch were idleitihied ats t'~pe 3.1 group B Coxsac kic of investigators bietween 1956 and the present\irtises. Fourteen oii tile strains were not necitraljied b\ limle.1

4. 14:1. I'M Thie siystem tlieN tusedi in reent. years\atiouls ant tseti horn the entermisius. llctles, and involvedi the dievelop~ment of a cloted strain of Detroit-li

adcumeoirul.s grouips. Of thtese strains two) were sttudied cells. These cells dleimonstratedl consistent suisceptib~ilityin dict aif stra in s 1 -62 andc 2-62). Bothi strlains wet e in the i r laboratories and1( rea sonala 1) (onsistent su s-11(111pa tiicgett fo r sick Iintg mitei an tic d lic not muiltiftply (cpt ibfilitN in i those of ot her workers, a It hochigl a least twoin c hitk vlinbr~ci. Thle\ (lid lnot show ain% lienaggluti tinatý - all) at te~ntits by .MorI) ka anid 1 ~fr antd Franiikli nitig ac(t isit\ alga ilst er~ thiroc(Nt C of mian, horse, li ic ken, a tici Siuc-faiti- 4

1 Were, ctinsmcessli ti il thle Lise of the cell lintigoose. slieep. ttontkc%, or gtlinea pig. Thec vircises were lot iso~lationi. Thie studiies by Rigfitsel andc his associatesstablfe in tile presetic o ci both ether atnd c fliorofortli. flint C pros icecl consiceleable itiformatioii. both ini labora-

A omtioiil anlt igeli of strain f-62 was iioted %%isl it icte otnt- an iil in ical dectaifs, onl thle fbelasiot of thfei r isolates.K 3 sit ts isolated iti Noscow. The oithet 12 notinipable Thlici hase been licllieotis relMorts of studcies by fornmerst rAnti were stimiflar to nstra ins 1-62 anid 2-62 iii their associates of thle grocup icl c ind ting Sic hnidet. Tays-or. Mc-11111iorec a nd \iccpa thici elfecct s. Futirtiher studcfies ott t I ese (a uglies and NIciirf iemad and NI ti irleacl and Sch nei-agents a i c t their reflationtlshipi tci I epa citi is -at )Ci ug dlci 12 1,11in ate still ac~t isl% etigagecd in isolat icin a ttemtslt.fK-rforiiicci. \lote rec-encl\. Cole i-i a/Y, lelicireci fromt Acistralia on

J lie repor t cof Sfia rla i 0 alI.""' swas iieiitcioined earl hie. cite cisc of lDecroi c-li c-ell linte for svircal isoliatic)I in neonatalShes c ietko. Daiiileiclietiko. aiiic Netefitsal i7ý isolated liset disease, as iwell ats in 28 sera of pattients acdmittedI tolilt c'le \topa ii ogci tc agents frcimi Ipat ieiitcs swith inllct iocts Fai tfielcl H-osp ital, V'icctria, .Austtsailia, with itilie diiagnosishiepatcitis fIm incants of tie Dec mi c-6 cell linte. A ll agentics of iiifec-cintls liepa cit is.Lt Of thle seta examiiiitecd. 27 werewic-itlsod icieatin atlt'C for 34) minuittes aiic wsere treportecd as lx)sitise with this cell line. A re~cent reportno sis t t td I ile effects oi either. Thie% cfid nocu by Ferris and Acol

4 described experiments chat usedssss iletivigglocit tia Ii g a hi Ii c aga in st ;kit\ oi ctie coded sera. aniic t fe Detroit-li cells sfiowedl cicopa cliic

er~tliroic 'des testecd. ' I lie c \topa iii i eflec t of thlese ageiits eflects alImiost ais frecj uen thI ii tilie cotolcrt sera as illlit tissue c cltulre was iicitralimcl hi cue sera cof tile sera frcnti patienits witih itifecticucs hepatitis. Crcoss and(it isa lesci Clt I a Iicti Is. bi11l l o iib sera o f known tisira I IartitioWn': alsoi repored a siml ar t~ iw of nolispedfci

ig(-dtI lic-Icitg ing toi the etiterociruts ir ac ettoirits ci-copathiti -eliecit itt cil tutres oh Boil ns Ii cil l m itilnggrocip. epic leli a I(cellIs. Thle exact caucsc of ctie c-eli ilar degeuiera-

NIcic Il til i e wcirk l etli nittiec \ Alt~~itericati Ii est iga cots c ion cobserv-ecd ii both (cell Ilintes wsas riot evicdentc.hias alt cal isIbeen cleso if ed inI stucieis i itc Ic isolates it len- El -A lii. SmitihtI, antdc Biesele'1 h Iave telpnt ed oltt h leti ficd itas ktnowtn i ta I ageints. Th iis is probail () itile to tilie passage otf a ii agenti i sol ate flrcnt tinfect ious Itepa tic isv tell sit an ti con tilt iintg ioik ini attemitpciting itc) icien 65-i ii materia I ii lcikocs ce ccii ctires: iii ectecl ccii cres demvon-illic i sioifates. Ili weser. tihIcre remnaill seseral Uniteidcc si at ec chit niosomte btca kage. No mentioin was macde ill.Scat c-s i soilaticl iii t cl ll its Ii i whid atI;gentis hia sc not tilte citigioaf teptirt conciniccliig tilie agent itself a icl[ itsbec-itce n dssi fic-t . Bol in oi~j* rc itrcec thi e rec\-ir\- cif poi ss ible- ila t ioutslip to ot bet iscla tes, or ctie pal ientitssc-rii IllIic-ai iiiIis i solmaIcs friotti so~ini it ters intocucIate wcc s ith sc-la firomt whIoin cthe agenit was rc ietmerel - el Ia arid

It Ni III 41 i sa jicicil (Thiis wais Iiic( it enti tca I licol inrg"' ha15 I a elsc iibed cthe stulti essioi cof let ikcuc te tiiicc-huhil ihicl I ()Nallc\ . (1i 1 a/ irecs ct tule A-I agenti. sis bohiti itt ptaticents siliilciirg frtcui iilifc ciocts Ilicatitis

Li 1lt tic-s tlievd its a .1lvi np/anita ga/itimpIil 11m,, 1:1 Thie iiic iii Ititnial Ictikix tes exptosedl to setitn fiott patient.%5 il .i''cIlts wc-crc itiitiail[\c- isc-rc-i ill iistic- c ilttie wiith ilt( cdiscase. iThese rplejdts. alcitcttigi (Ilie%. dco notofi iIltiall cling clIts. hilt afict pa~ssage iin tisstic. cilcutcl c cmiil itite altlitvc iabltl to ocui knowslecdge of cthe etiologictihti grew itt a wicic slic Iritt of ccll litnes. lit oscr l10t agenIts. iiaus pimsitlec partial inisighic intoc sonic of tile'ilic-ilfpis. Boint ,I a %l cIv nut able to isolate alt agent poissiblel fctifc s ouf agentis assutc atecl with intetiecciucs lielta-diitcc ls hlicut 111c sdiliit c- ciitlt- A-i aigc-nt. Latcc itI-lns tilts.Its Boini if ii Hia liiin gc-I - P1uttc-c an ti Al-~ sci c-I anld 6% .Atic-r a rc-crc siefiv-l e res1ic- ciif ilte literatutre ptub-Bl'uilu. ( .lusc. Alsesci - andiLci 11 11a;tl e iigisc- itcitc cticiieci lisliclto Iii dae. sescral fc is Itaic c-nic-tgec front all ofcs-ilt cit icuis so-iuiligic leicss 10101i \i cc-c dcicschitc citli thc sttiiiics rcliitocle. 'lue first cii diics is iliac at widleilulist. sc f Ilic isolatecs Ititli oluiltis difliltlc-ci gloilp~ iss arlcc-I of ;igctnts its ftcti isolatced fronti cascs of ill-.i1litigcll. I lIt-, fistis was itici toic-cl Suilll a c ittiplc- fec tutu5 iicpauiitis tlittgilg friiti hItl %ell knotwn virncissIliil fIvi cull lustan ii di ci sci iutitld c ~t Im-1 16 1 C citae toluuusc flt pit-cliioiisl dlassilaci. lIihc--lctre. 'w-hethterc tugohlticiti S Ict lic- luttci stiiii sliggcstc-t a1 lulaticitishiii tilt-c aiinsc- ci hlmitiaitis. Ilkc- tihe i itnittiir told. is lilcteal

M86 HERSEY AND) SHANW LABORATORY INVESTIGATION

in nature ar whether tile etiologic agent is one or et al.."' 2 thle CHAI viruses of Spies,'60 thle C5 fraction ofmo~re of tile specific agents, the hepatitis problem re- the Motol virus of Kubelka,~"l thle viral agent of Es-mains unsolved inasmuchd as most work has failed to sen,34 andi the A-2 (CW) plaque virus initially recoveredfulfill thle classic Koch's postulates. Moreover, there by thle staff at the Armed Forces Institute of Pathology,have been few attempts to correlate viral agents and Washington. D. C., and reported by Shaw and Banks.""'patients sera. obtained by different investigators, with Although the particle size might be similar to any num-one another. In spite of this difficulty, certain inter- ber of viral agents, it has provided a certain uniformrelationships have become apparent. For example, the criterion of size for a number of isolates to date.recent reports b) Liebhiaber, Krugrnan, McGregor, and It has been an adamant requirement that each newGiles"'- and by Prose, Balk, Liebliaber, and Krug- viral agent, recovered in association with this dlisease. bleinan, 140 concerning an agent (1%'B) which had been pre- thoroughly characterized and, if possible, classified.viously isolated and described"'0 an(1 which has now been Much of the confusion that exists with regard to etio-identified as a myxovirus, might have been of only passing logic agents has existed in subsequent susceptibility stud-interest. However, this particular agent was found to have ies in animials, in that such passages have possiblyan antigenic relationship to a type 5 simian virus. It activated a latent virus from thle animal lios~t. The lirol)-Should now be recalled that in an earlier report (Hsiting, lent of relating the aninmal host to the originall-, isolatedlsacson. and McCoilluM 4) inl 1962. it was noted that an virus and to the hitman disease syndrome has beemi con-agent (DA) was recovered from a fatal case of in- siderably diffictult in resolution. The pursuit of the etio-fectious hepatitis. Thle DA agent also was immutno- logic agent must be encouraged, not only under thlelogically related to type 5 simian viruis. There was standards and accepted principles arnd t~chniiques ofgood evidence to show that the DA virus did not viral immnta ology but also within the framework of neworiginate from thle simian tissue cultures subsequently conceptls evolving from stutlies of molecuilar genetics,used in the studies of Hsiung ei al. More recently, which have enlarged the comprehension and control ofKubelka et al.9ss reported that the hemnaggltitinating fac- the mnechanismus of viral and host cell interaclionls.tors of thle Motol virus were antigenically related to Mutch wosrk still remains to be (lone, in relating othertype 5 simian virus and( to the DA virus. Thsese reports, properties (of agents recovered in association wi6th viraloccurring independently on agents isolated and sttudied hepatitis, which the atithors hope will soon ble lorthi-over a wide range of time, have demonstrated ait least coilming.

a certain antigen in common among these agents re-coveredl in association with cases of human infectiotus A'tepled for philklitahirlm Jill% Ili. I!Nis.

hepatitis. Of course, thle plos..ibiity of simian virus con- REFRENCEStansinants or the activation of latent simian viruses in 1 l~pto amso i NItpoonlCpge, ..Althe tissue cultures cannot be precluded in every case, but r*c~ioliolop. eullitorl -in fll .%khanola. 1'. Wt. ()rganihoig COUli-in two of these reports this would appear to be highly iniute of the ongRiesm p)fil. 9-397. Nloscow, tIiioi ol tileunlikely. Auilisio, Wong. and Mforris," on the other Stniel Socialist Repiublit 1%116.hand, have reported that examination of pairedl sera for 2. Anauine. V. A. Absa. P. %1.. N~ai-kis. S. V.. flaijuski%. 1. E-.

Slihbiles a. V. D., anid St itrolaiiosa. V. (.. Stud% of tile role ofItype .5 simian virus-neutralizing antibody fronm 50 cases SAtet iussi ntioi eaii.Ii,.Alt.I:81.IW7

of hepatitis, contractedl by United States troops in sev- 3. *snio. V. A. Kaeriii N. V.. Nariskis. S. V.. and Ilaiiiiskis.eral geographic areas of thle Far East, showed that 41 1. F. Properliel of %iiiis isolated toim tile feces of iatikcitis will,

gave negative titer results in diltutiosns of 1/4I. Tile re- epidlemic hepatitis (Straini K3). t'op. Ill to~ 8: 217. P1%.3.maining nine paidsera shwe onl tier frm18 to 4. Aniaties. V. A.. a~e A. K., Xarski%. S. 1'.. ltatiiikis. 1. F..

1/3 inacttpaned shwdonvaecn ea Alyit-r frotalcn Kaserii. N. V.. and E%,tigneesia, N. A. Siud% oni tile actiologv1/3 i aut ad onalscnt er. ulsi c a."co- f Itokn eptei h-piiaiis. Studl. Croif- Inj,'iml o bior ]oI :

cltided that on the basis of their data it was reasonable 261, 19613.to avitume that type 5 simian virus was not of etiologic 5. Aillerseli. 1'. T.. and Floliliiiui. S. I he elililogN of heupatitisimportance in the hepatitis cases examined in their epiilcnica II. (Epideinit: jaundiice). Arlot .tte,. stood,. Q4 4917.

stud. Itis tis o colatio of ata rom3idesuyftithstpe ofclaino aafo iey 6. Arakawa. S.. Motiaolka. IT.. Kanecko. IT., Ss-ki. I .. and Got.,), X.

divergent sources that this review has attempted to stim- Exiperimnenial stiidies onl hepalitis infteiiixa. 1, (.illiisatiii (,IWalte by d ti ug old. new, anti extensive foreign entries so %irtis in (lie heni's egg anid fixatioun into tilie iiiloise bli al. la'..that th~ose investigators most familiar with the field 1 iiFbil 4 (4): 371, 19136.

might reevaluate ouir psresent knowledge and dhiscern 7. Ahison. R. W5., Casio. It. C_. and! liaminlon, Ss . NSl I. Adlcoo.si riisassorialedl defect ise tints paritic-les. .Sciroff , iy4: 7,14. 1%.`,.

previously ttnnoticedh similarities. 8. Aillisio, C. G... Wonig. It. C., aill Niorli i. J. A. NeoiaiiiiljiigAnother common factor which has emerged fronm the al iiiiodlies against sililianl %irtises 55" and! 55.. in, hitni1an s-ia.

work of numerous investigators has been the size of the Proc. Soc. MFxj Iliol. Aied. 117: 6. 1%114.

particles associated with thle inclassified agents that 1) Italiwlieri. Kt., Fiaschi. E.. Nan aralli. R.. S urn . A ..... tti. Papa.have now ))een recovered fromn human cases of infectiotus G. hltiqu liii otilie etioloigic agetit of humani %iial hlIwimmis.

Au..lutSur. .,Najtia V 47M1. 1%13.hepatitis. In reviews of tile s1tudies, which involved oh.- it0. Bang. F. IS. I1 he griowthI of organl ctlult, of15i lii ei aiiil theirser-ahion by electroun mticroscop~y of tile isi)lated agents, iuse ill the siinil of hepatitis. Iin Iicjsstitt% to.,tomr% Ilivnir

at least six investigators have repo~rted a viruis-like par- Ford Hospital Iiilernatioinal S%niplisioimi. t~liirsl h% Ilaillali.to~~~~ W()Ai imee. ~ ~ I.~ Loripplo.G. A. %fateer. .1. It., andBl ionoin. 1). 171.tidle with a size ranging from 180 o2WAi -aer. Rnstoii. Litile. Briiwn anl Co~.. 1957.

Their reports mi I tded t he AR- 17 isolate oh R igh tsel 11. Iiatig , t. It.. anl 5Sarwitk. A.Airinipis it) jirislut e destitittic ioptI ii.,14 th Ile R, Vs

9 , V6, a nd I 63S strains oSf I as/ln of cells In hepatitis siruis inl iisscic cuiltuv tire tS .r (ituo

Vol. 19, No. 5,1968 VIRAL AGENTS IN HEPATITIS

tile Laabaaaatlut- Pt opagatfioti and Deltection ofat he Agenta ofa 37. Essen, K. W., ant I embke. A. Neue experimentelle StudirnJieflaltist (Puhliaatioln No. 322, p). 10). Washington. 1). C;.. Na- ujier tlas Virus (ter Hepatitis Epidemica . Schweiz. Z. A11g. Path.tional At~adems% of Sntinc-s. National R~ewarci Coutncil. 1954. 16t: 289. 1953.

32. iBarunski%, 1. F. Anaiattac V. A.. and Sinaiko. G.. A. Serological 38. Esseat K. W., and 1xemhke. A. Cher dten Erreger der Hepatitisstudsl ot sirtus strains isolated from patients Willi epidemic Epialernica. //I. isakt. (Opig.)/ 159: 387. 1953.hepatitis I tiatkans disease). A. .llikrobial. 6: 120. 3964. 39. Essen, W., Lernbke. A.. and Schiecht, S. Uher dtenl Erreger tier

111. Hat inskis, . F..I and Kastrtn. N. V. A studN onl the nature of Hepatitis Epidemica. /iichrungirversuche itu betrureteln Huih-aI factor respontsibale for sirsceptibilit% atf cells to a %irus iso- iterei. ibi. I/ak/t. (Orig.) 187: 379, 1962.Waedl fraoni pattrnts Wi th atiitimots hepatitis. Voap. Vicra. 9: 219, 40. Esisent. K. W., JLemnbie, A., and Schleaht, S. On the pathogen ofI '(it-. epiudemnic hepatitis. Electron microscope studies onl adsorption

14. Roggs. J. D).. Capps, R. B.. Weiss. C. F.. andt l Mcean. 1. W., of simus particles on er~throc~te fragments. AtJ. /a/at. (Olig.)Ji. St atuis report onl I isste-c tilt tire ctultis ateti hepatitis s irus. II. /88: '297. 3963Cinicital trials. 1. .4. At. A. 177: 678, 1963. 43. Essen, K. W., l.etnbke, A., antI Sictlecht. S. Ftttersuchungen uiber

I . Kohnta V. S., Alseser. J. B., Berger, J. S., and Jars is. T. B. eittrn Hatatfest hei Patientett mit Hepatitis Epitiemica. ZAi.Stutd its ott sattatt aitt itnfections hepat it is s iruse% tif muail. I. latea. I Ouig.) 189: 34, 3963Preluitiitals report titl thv iso~latiti tof seruml and ittfectliaus 42. Bsants. A. S. Attemtpts tat t ralsiltit thie sirts ttf Itumaan hepatitishtepatitiis s irtses froma ttatt itt tissute cultutre. 1wamt if ioiot 1: tat pt ittates otherit thtan ttattl. lIt Sy niptutita eal thle Ietbota,-:160. 3961. tO,. V Iolultgitfiota tand Detectciaon of f/ae Agetalh of Hefiatits ( pub-

16i. Iloliti. V. S.. Biaattiatger. G. I'.. Pardee, J ., andt A Iss r. J. B. Illat iont Nto.322, 1t. 59). Washtingtotn, 1). (.. National Acatdemytsttatlts ttt tie si mutt antt ainfc-tiotts Itepat itis siruses otf mail, otf Sictiett-s, Na tionaal Resea-ara C:atul, il').4.II. ( antplt ttcutttitxaititl sttuties Willi %cr itti anal infetlacions 43. Ielst-tielal. A. 1). Persatnal communicttttatiotn. lt.hpitt i Ittlis51ttrses TS I i t it ft an 3: 455. 1963. 44. Evt-ris.A..,anl Ctale. R. A. Detroiatt 6t ccllIs anda infectis v lepa.

17 . Bolin. V. S. C hase, , S. Aiseser. J. It.. attd %final. F. 1). X iral tIlis. Aled. j. .tauil. i53(2): 1213, Mtir).

Ithep atits at-st. fed Pt1

2~t 4: 1.59. I1965. 45. F-rantklint, A. E.. antd Sincelair. J. C. 'I isstue ctultuor- s1tuies WilliIS. Ruthnitt It. K. anda Shirt is. Mi. Ilisstue aul tat t isolationt ttf ittfer titas Itepat itis spec imetns. Cnattd. J. liartiohaaio. 5: 5li7. 1959.

it atttis~stIihla agenits frtom tinf-ct ious hsepat it is patienits. Gittutr. 46. I-ilisais a K. Sttudit-s ott thle inlfecttaious hepatiatis it, ()kaama1. Pult la/ alrtlth 55 '14.I 394. prc-ft-c tre, pa rticulacllv ott its si ralogaral recarIitins. 1. Stutdies

14) litaI ilgltatti11 1 .. tna Mt Kee. A.-1'P. A.' Itetalssiat a.Ssociate,1

tt [lite ttoloplelttt-ltt fixat iont test. J. led. A,%isaaitiattn etf O/au-usilt tss att IV, nts satatisl' Blttl. Ipota. 101, - %.". wattta by) i): 959, 3957.

21. Iilapt /.-. ( .- targiatles. J.. atta Ittrwit -oIili awa. 1 . Catanparat ise 47. F tjissat a K, Stutdies ttit atie intfcctittus hepatitis it, (La~atiaaitit it gi siutits tn it(lit- p lttagrtii its otf sirtses. Mlatail pIt fc~t ntt particutlar[% oitt its seratlagi a I reactit ias. If. S talices

I1. Stlieratwa andi atel-itsi rtsr-s attt ttf (I tapatIttt(gctdt agents otn tilt, netit t tatitato Ic-st. I. Mled. A.orta. Okaatvaita by J4): %9,istlalt- i tatt c~ases ofi epitltltini hetpatitis. .4la Alet id. Pol). 3: 137. 391719621. 49. 3-tjiaisat K. Stutaieis tott Itit- inifect ittts htepat itis itl ()a~ atna

"I1. ( tlt-. R. A. Viral atgentts ft atat pat ientls Witlit intiftca ish-ep-atitis. prt fituttrt part actlarlm oiti its seratlogical reactionas. HL1 Studties.Au,a'i /. Ftsp. Ilit,). Aled. Se i. 03: 497. I 965. tilt I(lie ainfet ionh piorettttiag lest. 1. Alt-c. AsAsOt. i/avaiv nca bt9 j4):

22 tCilt- _ . . Dlatiks. I). M3.. saol Camtpbell. P. E. Hlepatitiis %irts 9l79. 1957.ill o malt- tal IaIis a-Y disease-. Leaw, 1 1: I1368. 396i5. 49). a td, S. a(tidt Alitu. K. Studie-s oil the itepatitis thims. lit Ielejaa

23. (Itis.(. K-. and1

Maiarntion. It. P. C I(:tl ttltiare andt itifectitits titti. fiotltle-, i lt-trs Fordl Hoispital Inltaernat itonal Ss tipo.hetpatilts._ litil. /. .. 4oI. 53 (2): I121I4. 3960i. sin .tatt tle it-i %t Hartmana. F. W.. IAoC ippot. (..A.Mateer-. j. B.,

21. hais i. E. V.. Isttlani itt if s irast-s Iyrata thiltlrta sitlt infec-tiotus sand ilariia. J.. p) Ili9. Botstotn. L~ittle, Blorwnt andt Cit.. 1957.Ittpa itisii. Siitrtr 133: 20.59. 396Il. 50. G ilIts. J1. 1'.. I ichtltaltt-. HI., Kintgmatni. S.. anit I ataimcere C.

2',. lcit-iilttil.fý 1., i In Holotrits. %'. Ail. Epitlr-ai itlogN atai e- tiology EarIh snri a w ;li Ili[ si taria itt itnftectitns htepatIitis. Vipofogs- 24:tal %ir~IIialheaitis. Ptttgr. Uiwe, D)i,. 2: 373. 36.(107. 19(%4.

216. Dolit-ttI tl. IV. Hottlttis. A. WI.. Caspp" R. It., ata Paoppt-r. If. '.5 HaaIsgt-tt E.. aiti W, Willirski -S~bcitel. K. / iry Fi gt- (ltr A titlogit-saioiv% li- it ilt- ti aimtntissii tait t f i aital % ira Ihepat itis lto ntarmt- tder Epidendaischit-t I ebt-rr-at tetttttag. Atila t. I/mg. flat/a 1 / 20(1 .,et ittittkts s I. I manastitissiatti t di isease. strisal passages. aitt tie- 1950I,sa ipllol itoitl list-I le-sionis. 1t. I:satled. 1 25: 6(73. 39%7. 12. Haw.s K.. Ltslaissagi. Y.. ()la~ aslti-lA. arinia. 1'.. Tsuchia.i

- I, ltittat. %.. KIi isit-ch taotld 3pitt-ivitioltgistli hr 1t-lachtttaigenl L. Stagis slit -1 .. Hatatti. S., andt Vastakachi. H. Studties oat alit-i gca-ltatIli-itt .. tallc-It-at suit Hlepat it is Eptide-tait a lit-tat %at sirtas istilalt-il florat a ses re tast- ii eptidemttit liepatitis. A pious -1, W-im~~,:t b: 532. 191 Iil at-a alsat is r agett tof t-pilettuit het-pat itis. I' tc. bill. Aotei

2S. Iii.a NE.3k.. kills. A.. Wl.. Blantbaida . M1. C.. 1-acquhaltt . 1.).. 2.N (5: 270,1. 912.Sitokts. I . it.. .auta lt-ilt-. %%. Studties tau t it- agetI (it inftcttiouts 531. llara. K.. kasaissagi. 1'.. laititaa. N'.. Oltaassii.A.'I salhisa.

catlial it. !I. I lit idist-asa- 1italolwc-t inl htum~tan sttluteels Ints it- I .. Stagissatta. I .. Nalsati. K. Abe. H.. fiala. "I, antd Osaitta. T.agv-i al it '11 lt-il iii tissiat a atltllr t- airot-attlts att-a blltas eggs. Slitdits tilttilt- cauatsiit 5 irtas ttf epitdemtic hepatitis. II. -titihtci1. 1 sp. lit-if. 1,': 293. 191,41. isoalsiatio aof astwo straits horaith bfor d lit ttl ai npialt-tic Itt-patiiiis.D2 uw. itsL. F.. M.. .-thung. (. .. avitd WcSiitek. E. Ulter ilas Virus tit-r aut hasatitaio oif filt first %train t)r-poarted ill tile livesions paper.11upatil,ii El'i'lultlt'a. Z. t1,10 m ttilaielso~ti 1'/. /11): 1129. 1943. l'oac. ttap. A4,,d. 2S 11): 533t. 19.72.

Iti1. 3-aloiiatal. Etitiltigia al poivitllttt of iii lulhullsn hiepatitis. tllf. 54. Ilara. K.. Kashiwagi. Y., and 'Isuchiss. 1. Studties ont thet

I t~a lo I~ 21(i.365.a tLausali ist-Siris of tsitlernii htepat itis. M1. Skin Itest (iml upitrlic-i-11. Vl- \lit. 0 . S.. Sitilih. 3'. \L aitld Bitsele. C.J.(IraatatliNAItal litpa~tiiis Witli alt anttigenilIteitatea fritltil ht- spleei aif mitce

hicaks ill homititi ljttutost- a itlitivts indtaia a-t is si agentt iii ~ illt tlat-i' Wsilli Outia sti-aii aof s icms. Paicr. Itta-p ad. 2Q (2): 83.tlt taiat tat iii la-ti i tatafols tilipa Ii us htltieitI s ltia ,yifaaa fI. taa) 39,153.

1217: 2S5. ItfS 5.Iaiasiii. R. I iattsiiiissiltl oif iatlectiitts ht-patitis lit itttt-biittint32. FoittiI. -1 . iI.. 'alt Rtaui-tt. C. i . atti Nagl-i, F I. 1'. Cox- jiriiaiu-* lIt littatlt,lA 1s 1 .%tivt- irett Rt-patII N.o. 26. 31. 24l).

sit kit- \11t s Iits ittlat-oiatalli ttioog itlatliclills itt.3atilis taon- Ailsaita. (wor-tgia. Cttatattiiuitalit Ilist-ase Cueiti-. 0668.

Ia~t,. (iatiid. it-al. A.4siai 1. 93: 74(1. I9615. lifi. IKt Iitait. t. W ~.. ItA ' ilt~iat. (.-SA.. -t-cr. 1.1 R. math Baioti.n'13. Frittl-i. J. F - (s 'loat;tlttloug of s itis illica hllsn (paltaiiIltl 1. ulisiitt-s. lle/alitfila Iloataala- s. II Fo-nl -ttti Hospital flter-

tI4-l-at- tol 11ii t lsit- iliirt- sltilies) . titla. i/a-i- Ali oaf,,a. S: nastiontal Ss uiiptsititt. Roastona. Little-. Biaas anda Coa., 391,7.$7.3. I'TS. 57. I ints 1 \%a--I .- V ., Low t. G . ).. and Eitela-i otk. M.i P. Silenas-

11. I-seiu. K. Wt. Pea-rsiial taitiiii -lo lliliCo. Pilifi. silus ill blooud a itt irtait cast-s of iiufrctiouts ha-Iptaili%. St-it-ut11. F~sco-i K. W.. - atit 3 aithika A.- K. on dea lr Erreger dear I pl-aIitis I 52: I 390. 19646.

Epaideutoiia als ialt-oituiihvit ga-lit-i /:/it. It ia/it. 27- 7-149. Pr19. I58. Ilist-kit i. R/. I.lt-ainhsit; itNa-pti.Sit-ic / Sat: 92. 3901.'W. E%%a-i. 1. WI.. antI Lt-itlikt, A. /trit taltiogit- tlr lIat-3tifilis FIi[ 19. Halt I, MI. If., maul Sit-tit. R. A. Victusas isollate-d Itatn achildiren

devoitn t. Z. sfAho. lit-alW. 115:57. I'-19. as-lft iiic(a-iiaai ho-aetiiiii. All.4 1n- . -flaatt-motaf.5 81 4111 1966.

370 HJERSEY AND SHAW LAliORATORY INVEIt (CATiON

60. H-asashi. K.. Kawasoe. H.. anid lakahashi. K. Epidemiiologic ccphalohepatoun'ocardifis) lccurritng ill the trewitorit piritxl

obsen rations; onl the outhIreak of( infectious hepatitis ill Ole-to. antI due ito tit fct ion with1 (;oxvAckie iitus group B. Ev itente ofcho. Nagasaki Prefecture (Japan) and anr out line of recent re- intrauterine infecttion withI this agetit. Pedialtpit3 22: 857. 11151.ports on the isolation of the infectious hepatitis %iruses (atn. 85. Kiinu ra. R.. and Hiotta. S. Expi-rineicrtal studies of irife~t tolts

tral %irus). 1Endoinir flii. Bull. Nagasaiki I tin. 6 (2): 81,t. 194. hepatitis. 'I ratsisissioti of a %i cal agent fronm hittnalt hepatit is61. Henle. W%. Ilissue culture studies relatedl to hepatitis. lit into livens of white ruice. Acta Sc/i. %led. Vi ,/t. Ajoto it0 (2):

SVnipositei al the I-ttbttrtlors- Pi opaga/.on and Deritcion of I133, 9q52.the Agent of/ Hejltatils (publication No. 322. p. 21). Washing. 86. KiSeII.a. N. V. Contribtutiotns ito (he eliology of epiiletttii hepa-ton., 1). C.. National Aca(Ieln of Sciences. National Researchs (iti it% 'Ii urktttenia. Illt A ot tacrt oif Pajwpes, IN oinlemtiatonat

Council. 1954. Congress fori Micwhaoilopr. edlitor-in-chief. Stakhanota. V. MI..

62. Henle. W. Review of antitepts at adaptatiotn of hPlipais iteirts Organi/ittg Committiee of the Conigress. p. 393. Moscow, I .S.S.R..to the chick einhrso. Ini S~vaituilo ti onl the Lii /olatoll Plo/ta- 1966.ga/ion and Detection of the A4gent of Heptititis (Puiblication No. 87. Kivachko. N. S.. atni (.alko. N. Y. Soune findinigs about culti-322. p. 2V). Washitngton. 1). C.. National Academyv of Sciences. vat ioti of itnfectious hepatitis %irus itt tis-tue t'tltures. Vlop.National Research Counctil. 1954. I'lir., 9: 735. 19164.

63. Henle. W.. Harris. S.. Hettle. G~.. Harris. I. N.. M~ake. MI. E.. 88. Kohtler. H.. atti Apodlata. J. Chter dfit- E-kermitttg %(oit Ademio-Mlangotldl. 1. antd Stokes. J., Jr. Studies ott the agent of itt e.-. v irisitifeltitittiei itt Stltweitivtiierett. Prirnairk ult urei ttiit hii etious hepatitis. 1. Propagat iotn of the agenit inl tissue ctultiute H istitlogisclser Veifahre-.t . I/. MOa . (0rig.) 144: 338. I 966i.anti in the embr~otatedl here's egg. 1. Exp. M~ed. 92: 271, 195-0. 89. Kollei. [I., Apodlara J., attd Springer. 1). Chite die titebritalige

64. Heriberg. K. Hler Kariarietttogel als Versuthist ier itt d~er Ilepa- Isitlierutg bislter btrimi %lerttshettiti nbekiamimi r Atlenimiiitei austitis Contagiusa ?orsch utg. AMill. llschr. 22: 676. 1943. Patientteti ittit Virmnstepati tis tinit H ilfte tisotlogischer V'eifalt ri-t.

65. Herilseig. K. i'beitragungyersutche ait Kaisarietisogeli itit i/it. liakt. (OP g.) 20(2: 11,. 1967."Hepa tit is-Cotitagiosa Mlaterial mrit einitns AuhIatsg). Der Kaita- W(. Kntrin lit. R. S.. andi Patttletkut. IL A5. Stud t fl fiftlie pat hogeicsi.

rieti..gCl als V'ersuch-itier. bI/i. Itakt. (010g.) / 1/: 81. 1944. its oIf irtiseq isolated foron itifettious hepatitis patients ont66. HilIlis. W%. 1). Art(tiltbreak iif ittfectioius hepatitis altioig tlt mt- t isstte cultuore. white mtice audtichick ettlruh ts. Betao. A/ps!. 4S:

paitiee handlers at a Untitedl Statesi Air Forece Base-. Allies. . 8217. 1967.Uvg. 7 7: 316. 19.61. 91. Kitwalm~kowa, J.. Pri/slvskiewvict. /., setudrawa. J.. Gectigi-

67. Hlillis. W. 1). Iiestructiont of cftitpanitee kidnley tells by~ seva ailes. J., Porwit -BIolsriwa. Z. FExpe itnieital sitidics of epkidentitfront patient-i with acute iitfeittituts hepatitis. Plt o. Soc. Exvp. Ittpat itis. I1. 1 he il. tatits otf v irein ia atnd of the des eltpitteitlhit/o. Mled. 10,8: 813. 196il. ofi atiatotiiipalithligical lesionis lin it-e Us'ers (itf titic inflected

&K. Hillis. W. 1). Viral he-patit is assisiiated with stib. hunat pt-i- with the itouse-y irilltt t 5ariaist itf the NMutotI irns. Alcta Med.mnates. Traw~fujiton ?: 445. l%63. Pot. 4: 281. 1963.

69. H illis,.U". I). Topics itt clinical titetitinit. etiotlogy iof -irial hepa - 912. Kiiwaltmkowa. -J.. Sir/utiraiva. J .. L askoiwski, A., artidl i-ittitala.tins. johit Hopkins Mled. 1. /120 (3): 176. 1967. %I1. ExperimenrttalI studilies oil hit-paliiiis epideittica. Ill. %Nltpho.

701. HilIlis. W4. D).. andl Iaig. F-. hB. The ctmldi atitat (Ii htumana ciii- logic le-siotis iii tire sple..iis andi lh itph notises (itl mice iiifcteicl

bnvonic liver cells. Exit. 0c/I I/es. 26: 91. 1402. with Motol virus. Aria %flel. Pill. 7: 191,. 1966.71. Ho~in. NI. 1). Preseutt otf %rtail '.iru-.-like particles itt sari- 93. Krugenaii. S., Ward. R., anid (iles. J. P. The natural Itisttis it

1

ous atletnt'irtis tspe 2. 5, 7 avid 12 preparm~ionts. Fed. Ptcoc. 2/ inokctious htepatitis. liner. J. MIed. 32?. 71 7. 1962.(2): 248. 1965. 94. Krugmtan. S.. Wardl. R.. Gilt-S. J. P.. I41oattvk%, 0.. Andi jatolis.

72. Hog/an. MI.I11.llatkliiw. N. R.. Rafajko. R., andl Rotwe. W. P. A5. MI. Itnfectiotus hepatitis. Illetectiiot itf tirii- during (ilie ill-(Amparat i'.e studie-s of adt-nt-assixriat-it s iriuses. Btart. Proc. 109 t colltationt pt-rind andI iii th ica lly itiappaeivt iiifeitutio.Yet1966. I-ng. J. liedl. 2b/: 729. 195.!)

7M Htioier. 1). MI.. atnd Newtonu. W.AJr. Serioius Coxsackie ittfec- 95. KIs-ibeka. V. Our knittileige abtout Xottitl %irus' isolatedl fropttutu in itt antsu andi t hiltlreti. Ms iwartihis. uevneuitgoenicpta lit k. itsfect iouis httpat iti-i. 7'. Gm tettil. 7bu: 2301. 1%64.andl hepatitis. Attie. 1. Dm/s C/i/ld. 96: 251. 1958. 96. Kuhielka. V. personial tinmmotltiiciitt, Milli.

74. Hsiting. G. W). saiclioti. P.. antI NMcol kiln. R. W%. Stutdit-s (tf a 97. Kuhtelka. V.. Salayik, K.. anth Situsek. 0. Ciritttilitioiitot it ileinrs so.. rs isolated front hutnait Ilotlit. 1. Isotlat ion antd protp- it ibh tf intft-ctious htepatit is. Pri-lintito o rt-ptiit. Zile. Ila/ierties. J. Jasttotittl. 5.: 2844. 1462. (Oltig.) 171: 4011. 1958.

75. Jeze~quel. A. W5/. atnd St-iner. .1. W..An li-clttron mticrotscotpic 98. K ubi-Ika. V.. Sltitit. E.. ;Iiin Stiliita. NM. Virus %lotitl isolaiedlstudy of tostle culturi- cells inlfectid with a tranismiissibile agenit front cases (tf htuitaui hepatitis, lei A/,%It rttt.% of Parpets - IX lIn-

isolated front hepatitis patients. Ga../rttotettioltg-s 48: 5104. 19.5. ternattttio/a Cuitigi ci fop Mtitt tietio/ugi.. (4hitor-itt -chlt i. Stak-76. JMi-quel. A. MI.. antI Steitner. j1. W%. Progit-is lit the study . itf Wialma. V~. %L. 4)rgatuilitig Cttmmttittee- it tf file tngtvss. p. 39.3.

transmnisslible agenits iisolated frontu patiet-ns with htepatitis.. fOS(OW. U.S.S.R., 1%66.Gast ruintet ohtgV it): 411, PWi ( . 99. Kur iu hi 1. Studieis otil thlt pat hiigeuu i agent oIf itt It I(tilts lulia-

77. jietjtiel. A. MI.. andl Stci-ict. I. WI. siuttie tilt rasi rutuittral atul tiltis iii ((Las ina prefemotvn. ( firitt iofetifi lIi i ntilt,- ttito...1

histiwhernical aspi-ct-i4 of (Oisat Lit- sittesi i-Il intitratt itts. Labt. 1. Med.i As sop Okeutsii tuit (7): 16153, 1917.Irn-est. Ill: 10155. 1%66. 1(81. I .pitiltitill K, Ileltatitis a-mituiated %%itlt ECH %it)yiu-, spi- 6i.

7M. Jittke. R. V.. Ki-all. 1). IW. L~eak. P. I.. Setatle%. N. F. artid Ann, .1itd tij si. /Ill/. htiti. /4: 34'2, 1966.

%vaht-rs. MI. N. I. Ilt-patitiý-iv-ttlu hahitis itn htiluaits with) leo. ]lei. - ptioliunit. K. llepatilis awntsiaedl %%ite (ll ( sit tius, ts~i 6i.%irus inifecittion .4cc/. Illicit. M5eed. lC/lictgu) 1/;: 811, 1%14. lit 4/ulmowi (Ittf Palw-u . IN hint-p ttili.ntt tuingurift, Im Me,-

Vk1 Kai haiti / . I1. (hIt. Propagat ion allt1

Ittotewtits (If lut-pat it is /uit.og)vdi-il ttt -tr-iii.Iuf. Stiakhta tiia. V. %I.. ( Itgait huig Cotrtt

Nats ti/tite 11,,tdtit) 24(8:M15, I (165. mitt e t I tf Owh Contlgresvs. Il. 3912. Mlustoitw. I SS . 11110i.

A 1. Karnon, 1 ). 1.. FEtkeit -(. G . L.a Mirt. le. L ., HIjls n-n. Y. S.. W12. I 6J I, Wil itt. F. K ie. 111(1 .,I. Pitt-, \IL Ablit ltltn , A.. attil

81, kdasul1. .. I ota. V-. artid Kttiglt. V. nfetiftitn iti f -iuillttter-s 207. 126. V16

1963. kitl L. Kwa.~ L ., 1-Iir.i P.. atil tud t- Bd .V P~ Es Itt-itii al I tital-

92. Keritn /raie. K. Fspeticitttt al I sI it s Ont pidluittti Itipa liti-i. I. its ti eptideic tIitn-pal itis,. SK Fpitr-ut 5 tit isuilaitiin it tilt- %thesit-.

-1 he rotle oIf soment % iitsime isolaed ltot-t Ith- eases. IUt/t I/ul 's 7: Vlot. Ilit% uS: 221. 1(114MW82. 1962. 10(1. I zvt.J.. Ituttt, NI. lit-p. W). Abltalt.int. A., iliutt. F., leittutt

93. Kiltiick. S. (:utre-tt -tatits of Coiukaici auth ECHOt 5 itiitse iti kmt. I.. Kavat. L.. (.itugui. P.. andt Pwdcl. A. Fspeivitttetial (in

Itanitot dJisease. Pp.igt . Siet. Voluti. h: 27, 141A4. %etst igat ittir itt u-;iduilttt it -pat itis. At-itllipts at is. ilatiitt of it844. Kiltrii k. S.. andu flelttitwhlke, K. Ssi-'%Y g-ttaIutl uise-asi It'l- l ites. %Itid Cet-toi. atItu lfui obicttltt. 111, ll 1(1W2.

V-Li [ I, No, -P. I'ioli VIRAl . .\GINIFS IN IIEPA 11115 571

Il0S l"s~. J.. Pciv. r L Ni lltp15. D" rhbcalcccc A.. Iklaici. E_. (.%okpi g i Uc limt'--tigat iccc. ccc ite.pit it ii '-1uiilcccica % us-. Bu ll. full Afar.P., Ilmccickcs. I ha iva L. aicci Iti-dc. A5. Isoclationc ofl lice 'citcc. lied. (,caccc/ Q: 37, 0ii5cx(if inflctioaus. he.pct it is Onc S:/e. S: 4.1. I 1%2. 12 ii. Nlccs i. J. I as Ici, K, a cc J uskiewit/ i,. Attemipts toc dis-

I I Ili. Liechhalrct. IT, K c cgcintc S_ (,ile%, J. P.. icd ccl It-it* gccr. 1). Ni. cclvi dl 'cli1ws ()I he.patitis. c pilclinac fiic adljitdtoit Houilds of thickRrcco c-rý ccl c tpachic agentis hocmi patienits willh cintucitions ec'uicic andcillcc tI.uc itilutcc flucid%. li ineal,.. ci like electron,hepatitis. 1ý[lioalct andc proipagationl icc culturecc I)(liil~liac clip- Hu(ilc-ccp I(P(1C lauI"Pcchilu icc,1 .tie-d. IccPop, Gdactsk 7: 45, 1956.mcid lcing fiicluhaii s% (W5I Ill). Visacc/cgl 24: W.1(S, 164. I127. ui cliicadEE. an Sc hne id eri c, I. Islclatiicn ofl canididate i

107. Lielihilies. IT. Kcingmiani S.. Nit(.regcc. D)., andc G.iles. J. P. ruse icc hepatictis. Alici Sled. 47: 17, 1965.Sc cluiii of a liiixcim ii. cs c-cc-cld fronti pat iet-s cil c in i cfctcious 1 2K. Ni iracka. I . Epc ccinicina s t udies. clit epicdemcic hepatitis. 11.hi-pacifii.1. I.olaic on u acndi Icaici cli-iac icc, J. JK-p. Aled. 122: I ccl icitm ofl mici t Iemici i i acnd temlpera rtire oic the fixed virus.

1131.,l1%.c. lap. 1. fliar1. 13 (10)): 5112, 1958.S1014. MacCall ciii F. 0. R ie.. ic ci alitcclii,.. to tc acscinit hepatitcis toc I 29. Mi icacikc. I . Expeciccetlc sii tudlies oci epicdeimic hepatitis. 111.

otiher acnnuccal itici acid sc..iev cit mecelc Eurccpeani effort% acc hum uccc-secciccgic ieactciccc ccf tilae icuiccic of epidemic hiIepatitisproipagat icc of clciisilt- ,..ti. Ici .%vccipucsc cci thi /e icLaboctaccv pitt icit%. /. Jccp. A cit. icif. Dii. 32 ill): 8(63, 1959.Mc cipcgictiic.cc told lric/cc-iicc of l/ict, Agentf cf lcpatcii: (Pualiic 1t 0i. NI ccacikic I . fxpeiiuicccc al suiceii ts ccc, epidlemic hepatitis. IV.Pioin Noc. 322. p. ")8. W~ashinigtoni 1). C;., Naticonal Acaclemvc of Hi..itliallcclccical cclii, iiicclin cclfi lce lier of mnice incciulatedlSt ielcce.. Nat ioncal Resea rch Coucilcc . 19154. withi t he' fc ixed 'cittc. io* ick,, Meld. 1. 76 (3): 139, 1959.

li0t. Nlalhai-cic- VL. Hai ivic. R.. acid Sm ithc A.A If. Aui cititliceak (i 131. O gasa~haia c McN. Sui cii it-s cicc le pathicgeniic agent ofl infectiousic~eptiii mnciicgilas amoii~aiteci ciilt ECHO1 %inc. tvpe 4. S. ..- fc. lo~patifs icc ()La'aiiia lielt-Ic t 1. M.ulcctiplication of ililectiotiqAled. 1. ;1: 1261. 197.7. hcepatitis. %im% fit. iltc c- cliii I ~cuibm)c. /. .%ed. A iscic. Okccyamca 70

110). MflcCollumc. R. WS. lI if( clcc.isi- iciccicgit agentsiftiicl ial hipit. 7): 2.397. 1111S.lilti. .4cc,. J. Pu1, lcic Uric S3c5 : 163A), 1%3. 132. ( gicalcacic. WI Stwivc on'. clilt Iicc ho geicic agent iif infecitiocus

ill. Mit Kit-. A. P. Icc %ctccmicce of Artinccc cit fthe (Cccfienricc ont t/he ict~lcaitii fic Okasacic., Jitelic ouc. IL. Iciecliccus mroutes anti thebFiacihcg cf Incfecticus Hepcuctifi. (Cccc..ctec at trie (Cccmncictia- cicctiplicato (ici llt-1c Icic tcccs icepatitis vinus fic the chicklie lt, ecs Di w(enter, Nuts 2R-251. Atlcantia. (.t-iingia . I1962 eihiclc /i. jliecdA ii . ( vi h ica 7(1 (7): 24015. I1958.

a 12 U iP MLeaci. 1, W5. I iSsue culture icc tilt isoilationi (ci (lice hepatitis 133. O'Malie . .1. Pi.. Nub-i, /. AItatile. Mi. f.. anti Harker. L. F.%irui. iii ilepcfilic I ,icfc .. . "teid Fcrdi litc-pital lInternatiomnal Icieccilica~tionc ccl lit-e AI agicc ais lipcc lhcuna guc/hisefnirumc.%%sicpcnicilc. edlited 6ci Hartmcacn, F. WS.. IciGrippic. G;.AMa- PI'c. No.d ..lcd. Acrt. 1 %A. 54: 8915.195tctir. J B_. acid Bacrrocn. J_ 1). 153. Boiston.i LittleIi. Bronic acic Coc.. 1.31. O'Mlt Ic. J. 1 '.. Nle~ci. I. Mi., Jr.. a ccc Sitaiclel J. E. Antibcody19157. inc Iccpatitis pat ienct% s aiciiic a ciecil isilateil sirici. Ploc..Nsac.

Ill McI caoc. 1. W5.. Jr. Eticilcgict cccl is ccf itifec Iiccnt acid serum Exp. /i l.cc Sled IllS: 200,i I 11051.hepac it.. Pc-ccPec .Ii, 1 c. 1 #).; 11(4. 1963. 131. Occiticka. R. Situdies onccirce susitpifrilcit ofr ici Itis ro hepatitis

114. Nh I ia c. I. W5 ,1. Jc-(liii ic signcificaccie ccl cectillt iciscitigaici s itch. 5 i ii .I itc iii. c- i cccic sicc acid delttic ciciat icic of inf ection,ifli hc1 itiis c ir cis. Pcccgca c MlSed. 15: 491(. IPloct. tilter. /. Med. .4 ....r. Okauui a cicctPQ (1): 191,. 19(57.

111 Nit-I a. B . andc aIincg. 1). 1. Imclxst citcosis. Supplessicnini 1cI36. ((cccukia. R. Srudiiiits cif lilt- scsicept ililit if fow lsc. cit hepatitislitric asxicc tie "isih mut 11cc itilci tilc.it-paliti... S~iryute 1 55: 90, s ni. 11. 1-speiitcccc-s ccf chrociic itiftecticci. J. lied. Aooc. Oka.[9%7. cccacby (1

5 ): 213, 1957.

1I Ili. Nh cit I. G.. S. Ricci ccl cc ticclpt.. to pci pagale thii ciltlo iiicl l- 37. I) Slitughousccci. WS. J., acid Blueclier. H. A.- Hepat itis assxacitteI

macn hepaticIII i lctcni cc.. Ini S)Ppo m an/ccicc cilt/c- Iabiaco pfcl cictht a ICcxsackLie 131, s irds infeciticon (Tucing late pregtcanc).Prccpagalczccc andcc fllit ec Iicc cf f/he A4gentf c ilcfccctfiti (Pibiccia - 1. .4. . .4. 7y 7 I1. 1%62.thon Noc. 3,22, p.

37). 55ahinlc ncc, 1). C.. Nactiontal Acicieniv ccf 138. Park%. W5. 1.. V~iss WI. R., acid Nielcick. J. L. Hepatitis studies

Siccic ci. Nat iconal Research ICounicil . 19141. tli main~ ticiset s. Barct. Mac. 150,. 1%8(.W1. Ni lit cci. I).. Bal-hi. C_. Catjal - N.. TDtiet enecci. R.. lcciestc c. -F., 1-39. Penttl. 0 . oilut tiel cicclccg cc infectious hepatitis. Khic. Med.

acid[ Pcpit. Ni. Fticiiogic i isc-t igait iic. icc cpt-1 ic ii hpac itis. I. l~ifec) 2: 7995. 14(47.S~t cc Isc of ,cc som itum ci'so.iic l ccin icccc-l cccfl ie.. fronti ipicic-ciic 1401. Procs-, P. H.. Balk.- S. D).. I ichihalcer. If., arid Krugmai. S. Stud-hcvpatii car stc-,s. Si ccl. Cc c.Icfec ,cc cc/udIa I?: M95. 1%62. scs. clt a 11 cxmiiiiirecic etc-cl fronti patcients with inf ectious

II1R. miit icic. (5.. Kariumc C .>(aja I. N*.. DlciciIcimmuce - f.. Icciescci - .. hi-patici... It. I-icne sit tccturtr acid vlectsncti circcscopic demcon-acid Pccpa. Ni. (cinipicauste sc-cclccgii al vit clits oflNiciswclcc ;tra in stricticc ccl inctccirccplucsmic intrncalI compimienec and viral lila-K.3 acid ..icaicc iscclactec frtcci piatientt of epidemtiic hepatitis icc mcecci formationi /. iulc. lred. 122: I1151. 1%65.oiur ccccc cim. Sf cccl. Cc-eec!. Iccfcc ccamrc cc/c. /b: 115. 19(61. 141. Pricikucc c.ickca - N. B. acnd Ncc'cikos a. N. a'. Properties of %iruses

1191. Nh rccicc. 0.. acid( Icctes-i T. r.\Iteracctiontif cc eciitai pripeccic~s (if whcichc were sc-paciated frontc t ire feces ol puatients with infectcoustlct- BF- itrcain (isoilactedc frccn a ca cif ct ipiclecit ihcpat iris) inc (lit- hepitilivi. Iap V'~Iicili /0. 62,2. I 1965.icoucrse(if adaiptaticn icc ii icaci emiccic ccccli clI cult-r. Re,. Rict. 142. R ilk icci. R.Aacid Mocrgan.c C . 'The hepatitis viruses. Ailed.Pyaccia ticcfcncciceo/cicc. 2: 45. 1965l. Clic .N.4 ccer. 47: 7759. 15(63

1 20 Mit roinc. 0.- acid Toccincc. -1. Ccccipccu isc- %e-rccligic ec-c-wccIcc-s 143. Righlitl. W. A.. Kcltsich, R. A. I'afor. A. R.. Bogi~s J. D., acidtin strincIic K3 Nicsccc andcc it-e icraicis isola~ctedc in tlire Rioumanciani MtcIcat. 1. WI.. ~Jr. Statuc. cepsiri cn risicie-cultr nr cultivatedlPc-ccplc-st Repiclii fronti epidicicc ihccpatcit is patic-nis. Revc. Retc- hepicicit %ir sici. 1. ascclccg.. lalioratcrv sctucdies. 1. A. AT. A.- 177:cccanicidacfccceccc 2: 121. 1965i. 671. 1%(1.

121. Nhc cirii, 0.. Icicii-cut. 'T., an ccl cpat. NM. Icccpe~tic-iv (cl the BF 144. tiglitwic W. A.. Ki tst It R. A. I~ ikichaný. M.-acid MicLean,%Iraicc imiclaited frocm a cast ccl ipic liciic Iccpacitiis in ci( 1coursic I. W. ]'.J I ltc-itilt cclitecc cult is aticic of cvccipathtigeic agentsccf adlaptaticon in iccranii c-civiccccci i-i-l c cnicicis. Arfccc Ceccef. friom patriet,.c cithl c liccc l heipatitis. Vifecice 124: 226. 1956.lccfraccicccclciccl. /6. 51. 1%15. 145. Sabin. A. It. Kiinctictgil E. R.. aicc Sligictd. R. ECHO type

122. Mf crnis, j . A. ReporIcIt cc I ic ic- cclI% a Icc-icvI 11its % cc atdIap)t I Ifcc%- incise 9i s ics cliiscit a ccclccgi ills icctricllei clinccial and epidecnio-

ccl human~ c hiecatit is icc A sIit I ti-ifc cc~-s ccci 11c-cc ic ancimaIs. Tit lo gic observai t cciin dintic g 19t1 7 hIc liiecccic icc Miliwaukee withýjmooii a Iifl~ptial"P ad h-e'iaI f iles cciccliidc cccten sioutar c iwascese aiscciateci with (ox~cscakic

tice A4genc ct a liepfaitiic c I i'cll catt cciNc. 322. p). 76). SWashing- a4n. ofiiti ECi O %trusIts . .4i r t. 1.c lhit 11(Chil-f . ci nihd. 9:17 98

lcd, 1 ). C.. Nat iccnal Sc ac cii cl ofSic ccc -. Natcional~ I tcst-arch V.F..I cciti, c-cdti ccc,,cci tacidi slung isils with emnphasis cn an(tiuncil. 19154. acgent cf Ic cli ;I iawc cc(if c iccici hpicictiic. 1. Lab. C/itc. Sled. 60:

123. Morris. J.A. Putumac~l ccltomluidil ittic. V967. Il016. 11162.124. Mlorris. A.. FhlKIsi-g B. I.. PccRI. W5. I . acid S%(Ic-id. P. A 147. Schitt. V-.. Bc-clotcc. ft.. lijsici. Ni. andi Scarek. Mi. Pathogenesis

Hi-patitiis asuicc tI-le cill c \c I c isci %filsi. group~ A, i pe I. .New ccf liict i atis i c he %ItrcitsNcc% c iccni icc-. T. Ga.vlroen,il 7: 2t9.Fricg. I. Sled. 267! 12301. 19162. 1%(4.

125. Moccn~ka. M., acid laclcci. AS At tempt~s cc uisc miticin fcltroici 6 Its1. Segagtti- K. V.5,tAcimii X .. ancii Nigric. N. Itimmunoclogical and

572 IJERSEY AND SHAW f.A!5ORATORY 3 NVrSTIOATION

irologacal studies an infectious hepatit is. !lfinens-a Pedant. I � I Sit Stud an.s, II. I-. 3x3 ,eiiitu-tttal soil spcsmtalseo�as i cal bria iii is262. 1961. iii primaws. liii,. I.nest. II: ISti4i. 1965.

349. Sengoku. A. Studies on the epidemic hepatitis sirus. j. Med. l�7. Smnetatta, H. F. The histopashology of the hepatitis epidemic ittAssoc. Okayavna 72: 229. 3959. Acera. (.hatta, 1962- 3963. Lupulslished obsersations, cited in

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