HLA antigens and immunoglobulin allotypes in patients with malignant melanoma

HLA Antigens and Immunoglobulin Allotypes in Patients with Malignant Melanoma J.P. Pandey, A.H. Johnson, H.H. Fudenberg, D.B. Amos, J.U. Gutterman, and E.M. Hersh

ABSTRACT: HLA antig,,,ns and Gin, A2m, and Km allotypes were examined in Caucasian pattents with malignant mdanoma. No significant associations were found fi, r any of the HLA antigens tested. Significant association was found with Gin(2), and the rdative risk for individvals with this markcr was calculated at 1.9. The data indicate that Caucasians positive for Gin(2) are almost twice as likely to develop malignant melanoma as those without this marker.

.q r R O D U C T I O N Patients with malignant melanoma generate humoral and cellular immunologic responses to melanoma antigens [.1,2]. Since immune r~.~sponsiveness in humans is related to the major histocompatibility complex [.3,4] and immunoglobulin (lg) allotypes ['5-7], it seemed of interest to ascertain whether the frequencies of certain HLA antigens and lg allotypes in this patient group differ from those in the normal population. Further, o¢ all malignant tumors, melanoma has the most variable course, including spontaneous remissiolls presumably because of immunologic factors, and is the s,~Pd tumor for which various forms of immunotherapy have been used most often [8]. In the past few years, conflicting reports regarding associations of both HLA antigens and Ig allotypes with malignant melanoma have appeared in the literature. An elevated frequency of HLA-A9 and deficiency of B5 have been reported [.9-1 l]. In other studies no significant deviations were foand [.12-17]. In ailotype studies, J6rgensen and L~d [18] reported a highly significant association between Gin(2) and malignant melanoma, but subsequent studies b'/other investigators have failed to confirm this observation [19-21]. In the present study, Ig allotyping was performed on serum samples from 89 Caucasian melanoma patients and 906 controls. Twelve different atiotypic markers were examined, more than in any previous published study. A significant association (p < 0.01) was found between Gin(2) and malignant melanoma. Fifty-eight of these patients from whom lymphocytes were available were typed for HLA antigens. No significa~atassociations were found for any of the HLA antigens tested.

From tin. D~partmentof BaJic and Clinical lmmundogy and Microbiology,Medical University of South Catalina, Cbarksteu, South Carolina, the Division of Immunology, Duke University Medical Center, Dud~am, l*4orthCatalina, and the 12elmrt~nt of Developmental Therapeutics, M.D. Anderson Hospital and TurnerInstitute, Texas Mrdkal Center, Houston, Texas. Addr¢ss r,qu¢~tsfor reprints to Dr. J.P. Pando, Departmen.', of Basic and Clinical Immunology and Microld*log~, Medical University of South Carolina, ! 71 Ashley At,enue. Charleston,SC 29403. Rccdved 1980.

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186

J.P. Pandeyet al.

MATERIALS AND METHODS Blood was collected from 89 Caucasian malignant melanoma patients at the M.D. Anderson Hospital and Tumor Institute, Houston, Texas. The average age of the patients was 52.3 yr, and the average time lag between the onset of the disease and the initial vLsit to M.D. Anderson~ Hospital was 20.5 too. Our standard hemagglutination inhibldon technique [2~] was used to detect Gm (1, 2, 3, 17; 5, 6, 13, 14, and 21), A2m (1 and 2), at~d Km(1) antigens. Of these patients, 58 were typed for 40 HI.A antigens by the two-stage cytotoxicity test reported as the "modified NIH technique" [23~ The HLA specificities determined were as follows: HLA-Aml, 2, 3, 11, 25, 26, 28, 29, W23, W24, W30, W31, W32, W33, W34, W36; HLA-BmS, 7, 8, 12, 13, 14, 15, 17, 18, 27, 37, 40, W22, W35, W38, W39, W,tl, W42, W49, WS0; HLA-C--W1, W2, W3, W4. Statistical significance of the results was d~.termined by use of a 2 x 2 condgency chi-square, and the strength of association was measured by relative risk (RR) using Woo|fs formula [24]:

RR =pd (1 -pc)

(1 - pd ) pc "

where pd and pc are the frequencies of the antigen in pattents and controls, respectively. A feint/re risk indicates how many times more frequent the disease is in individuals carrying the antigen relative to those lacking itl RESULTS As shown in Table I, no significant associations were found tdr any of the HLA antigens tested. After correction for the number of ~omparis~ns made, none of the Gm phenotypes was associ~ed with malignant melanom~1 (Table 2). However, analysis of the data with respect to individual markers showed the frequency of the Gin(2) allotype to be significantly ,acreased in patients (30%) as compared to controls (18%) (estimated rel~ive ~isk: 1.9). As expected, all patients were A2m(1). (The frequency of the A2m(2) ~ l e in Caucasians is only 0.015.) No signifgant deviation was found in the frequency of the Kin(l) allotype in the patient population as compared to the controls (8% vs 10%). Analysis of HLA antigens in Gm(2)-posidve and Gm(2)-negative mehnoma patients did not indicate any ioint amnciation. DISCUSSION Our results agree with the findings of others [12-17] in that we found no significant association between HLA antigens and nmlignant melanoma. These data do not lend support to those obtained by Clark et al. [10], who repotted a complete absence of HLA-B5 in ~ t s with malignant melanoma~ In our study, the HLA-B5 frequency was the ~me as m ~ c~mtrol population (14%). The increased frequency of Gin(2) in our parents is in accordance with results obtained by J6rgensen and Lal [18~ Indeed, in view of their findings, and since the incidence of maligtumt melammm is very low in Blacks, who almost universally lack Gin(2) [25], we expeg:ted a priori a rehtiomhfip between Gm(2) and melanoma in C~ucas/ans. For th~ tx~son, in our analysis of Gin(2) no correction was necessary for the mmtber of compu/sons made. Associations between vmious hefedit~V ~ and diseau~ can be divided into two cate/g,'t4es: those present in ~ ~ c y in ~ st~dk~d at the onset of disease and those present in high ~ ¢ y in p~ent$ studied later in the disease course. The former represent stugel~fib~ty, w ~ e ~ the latter

TABLE

1

HI.A Antigens A SERIES 1 2 3 11 25 26 28 29 W23 W24 W30 W31 W32 W33 W34 W36 B SERIES 5 7 8 12 13 14 15 17 18 27 37 40 W22 W35 W38 W39 W41 W42 W49 WS0 C SERIES Wl W2 W3 W4

H L A - A , - B , iand - C a n t i g e n f r e q u e n c i e s ( p e r c e n t ) ~n p a t i e n t s ( N ~ 58) c o m p a r e d w i t h n o r m a l c o n t r o ~ ~N ~ 8 5 5 L Melanoma

17 50 24 14 7 9 5 7 2 17 3 2 14 2 0 0

2~ ~ ~ g ~¢ 4 ~ ,4 $ 2 @

14 19 12 36 5 3 16 7 12 14 3 7 2 14 3 2 0 0 0 2

~ ~ 28~ 22 6 9

9 10 29 17

i@ 9 19 2~

"Data from the First HLA Workshop of the Amerkas, 1975. Edited by R.J. D ~ . DHEW Publication (NIH) 76-1064.

~I

$ ~@ 2@ * g4 "~ 19

'~ l

2

~

T.C

6~ < O.OI. l ~ l a t i v c risk ~ 1.9,

6 3 1,27

Gin(l,17;21) 8 4 3,47

Gin(l,2,17 ;21) 36 47 4,39 ~

Gm(3;5,13,14) 28 31 0.28

Gm(1,3,17;5,13,14,2 I)

22 14 3.90"

Gm(1,2,3,17;5,13,14,2 I)

Distribution of Gm phenotypes (percent) in melanoma patients (N = 89) and normal controls (N = 906)

~# < 0.05 (~u~ c,~rrt, ccvd).

Mehaoma Ctmtroi C~i-~quared (1 d,f.)

TABLE 2

7,48 b

30 18

Gin(2)

HLA and Ig AHotypes in Malignant Melanoma

189

probably indicate resistance co death from the disease. For example, in Hodgkin's disease the presence of HLA-B8 is associated with increased surv/val time [26], whereas A w l 9 and/or B5 are apparently associated with decreased survival [27]. Our results suggest that Gm(2) is probably ass~a:iated with susceptibility to melanoma rather than resistance to death, since (1) the a v ~ time lag bet-~',¢eenthe onset of the disease and when the patients w*ere first seen at the M.D. Anderson Hospital was only 20.5 mo, and (2) the numbers o f p m i e m , in the various stages were similar to those seen in most un/versirlr centers. (In malignant melanoma, unlike, for example, lung cancer, it is relatively easy to establish the time when the lesion first appeared, since it is easily visible by the patient.) This may explain the discrepancy between our results and those reported by Schultheis et al. [19], Davean et al. [20], and Walter et al. [2i~ since their patient populations may have included a high prolx~rtion of longterm survivors, and it is highly unlikely that the same genetic marker would be associated both with susceptibility to the disease and long-tezma survival. In summary, our data indicate that Caucasians with the Gin(2) a/lotype have an increased (almost double) risk of developing malignant melanoma. ACKNOWLEDGMENT We thank Charles L. Smith for editorial assistance, and Beth Phifer and Vennie Mo~re for technical assist'~nce. Research was supported in part by USPHS Grants HD-09938 and CA-25746. This is publication No. 379 from the Department oi~"Basic and C[in/cai Immunology and Microbiology, Medical University of South Carolina. REFI/;RENCES 1. Ferrone S, Pel egrino MA: Antigens and antibodies in malignant melanoma, in: H Waters, Ed. H mdbook of Cancer Immunology. Vol. 3: Immune Status/n ~ e r Treatment and Prognosis--Part A. New York, Garland STPM Press, 1978. pp. 291-327. 2. Mastrangelo l~IJ, Bellet RE, Berd D: Immunology and immunotherapy of huma~ cutaneous malignant melanoma. In: WFI Clark JR, LI Goldman, MJ Mastam~geio, Eds. Human Malignant Melanoma. New York, Grune and Strarton, 1979. pp355-416. 3. Snell GD, Dat sset J, Nathenson S: Histocompatibility. New York, Academic Press, 1976. 4. Svejgaard A, I-lauge M, Jersild C, Platz P, Ryder LP, Staub Nielson L, Thomsea M: The HLA System. New York, S. Karger, 1979. 5. Mackay IR, Wells JV, Fudenberg HH: Correlation of Gm allo~Jpe, a n ~ y response, and mortality. Clin Immunol Immunopathol 3:408, 1975. 6. Nero S: Association of typhoid fever and response to vaccination with polymorphic systems in man. Prot Biol Fluids 22:649, 1975. 7. Pandey JP, Fudenberg HH. Virella G, Kyong CU, Loadholt CB, Galbraith RM, Gotschlich EC, Parke JC Jr: Association between immunoglobulin alk~pes and immune responses m HaeTt:ophilus influenzae and meningococcus p o l y s a c c ~ Lancet 1:190, 1.979. 8. Math6 G: Cancer Active Immunotherapy. New York, Springer-Verlag, 1976. 9. van Wijck R, Bouillenne C: HLA antigen and susceptibility m malignant melamine. Transplantation 16:371, 1973. 10. Clark DA, Necheles T, Nathanson L, Silverman E: Apparent HL-A5 defic/e~y/a malignant melanoma. Transplantation 15:326, 1973.

190

J.P. Pandey et al. 1 I. Braun WE: HLA and Disease. l~xa Raton, FL, CRC Press, 1979. 12. Cordon AL: HLA and malignant melanoma. Lancet 1:938, 1973. 13. Takasugi M, Terasaki PI, Henderson B, Mickey MR, Menck H, Thompson RW: HLA antigens in solid tumors. Cancer Res 33:648, 1973. 14. Singal DP, Bent PB, McCuiloch PG, Blajchman MA, MacLaren RGC: HLA antigens in malignant melanoma. Transplantation 18:186, 1974. 15. Lamm LU, Kissmeyer-Nielsen F, Kjerbye KE, Mogensen B, Petersen NC: HLA and ABO antigens and malignant melanoma. Cancer 33:1458, 1974. 16. Bergholtz B, Brennhovd I, Klepp O, Kaakinen A, Thorsby E: HLA antigens in malignant melanoma. In: GP Murphy, Ed. HLA and Malignancy. New York, Alan R. Liss, 1977, pp. 175-180. 17. lqathanson SD, Park MS, Drew Sl, Morton DL, Terasald PI: First and second B-lymphocyte antigen expression in malignant melanoma. Transplant Proc 12:118, 1980. 18. J6rgensen G, Lal VB: Serogenetic investigations on malignant melanomas with reference to the incidence of ABO system, Rh system, Gm, lnv, Hp and Gc systems. Humangenetik 15:227, 1972. 19. Schultheis W, Peter HH, Dekher H: Gin(l) and Gin(2) immunoglobulin ailotypes in patient with malignant melanoma. Humangenetik 28:177, 1975. 20. Daveau M, Pavie-Fischer J, R/vat L, R/vat C, Ropartz C, Peter HH, Cesarini Jl~, Kourilsky FM: lgG4 subclasses in malignant melanoma. J Nati Cancer lnst 58:189, 1977. 21. Waiter H, Brachtel R, Hilling M: On the incidence of blood ~oup O and Gin(- 1) phenotypes in patients with malignant melanoma. Human Genet 49:71, 1979. 22. Vyas GN, Fudenberg HH, Pretty HM, Gold ER: A new rapid method for genetic typing of human immunoglobulins. J Immuno1100:274,1968. 23, Amos DB, Pool P, Grit: J: HLA-A, -B, -C, and -DR. In: NR Rose, H Friedman, Eds. Manual of Clinical ~mmu,ology, 2nd ed. Washington, DC, American Society for Microbiology, 1980. 24. Woolf B: On estimating the relation between blood group and disease. Ann Human Genet 19:251, 1955. 25. Grubb R: The Genetic Markers of Human lmmunoglobulins. New York, Springer-Verlag, 1970. 26. Falk J, Osoba D: HLA antigens and survival in Hodgkin's disease. Lancet 2:1118, 1971. 27. Falk J, Osoba D: The HLA system and survival in malignant disease: Hodgkin's disease and carcinoma of the breast. In: GP Murphy, Ed. HLA and Malignancy. New York, Alan R. Liss, 1977, pp. 205-216.