[Identification of LD determinants by mixed lymphocyte culture test (author's transl)].
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Alleles↗
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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↗
The mixed lymphocyte culture test.
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Graft Survival↗
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↗
The immunosuppressive mechanism of azathioprine. I. In vitro effect on lymphocyte function in the baboon.
The effect of azathioprine on in vitro baboon lymphocyte function tests was evaluated using the mitogen stimulation test, the mixed lymphocyte culture test, the migration inhibition factor test, the cell-mediated lymphocytotoxicity test and the antibody dependent cell-mediated cytotoxicity test. It was found that azathioprine inhibited phytohemagglutinin and Concanavalin A stimulation at lower concentrations than those required to inhibit pokeweed mitogen stimulation. It inhibited the MLC reaction with as little as 0.2 mug/culture in the microculture system. Azathioprine had no effect on (a) the release of migration inhibition factor, (b) the cell-mediated lymphocytotoxicity assay if presensitized cells were used, and (c) the antibody-dependent cell-mediated cytotoxicity assay. However, azathioprine inhibited CML if it was added during in vitro sensitization and induction of killer cells. These in vitro results suggest that azathioprine inhibits those reactions which require cellular division.
Animals↗
[Mixed lymphocyte culture (MLC test) in HLA identical relatives and between HLA identical unrelated subjects (author's transl)].
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HLA Antigens↗
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↗
Evidence for subtypic determinants in the HLA-DW3 cluster.
This study was undertaken to get more insight into the previously suggested heterogeneity of the HLA--DW3 cluster. Preliminary evidence of DW3 heterogeneity was derived from results of intrafamilial mixed lymphocyte culture tests (MLC) where cells of apparently homozygous offspring revealed unexpected stimulations of one of the parents' cells. Therefore, 15 different homozygous typing cells (HTCs) of DW3 specificity were tested against 43 HLA-DW3 heterozygous individuals. The response patterns of the 43 HLA-DW3 heterozygous cells toward 13 HTCs leads to the definition of at least three groups of DW3 stimulating cells. According to these patterns, four groups of responding cells could be classified. These results were confirmed by a MLC checkerboard experiment running all DW3-HTCs against each other. Discussing all possible explanations for these observations, the authors conclude that the existence of DW3 subtypes having some properties in common is the most likely interpretation of the results obtained. Family segregation studies will be needed to define the genotypic situation of the DW3 cluster.
Antibody Specificity↗
Lymphocyte-defined loci in cattle.
Using the results of all paired one-way mixed lymphocyte culture tests on families of half-sibs, we have established that the lymphocyte-defined system in cattle contains a minimum of two loci. The methodology presented is applicable to studies of the lymphocyte-defined systems of other species.
Alleles↗
The distribution of HLA-d (MLC) locus determinants in the Czech population.
The distribution of 8 HLA-D determinants in the group of 83 unrelated individuals of the Czech population was ascertained by means of homozygous typing cells using the mixed lymphocyte culture test. In the panel tested, the gene frequencies of HLA-D determinants, the distribution of HLA-D phenotypes and the magnitude of the linkage disequilibrium between the determinants of the HLA-B and HLA-D loci were determined.
Czechoslovakia↗
T-and B-lymphocyte chimerism in the marmoset.
Marmosets are natural blood chimeras, this condition resulting from the high frequency of fraternal twinning and the consistent development of placental vasular anastomoses between the two embryos. Identification of chimerism by sex-chromosome analysis of cultured blood lymphocytes provided a means of determining the proportion of chimerism among T and B lymphocytes. Peripheral blood lymphocytes were enriched for T or B cells by filtration through a nylon column (yields greater than 95 per cent T-cells) or inactivation of T lymphocytes by treatment with a goat anti-marmoset thymocyte antiserum in the presence of complement (yeilds greater than 95 per cent B cells). Mitogenic stimulation of these separated, enriched cell populations yielded metaphase plates which could be scored for percentage male and female cells. Tests on five different blood chimeras showed the T- and B-lymphocyte chimerism to be the same. Stimulation of blood lymphocytes with cells from another species of marmoset in a mixed lymphocyte culture test revealed the chimeric T-cell response (i.e., host and co-twin cells) to be similar to that obtained with a mitogenic lectin. The demonstration of equivalent T- and B-cell chimerism in these animals suggests derivation of these cells from a common stem cell pool and the response of both T-cell populations to an antigenic stimulus in proportions similar to their percentage chimerism suggests complete immunologic tolerance exists in this species for co-twin histocompatibility antigens.
Animals↗
Diminished mixed lymphocyte response in ankylosing spondylitis.
A diminished mixed lymphocyte response was reported by Nikbin et al. (1976) among patients with ankylosing spondylitis, their asymptomatic relatives and also normal controls carrying the B27 antigen. In the present communication, the responses in 48 ankylosing spondylitis patients and 45 controls were examined in mixed lymphocyte cultures tested against a 'standard stimulator' made up of pooled lymphocytes. A significantly diminished response is confirmed among the ankylosing spondylitis patients, but not in the control group carrying the B27 antigen. The diminished mixed lymphocyte response therefore appears to be more directly associated with the disease than with the B27 antigen, and possibly represents a specific T-cell defect associated with the pathogenesis of the disease.
Female↗
[Responsiveness of lymphocytes to stimulation by pha and autologous blasts in acute lymphatic leukemias during remission (author's transl)].
The paper deals with the question how far the lymphocytes of acute lymphatic leukemia patients in remission can be stimulated by PHA and autologous blasts in the MLC-Test. The lymphocyte stimulation by PHA showed considerable variation dependent on the phase of therapy. A significant reduction of the responsiveness to PHA stimulation was found during the prophylactic reinduction treatment with Vincristin and Prednisolon compared with the results during maintenance therapy. The examination of lymphocyte stimulation by conserved autologous blasts in the mixed lymphocyte culture-test yielded positive results in one of three children at the beginning of remission and in 2 of 3 children during the consolidated remission period.
Child↗
Lymphocyte transformation in vitro in patients with immunodeficiency diseases: use in diagnosis, histocompatibility testing and monitoring treatment.
We have used lymphocyte stimulation in vitro to characterize the degree of cell-mediated immunodeficiency in different patients. The effect of treatment of patients with immunodeficiencies is illustrated by lymphocyte transformation in vitro before, during and after therapy (eg bone marrow transplantation of severe combined immunodeficiency and transfer factor treatment in Wiskott-Aldrich syndrome). The mixed lymphocyte culture test has been used for selection of bone marrow transplant donor for a patient with CID when an HL-A identical sib was not available. The results of the transplantation in this patient with graft from a donor who differed from the patient in respect of both HL-A haplotypes (but MLC identical with the patient) is reported. The MLC data on another family with 2 children with Nezelof disease are reported. The data indicate that one of the patients could be a chimera after intrauterine grafting of maternal immunocompetent cells.
Adolescent↗
Antibody-dependent cellular cytotoxicity and chronic renal allograft rejection.
Antibody-dependent cell-mediated cytotoxicity (ADCMC) was studied retrospectively in 20 long-term (1.3-11 years) renal allograft recipients. The serum of all seven patients having persistent proteinuria greater than 1 g/24 hr exhibited positive ADCMC activity to donor lymphocytes. All 11 patients having a negative ADCMC test had normal levels of urinary protein (mean + 0.28 +/- 0.06 (SE) g/24 hr). Two patients had a positive ADCMC test but had normal urinary proteins and no evidence of chronic rejection. In the ADCMC positive group, the mean serum creatinine values were significantly higher and the mean creatinine clearance values were significantly lower than in the ADCMC negative group. There was no significant positive correlation between the presence of ADCMC and the number of HL-A mismatches identified or the response to donor lymphocytes in mixed lymphocyte culture tests in the two groups. Two patients had transplant nephrectomy and histologically showed both glomerular and arterial lesions of chronic rejection. Post-transplant ADCMC activity showed a significant correlation with proteinuria, decreased creatinine clearance, and elevated serum creatinine, and suggests that the chronic rejection syndrome may be related to ADCMC activity in the recipient.
Antibody Specificity↗
[Immunological examination of lymphocytes in patients with sarcoidosis and bronchial asthma (author's transl)].
Blood lymphocytes of 31 patients with bronchial asthma and of 20 patients with sarcoidosis were isolated and investigated for the following surface markers: E-receptors (E-rosettes, a T cell characteristic), Fc-receptors (EA-rosette assay), C3-receptors (EAC-rosette assay), surface immunoglobulins (immuno-fluorescence technique). In addition, the reactivity against allogeneic lymphocytes was tested in mixed lymphocyte culture (MLC). MLC properties were defined by homozygous reference cells (so-called LD-typing). HLA-typing was performed simultaneously. The results, so far, indicate in both disease groups a series of remarkable alterations of the lymphatic subpopulations. A marked diminution of circulating T lymphocytes was encountered in some cases. The immunological results are discussed in relation to pathogenetic aspects of the diseases.
Asthma↗
Prelusive skin grafts in live-donor kidney transplantation.
In recipients receiving oral azathioprine (50 mg. a day) the fate of skin grafts taken from live human prospective kidney donors correlates well with the outcome of kidney transplantation from the same donors. Correlation between the fate of skin grafts and HL-A serotyping is not nearly as good except in the case of complete HL-A identity. There is little or no correlation between the mixed-lymphocyte-culture test and the fate of skin grafts and the outcome of kidney transplantation. When the degree of compatibility does not exceed one haplotype the results of live human-kidney transplantation can be significantly improved if the fate of a preliminary skin graft from the prospective kidney donor is used as an index.
Azathioprine↗