Blood, Vol. 52, No. 2 (August), 1978 261

BL(IX)DThe Journal of

The American Society of Hematology

VOL. 52, NO. 2 AUGUST 1978

M onocyte-mediated Inhibition of LymphocyteBlastogenesis in Hodgkin Disease

By Geraldine P. Schechter and Frances Soehnlen

Mononuclear leukocytes isolated from theblood of previously treated patients with

advanced active Hodgkin disease con-tained high concentrations of monocytesand showed poor lymphocyte blastogene.sis to mitogens. In five of eight patientswith disseminated disease, blastogenesisbecame normal or improved markedlywhen the leukocyte suspensions were de-pleted of monocytes before culture. Addi-tion of autologous macrophages to themonocyte-depleted lymphocytes resulted

in a reappearance of the inhibition ofblastogenesis. Monocyte inhibition wasassociated with the presence of activedisease, lymphocytopenia, and lowlymphocyte/monocyte ratios in the periph.eral blood. The role of previous treatmentis uncertain, since inhibition tended todisappear when the patients were re-treated. Inhibitory monocyte-lymphocyteinteractions may be one of the causes ofimpaired cell-mediated immunity inHodgkin disease.

M ANY LABORATORIES have found in the last decade that monocytesand macrophages potentiate the lymphocyte blastogenic response to

mitogens and antigens in both human and animal lymphocyte cultures.’ Re-

cently, however, there have been a number of reports of suppression of lymph-

ocyte transformation by macrophages or adherent cells from animals thought

to have “activated” macrophages. Rodents bearing tumors, undergoing graft-

versus-host (GVH) reactions, or treated with BCG or Corynebacterium parvum

have been found to have “suppressor” macrophages.68The present report describes evidence for analogous monocyte-mediated

suppression of lymphocyte transformation in some patients with advanced

Hodgkin disease. Severe impairment of lymphocyte blastogenesis to phyto-hemagglutinin, pokeweed mitogen, and antigen could be reversed by depletionof monocytes from the mononuclear cells of these patients.

From the Veterans A dministration Hospital and the George Washington University School of

Medicine, Washington, D.C.

Submitted November 9. 1977; accepted March 21, 1978.

Supported by the Medical Research Service of the Veterans Administration.

Address for reprint requests.’ Geraldine P. Schechter. M.D., Chief, Hematology Research Section,

Veterans Administration Hospital. 50 Irving St. N. W., Washington, D.C. 20422.

© 1978 by Grune & Stratton, Inc. 0006-4971/78/5202-0401$02.00/O

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262SCHECHTER AND SOEHNLEN

MATERIALS AND METHODS

Patients and controls. The leukocytes of I 2 adults with Hodgkin disease and 12 healthy adults

were studied. The extent of disease in the patients was determined according to the Ann Arbor

staging classification.9 Nine of the patients had advanced Hodgkin disease (stages IIIB, IVA,

JVB), and three had localized disease (hA, IIB, and a late marginal recurrence after radio-therapy in a stage IIIB patient). (The patients are listed individually in Table 2 and are denoted

by the same numerals in the later tables.) The patients with advanced disease had been previously

treated with radiotherapy and/or chemotherapy. The interval between the completion of any

previous therapy and the time of initial study ranged from I mo to 3 yr. the median interval

being 5 mo and the average 10 mo. All but one patient (No. 9) had active disease at the time

ofstudy. Two patients (Nos. I and 3) were studied during periods of both remission and relapse.

The patients gave informed consent for the studies: the normal subjects were hospital em-

ployees who were paid volunteers.

Each ofthe patients was skin tested with 0.1 cc ofat least two of four antigens, including mumps

antigen (Lilly), streptokinase-streptodornase (Lederle), Candida albicans (Hollister-Stier), and

intermediate strength PPD tuberculin (Connaught). All of the patients with advanced disease

and two of the three patients with localized disease were anergic to these antigens.

Preparation ofleukocyte cultures. A heparinized blood specimen from each subject was divided

into two parts. One part was incubated with carbonyl iron powder (General Aniline and Film,

Dyestuffand Chemical Division, Linden, N.J.), 150 mg to 10 cc blood at 37’C for 60 mm. There-

after the two portions of blood were treated identicalLy. The mononuclear cell fractions from

each were isolated by Hypaque-Ficoll density gradient centrifugation using the method of

B#{246}yum.10 The monocyte-rich lymphocyte suspensions are denoted in the tables as Hypaque-

Ficoll (HF) cells and the monocyte-depleted lymphocyte suspension derived from the carbonyl

iron�treated fraction are called the iron Hypaque-Ficoll (FeHF) cells. Monocyte-depleted

lymphocyte suspensions were also prepared by passage of leukocyte-rich plasma over glass bead

columns as previously described.3 Differential counts were done by phase-contrast microscopy

using a hemocytometer. These counts were checked in selected instances by examination of Wright-

Giemsa- stained smears and preparations stained for nonspecific esterase activity using a-naphthylacetate for the substrate.11

The stimulants incubated with the cells included phytohemagglutinin (PHA) I ,�g/ml (Bur-

roughs Wellcome, NC.), pokeweed mitogen (PWM) 40 �g/ml (Grand Island Biological, Grand

Island, N.Y.), and streptolysin 0 (SLO) 0,010, (Difco Laboratories). The monocyte-rich and

-depleted cultures were adjusted to contain the same number of lymphocytes. The PHA-stimulated

cultures contained 200,000 lymphocytes/well. Cultures with other mitogens contained 400,000

lymphocytes/well. In certain experiments, cells were irradiated with a cobalt ‘y source as pre-

viously described.3 The cells were cultured in flat-bottomed microtiter plates (Falcon, Oxnard,

Calif.) in 10#{176},autologous plasma and 90#{176},RPMI-1640 with HEPES buffer (Grand Island Bio-

logical) in a final volume of 200 zl. The plates were sealed with a plastic adhesive cover and in-

cubated at 37’C. The culture period depended on the mitogens used: PHA cultures were incubated

for 3 or 4 days, PWM and SLO cultures for 4 or 5 days. For the final 4 hr of incubation I �.sCi

of 3H-thymidine (5 Ci/mole) (Amersham-Searle, Arlington Heights, Illinois) was added to each

culture. The cells were then deposited on a glass fiber filter disc with a multichannelled cell har-vester (Model M-12, Biomedical Research Institute, Rockville, Md.), and scintillation counting

was done.

Experiments were performed in triplicate, and individual cultures were generally well within

10#{176},of the mean. Cultures selected for microscopic examination were grown in wells containing

a glass cover slip that was removed at the end of the incubation period and stained with Wright-

Giemsa.

The ability of the lymphocytes in the monocyte-rich and monocyte-depleted suspensions to bind

sheep red cells (E rosetting) was examined by the method of Bentwich et al)2 using heat-

inactivated AB serum (Grand Island Biological) and an overnight incubation at 4#{176}C.The

absolute peripheral blood lymphocyte and monocyte levels were calculated from the white blood

cell count (Coulter Counter, Model S) and the percentage of lymphocytes and monocytes de-

termined from a Wright-Giemsa-stained smear. The data were analyzed according to the t test

for independent samples.’3

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Table 1. Monocytes (%) in Mononuclear Cell Suspensions Isolated from Peripheral Blood

Subjects No.

Monocytes (%)

HF FeHF

Normal controls

Hodgkin disease

Active

Advanced

localized

Inactive

12

8

3

3

12.6 ± 3(6-16)

38.5 ± 4.3 (23_61)*

16.6 ± 5.8 (10-20)

19.8 ± 4.5 (14-25)

1.3 ± 0.5 (1-2)

5.5 ± 5.0 (1-14)

2.0 ± 1.0 (1-3)

1.8 ± 1.0(1-3)

by Hypaque-Ficoll centrifugation

incubation of whole blood with

RESU ITS

MONOCYTE INHIBITION IN HODGKIN DISEASE 263

Mean ±SD, range in parentheses. HF, mononuclear cells prepared

of whole blood. FeHF, mononuclear cells depleted of monocytes by

carbonyl iron powder prior to Hypaque-Ficoll centrifugation.

*p <0.001 compared to normal controls.

Characteristics of the Isolated Leukoc�’te Suspensions

The percentage of monocytes in the mononuclear cell suspensions from theHodgkin disease patients and normal controls is shown in Table 1. These per-centages were determined by phase-contrast microscopy differential counts of

the cell suspensions cultured in the experiments reported below in Tables 2 and

3. Patients with active, advanced disease had the highest mean concentration ofmonocytes (39#{176}c)) in the Hypaque-Ficoll leukocyte suspensions, which wassignificantly different from the mean (l3#{176}.,�)of the normal controls (p < 0.01).

Table 2. 3H-Thymidine Incorporation of PHA-stimulated Monocyte-rich and

Monocyte-depleted Lymphocyte Cultures

Patients’

3H-Thymidine Incorporation (DPS)t

Monocyte-rich Monocyte-depleted Inhibition Ratio(

Advanced disease

1. IVB relapse 100 4450� 44.5

2. IVA relapse 170 3110� 18.3

3. IVB relapse 200 770� 3.9

4. IlIB relapse 470 2790� 5.9

5. IlIB relapse 1050 5290� 5.0

6. IVB relapse 850 760 0.9

7. IVB relapse 1960 2530 1.3

8. IlIB relapse 3100 2720 0.9

9. IVB remission 2070 3075 1.5

localized disease

10. IIIB marginal recurrence 2970 2320 0.8

11. IIA untreated 5230 4940 0.9

12. IIB untreated 4240 2780 0.7

Normal Controls(n = 12, mean ± SD) 4600 ± 2690 5310 ± 2050 1.4 ± 0.7

*patients 2, 5, and 7-10 had nodular

Hodgkin disease.

tDisintegrations per second. 3H.thymidine incorporation of cultures without PHA was 60 DPS in all casesand 20 DPS in 85% of the cultures.

�Ratio of the 3H’thymidine incorporation of the monocyte-depleted culture to that of the monocyte-rich

culture.

§3H-thymidine incorporation of the monocyte-depleted culture was significantly greater than that of

monocyte-rich culture.

sclerosis Hodgkin disease; the remaining had mixed cellularity

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264 SCHECHTER AND SOEHNLEN

Table 3. Consecutive Studies in Three Patients with Active Hodgkin Disease and MonocyteInhibition of PHA-stimulated Lymphocyte Blastogenesis: Correlation With

Monocytes (%) in Monocyte-rich Cultures

Patient Exp.’ Monocytes (%)

3H.Thymidine Incorporation (DPS)

Inhibition ratioMonocyte-rich Manocyte-depleted

it 1

2

3

4

5

6

26

61

56

25

44

56

170

100

150

2050

140

50

2370

4450

3450

2030

3450

5170

13.9

44.5

22.8

1.0

24.6

103.4

2t 1

2

3

4

45

43

32

36

170

210

1300

1360

3110

1850

2630

1550

18.4

8.8

2.0

1.1

4t 1

2

3

23

32

54

470

1530

1260

2790

7080

2040

6.0

5.6

1.6

*These experiments were performed during a 4-9-mo period for each patient. The percentage of

monocytes in the monocyte’rich Hypaque.Ficoll-separated mononuclear cell fraction is given. The in-

hibition ratio is the PHA-stimulated 3H-thymidine incorporation of the monocyte-rich culture divided by that

of the monocyte-depleted culture.

tPatient 1 was in untreated clinical relapse during experiments 1-3. He was then treated and had a

short-lived response at the time of experiment 4. Experiments 5 and 6 were done when the patient relapsed

again. Patients 2 and 4 had active disease at the time of each study. Experiments 3 and 4 in pa-

tient 2 were done 3 mo after local radiotherapy and 1 mo after combination chemotherapy. Patient 4

had received combination chemotherapy 1-3 mo before each study.

Patients in remission or with localized disease had only a slightly higher mono-

cyte concentration (mean 18%) than the controls. Treatment of blood with

carbonyl iron before Hypaque-Ficoll centrifugation consistently depleted the

monocytes in the cell suspensions of both the patients and the normal controls.

Passage of leukocyte-rich plasma over glass bead columns also depleted mono-

cytes to less than 2%.

The phase-contrast microscopy differential counts of the leukocyte suspen-

sions correlated well with differentials based on Wright-Giemsa-stained smears

and preparations stained for nonspecific esterase activity. Leukocyte suspen-

sions from patients with advanced disease also contained occasional myelo-

cytes, atypical lymphocytes, and cells resembling promonocytes. These cells,

however, were not depleted by carbonyl iron treatment. The cells that were

depleted in all cases were nonspecific esterase-positive cells or mature mono-cytes. The numbers of T lymphocytes determined by E rosetting in the mono-cyte-rich and -depleted leukocyte suspensions from the patients were notsignificantly different in eight experiments (mean ±SD: 36% ± 19% and39% ± 19%, respectively.) However, the mean percentage of T lymphocytes in

the patients was significantly lower than the percentage of T cells in a group ofnormal controls (58% ± 10%,n = l5,p < 0.01).

The survival of lymphocytes in the two preparations was assessed by countingthe number of lymphocytes that excluded trypan blue after being cultured for 3days without mitogens at 37#{176}Cand comparing that number to the original

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MONOCYTE INHIBITION IN HODGKIN DISEASE 265

number of lymphocytes planted. In an experiment done in triplicate with the

cells of a patient (No. 1) with advanced Hodgkin disease, there was no differ-

ence between the percentages of lymphocytes surviving after 3 days in the

monocyte-rich cultures (26%-30%) and in the monocyte-depleted cultures

(28%-31%). However, the number of viable lymphocytes from the patient’s

cultures was halfthat ofthe cells ofa normal subject (53%�6l%).

Lymphocyte Blastogenesis in Monocyte-rich and Monocyte-depleted

Leukocyte Cultures

The effect ofremoval ofmonocytes on blastogenesis was studied. The patients

with advanced disease generally had depressed lymphocyte transformation in

response to PHA in the monocyte-rich cultures (Table 2). However, monocyte

depletion with carbonyl iron was associated with a marked increase in 3H-

thymidine incorporation by the cultured mononuclear cells from five of eight

patients with active advanced disease. In four of the five patients the incorpora-

tion rose from being severely impaired to levels within the normal range. In con-

trast, there was no difference between the thymidine incorporation of the

monocyte-rich and the monocyte-depleted cultures from patients with localized

or inactive disease or from normal controls. The highest inhibition ratio (ratio

of3H-thymidine incorporation of monocyte-depleted to that of monocyte-rich

cultures) for a normal control was 2.6, and the median for the normals was 1.0.

Monocyte depletion had no effect on the 3H-thymidine incorporation of the Un-

stimulated cultures from either the patients or the controls.

The PHA response of the patients’ cultures was also assessed morpho-

logically. The monocyte-rich cultures from patients 1 through 4 contained

many macrophages, few lymphocytes, and only occasional blasts (Fig. 1). In

contrast, the cultures prepared after monocyte depletion showed masses of

lymphoblasts and few macrophages, similar to PHA-stimulated monocyte-rich

cultures from normal individuals (Fig. 2).

Longitudinal studies of the lymphocyte reactivity of the patients showing

monocyte inhibition were performed (Tables 3 and 4). During periods of active

disease the studies showed high monocyte concentrations in the mononuclear

cells prepared without carbonyl iron pretreatment and frequently showed

evidence of monocyte inhibition (inhibition ratios >3) of PHA-stimulated cul-

tures. After chemotherapy and during periods of remission, monocyte concen-

trations in the mononuclear preparations tended to decrease and evidence of

inhibition to disappear. However, in not all patients was a direct relationship

between the monocyte concentration and inhibition shown (see, for example,

patient 4, Table 3), suggesting that a high concentration of monocytes was not

the sole requirement for blastogenesis inhibition. Inhibition of PWM- and SLO-

stimulated cultures was also seen and reversed with monocyte depletion. A

representative experiment from patient 3 is shown in Table 4.

It is well known that patients with widespread Hodgkin disease often have

lymphocytopenia’4 and monocytosis.’5 Therefore, the peripheral blood lympho-

cyte and monocyte levels of the patients with monocyte inhibition were com-

pared to those without (Table 5). Both the lymphocyte levels and the lympho-

cyte/monocyte ratios were significantly lower (p < 0.025 and p < 0.01,

respectively) in the patients who showed monocyte inhibition than in those who

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- -�

--

‘:.

Fig. 2. PHA-stimulated monocyte-depleted Ieukocyte culture from patient 1. Masses of Iympho-

blasts and rare macrophages are seen. Wright-Giemsa. x 630.

266 SCHECHTER AND SOEHNLEN

Fig. 1. PHA-stlmulated monocyte-rich leukocyte culture from patient 1. Note increased num-bers of macrophages and the paucity of lymphocytes or lymphoblasts. Wright-Giemsa. x 630.

-� : , - #{149}-#{149}-.

H ‘#{149}�‘

#{149}:-� �

� �

1:.’ ‘�

a -

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MONOCYTE INHIBITION IN HODGKIN DISEASE 267

Table 4. Activity of Hodgkin Disease and Monocyte Inhibition of Mitogen- and

Antigen-stimulated Lymphocyte Blastogenesis

atient No. 3’ es (%)

3H-Thymidine Inc orporation (DPS)

P Monocyt PHA PWM SIO Control

(1) Remission HF�

FeHF�

20

2

1390

920

1040

780

570

210

30

7

(2) Relapse HF

FeHF

56

2

200

770

40

1350

40

640

4

7

(3) After Therapy HF

FeHF

18

1

760

900

-

-

-

-

8

3

*The first study (1) was one of two that showed similar results. The patient had last received chemo-

therapy 30 mo previously. Relapse occurred 6 mo later, when study (2) was done. Study (3) was done 2 mo

after a 5-mo course of combination chemotherapy, at which time there was no evidence of progressive

disease.

tSee footnote to Table 1 . HF, monocyte-rich, FeHF, monocyte-depleted mononuclear cells.

did not. Although a moderate absolute monocytosis was generally noted at the

time of monocyte inhibition, the mean level of monocytes was not significantly

higher than when monocyte inhibition was not found. Therefore the most

relevant index of monocyte inhibition appears to be the ratio of lymphocytes to

monocytes in the circulation.

A number of experiments were undertaken to ensure that the monocyte in-

hibition was not due to trivial reasons, such as sequestration of stimulant by

macrophages, and to confirm that the suppression was mediated by monocytes.

Increasing the PHA concentration from 1 to 100 zg/ml did not improve the

blastogenesis of the monocyte-rich cultures in the patients with reversible sup-

pression. Decreasing the number of cells per culture while holding the lympho-

cyte/monocyte ratio constant did not change the 3H-thymidine incorporation in

three experiments. Depletion of macrophages by another technique, passage

over glass bead columns, also resulted in monocyte-depleted (<2%) lymphocyte

suspensions with an increased blastogenic response to PHA.

In an effort to rule out “overcrowding” of the cultures as a cause of monocyte

inhibition, the role of monocyte concentration was tested. Mixing the monocyte-

rich and -depleted cell suspensions so that the monocyte concentration was

halved while the lymphocyte concentration remained constant still resulted in

Table 5. Peripheral Blood Lymphocytes, Monocytes, andLymphocyte/Monocyte (L/M) Ratio

Monocyte Inhibition’ Lymphocytes/al Monocytes/tI I/Mt

Present 590 ± 230 1220 ± 380 0.6 ± 0.4

Absent 1290 ± 600 870 ± 720 1.9 ± 0.9

p value <0.025 NS <0.01

*3H.thymidine incorporation of PHA-stimulated monocyte-depleted cultures three or more times greater

than that of monocyte-rich cultures. Mean ± SD of peripheral blood absolute lymphocyte and monocyte

counts and lymphocyte/monocyfe ratio is given for five patients at the time of six experiments showing

monocyte inhibition and ten patients at the time of ten experiments without monocyte inhibition.

tl/M (mean ± SD) for seve’s normal controls was 4.4 ± 0.9, which was significantly different from bothgroups of patients (p < 0.001).

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268 SCHECHTER AND SOEHNIEN

Table 6. Effect of Addition of Adherent Cells on PHA

Response of Monocyte-depleted Leukocytes

Cells

Monocyte

Inoculum perCulture

3H-Thymidine

Incorporation(DPS)

Patient 1

HF’ 300,000 60

FeHF’ 2,000 2010

FeHF + adherent cellst 42,000 830

Patient 4

HF 265,000 1260

FeHF 2,000 2040

FeHF + adherent cells 30,000 710

FeHF + adherent cells 100,000 770

Normal Control

HF 20,000 3290

FeHF 4,000 4460

FeHF + adherent cells 34,000 4440

FeHF + adherent cells from patient 4 34,000 3190

lymphocyte number was constant at 200,000/culture. 3H-Thymidine incorporation of the allogeneic mix-

ture without PHA was 13 DPS.

‘See footnote of Table 1 . HF, monocyte-rich, FeHF, monocyte-depleted cells.

tAdherent cells (>99% monocytes) were prepared by allowing HF cells to adhere to a plastic surface for

1 hr. after which the nonadherent cells were washed free. The adherent cells were harvested by scraping

cells off the surface with a “rubber policeman.” Adherent cells are autologous, unless otherwise stated.

marked inhibition of the PHA response. The inhibition persisted after irradiat-

ing the monocyte-rich cells with 3000 R.

The effect of the addition of purified macrophages to monocyte-depleted

lymphocytes was tested. Preparations of adherent cell consisting of >99%

macrophages from patients 1 and 4 reduced the PHA-stimulated 3H-thymidine

incorporation of their autologous monocyte-depleted lymphocytes by greater

than 50% (Table 6). On the other hand, adherent cells from a normal indi-

vidual had no effect on the proliferation of his autologous lymphocytes at

similar lymphocyte/macrophage ratios. Furthermore, the adherent cells of a

patient whose macrophages inhibited his own lymphocyte transformation had

little effect on the response of normal lymphocytes (Table 6).

DISCUSSION

Impairment of lymphocyte blastogenesis of patients with Hodgkin disease

has been known for more than a decade,’6’8 but the mechanisms involved are

not well understood. Twomey et al.’9 noted that mononuclear cells from pa-

tients with Hodgkin disease suppressed the response of normal lymphocytes

in the mixed leukocyte culture. This effect was at first attributed to a T sup-

pressor lymphocyte; however, experiments recently reported suggest that the

suppression is mediated by monocytes.2#{176} In the present study we have shown

that leukocyte suspensions prepared by density gradient centrifugation of the

blood of patients with symptomatic advanced Hodgkin disease contain high

concentrations of monocytes that inhibit the lymphocyte blastogenic response to

PHA, PWM, and streptococcal antigen. In five of six patients with severe im-

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MONOCYTE INHIBITION IN HODGKIN DISEASE 269

pairment of blastogenesis in monocyte-rich cultures, there were striking in-

creases in 3H-thymidine incorporation and morphologic blastogenesis when the

leukocyte suspensions were depleted of monocytes prior to culture. The

monocyte-rich and -depleted leukocyte suspensions did not differ in lymphocyte

viability or in number of E-rosetting lymphocytes. Marked inhibition was

noted even in mixtures of monocyte-rich and -depleted cells where the number

ofmonocytes was decreased by half, suggesting that nonspecific inhibition due

to overcrowding was unlikely. Irradiation of the monocyte-rich population did

not affect the inhibition, suggesting that it was not dependent on a proliferative

event. Finally, addition of isolated macrophages to autologous monocyte-

depleted lymphocytes again caused inhibition of blastogenesis.

The inhibitory effect of isolated Hodgkin disease macrophages even at low

monocyte/lymphocyte ratios suggests that the phenomenon may be due to a

qualitative difference between these cells and normal macrophages, perhaps be-

cause of activation. Although recent evidence favors the idea that the Reed-

Sternberg cell is of macrophage origin,21’22 the morphology of the circulating

monocytes does not suggest that they stem from the malignant clone. Activa-

tion ofpatients’ monocytes by the systemic disease seems a more likely basis for

the difference from normal cells. Increased macrophage activation measured by

a number ofparameters has been reported in patients with Hodgkin disease.2325

Alternatively, the inhibitory effect of the Hodgkin disease monocytes may be

primarily quantitative. The patients’ lymphocytes may have an increased sen-

sitivity to the high number of macrophages introduced into the cultures owing

to the altered peripheral blood lymphocyte/monocyte ratio associated with ad-

vanced disease and extensive previous treatment. Previous treatment alone is

unlikely to cause this phenomenon, since inhibition tended to disappear with

treatment of recurrent disease.

The fact that the PHA response of normal lymphocytes was not affected by

similar numbers of autologous macrophages or Hodgkin disease macrophages

also favors the idea that abnormal lymphocyte function plays a role in mono-

cyte inhibition. We have preliminary data that high concentrations of normal

macrophages (three times higher than that required to suppress the PHA blasto-

genic response of Hodgkin disease lymphocytes) can inhibit antigen-stimulated

blastogenic responses of autologous normal lymphocytes. Laughter et al.2#{176}also

have shown that large numbers of stimulating mononuclear cells inhibit re-

sponding allogeneic cells in the normal mixed leukocyte culture, which they

ascribe to a monocyte effect.

A recent finding of Goodwin et al.26 may account, at least in part, for the

monocyte inhibition noted here. They found that the hyporesponsiveness of

PHA-stimulated lymphocytes from Hodgkin disease patients could be reversed

by prostaglandin synthetase inhibitors and that increased levels of prostaglandin

E2 (PGE2) were present in the cultures. When glass-adherent cells, presumably

monocytes, were removed, prostaglandin production decreased. We also found

large levels of PGE in the unstimulated monocyte-rich cultures of patient No. 1

that increased further after stimulation with lipopolysaccharide.27

The relevance of the monocyte suppression in vitro noted here to the well-

documented deficiency in delayed hypersensitivity28 of patients with Hodgkin

disease is intriguing. All of the patients with suppressive monocytes were

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270 SCHECHTER AND SOEHNLEN

anergic, but a number of our patients who did not exhibit monocyte suppres-

sion in vitro were also anergic. Furthermore, we found that monocyte suppres-

sion is not limited to Hodgkin disease. We have shown suppression of the re-

sponse in vitro to tuberculin in monocyte-rich cultures from patients with active

tuberculosis who retained delayed hypersensitivity to tuberculin �29

Recently, Markenson et al. also reported a similar phenomenon in vitro in

patients with systemic lupus erythematosus; however, they did not correlate

their findings with the patients’ capacity to mount delayed hypersensitivity re-

actions.30 Similarly, Zembala et al. also recently showed adherent cell inhibition

of lymphocyte transformation from patients with advanced cancer.3’

It is clear that cellular interaction between monocytes and lymphocytes must

be evaluated further before impaired lymphocyte blastogenesis is attributed

solely to an intrinsic defect of the lymphocyte or macrophage. Moreover, the

aberrant cellular interaction noted here could be responsible for, or be a resulL

of, impaired immunity in Hodgkin disease.

ACKNOWLEDGMENT

We thank Dr, Joost J. Oppenheim for his critical review of the manuscript.

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1978 52: 261-271  

GP Schechter and F Soehnlen diseaseMonocyte-mediated inhibition of lymphocyte blastogenesis in Hodgkin 

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