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Biomedical subjects

L M Vaessen

Publications and source records attributed to L M Vaessen.

At least 73 records · Page 4Linked to original sources
PubMed1992

The influence of HLA-mismatches on phenotypic and functional characteristics of graft infiltrating lymphocytes after heart transplantation.

We studied the influence of HLA mismatches on T lymphocyte cultures that were derived from endomyocardial biopsies (EMB) from 118 heart transplant recipients. From patients with DR mismatches, the majority of the EMB-derived cultures were dominated by CD4, while in patients without DR mismatches, CD8 was the predominant T cell subset. The majority (75%) of the cultures were cytotoxic against donor antigens. A significantly (P < 0.005) lower proportion of the cultures showed cytotoxicity (36%) against HLA-A antigens when compared to HLA-B (53%) or HLA-DR (49%). A dose effect phenomenon was detected for all HLA antigens, including HLA-A: a higher number of A, B or DR mismatches resulted in a higher number of cytotoxic cultures directed against these antigens. B and DR matching had the greatest influence on 6 month freedom from rejection. Both our experimental and clinical data indicated that HLA matching played a role in the immune response against a transplanted heart.

Biopsy↗
PubMed1992

Monitoring of cardiac graft recipients: comparison of in vivo activated, committed T lymphocytes in peripheral blood and in the graft.

The proliferative and cytotoxic capacity of peripheral blood lymphocytes (PBL) and the cytotoxic activity of lymphocytes propagated from endomyocardial biopsies (EMB) towards donor cells was used to identify in vivo activated, committed T cells. A series of 39 PBL samples and 38 EMB simultaneously taken from 20 patients after heart transplantation was cultured in interleukin 2 (IL-2) conditioned medium. The cytotoxic capacity of these cultures against donor cells was tested in a 4-h chromium-51 release assay. From a comparable patient group, 224 samples were evaluated for donor reactivity by a primed lymphocyte test (PLT). Analysis showed that PBL cultures hardly ever contained committed cytotoxic T lymphocytes (cCTL, 2/39) or committed proliferative T lymphocytes (cPTL, 1/224). In contrast, significantly more EMB cultures (17/38, P < 0.001, chi2 test) demonstrated donor-directed cytotoxicity. This was especially found during rejection (11/17 vs 6/21 without rejection, P = 0.05). These results show that after heart transplantation, committed cells are mainly found in the graft.

Cell Culture Techniques↗
PubMed1992

Acute rejection in heart transplant patients is associated with the presence of committed donor-specific cytotoxic lymphocytes in the graft but not in the blood.

In vivo-activated, committed donor-specific cytotoxic lymphocytes (cCTL) can be propagated and expanded from endomyocardial biopsies (EMB) in IL-2-enriched medium especially during an acute rejection episode. We report here our efforts to detect these cCTL by the same technique in peripheral blood at the moment of rejection and when no rejection was diagnosed. During or just before rejection, significantly less frequent (P less than 0.01) donor reactive cCTL were found in PBL samples (two out of 20) than in the simultaneously taken EMB samples (13 out of 19). Donor B-LCL and/or third-party B-LCL were lysed by 15 PBL samples. Inhibition studies revealed that this lysis was due to LAK-like cytotoxicity. The results show that peripheral blood does not reflect intra-graft events, which is probably the reason for the irreproducible results of diagnosis of rejection by monitoring immunological parameters in the peripheral blood.

Biopsy↗
PubMed1991

Phenotypic and functional analysis of T cell receptor gamma delta-bearing cells isolated from human heart allografts.

Endomyocardial biopsies from human heart transplant recipients were investigated with respect to the occurrence of in vivo activated, alloreactive TCR-gamma delta+ cells. More than one year after transplantation, 30% of the biopsy-derived T cell cultures contained TCR-gamma delta+ cells, whereas in the first year after heart transplant in only 8% of the culture TCR-gamma delta+ cells were found. Such an increase of TCR-gamma delta+ cells was not observed in the peripheral blood of the patient. In most biopsy-derived cultures, the gamma delta cells were delta-TCS1+. No donor-specific cytotoxic activity could be demonstrated for TCR-gamma delta+ cells tested, whereas non-MHC-restricted cytotoxicity was found in several cultures. The occurrence of nonalloreactive TCR-gamma delta+ cells late after transplantation, when acute cellular rejection episodes are rare, suggests a role in the down-regulation of the allo-Ir.

Cells, Cultured↗
PubMed1991

Alloreactive lymphoid infiltrates in human heart transplants. Loss of class II-directed cytotoxicity more than 3 months after transplantation.

From 535 endomyocardial biopsies (87 heart transplant recipients) 283 cell cultures could be generated. All cultures tested contained T lymphocytes and in most cases CD4 was the predominant phenotype at any time posttransplant. A significantly higher proportion of CD8-dominated cultures was found among cultures from biopsies without myocytolysis. In the first 3 months post transplant 57% of cultures showed cytotoxicity against both class I and class II mismatched donor major histocompatibility complex (MHC) antigens, changing to an incidence of 33% at greater than 90 days. This proved to be due to a significant decrease in the number of cultures with human leukocyte antigen class II-directed cytotoxicity. This study shows that early after transplantation a heart transplant is infiltrated with activated donor-specific cytotoxic T cells which recognize a broad spectrum of mismatched donor MHC antigens, and that in time this spectrum becomes more restricted.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1990

Rejection prophylaxis with sequential OKT3 and CSA after kidney transplantation.

Sixteen kidney transplant recipients received the IgG2a anti-CD3 monoclonal antibody OKT3 and azathioprine as rejection prophylaxis during the first two postoperative weeks. Concomitant immunosuppression consisted of low dose steroids while cyclosporine A therapy was instituted on day 12. Side effects included fever, bronchospasm, hypotension and diarrhoea. OKT3 caused T cell modulation resulting in CD3 dim+, CD4+ or CD8+, CD5+, WT31- and 11F2- cells. Anti-OKT3 antibodies were found in approximately 50% of the patients. The protocol induced a 100% patient and graft survival and a 81% actuarial freedom of rejection at 18 months. It prevented CsA associated nephrotoxicity in the direct postoperative phase. These beneficial effects outweighed the side effects of OKT3.

Adult↗
PubMed1990

gamma delta T-cell receptor-positive T-cell clones derived from human heart transplants do not show donor-specific cytotoxicity.

gamma delta T-cell receptor-positive T-cell clones were obtained from three endomyocardial biopsies taken from two patients. Clones obtained from two biopsies from patient A, one taken during rejection and one after resolution of this rejection, were WT31-, 11F2+, Ti gamma A-, delta TcS1+. Clones from patient B grown from a biopsy without histologic signs of rejection were either WT31-, 11F2+, Ti gamma A+, delta TcS1-, or WT31+. All gamma delta T-cell receptor-positive clones had cytolytic potential but did not demonstrate donor-specific lysis.

Biopsy↗
PubMed1990

Vascular thymus transplantation in rats. Technique, morphology, and function.

A new method of thymus transplantation is introduced, in which the graft is directly connected with the recipient's vascular system. This procedure was used both in euthymic rats and congenitally athymic nude rats. At all tested intervals after transplantation thymus grafts hardly differed from the recipient's own thymus in immunohistology and lymphocyte yield. In athymic nude rats, T cell-dependent immunity, tested by mitogen- and alloantigen-induced T cell responses, as well as by antibody production and delayed-type hypersensitivity after ovalbumin administration, showed that vascular thymus grafts could generate T cell functions to euthymic control levels. We conclude that the technique of vascular thymus transplantation represents a valuable tool, either in fundamental research on thymus function, or for the purpose of immune (re)constitution.

Animals↗
PubMed1990

Quality control of anti human CD3 and CD4 monoclonal antibodies.

By testing two monoclonal antibodies (MoAb), we tried out two methods, which might be suitable for quality control of anti human T cell MoAb. RIV6 and RIV9 were investigated by flow cytometry and Enzyme-linked Immunosorbent Assay (ELISA). The characteristics, specificity and biological activity of RIV6 and RIV 9 were determined and compared respectively with other CD4 specific MoAb, OKT4, OKT4a, LEU3a and the CD3 specific MoAb OKT3 and LEU4 by flow cytometry. An ELISA was set up to quantify mouse IgG3 antibody. Specificity, plate variability and day to day effects were determined. The results illustrate that ELISA and flow cytometry are valuable tools in the quality control of these antibodies.

Animals↗
PubMed1989

Changes in immunological parameters after conversion from cyclosporine A to azathioprine in renal transplant recipients.

Long term CsA therapy did not interfere with the basal levels of natural killer (NK) activity in stable cadaveric renal transplant recipients. However, 3 months after changing immunosuppressive therapy from CsA to AZA, NK activity was significantly decreased (36 +/- 25% vs 19 +/- 15%, P less than 0.01). Following in vitro exposure to IFN-gamma an increase in NK activity from 36 to 44% (P less than 0.05) could be induced during CsA therapy but this was no longer observed after conversion to AZA (19 to 22%, N.S.). A prominent decline in the number of NK cells expressing the surface receptor for the Fc portion of IgG was also found postconversion. The IFN-gamma production capacity after mitogen stimulation of unprimed lymphocytes was more depressed during CsA than during AZA therapy (median 25 vs 80 U/ml 10(6) cells, P less than 0.05), suggesting a reversible inhibition of CsA on lymphokine production. Despite the better IFN-gamma production capacity, both the activity, inducibility and number of NK cells were significantly lower under AZA therapy than under CsA therapy. These findings indicate that CsA exerts its immunosuppressive action without an important interference with NK activity. Monitoring mononuclear cells showed a decrease in absolute numbers of all phenotypically distinct cells studied after conversion. The prominent decrease in CD 8 cells resulted in an increase of CD 4/CD 8 ratio.

Azathioprine↗
PubMed1987

Implantation of cultured thymic fragments in congenitally athymic (nude) rats. Influx of lymphocytes and dendritic cells.

We studied the early events in athymic immunoincompetent rats after implantation with cultured thymic fragments (CTF) under the kidney capsule, with special emphasis on the settlement of lymphocytes and non-lymphoid RT1 class II elements. At 2 weeks after grafting, tissue under the kidney capsule comprises strands of keratin-positive epithelial cells from the graft, without immigrant cells. At 3 weeks, the CTF graft is populated with lymphocytes and with non-lymphoid RT1 class II-positive cells expressing the recipient haplotype (allogeneic combinations). Part of these cells bear determinants recognized by an anti-rat dendritic cell antibody. At 4 weeks the graft exhibits a completely restored thymic architecture. At the periphery, the first indications of T-cell competence generated after CTF implantation are observed 6 weeks after implantation. At 18 weeks, the peripheral thymus-dependent immune system is almost completely developed. This includes in vitro alloreactivity, even to the donor RT1 haplotype of the graft. But skin grafts of the allogeneic CTF donor haplotype are not rejected. Thus, a state of in vivo tolerance is induced under the influence of grafted epithelium, which is not due to a specific deletion of alloreactive cells. We conclude that CTF regain their original thymic architecture between 2 and 4 weeks after implantation in (allogeneic) athymic nude recipients, and that only after this restoration does peripheral thymus-dependent immune competence start to develop.

Animals↗
PubMed1986

Implantation of cultured thymic fragments in congenitally athymic nude rats: ignorance of thymic epithelial haplotype in generation of alloreactivity.

We studied the development of thymus-dependent immunity in congenitally athymic nude rats after implantation of cultured thymic fragments (CTF), particularly the development of in vitro alloreactivity in allogeneic combinations. CTF of DA (RT1a), PVG (RT1c), and RP (RT1p(u,1] origin were implanted in nude rats of WAG (RT1u) origin. In analysis 14 to 18 wk later, all recipients exhibited thymus-dependent immunocompetence assessed by (immuno)-histology of lymphoid organs and responsiveness to in vitro concanavalin A stimulation and in vivo ovalbumin immunization. Control nude animals were unresponsive. Also, in vitro alloreactivity was observed, measured by mixed lymphocyte reaction and cell-mediated lympholysis. The alloresponse to the allogeneic CTF donor haplotype was as to a third party, but that to the recipient was negative. The CTF before implantation were devoid of lymphoid elements and revealed epithelial-like cells as the major component. Cells in CTF showed expression of RT1 class I and class II antigens. CTF at autopsy had the architecture of a normal thymus. In immunohistochemistry using haplotype-specific antibodies, lymphocytes showed RT1u class I expression as in the normal WAG thymus. In the cortex-like area of CTF, stromal cells revealed class I and class II haplotype expression of the donor thymus, but in the medulla-like area, class II haplotype expression was that of the recipient WAG rat. These data indicate that after implantation in nude rats, CTF become populated not only with lymphoid elements, but also with stromal components from the recipient. In induction of thymus-dependent immunity, these acceptor-derived stromal (dendritic) cells may be involved in generation of allospecificity; class I and class II haplotype expression by the donor cortex (epithelial) compartment is ignored in this process.

Animals↗