Presentation of human minor histocompatibility antigens by HLA-B35 and HLA-B38 molecules.

Abstract

Cytotoxic T lymphocyte (CTL) clones specific for human minor histocompatibility antigens (hmHAs) were produced from a patient who had been grafted with the kidneys from his mother and two HLA-identical sisters. Of eight CTL clones generated, four recognized an hmHA (hmHA-1) expressed on cells from the mother and sister 3 (second donor); two recognized another antigen (hmHA-2) on cells from the father, sister 2 (third donor), and sister 3; and the remaining two clones recognized still another antigen (hmHA-3) on cells from the father and sister 3. Panel studies revealed that CTL recognition of hmHA-1 was restricted by HLA-B35 and that of hmHA-2 and hmHA-3 was restricted by HLA-B38. The HLA-B35 restriction of the hmHA-1-specific CTL clones was substantiated by the fact that they killed HLA-A null/HLA-B null CTLs (3,[9][10][11][12]. Other hmHAs were also detected by CTLs (2, 4-8). As in the mouse, responses of these CTLs were restricted by major histocompatibility complex class I antigens. Only a limited number of HLA allospecificities have been reported as restriction molecules in the recognition of hmHAs. Among them, HLA-A2 and -B7 were two major ones, although there are examples of restriction by 13). These studies strongly suggested that certain HLA-A/B molecules can present hmHAs to T cells.In the present study, we attempted to generate CTL clones recognizing hmHAs from a recipient of multiple renal grafts and to characterize in detail CTL clones and corresponding hmHAs. Furthermore, the restriction elements in the CTL recognition of hmHAs were investigated by using Hmy2CIR cells transfected with HLA class I genes.MATERIALS AND METHODS Cells. B cells from the patient and the second donor (sister 3; S3 in Fig. 1) were transformed by Epstein-Barr virus (EBV). These transformed B-cell lines were grown in RPMI 1640 medium supplemented with 2 mM L-glutamine and 10% (vol/vol) fetal calf serum. EBV-transformed cell lines HVB-B5, LeolO, FS, CRB-087B, JBUSH, and AMAI were kindly supplied by F. Otani (Kitasato University, Kanagawa, Japan) and grown in the same medium. Peripheral blood lymphocytes (PBL) from six healthy staff members (Sug, YO, KT, YK, ST, and Tm) as well as those from the father, mother, sister 2 (S2), and S3 of the patient were stimulated twice by 0.2% phytohemagglutinin (PHA; Difco). The PHA-induced T cells were used as target cells for the CTL assay.Hmy2CIR cells expressing HLA-B35, -B51, or -Bw52 antigens or HLA-B51/35 chimeric antigens were previously generated (14-16). The HLA-Bw53 genomic gene has been cloned and transfected into Hmy2CIR cells (unpublished data). These cells were grown in the same medium supplemented with hygromycin B (0.2 mg/ml). Untransfected Hmy2CIR cells were grown in the medium without hygromycin B.Antibodies. W6/32 (17) and ME40.5 (18) HLA class I monomorphic, L243 HLA-DR monomorphic (19), antiLeu-10 HLA-DQ monomorphic (20), B7/21 HLA-DP monomorphic (21), OKT3 anti-human CD3 (22), OKT8 anti-human CD8 (22), OKT4 anti-human CD4 (22), and WT31 anti-human a,,3 T-cell re...

Polymorphism of human minor histocompatibility antigens: T cell recognition of human minor histocompatibility peptides presented by HLA-B35 subtype molecules.

“…and the cells were used for CTL assays. EBV-transformed cell lines from German individuals, EBV-transformed self-B cell line from a recipient of HLA-identical renal grafts (Pt cells) (7), EBVtransformed B cell lines from a donor (Dn-2) (7), and Hmy2CIR (CIR) cells were maintained in RPMI 1640 medium with 10% FCS. T2 cells expressing HLA-B*3501 (T2-B*3501) were previously generated (29), and they were cultured in RPMI 1640 medium with 10% FCS.…”

“…B*3501-restricted Human mH Antigen-specific CTL Clones. Our previous study (7) showed that four mH antigen-specific, HLA-B*3501-restricted CTL clones (NH-5.2, NH-5.3, NH-5.5, and NH-5.9) killed B cell lines derived from all of five individuals with HLA-B35. These findings suggest that the human mH antigens recognized by these CTL clones are very well conserved among Japanese or other populations and that only members of this family lack these antigens.…”

Transfected human class I gene product adequately assembles minor histocompatibility antigens

“…Similar to the naturally expressed A2 gene products, the transfected gene products associate with the miH peptide(s) creating an assemblage suitable for A2/miH antigen specific T-cell receptor recognition. Recognition of human miH antigens upon Implantation or by gene transfection of the required HLA antigens has been reported previously (Zier et al 1987, Yamamoto et al 1990). Our results are in line with both latter reports.…”

Adoptive immunotherapy against kidney targets in dog-leukocyte antigen-identical mixed hematopoietic canine chimeras

“…Yet, such organs, when transplanted into MHC-matched individuals, in the absence of immunosuppressive treatment, are promptly rejected (11,12). After rejection, cytotoxic T lymphocytes (CTLs) directed against the rejected organs may be identified (11,13). Here, we used a canine model of mixed-donor host-hematopoietic chimerism (14 -16) to explore whether marrow donor cytotoxic lymphocytes could be generated against chimera kidney that, upon infusion of such lymphocytes into the chimera, caused kidney damage, a finding that would be consistent with graft-versus-kidney effects.…”