Jerome Bill

A T cell receptor Vβ segment that imparts reactivity to a class II major histocompatibility complex product

We have identified in mice an allele of a new T cell receptor V beta gene, V beta 17a, whose product is bound by the monoclonal antibody KJ23a. Over 90% of T cell hybridomas prepared from V beta 17a+ T cells of SWR mice respond to allogeneic forms of the IE class II MHC protein, indicating that V beta 17a has an appreciable affinity for IE regardless of the other components of the T cell receptor. These results suggest a bias in the germ-line T cell receptor repertoire toward recognition of MHC proteins and indicate that the V beta portion of the receptor may form the most important contact points with MHC ligands.

The T cell receptor Vβ6 domain imparts reactivity to the Mls-1a antigen

Abstract A monoclonal antibody secreting hybridoma was established by fusing spleen cells from a rat immunized with a murine T cell clone, OI11, which has I-Ab restricted specificity for the male H-Y antigen and unrestricted specificity for the minor lymphocyte stimulating antigen, Mls-1a, to the mouse myeloma P3X63AG8.653 and screening for the capacity of the hybridoma supernatants to stimulate the OI11 T cell clone. An antibody (RR4–7) was found to be specific not only for the immunizing T cell clone but virtually for all T cells using the Vβ6 TCR gene product as part of their surface antigen receptor. When the expression of the Vβ6 gene in various strains of mice was analyzed, it was found that strains expressing the Mls-1a antigen contained few T cells expressing Vβ6-encoded TCRs. The majority of T cell hybridomas which expressed Vβ6-encoded TCRs were found to be reactive to the Mls-1a antigen. These data confirm the finding of H. R. MacDonald et al. (Nature (London)332, 40, 1988) that most TCRs encoded by the Vβ6 gene have a biased specificity for the Mls-1a antigen.

Beryllium Presentation to CD4+ T Cells Is Dependent on a Single Amino Acid Residue of the MHC Class II β-Chain

Chronic beryllium disease (CBD) is characterized by a CD4+ T cell alveolitis and granulomatous inflammation in the lung. Genetic susceptibility to this disease has been linked with HLA-DP alleles, particularly those possessing a glutamic acid at position 69 (Glu69) of the β-chain. However, 15% of CBD patients do not possess a Glu69-containing HLA-DP allele, suggesting that other MHC class II alleles may be involved in disease susceptibility. In CBD patients without a Glu69-containing HLA-DP allele, an increased frequency of HLA-DR13 alleles has been described, and these alleles possess a glutamic acid at position 71 of the β-chain (which corresponds to position 69 of HLA-DP). Thus, we hypothesized that beryllium presentation to CD4+ T cells was dependent on a glutamic acid residue at the identical position of both HLA-DP and -DR. The results show that HLA-DP Glu69- and HLA-DR Glu71-expressing molecules are capable of inducing beryllium-specific proliferation and IFN-γ expression by lung CD4+ T cells. Using fibroblasts expressing mutated HLA-DP2 and -DR13 molecules, beryllium recognition was dependent on the glutamic acid at position 69 of HLA-DP and 71 of HLA-DR, suggesting a critical role for this amino acid in beryllium presentation to Ag-specific CD4+ T cells. Thus, these results demonstrate that a single amino acid residue of the MHC class II β-chain dictates beryllium presentation and potentially, disease susceptibility.

Relationships between genes on human chromosome 11 encoding cell-surface antigens.

Genes encoding seven monoclonal antibody-defined cell-surface antigens have been regionally mapped on human chromosome 11, and compared to those of the AL complex defined by polyclonal antibodies using mutational analysis. MIC1, encoding W6/34 antigen, is probably identical to S1, previously mapped to 11pter-p13. MDU1 and MIC8, encoding 4F2 and TRA-1.10 antigens, respectively, are probably identical to S2(a4) and map to 11q13-q22. MIC9, which governs expression of 4D12 and 2E2 antigens, and maps to 11q22-qter, is not related to any of the five AL genes. MIC4 and MIC11, both mapping to 11pter-p13, may have some relationship to S3 and S1, respectively, but identity has not been proven.

Expression of human chromosome 11-encoded cell-surface antigens by DNA-mediated transfectants

DNA-mediated transfectants were isolated that expressed two of the cell-surface antigens encoded by human chromosome 11. These tranfectants were used to analyze monoclonal antibodies selected to recognize human cell-surface antigens expressed by a somatic cell hybrid containing 11 as its only human chromosome. Analysis of the transfectants, deletion hybrids, and mutants showed that the monoclonal antibodies recognized at least five different antigens, one of which we had not identified previously. A majority of the monoclonal antibodies recognized the a1 antigen. The use of cells from higher primates demonstrated that the a1 -specific monoclonal antibodies recognize at least two epitopes.

Identification of a mouse T-cell antigen receptor α-chain polymorphism by a Vα3.2 chain-specific monoclonal antibody

T-cell antigen receptors (Tcrs) interact with antigenic peptides associated with major histocompatibility complex (MHC) class I or class II molecules (Babbitt et al. 1985; Townsend et al. 1986) and this interaction plays a critical role in both T-cell antigen repertoire selection in the thymus and in the response to foreign antigens in peripheral lymphoid organs (Zinkernagel et al. 1978; Nikolic-Zugic et al. 1990; Sha eta! . 1990). The Tcr is composed of disulfide-linked a and/3 chains which are encoded by non-contiguous variable (I0, joining (J), and constant (C) gene segments for ~ chains and V, diversity (D), J, and C gene segments for/3 chains (Chien et al. 1984; Hedrick et al. 1984; Saito et al. 1984; Yanagi et al. 1984). The potential repertoire created by gene rearrangement and N-region nucleotide addition for both and/3 chain Tcrs is estimated to be more than 1012. The locus encoding the entire Tcrb gene is mapped on chromosome 6 in the mouse and each of the known Tcrb-V genes in relationship with D, J, and C gene segments has been mapped within 450 ldlobases (kb) of DNA (Epstein et al. 1985; Roehm et al. 1985; Wilson et al. 1988). Although it has been demonstrated that the deltion of either several Tcrb-V gene segments or part of D and J, and one Tcrb-C gene segment exists in some strains of mice (Behlke et al. 1986; Lai et al. 1987), only a limited polymorphism for the known Tcrb-Vgene family has been demonstrated (Wade et al. 1988; Klotz et al. 1989). The Tcra gene is located on chromosome 14 in the mouse and spans more than 370 kb of DNA which also contain the Tcrd gene segments (Dembic et al. 1985; Kranz et al. 1985; Chien et al. 1987). Restriction fragment length polymorphism (RFLP) analysis of the Tcra locus using

Molecular cloning of MER-2, a human chromosome-11-encoded red blood cell antigen, using linkage of cotransfected markers

We report the molecular cloning of a human gene MER-2located on chromosome 11 that encodes a cell surface antigen which is polymorphic on red blood cells. An essential element of the cloning strategy was cotransfection-induced linkage of pSV2-neo, which encodes resistance to the antibiotic G418, to the human MER-2gene. An important feature of the pSV2-neo construct is that the same gene (the transposon, Tn5) that encodes G418 resistance in eukaryotic cells confers neomycin resistance in bacteria. Chinese hamster ovary (CHO) cells were cotransfected with pSV2-neo and genomic DNA from a CHO ×human cell hybrid containing a single human chromosome (chromosome 11). Transfectants expressing both the human MER-2gene and G418 resistance were isolated by selection in the antibiotic G418, followed by indirect immunofluorescence using the monoclonal antibody 1D12, which recognizes the MER-2 antigen, manual enrichment, and single-cell cloning. Genomic DNA from a primary transfectant positive for MER-2expression and G418 resistance was used to construct a cosmid library and cosmid clones able to grow in neomycin were isolated. Of 150,000 cosmid clones screened, 90 were resistant to neomycin and of these, 11 contained human repetitive sequences. Five neomycin-resistant cosmid clones containing human repetitive DNA were able to transfect CHO cells for G418 resistance and MER-2expression.