Decreased lymphocyte blastogenic responsesin patients with multiple basal cell carcinomaPatricia L. Myskowski, M.D., Bijan Safai, M.D., andRobert Alan Good, Ph.D., M.D.New York, NY

Fourteen patients with multiple basal cell carcinomas wereimmunologically evaluated by measuring lymphocyte blastogenicresponse to concanavalin A. Patient responses were compared to theage- and sex-matched controls. There was a statistically significant(p < 0.02) reduction in the mean peak lymphocyte stimulation in thepatient group. The possible etiology and significance of these findingsare discussed. (J AM ACAD DERMATOL 4:711-714,1981.)

Although skin cancer is the most commonmalignancy in Caucasians, the role of impairedcellular immunity as a possible contributing factorhas received attention only recently. 1.2 The higherthan expected incidence of epithelial carcinoma inimmunosuppressed humans suggests a role forimmune defects in the development of skincancer.>? Previous work with mice has demon­strated that exposure to ultraviolet light can alterhost reactivity against a skin cancer, as well asinitiate the cutaneous neoplasm. 6 The presentstudy was undertaken to investigate cellular im­munity by observing lymphocyte blastogenic re­sponses in patients with basal cell carcinoma(BCC).

MATERIALS

Fourteen Caucasian patients, ranging in age from 30to 79 years (mean, 60.4 years), who had multiple (i.e.,

From the Memorial Sloan-Kettering Cancer Center.

Supported in part by PHSH grants CA-19267 and CA-08748 from theNational Cancer Institute, Witty'S Fund, Chesebrough-Pounds,Inc., The Handmacher Foundation, Inc., and the Robert S, SinnFund.

Presented at a symposium at the Annual Meeting of the AmericanAcademy of Dermatology, Chicago, IL, December, 1979.

Reprint requests to: Dr. Bijan Safai, Memorial Sloan-KetteringCancer Center, 1275 York Ave., New York, NY 10021.

0190-9622/81/060711+04$00.40/0 © 1981 Am Acad Dermato1

more than seven) BeCs were studied. BCCs ranged insize from 0.5 to 3 cm. Patients with internal malignan­cies, serious medical problems, autoimmune disorders,or those on any immunosuppressive medications wereexcluded.r"? Three patients also had squamous cellcarcinoma (SCC) of the skin, and one patient had thenevoid basal cell carcinoma syndrome. All patientsdenied exposure to arsenic. Eleven healthy Caucasianvolunteers without skin cancer, ranging in age from 30to 73 years (mean, 56.6 years) were age- and sex­matched to the patients and were used as controls.

METHODS

Heparinized whole venous blood, 10 ml, was ob­tained from each patient and control simultaneously fordetermination of lymphocyte blastogenic response toconcanavalin A (Can A). A standard laboratory controlwas also studied each day, and each test was donetwice. The blood was diluted 1: 1 with sterile normalsaline, layered over lymphocyte separation medium(Bionetics, Kensington, MD), and centrifuged at 2,000g for 30 minutes. The lymphocyte-rich layer was re­moved from the interface, dlluted 1: 2 with normal sa­line, and centrifuged at 1,600 g for 10 minutes. Cellswere then washed with Hanks solution and centrifugedat 1,200 g for 10 minutes three times. Cells were sus­pended in RPMI-1600 (Grand Island Biological Co.,Grand Island, NY) containing 15% fetal calf serum(Grand Island Biological Company), 1% penicillin andstreptomycin, and 0.8% L-glutamine. The concentra-

711

712 Mvskowski et al

PATIENTS VS. AGE-MATCHED CONTROLS\2.

10

8

Q><6

~

4

2

O'--~=---:--------::-:~"""'-

Fig. 1. Mean peak lymphocyte blastogenic responses ofthe patients with multiple basal cell carcinoma vs age­and sex-matched controls.

tion of lymphocytes was adjusted to 3.3 X 105 cells permilliliter. Of the cell suspension thus prepared, 150 mlwere transferred to individual culture wells in microtiterplates (Cooke Engineering Co., Bloomington, IN).

To obtain a dose-response curve, 25 ml of con­canavalin A (Con A) in the following concentrationswere added to the wells: 20 jLg/ml, 40 jLg/ml, 100,ug/ml, and 200 jLg/ml. In control wells, 25 ml ofcomplete media were added instead of Con A. Cellswere then incubated at 37° C in a humidified incubatorwith 5% COl! for 72 hours, and then standard pulse ofC 14 thymidine (20 ml) was added for each well. Cellswere harvested 24 hours later in a Titertek Cell Harves­ter (Flow Laboratories, Rockville, MD) in Shatronfilters, air-dired, transferred to counting tubes, andcounted in a scintillation counter.

RESULTS

A markedstatisticallysignificant (p < 0.02) re­duction in the mean peak lymphocyte stimulation

Journal of theAmerican Academy of

Dermatology

was seen in the group of patients with multipleBCCs, as compared to the controls (Table I). Thedecrease in lymphocyte responsiveness at higherconcentrations of mitogen was comparativelygreater in the BCC group than in the control group.Fig. I shows that nine of the fourteen (64%) pa­tients with BCCs had lymphocyte blastogenic re­sponses thatwere lower than their age-matched con­trols and also lower than the lowest control value.Two of the BCC patients (14%) had blasto­genic responses higher than their age-matched con­trols, and two had values nearly the same as theircontrols.

DISCUSSION

Studies in patients with internal neoplasms haveshown a number of qualitative and quantitativedefects in cell-mediated immunity, some of whichhave been associated with increased recurrencerates of the neoplasm and shortened patient survi­val. 1O

-13 It is known that peripheral blood T

lymphocytes decline both in number and func­tion with advancing age and certain types ofmalignancy. 14-16 T lymphocyte defects havebeen shown to correlate with stage and prognosisin various internal malignancies, especially sceof the head and neck,17,18 Immunologic deficitshave also been observed in patients with malignantmelanoma, cutaneous T cell lymphoma, andKaposi's sarcoma. 19-22

However, evaluation of cellular immunity hasbeen done infrequently with the less aggressiveskin malignancy, BCC. Dellon et all studiedseventy-five patients with SCC, BCC, and Bow­en's disease and discovered that low peripheralblood T cell levels correlated with evidence ofdecreased lymphocytic infiltration around BCe. 1

Bustamente et al23found the percentage of T lym­phocytes to be greater around BCC and sec thanaround melanoma, and they observed an increasednumber of immunoglobulin-producing cells aroundbasal cell carcinomas. Immune parameters, asmeasured by positive conversion of skin tests,have been suggested to be of some prognosticsignificance in patients with BCC, basosquamouscell carcinoma, or sec and in situ epidermoidcarcinoma of the skin.2,j

Volume 4Number 6June, 1981

Decreased blastogenic responses with multiple basal cell carcinoma 713

Table I. Mean count per minute of peak response to different concentrations of mitogen

Concentrations of concanavalin AOverall mean peak re-

I I I 100 ILgfml I 200 ILg/mlsponse ± standard deviation 0 20 ILg/ml 40 ILg/ml

Controls 7,207 242 4,650 4,345 2,069 1,387(mean age::::: 56.6 yr) ±2,54l ±162 ±3,178 ±3,421 ±2,898 ±2,075

Patients 3,755 143 3,400 737 128 45(mean age = 60.4 yr) ±3,697 ±90 ±3,924 ±859 ±155 ±19

Significance p < 0.02 <0.1 <0.1 <0.001 <0.02 <0.05

In the present study we have demonstrated aqualitative defect (decreased blastogenic responsesto Can A) in peripheral blood lymphocytes in agroup of patients with multiple BCCs. It is un­known whether the decreased lymphocyte respon­siveness played a role in the induction of the BCesor occurred after their onset. Perhaps the neoplasticbasal cells interact with T lymphocytes to alter Tcell function or directly produce a nonspecificsuppressor cell response. On the other hand, ul­traviolet light may induce the impaired lymphocyteblastogenic response, which in turn permits prolif­eration of abnormal basal cells. Further studies arenecessary to clarify these speculations. The clinicalsignificance, if any, of these findings has to awaitfurther investigation.

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