Tissue Antigens (1983) 22, 167-169

B8, DR3 antigens and production of human leucocyte migration inhibitory factor (LIF) by mononuclear cells stimulated with concanavalin A (Con A) CALOGERO CARUSO and LMA BELLINA

Servizio di Immunologia Tissutale, Istituto di Patologia generale dell’Universiti di Palermo, Palermo, Italy

LIF release by Con A stimulated mononuclear cells was evaluated in 67 randomly selected healthy Sicilians typed for HLA antigens. The results show that B8 and/or DR3 positive subjects release less LIF than negative ones, suggesting that this immunological response might be controlled by HLA-linked immune response (Ir) gene(s).

Received for publication 29 November 1982, revised, accepted 25 February 1983

Antigen specific and nonspecific Ir genes MHC-linked have been demonstrated in ex- perimental animals (McDevitt 1981, Biozzi et al. 1980). Several investigators have suggested that there might be MHC-linked antigen specific Ir genes also in man, by de- monstrating statistical association between immune responsiveness to specific antigens and HLA specificities (Dausset & Contu 1980, de Vries 1979, van Rood et al. 1977, 1981). Moreover there is some evidence of the existence of antigen nonspecific HLA-linked Ir genes as well. The haplotype 88 , DR3 which is associated with several im- munological diseases would bear Ir genes al- lowing for a nonspecific increase in antibody production and decrease in cell mediated function (Ambinder et al. 1982, Dausset &

Contu 1980, Lawley et al. 1981, Legrand et al. 1982).

We have searched for B8, DR3-linked control of immune response on LIF release by Can A stimulated mononuclear cells (Bendtzen 1980, Rocklin 1981). To this pur- pose we have evaluated this in vitro antigen nonspecific correlate of cell mediated im- munity in 67 randomly selected healthy Sicilians typed for HLA antigens. LIF test was performed as previously described (Bellina & Salerno 1981). HLA-A, B and C antigens were determined by using the standard NIH microlymphocytotoxicity technique. Typing for HLA-DR, MB and MT specificities was performed with the use of the two-colour fluorescence test (van Rood et al. 1976).

Arkhmetical mean LIF values were

168 CARUSO AND BELLINA

Tuble 1. L I F release by Con A stimulated mononuclear cells from 88 positive, 8 8 negative, DR3 positive and DR3 negative subjects.

PhenotypeZ No. LIFrelease P5 subjects3 values4

B8-pos. 7 42.1k8.3 B8-neg. 60 59.922.5 DR3-pos. 13 46.3k6.6 <0,02 DR3-neg. 53 60.Yi2.5

Thc activity of the supernatants obtained from mononuclear cells ( 3 X 106/ml) stimulated with Con A (25 pgiml) was tested on chicken huffy coat leucocytes and the percent of migration in- hibition of targct cells (which expresses LIF re- lease) calculated (Bellina & Salerno 1981). All the subjects were bled at least twice on different days and the results of the different assays were pooled. In the normal Sicilian population the frequency of B8 antigen is 0.10 (Salerno et al. 1981) and that of DR3 is 0.15 (Caruso et al. 1983). All the subjects B8 positive were positive for DR3. 67 unrelated randomly selected healthy indi- viduals born in Western-Sicily from West- ern-Sicilian parents (42 males and 25 females, 21 to 59 years of age, mean age 29). The distribution of LIF values was close to the theoretical normal distribution curve since the 63% of LIF release values was included into X ?z lo and the 95% was included into X * 2 o (x = 58.0; u = 20.3). Data are expressed as the mean i standard error. Significance between positive and negative sub- jects by Student t test. The applicability of the t test to the data was verified by F test. P values were not multiplied for the number of possible comparisons since it was looked a priori for thc influence of these antigens.

analyzed according to HLA phenotypes. B8 and DR3 positive subjects showed the lowest values. The mean LIF values of B8 positivc, B8 negative, DR3 positive and DR3 negative subjects (all the subjects B8 positive were DR3 positive) are reported in Table 1.

B8, DR3 positive mononuclear cells re- leased significantly less LIF than B8, DR3

negative ones. The same was true for DR3 positive mononuclear cells on the whole. ( P values were not corrected for the number of possible comparisons, thus these results might need confirmation in a larger sample of sub- jects).

Our results demonstrate that mononuclear cells of B8 and/or DR3 positive subjects show a significant lower LIF production after Con A stimulation. This is likely due to the influ- ence of antigen nonspecific Ir gene(s) in lin- kage disequilibrium with B8, DR3 gene(s).

There is evidence, indeed, that B8, DR3 antigens might be linked to a system allowing for a nonspecific increase in antibody produc- tion and a decrease in cell mediated function. It has been demonstrated that B8 and/or DR3 positive subjects show a reduced macrophage catabolic activity (Legrand et al. 1982), an in- creased immunoglobulin production by peripheral B cells (Ambinder et al. 1982, Lawley ct al. 1981), a low spontaneous T cell proliferation (Ambinder et al. 1982) and a low lymphocyte blastogenic response to Con A (Lawley et al. 1981).

B8, DR3 antigens are frequently linked to diseases with immunological disorders (Dausset & Contu 1980). In agreement with the results obtained in previous studies (An- hinder et al. 1982, Dausset & Contu 1980, Lawley et al. 1981, Legrand et al. 1982) de- monstrating that immunological dysfunctions are more frequent in normal B8, DR3 indi- viduals, our observation, performed on a ran- domly selected population, show changes in LIF release due to B8, DR3 antigens.

Acknowledgments

We thank Prof. Jon J. van Rood and Prof. Alfred0 Salerno for helpful suggestions and criticism. This work was supported in part by grant no. 80.00573.04 from the Consiglio Nazionale delle Ricerche, Rome. The excel-

LIF PRODUCTION RY R8 DR3 SIJBJECI'S 169

lent technical assistance of Carlo Muratore is acknowledged. We wish also to thank NIH, Bethexla, Maryland, USA, for providing sev- eral HLA-antisera.

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262, 795-797.

Address: Dr. Calogero Caruso Servizio di Immunologia Tissutale lstituto di Patologia generale dell'Universita Corso TiJkory 2 11 90 134 Palermo, Italy