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The antigenic specificities, major histocompatibility complex restrictions and functional properties of influenza virus-specific proliferative cloned cell lines have been studied. These lines were specific for the H3 hemagglutinin subtype of influenza A viruses. By using a large panel of HLA-phenotyped antigen-presenting cells, it was found that the polymorphic structures, defined as DR1 and DR7 molecules, or closely associated structures, function as the restricting elements. We excluded for these lines a possible restricting role of supertypic specificities, known cross-reacting elements on DR molecules, or products of other loci in known linkage disequilibrium with the HLA-DR molecules. Such exquisitely restricted clones might be of great help in the class II typing of antigen-presenting cells. Their specific activity was stable for several months. This has allowed the study of some functional properties of these long-term-cultured cloned cell lines: interleukin 2 sensitivity and production, helper function in specific antibody synthesis and ability to stimulate in mixed leukocyte reactions.

Binding of D-Penicillamine (D-Pen) to human monocytes was examined by flow cytometry with fluorescent D-Pen conjugate. Cells from HLA DR1-positive healthy females bound significantly more D-Pen than cells from DR1-negative healthy females (P = 0.015), and DR1 was associated with the highest binding among HLA DR antigens. In contrast, DR4 was associated with the lowest binding in healthy females. A difference in D-Pen binding between healthy females who were DR1-positive, DR4-negative and those who were DR1-negative, DR4-positive was statistically significant (P = 0.026). Neither healthy females nor healthy males showed significant associations of D-Pen binding with HLA A, B, or C antigens, nor did healthy males show an association between strength of D-Pen binding and any DR antigens.

In a new series of RA patients the association with HLA-DR4 was again found to be highly significant. Also increased were the DR4 associated antigens DRw53 and DQw3. Hybridization of a DQ-beta probe to Bg1 II-digested DNA showed that a variant of DQw3, characterized by a 4.3 kb fragment and related to the serological specificity TA10, was markedly increased among DR4 positive RA patients. HLA-DR1 was also increased in RA and appeared to be independent of the increase of DR4. Although the DR1 association was weaker, it was observed in both patients with and without RF. In contrast, DR4 was found to be increased significantly only in the RF positive group of patients, as previously observed. Development of RA in multiple-case families was found to be linked to the inheritance of DR4 positive haplotypes. The possible role of risk factors associated with polymorphic markers of the T-cell receptor genes was investigated. An allelic marker associated with one of the variable gene subfamilies of the beta-chain was found to be associated with RA. The results suggest that susceptibility for development of RA is influenced by alleles of the T-cell receptor in conjunction with the class II HLA factors with which the T-cell receptor interacts in the course of the immune response.

The SDR1 monoclonal antibody reacts only with cells which express the HLA-DR1, 2,w6 or w8 allogeneic specificities. Two-dimensional nonequilibrium pH gradient/sodium dodecyl sulfate polyacrylamide gel electrophoretic analyses of SDR1 immunoprecipitates from [35S]methionine biosynthetically labeled cells revealed the typical pattern of Ia antigens, namely two polypeptides of about 34kDa and 29kDa (designated epsilon and beta-3) as well as the "basic invariant spot" of about 31 kDa. The epsilon and beta-3 polypeptides were only weakly represented in similar analyses of immunoprecipitates performed using a monomorphic HLA-DR monoclonal antibody, TDR31.1. The epsilon and beta-3 polypeptides of B lymphoblastoid cell lines homozygous for HLA-DR2 and w6 were structurally polymorphic as judged by two-dimensional gel analyses. This polymorphism was independent of the HLA-DR specificity. It is concluded that the SDR1 antibody recognizes a polymorphic set of Ia antigens that are coded by a locus other than HLA-DR. These antigens probably also express the MT1 (DC1, LB12) alloantigenic specificity and are most likely the human equivalent of the murine I-A subregion antigens.

Une epreuve de compatibilite croisee negative en utilisant le serum autologue du receveur traite par le dithioerythritol peut identifier un groupe de malades qui peuvent etre transplantes avec un minimum de crainte de rejet hyperaigu ou de necrose tubulaire aigue. D'autres parametres permettent de mieux definir ce groupe de candidats a la transplantation presentant un anticorps antilymphocytotoxique IgM

Stimulation of human lymphocytes with concanavalin A (Con A) resulted in variable lymphokine responses as indicated by factors inhibiting macrophage migration (MIF) or stimulating macrophage migration (MStF), or resulted in negligible responses. These responses were consistent for a given individual when repeated after several months. MIF responses were observed more frequently than MStF responses in patients with renal failure who had demonstrable alloantibodies. MStF responses were statistically associated with the presence of HLA DR1 antigens in patients with renal failure and two separate groups of healthy individuals, while MIF responses were associated with DR7 in the three groups studied. There was no correlation between immunoglobulin allotypes and lymphokine responses. These results suggest that lymphokine responses to Con A are indicators of nonspecific immunological responsiveness and are influenced by genes associated with the major histocompatibility complex.

Aucun gene HLA seul n'a ete identifie comme conferant un risque de maladie. Distinction et definition des sous-types DR4. Role de la 3eme region hypervariable de DRβ1. Relation entre risque de pemphigus et DW1eme et DRW6eme, et relation entre susceptibilite a la polyarthrite rhumatoide et un groupe d'epitopes trouves dans les sous-types non DW10 de DR4

Previous testing of patients with rheumatoid arthritis showed that one HLA-D type, Dw4, occurred more frequently than in normal controls. B-cell alloantigens closely related to HLA-D can now be identified by a simple serologic procedure. Using this test, I studied 80 white patients with erosive, rheumatoid-factor-positive rheumatoid arthritis. The B-cell alloantigen HLA-DRw4 occurred in 70 per cent of 54 patients, as compared to 28 per cent of the 68 normal controls (P less than 10(-5)). Both groups were also tested for the HLA-A, B and C antigens and for HLA-D. HLA-Dw4 occurred in 54 per cent of the patients and 16 per cent of the controls (P less than 10(-5)). Small differences observed in several of the HLA-A and B antigens were not statistically significant. The results indicate that rheumatoid arthritis in whites is associated with genes of the HLA-D region and that immunogenetic factors linked to HLA have a role in its pathogenesis.

Fifty-two patients with thyroid epithelial cell cancer were studied for evidence of association with human leukocyte antigens (HLA). Twenty-eight patients (53.8%) and 19.4% of 160 controls were HLA-DR1-positive, conferring a relative risk of 4.85 (chi 2 = 21.3, P less than 0.0001). HLA-DR1 was increased in all histologic types of thyroid cancer. Interestingly 10 of 12 patients with metastatic disease were DR1-positive compared to 18 of 41 patients without metastases (relative risk = 6.1, chi 2 = 4.7, P less than 0.05). This study suggests that major histocompatibility complex-linked gene(s) determine susceptibility to and the biologic behavior of thyroid cancer.

DR1 was defined at the Seventh Histocompatibility Workshop by five sera [1], and at the Eighth Workshop it was clearly identified by 10 sera. There was no evidence for a split of DR1 [2]. At the Ninth Workshop, the frequency of DR1 was 14.7% in Caucasians and 9.45% in Negroes.

Of the HLA allelic associations with insulin-dependent diabetes (IDD) reported to date. DR3 and DR4 have been the most positive and DR2 the most negative. In 952 Caucasian proband patients reported here, only 57 or 6% had no DR3 or DR4 alleles. When these 57 patients were compared to 249 Caucasian controls similarly lacking DR3 and DR4 antigens, there were excesses of DR1 (P = 0.13) and DRW8 (P = 0.01) and deficiencies of DR2 (P = 0.03) and DR5 (P = 0.03) in the patient group. The most common phenotype in this group of patients was DR1/DR7 (12.3%). Only four DR-homozygous patients involving alleles other than DR3 and DR4 were found by genotyping, and all were DR1 homozygotes. Among 506 patients wuth DR3/DRX or DR4/DRX phenotypes, DR1 was more frequent (P = 0.001; Bonferronni P = 0.006), and DR2 (P = 0.001) and DR5 (P = 0.001) less frequent than 243 HLA-matched controls. Of 187 patients with a single DR3 and no DR4, DR1 was more frequent (P = 0.02), with DR2 (P = 0.001) and DR5 (P = 0.02) less frequent than 94 HLA DR-compatible controls. Among 319 patients with a single DR4 but no DR3, DR1 was again more frequent (P = 0.01) and DR2 (P = 0.001) and DR5 (P = 0.001) less frequent than 149 HLA-matched controls. We conclude that DR1 is an additional risk DR allele for IDD to that of DR3 and DR4, and DR5 an additional protective DR allele to that of DR2.

Among individuals with rheumatoid arthritis the presence of the polymorphic Ia antigen epitope detected by the monoclonal antibody 109d6 is more strongly correlated with disease susceptibility than are other specificities, such as HLA DR4, DRw53 (MT3) or the antigenic determinant, defined by the monoclonal antibody 17-3-3S. The cells of 93% of Caucasian and Hispanic patients react with the 109d6 reagent. As was the case in normal individuals, all DR4-positive patients express the 109d6 determinant; however, 26% of those with rheumatoid arthritis have the epitope recognized by antibody 109d6, but lack the specificity DR4. Of these, one-third expresses only HLA DR1 and DQw1 (MT1, MB1) determinants. Studies of family members reveal that here the determinants 109d6, DR1, and DQw1 are encoded by the same unusual haplotype. In certain other individuals with rheumatoid arthritis who express DR4, DRw53, and the 109d6 determinants, family studies show that the 109d6 epitope is encoded not only by the haplotype specifying DR4 but also by the opposite haplotype that does not bear the genes for DR4. This suggests that homozygosity for certain Ia epitopes is relevant to determining the disease-susceptibility state. These studies emphasize the utility of monoclonal antibodies as reagents for the recognition of Ia epitopes that are more closely involved in the determination of disease susceptibility than are allomorphic molecules detected by conventional typing alloantisera.

Major histocompatibility complex (MHC) class II molecules, which function to restrict the antigenic recognition of T lymphocytes, are important genetic factors in the susceptibility to seropositive rheumatoid arthritis (RA) (1–4).By utilizing alloreactive human T-cell clones we have defined disease-associated T-cell epitopes encoded within the HLA-DR subregion of patients with RA (2). Interestingly, such T-cell clones can recognize these T-cell epitopes outside of the HLA-DR4 haplotype, partially overcoming the incomplete genetic linkage between DR4 and the disease. We have now generated a panel of T-cell clones that identify multiple T-cell recognition sites shared among the DR molecules of HLA-DR4 and DR1. A segment of 15 amino acids within the third hypervariable region of the HLA-DRβ1 chain is of primary importance for the formation of these clustered T-cell epitopes. Within HLA-DR1 the T-cell clones identify a cassette of polymorphic sites identical to the third hypervariable region of a DR4 allele. The finding that the cross-reactive T-cell determinants are frequently expressed on antigen-presenting cells of RA patients suggests that these polymorphic structures are of particular relevance for T-cell-HLA interaction in RA.

Forty-nine Israeli Jewish patients with rheumatoid arthritis (RA) were studied for their HLA A, B, C, DR antigen frequency. A significant increase in HLA Aw31 and HLA DR1 was observed (p less than 5.10(-5) and p less than 5.10(-3) respectively). 45% of Aw31 positive patients were sero-negative for rheumatoid factor, while most HLA DR1 positive individuals were seropositive. DR5 was found to be significantly decreased (p less than 5.10(-4)). Contrary to previous reports, DRw4 was found to be within the range of antigen frequency of the controls (34.7% vs. 32%). It is suggested that in our population of patients Aw31 and DR1 and not DR4 are highly associated with adult onset of RA.

Sixty three Caucasian patients with either melanoma, keratoacanthoma or squamous cell carcinoma were human leucocyte antigen (HLA) typed. The regressing tumour groups were compared with their non-regressing counterparts, and the patient groups were compared with a control Caucasian population. Melanoma patients showing histological regression were more likely to be HLA-B22 positive, and HLA-B27 and -DR1 negative, than those without features of regression. When compared with a control population, the group of melanoma patients were more likely to be HLA-B22 positive. Comparison of the group of keratoacanthomas, a self-regressing tumour, and the group of squamous cell carcinomas, a non-regressing tumour, did not show any significant differences in HLA typing.

Introduction We used DR1 transgenic mice and covalently linked DR1 multimers to characterize analog-specific inhibitory T cells in collagen-induced arthritis (CIA). Because of the low numbers of antigen-specific T cells in wild-type mice, functional T-cell studies in autoimmune arthritis have been challenging. The use of T-cell receptor (TCR) transgenic mice has provided useful information, but such T cells may not represent the heterogeneous T-cell response that occurs in natural settings. Our focus was to develop tools to identify and characterize the population of immunoregulatory T cells induced in wild-type mice by an analog peptide of CII259-273, which contains amino acid substitutions at positions 263 (N) and 266 (D) (analog peptide A12). Methods DR1 multimers, developed by loading empty class II molecules with exogenous peptide, provide a method for visualizing antigen-specific T cells with flow cytometry. However, the low binding avidity of A12 for the major histocompatibility complex (MHC) made this strategy untenable. To overcome this problem, we generated DR1 multimers in which the analog peptide A12 was covalently linked, hoping that the low-avidity analog would occupy enough binding clefts to allow detection of the responsive T cells. Results Staining with the tetramer revealed that A12-specific T cells were readily detectable at 10 days after immunization. These CD4(+) T cells are a highly selective subset of the TCR repertoire and have a limited clonality. Analysis of cytokine expression showed that cells detected by tetramer (A12) expressed primarily suppressive cytokines (interleukin-4 (IL-4) and IL-10) in response to collagen, compared with control cells. Although they did not express Fox-p3, they were extremely effective in preventing and suppressing inflammatory arthritis. Conclusions In summary, our studies showed that the use of covalently linked multimers allows characterization of analog-specific T cells that are otherwise difficult to detect. The suppressive character of the analog-specific T-cell response suggests that these cells attenuate autoimmunity and differ significantly in phenotype from the inflammatory T cells predominantly found in arthritic joints. Such reagents will become powerful tools to study T-cell responses in RA patients in upcoming clinical trials.