Specificity of alloactivated human T lymphocyte clones in secondary proliferation, cell-mediated lympholysis and interleukin-2 release.

Alloreactive cytolytic T cell clones were generated from mixed leucocyte reactions (MLRs) between unrelated individuals. Two clones exhibited proliferative responses specific for class I HLA antigens B21 and B17. They were also found to be cytolytic toward cells bearing these HLA-B antigens as measured in cell-mediated lympholysis (CML) assays.

Heterogeneity of human T-lymphocyte clones generated from autologous mixed-lymphocyte cultures.

To assess the heterogeneity of T cells activated during the autologous mixed-lymphocyte reaction (AMLR), a cloning procedure based on the soft agar colony assay was developed. Supernatants of allogeneic MLR cultures were used as a source of interleukin 2 (IL-2) to generate two types of colonies: upper and lower colonies. Both types of colonies were expanded in long-term cultures using supernatants of phytohemagglutinin (PHA)-activated lymphocyte cultures.

Serological and cellular definition of a new HLA-DR associated determinant, MC1, and its association with rheumatoid arthritis.

A new serological HLA-DR associated determinant was defined by a cluster of six B cell alloantisera. This determinant, designated as MC1, was strongly associated with DR1 and DR4 and appeared unrelated to MB, MT, and SB. The antigen frequency of MC1 was 47% in Caucasoids and 43% in Blacks.

The complexity of HLA-DS molecules. A homozygous cell line expresses multiple HLA-DS alpha chains.

Ia molecules expressed by an HLA-DRw6 homozygous cell line were immunoprecipitated with anti-Ia allosera and monoclonal antibodies and analyzed by 2-D gel electrophoresis. The DRw6 homozygous cell line was shown to express two DS beta chains; this observation extends our previous finding that a DR5 homozygous cell line expresses two DS beta chains and suggests that the expression of at least two DS beta chains by DR homozygous cell lines is a generalized phenomenon. The data presented here document for the first time that a DR homozygous cell line expresses at least two DS alpha chains.

Monoclonal antibody analysis of responder and stimulator cells in the human autologous mixed lymphocyte reaction.

Responder and stimulator cell subpopulations in the autologous mixed lymphocyte reaction (AMLR) were determined with the OK series of monoclonal antibodies. Mitomycin-C-treated, monocyte-enriched cell populations were used as stimulator cells in the AMLR. Treatment of these monocytes with either OKM and/or OKI monoclonal antibodies and complement resulted in a marked loss of ability of these cells to act as stimulators in the AMLR.

Linkage disequilibrium of HLA-SB1 with the HLA-A1, B8, DR3, SCO1 and of HLA-SB4 with the HLA-A26, Bw38, Dw10, DR4, SC21 extended haplotypes.

Homozygous typing cells from 13 normal HLA-A1, B8, Dw3, DR3 and five normal HLA-A26, Bw38, Dw10, DR4 individuals were typed for the following markers: HLA-SB, MB, MT; complement proteins BF, C2, C4A, C4B; and GLO. Ninety-one percent of A1, B8, Dw3, DR3 homozygous individuals (HI) tested were homozygous for BF*S, C2*C, C4A*QO, and C4B*1 (SCO1 complotype), which indicates that the SCO1 complotype is in linkage disequilibrium with the A1, B8, DR3 haplotype in randomly selected normal populations. Sixty-seven percent of HLA-A1, B8, Dw3, DR3, SCO1 positive HI also expressed SB1; since the frequency of SB1 in random Caucasian populations is 11.

Dissociation in expression of MB1/MT1 and DR1 alloantigens in mutants of a lymphoblastoid cell line.

Cytotoxicity tests with alloantisera were used to study the expression of HLA-D region antigens in HLA-DR-null mutants of a human lymphoblastoid cell line. The initial cell line contained just one copy of the MHC as a haplotype that included DR1 and MB1/MT1. Gamma ray mutagenesis of the single haplotype cells followed by selection with complement and an anti-DR monoclonal antibody were then used to isolate DR-null mutants.

Immunogenetic analysis of the HLA complex with alloreactive T cell clones.

Recently developed technologies to clone alloreactive human lymphocytes have provided a powerful tool for the immunogenetic analysis of the HLA region. Alloreactive T cell clones can be used as specific reagents for HLA encoded cellular determinants detected in secondary proliferation assays such as the primed lymphocyte test (PLT) and, in cell-mediated lympholysis (CML). Lymphocyte clones can also be used for functional assays (e.

Bone marrow transplantation for severe aplastic anemia using a phenotypically HLA-identical, SB-compatible unrelated donor.

A three-year-old boy with severe aplastic anemia (HLA-A1,B8(Bw6), Cw7,DR3, MB2, MT2, SB4/A1,B8 (Bw6), Cw7,DR3,MB2,MT2,SB-) received a bone marrow transplant from a phenotypically HLA-identical, SB-compatible female unrelated donor. This donor was selected from eighteen HLA-A1,-B8,-blood donors after extended serotyping, mixed leukocyte culture testing and secondary proliferation assays with primed lymphocyte typing reagents specific for SB. Although patient cells proliferated well as responders in MLR, their stimulatory capability was greatly impaired.

A linkage study of the HLA region using C-band heteromorphisms.

A linkage study was performed to establish the map distance from the HLA region to the centromere on 6p. In this study, seven families were investigated for the segregation of their HLA types and a polymorphic C-band marker on chromosome 6. Four out of 29 children were found to be recombinants.

Primed lymphocyte testing specificity of alloreactive lymphocyte clones for HLA-B locus determinants.

We have generated two alloreactive T-lymphocyte clones that recognize the products of the HLA-B locus. The primed lymphocyte testing (PLT) reactivity of clone HJ21 was monospecific for the HLA-B antigen Bw49, a subtype specificity of HLA-Bw21. The PLT specificity of clone RD3 was strongly associated with HLA-Bw21.

Detection of non-HLA-DR determinants by alloreactive lymphocyte clones.

Using the soft-agar colony assay, we have generated three MT3-associated clones: HJ1, HJ13, and HJ39, from an MLR combination of two unrelated individuals. Another clone, HJ37, appeared to recognize a novel HLA-D determinant. PLT inhibition studies with monoclonal anti-Ia-like antibodies (Mab) were conducted on clones HJ1, HJ39, and HJ37.

Detection of HLA-DR associated PLT determinants by alloreactive lymphocyte clones.

This study deals with alloreactive T-cell clones which recognize cellular determinants associated with HLA-DR antigens. Two clones, CB55 and DS56, exhibited a PLT specificity that was perfectly associated with DR5. On the other hand, clones CB7, DS1 and HS1 showed PLT reactivity with approximately one-half of the DR5 positive cells and none of the DR5 negative cells, whereas clone MD4 largely reacted with the other half of DR5 positive cells.

Three Ia species with different structures and alloantigenic determinants in an HLA-homozygous cell line.

Three distinct molecular subsets with different structures and alloantigenic determinants were identified in human Ia antigens from cells of an HLA-Dw7 homozygous cell line. The subsets carried DR7 specificity, BR4X7 supertypic specificity and MB2 supertypic specificity, respectively, and were immunospecifically separated by the use of operationally monospecific alloantisera. These specificities showed HLA-linked segregation in families and they were distributed in the population according to different but partially overlapping patterns.

Mitogen- and alloantigen-stimulated blastogenic responses of dinitrophenyl-modified human lymphocytes.

The present study was designed to investigate alterations in the blastogenic response of dinitrophenyl (DNP)-modified normal human lymphocytes. Cells were isolated from heparinized blood and treated with dinitrofluorobenzene (DNFB) at ratios ranging from 10 to 10 molecules per cell. Unmodified and DNP-modified lymphocytes were cultured in the presence of Concanavalin A, pokeweed mitogen or phytohaemagglutinin (PHA).

Heterogeneity of cytolytic and suppressor clones of alloactivated cells generated from soft agar colonies.

Long-term cell cultures (or clones) were developed from soft agar colonies of lymphocytes alloactivated in mixed leukocyte culture reaction (MLR). Two types of colonies were identified: upper colonies that grew on the agar surface, and lower colonies found within the agar layer. Virtually all cytolytic clones originated exclusively from the upper colonies.

Association between HLA-D region antigens and disease-free survival in childhood non-T, non-B acute lymphocytic leukemia.

The frequency of three serologically defined HLA-D region antigens--DR, MB, and MT--was determined in a group of 74 children with non-T, non-B acute lymphocytic leukemia (ALL). Statistically, there were no significant differences in the frequency of any antigen in these ALL patients as compared with a panel of 85 normal controls. However, significant differences in HLA-DR frequencies were observed between patients who relapsed or who remained disease-free during a 30-mo period of chemotherapy.

Demonstration of a third structurally distinct human Ia beta chain by two-dimensional gel electrophoresis.

Previous studies have indicated that HLA-DR homozygous cell lines express two Ia alpha and Ia beta chains that combine to form at least two Ia molecules. This report demonstrates by two-dimensional gel electrophoresis the existence of a third structurally distinct human Ia beta chain on DR2 and DR5 cell lines. This suggests that at least five separate genes control the expression of Ia molecules on HLA-DR homozygous cell lines.

Effect of HLA Bw4/Bw6 compatibility on platelet transfusion responses of refractory thrombocytopenic patients.

Platelet transfusions from donors matched for cross-reactive antigens have been shown to be effective in providing hemostasis in alloimmunized thrombocytopenic patients. A significant number of these transfusions, however, fail to provide posttransfusion platelet recoveries. We investigated incompatibility in the Bw4/bw6 system as a possible explanation for these failures.

Molecular relationships of the human B cell alloantigens, MT2, MB3, MT4, and DR5.

The human class II, HLA-linked, B cell alloantigens include the HLA-DR, MB, MT, and Te determinants. Interest in the molecular relationships of these antigens has recently intensified because of their homology to the murine Ia antigens and their possible importance in disease predisposition and transplantation. We have used alloantisera with carefully defined immunochemical as well as serologic specificity, and two immunochemical techniques, sequential immunoprecipitation with analysis by SDS-PAGE and two-dimensional gel electrophoresis, to explore the molecular relationships of the MT2, MB3, MT4, and HLA-DR5 antigenic determinants.

Association of PLT specificity of alloreactive lymphocyte clones with HLA-DR, MB and MT determinants.

The HLA-D region encodes for several serologically defined systems, including DR, MB, and MT. The antigens of MB and MT are strongly associated with two or more DR specificities. The purpose of this study was to determine the role of MB and MT antigens in lymphocyte alloactivation.

Serologic studies in a family with heterozygous C2 deficiency.

Twelve family members of a patient with systemic lupus erythematosus (SLE) and heterozygous deficiency of the second component of complement (C2) were studied. Histocompatibility (HLA) typing was determined for A, B, and DR and MB antigens. Serum samples were tested for a variety of antinuclear antibodies (ANA), lymphocytotoxic antibodies and rheumatoid factors, and C2 levels were determined by hemolytic titration.