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Transitivity of response in the mixed lymphocyte culture test.

We present a model of mixed lymphocyte culture (MLC) response which assumes one specificity per locus. It also assumes that an animal A will fail to stimulate animal B if, and only if, the set of specificities possessed by A is a subset of the set of specificities in B. The last assumption implies that non-stimulation is transitive; that is, if A does not stimulate B, and B does not stimulate C, then A will not stimulate C. The inclusion of antigenic sets can be used to partially order the animals in a hierarchy. Partial ordering can detect multiple lymphocyte-defined (LD) loci with relative ease; it indicates the number of antigens present in particular individuals; and it detects exceptions to the rule of transitivity which may expose immune response genes, minor loci, or other mechanisms that affect MLC response. This analytical procedure is most useful when testing half-sib families or hybrids sharing a common parental strain. We have applied this procedure to the MLC in cattle half-sib families and found that the data strongly support the existence of at least four LD loci.

Animals↗

Response of HL-A identical unrelated individuals in the mixed lymphocyte culture test.

The immunological responsiveness as measured in the mixed lymphocyte culture test has been studied in 13 pairs of HL-A identical unrelated individuals. In all combinations stimulation occurred and it was frequently of a similar magnitude to that observed against non-HL-A identical subjects.It is postulated that another locus, adjacent to the HL-A locus, is also responsible for the non-stimulation observed between HL-A identical siblings, and that observations made in the related situation should not be transposed to the unrelated situation without some reservation.

Cytotoxicity Tests, Immunologic↗

Accelerated mixed lymphocyte culture test potentially useful for cadaver organ transplantation.

The mixed lymphocyte culture (MLC) test accurately predicts the strength of rejection of transplanted organs. Ironically, the MLC test can seldom be used in cadaver organ transplantation, where it would be most useful, because the test takes 4 to 6 days to get results. We have developed an accelerated MLC test using precultured cells, which gives interpretable results in 24-48 hr. Precultured cells become responsive to T cell growth factor, reflecting the generation of lymphoblasts during preculture. When 7 consecutive pairs of subjects were compared in the fast and conventional MLC tests, the response at 48 hr in the fast MLC was significantly correlated with the response in the conventional MLC test on days 5 and 6 (P less than 0.01). Precultured cells can be frozen in liquid nitrogen without loss of accelerated responsiveness and significant proliferation is also observed within 24 to 48 hr using frozen cultured cells. Thus, preliminary culture followed by cryopreservation provides the basis for a clinically useful fast MLC test for cadaver organ transplantation.

Animals↗

Rapid mixed lymphocyte culture testing by analysis of the insulin receptor on alloactivated T lymphocytes: implications for human tissue typing.

Responses in the mixed lymphocyte culture (MLC) are traditionally evaluated by measurement of DNA synthesis or blast transformation. However, these events occur too late in the MLC to permit prospective matching for cadaveric renal transplantation. Presentation of allogeneic cells to the T lymphocyte within the MLC results in the emergence of an insulin receptor pharmacokinetically similar to that on other tissues such as fat, liver, and muscle. Intrafamilial MLC were studied by simultaneous assessment of DNA synthesis and insulin receptor binding. In 68 studies from seven families that provide examples of two haplotype identical matches, haplo-identical matches and total haplo mismatches, the presence of an insulin receptor correlated in every case with a positive MLC as estimated by [3H]thymidine incorporation. A quantitative relationship existed between the strength of the MLC and the amount of receptor binding. Based on analysis of cells from several families in which crossover events were known to have occurred, the appearance of an insulin receptor always corresponded with a mismatch at the portion of histocompatibility leukocyte antigen (HLA) chromosome bearing the D region. Finally, it was demonstrated in each of 30 cultures that insulin receptor emergence occurred significantly before detectable DNA synthesis, as early as 24 h after the initiation of the MLC, well within the time-constraint limitations for renal preservation. Appearance of the insulin receptor on activated lymphocytes may be a more rapid measure of mixed lymphocyte responses, and should permit prospective matching for cadaveric renal transplantation.

Cadaver↗

Prognostic value of HL-A typing and mixed lymphocyte culture tests in kidney transplantations from living related donors.

In 45 recipients of intrafamilial kidney transplants, the one year survival of sibling grafts was 95% and of parental grafts 71%. HL-A compatible grafts had a one year survival of 94% compared to 75% of HL-A incompatible grafts. At the same time, the survival of MLC compatible grafts was 100% and that of MLC incompatible grafts 74%. These differences are not statistically significant but suggest a prognostic importance of MLC, as well as HL-A matching. MLC compatibility might be more crucial than HL-A compatibility. MLC test performed after transplantation when the recipients were on maintenance immunosuppressive therapy, did not show reduced reactivity of recipient lymphocytes as compared to tests performed prior to surgery.

Consanguinity↗

Antigens associated with acute leukemia detected in the primed lymphocyte test.

Mixed lymphocyte culture (MLC) studies of families with several leukemia patients, all potential bone marrow transplant recipients, demonstrated that cells from acute myelogenous leukemia patients (5 of 5) and acute undifferentiated leukemia patients (1 of 4) in relapse stimulated autologous lymphocytes as well as lymphocytes from siblings known to be identical at the major histocompatibility linkage group. In the patients studied, the blast transformation induced by leukemia cells was not detectable when the patient was in remission. Stimulation by leukemia cells also elicited increased responses of the lymphocytes from normal haploidentical siblings, parents, and unrelated individuals as compared to stimulation by normal allogeneic cells or leukemia cells of patients with leukemia in remission. The primed lymphocyte test (PLT) was used successfully to establish HLA-D identity of the leukemia patients and their respective HLA-identical siblings, despite high percentages of circulatory blasts. Utilizing lymphocytes from normal siblings primed against the leukocytes from an HLA-identical sibling with leukemia, we also presented results of PLT's which suggested that the stimulation induced by leukemia cells in MLC was produced by leukemia-associated antigens.

Acute Disease↗

Pretransplant monitoring of donor-recipient compatibility.

In these studies, pretransplant testing of donor-recipient compatibility was performed in 10 related donor and 26 cadaveric renal transplants using a variety of cell-mediated immunity tests. Mixed lymphocyte culture results did not correlate with acute rejection (AR), acute irreversible rejection, or chronic rejection (CR). Lymphocyte-mediated cytolysis also did not correlate with AR, acute irreversible rejection, or CR. Cell-mediated lympholysis correlated with AR but not with acute irreversible rejection or CR. Antibody-dependent cell-mediated cytolysis (ADCMC) was positive pretransplant in 13 (36%) of the recipients. Of the positive patients, 4 had early severe AR and 9 developed typical CR. Of these 13 patients, 9 or 69% lost graft function to rejection whereas only 3 of 20 (15%) of ADCMC-negative patients lost graft function because of rejection (P less than 0.05). In summary, cell-mediated lympholysis testing demonstrated a capability to predict AR episodes. The most useful pretransplant monitoring assay in this patient series was the ADCMC assay. A positive ADCMC against donor cells pretransplant indicates a relatively poor prognosis for long-term graft survival.

Antibody-Dependent Cell Cytotoxicity↗