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PubMed1980

[Cross test with B-lymphocytes in transplantation immunology].

Cross-matching procedure with B-lymphocytes may facilitate the detection of cytotoxic antibodies. This finding suggested a reliable use of the test in immune monitoring after kidney allograft from living donor. The possible enhancing role of B-lymphocyte antibodies was also discussed.

B-Lymphocytes↗
PubMed1995

The influence of MHC polymorphism on the selection of T-cell determinants of FMDV in cattle.

There is a quest for the development of a new generation of vaccines consisting of well-defined subunit antigens. For a number of practical reasons it is attractive to develop vaccines on the basis of synthetic peptides. However, their efficacy may be limited by genetic restrictions imposed on T-cell recognition via major histocompatibility complex (MHC) polymorphism, as shown by many studies using inbred animal species. To study the effect of MHC polymorphism in an outbred species, we selected four cattle homozygous for different A-DR-DQ haplotypes, and another four cattle which shared one haplotype in combination with a haplotype of one of the MHC homozygous animals. We analysed responses to synthetic peptides comprising defined T-cell epitopes of foot-and-mouth disease virus (FMDV) in this selected group of FMDV-vaccinated cattle. This analysis shows that even in outbred animals. MHC polymorphism influences the responses to synthetic peptides. Interestingly, one of the peptides, VP4[20-34], was recognized in association with at least four different MHC haplotypes. Fine specificity analysis of this peptide revealed subtle shifts in the core epitope recognized. All peptides that induced lymphocyte proliferation in vitro were found to induce a T-helper type-1 (Th1) type of response, irrespective of the MHC haplotype involved. Together, these data support the notion that individuals carrying distinct MHC types can be vaccinated successfully by vaccines that include only a limited number of peptides. In the design of a peptide vaccine against FMDV we suggest inclusion of the highly conserved VP4 sequence 20-34.

Amino Acid Sequence↗
PubMed1976

[Contributions to the mechanism of the Witebsky-test (author's transl)].

The assumption that immune and normal alloantibodies of the ABO system could be distinguished by partial neutralization with soluble specific blood group substance (Witebsky test) has been reexamined. The following results were presented: 1. ABO alloantibodies in both normal and immune sera as well as their IgG and IgM preparations were inhibited by their homologous soluble specific blood group substances. 2. Anti-A and anti-B immune antibodies as well as normal antibodies of the IgG class were found to be strong hemagglutinins in a saline medium; therefore they have to be called "complete" hemagglutinins as have anti-A and anti-B antibodies of the class IgM too. 3. ABO alloantibodies in both IgG and IgM preparations were able to form precipitation pellets with their homologous soluble specific blood group substances. IgM revealed a stronger precipitation power as IgG. 4. Agglutination reaction in saline was inhibited by the 50-200-fold group substance concentration needed for a optimal precipitation reaction, whereas agglutination of enzyme treated erythrocytes or red cells tested with antiglobulin (Coombs) sera was inhibited by a 20,000-80,000-fold concentration of the blood group substances. 5. Soluble antigen-antibody complexes, prepared from solubilized precipitates or from Witebsky test mixtures using chromatography, ultracentrifuge or ultrafiltration for separation were able to agglutinate erythrocytes in the antiglobulin or papain test. Following conclusions were drawn: A. Soluble antigen-antibody complexes are the main component leading to a positive Witebsky test. B. The mechanism of the Witebsky test as it has to be assumed in respect to our findings do not allow the distinguish immune and normal alloantibodies resp. IgG and IgM alloagglutinins in the ABO system.

ABO Blood-Group System↗
PubMed1984

Ninth International Histocompatibility pre-workshop testing of Dw6 HTCs. Two subtypes of Dw6.

In the scope of the cellular part of the 9th International Histocompatibility Workshop, the offered homozygous typing cells (HTCs) of several specificities have been screened in a pre-Workshop. In Leiden and Seattle all HTCs typing for "HLA-Dw6" have been tested. This implied that in both laboratories mixed lymphocyte culture (MLC) matrices were performed between the Dw6 HTCs and that all HTCs were tested as stimulator cells against a panel of heterozygous responder cells. The results clearly demonstrated that "HLA-Dw6" as defined by the different participating laboratories can be split into two major groups, Dw6.a and Dw6.b. This observation confirms and extends previous reports. None of the 9th Workshop B-cell sera could discriminate between the two presently described subgroups of HLA-Dw6.

HLA-DR Antigens↗
PubMed2006

Suppression of (5R)-5-hydroxytriptolide (LLDT-8) on allograft rejection in full MHC-mismatched mouse cardiac transplantation.

BACKGROUND: (5R)-5-hydroxytriptolide (LLDT-8) is a new compound derived from triptolide, which is the major immunosuppressive fraction of Tripterygium wilfordii Hook. F (TWHF). Studies in vitro and in vivo have demonstrated that LLDT-8 had potent immunosuppressive activities. Here we tested LLDT-8 in major histocompatibility complex (MHC)-mismatched cardiac transplantation and investigated the mechanisms underlying the prevention of transplant rejection. METHODS: LLDT-8 was administered orally to recipients in Balb/c to C57BL/6 murine cardiac transplantation model. Allograft survival after transplantation was recorded in recipients. The T cell immunity and cytokine production were observed. Histological analysis was performed. The chemokine and its receptor were analyzed by reverse transcriptase-polymerase chain reaction on cardiac graft RNA. RESULTS: LLDT-8 administered orally significantly induced the survival prolongation of allogeneic cardiac graft. Histological results showed that LLDT-8 well preserved myocardium and significantly reduced infiltration of the graft with inflammatory cells. LLDT-8 decreased IL-2 production in recipient splenocytes stimulated by concanavalin A (ConA) ex vivo. LLDT-8 significantly inhibited the immunoreactivity of recipient to specific donor alloantigens, but preserved immunity to third-party alloantigens and mitogen. However, the flow cytometry analysis of the proportion of CD4+, CD8+ T cell subgroup in recipient spleens showed LLDT-8 had a normalizing effect on the splenic lymphocytes population. LLDT-8 decreased CC chemokine receptor 5 (CCR5) and their ligands macrophage inflammatory protein 1 alpha (MIP-1alpha) and beta (MIP-1beta) mRNA expressions in allografts. CONCLUSION: The results outline the great potential of LLDT-8 as a therapeutic tool in transplant rejection.

Animals↗
PubMed1993

Limiting dilution analysis of the allo-MHC anti-paternal cytotoxic T cell response. II: Recurrent spontaneous abortion and the effect of immunotherapy.

Using limiting dilution analysis (LDA) we determined anti-paternal cytotoxic T lymphocyte precursor (CTLp) frequencies in the peripheral blood of 10 women with unexplained recurrent spontaneous abortion (RSA) before and after immunization with paternal lymphocytes. The women and their partners were HLA tissue-typed and none of the women had anti-paternal cytotoxic antibodies (APCA) before immunization. All other known causes of RSA were excluded. All 10 women were found to have high frequencies of specific anti-paternal cytotoxic T cells before immunization (range 1/1030 to 1/9574). Splitwell analysis showed that these cytotoxic cells were specific to paternal MHC antigens. These frequencies rose significantly following immunization (range 1/683 to 1/4652). The cytotoxic T lymphocyte frequencies against an HLA-mismatched third party varied from woman to woman, but were not affected by the immunization. The LDA data conformed to single-hit kinetics, indicating that only cytotoxic T cells were limiting in the assay. Our data are in sharp contrast to the previously held view that women with RSA may be hyporesponsive to paternal MHC antigens. Immunizing such women with paternal leucocytes further sensitizes them. These findings cannot be reconciled with a favourable outcome in the treatment of RSA with immunotherapy. We would argue that this treatment is at best of unproven value, and may even be harmful. That these women may sometimes have successful pregnancies following immunotherapy testifies to the effectiveness of the classical MHC antigen-deficient trophoblast as an immunological barrier between mother and fetus.

Abortion, Habitual↗