Rabbit complement in the lymphocytotoxicity test.
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In Indian patients with Multiple Sclerosis it was observed, upon testing for serum determinable histocompatibility antigens, that HLA-B12 antigen was present in excess (77.7%), in comparison with normal controls (13.8%). In fact, in the sub-group of 'clinically definite' patients, the B12 antigen excess was remarkable (84%). These preliminary findings seem to point to a different immunogenetic profile of Indian MS patients in comparison with Western and Japanese series. Relevant published information relating to the HLA-B12 alleles in disease processes is discussed in order to provide a basis for further work.
BoLA w6 is one of 16 specificities agreed upon at the 2nd International BoLA Workshop and many laboratories have produced sera reacting with this specificity. This paper presents evidence for at least four sub-groups in w6. Three Edinburgh sera showing identical reaction patterns in the 1st and 2nd BoLA workshops have been studied by absorption. One serum contained a single antibody population reacting with an epitope common to all w6 positive cells. The other two contained antibodies against the same common epitope, antibodies to epitopes on four sub-groups and an antibody reacting only with one subgroup w6.1. Another Edinburgh serum contained two populations of antibodies. One reacting with the common epitope and one with an epitope on two or possibly three subgroups. Immunisation between w6 positive individuals produced antisera to two subgroups without producing antibodies to common epitopes. At least two additional subgroups are likely to exist. These results indicate the presence of specificities unique to individual subtypes coupled with epitopes common to some and all w6 subgroups.
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The development and persistence of leukoagglutinating and lymphocytotoxic alloantibodies were systematically studied in 54 patients who had received single massive blood transfusions because of open-heart surgery. Of these patients over 95% were sensitized when tested for both leukoagglutinins and lymphocytotoxins, whereas 74% were found to be immunized when searched for cytotoxic antibodies only. The optimal time for detecting antibodies was 2 weeks after blood transfusions.
In a prospectively designed comparative study, HLA class I and class II typing was performed after B/T lymphocyte separation by nylon wool (NIH technique) or using immunomagnetic beads (IMB technique). Samples from 70 healthy individuals (group A) and from 47 patients with potential typing problems (group B) were examined. Complete typing failures occurred more frequently with NIH than with IMB. Except for class I in group A, the rate of correct decisions for antigen assignment was significantly higher with IMB. For class II serum reactions, specificity, predictive value and efficiency were superior for IMB, while sensitivity appeared higher for NIH. This can possibly be explained by a prozone effect which was observed in 9 of 36 titration experiments only with the IMB technique. Correlation of both methods in 104 crossmatches prior to kidney transplantation was poor using peripheral blood lymphocytes, but excellent with spleen cells. In addition to the important advantages of the IMB technique for potential problem cases, especially for class II, typing time is reduced to one third, which is of particular clinical importance in kidney transplantation. However, lymphocytotoxic crossmatches prior to transplantation, if performed by IMB, should be carried out using different serum dilutions or with donor spleen cells instead of peripheral blood lymphocytes.
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Human peripheral blood lymphocytes (PBL) were evaluated by their responses to phytohemmagglutinin (PHA-P), concanavallin A (con-A), and pokeweed mitogen (PWM), both before and after treatment with an antiserum against human thymic lymphocyte antigens (HTLA) that had been made T-cell-specific by multiple absorptions with immunoglobulin EAC-positive lymphoblast cell lines (B cells). Cells treated with HTLA were examined for their ability to react in a mixed lymphocyte culture (MLC) and to form killer cells in a cell-mediated lymphocytotoxicity (CML) system. Sensitized cells were also examined for their ability to respond to purified protein derivative (PPD) by blastogenesis, migration inhibitory factor release (MIP), and lymphotoxin (LT) production, both before and after treatment with HTLA and complement. The HTLA was in itself highly stimulatory to PBL. However, with the addition of complement and subsequent cell destruction, a marked decrease in its stimulatory response was noted. PBL treated with HTLA and complement exhibited marked inhibition of responsiveness to con-A with little decrease in PHA-P -OR PWM stimulation except at very high concentration of HTLA. MLC reaction was inhibited only when responder cells were treated with HTLA + C'. Treatment of stimulator cells with HTLA + C' did not significantly alter the MLC response. The HTLA + C'-treated cells failed to form killer cells in the CML reaction and inhibited PPD-induced blasto-genesis from PPD-sensitized individuals; however, treatment of sensitized cells with HTLA + C' had little effects on the release of MIF and LT. It is suggested that subpopulations of T-cells carry surface antigens that bind with this specific antisera, and that the con-A-responsive cells, the responder cells in the MLC, and killer T-cells comprise a separate subset from cells responding to PHA-P or PWM, OR THE MIF-and LT-producing cells.
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Systemic autoimmune disease (AID) can be controlled with conventional therapies in most patients. However, relapses are common, leading to progressive disability and premature death. Nonmyeloablative conditioning and allogeneic bone marrow transplantation (BMT) could be an effective treatment for severe AID, because of mild toxicity of the conditioning and the potential benefits of donor chimerism. We examined the effects of this treatment in experimental autoimmune arthritis. Our results demonstrate the induction of complete donor chimerism and significant suppression of disease activity. No clinical graft-versus-host disease (GVHD) was observed. The beneficial effects were most likely caused by the elimination of plasma cells producing pathogenic autoantibodies, because these antibodies disappeared rapidly after BMT. Although this type of treatment was effective in organ-specific T-cell-mediated AID, the present study provides convincing evidence that nonmyeloablative conditioning and allogeneic BMT can effectively treat severe B-cell-mediated AID with a systemic inflammatory component.
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Previous studies of the rapid rejection of MHC-disparate lymphocytes in rats, named allogeneic lymphocyte cytotoxicity, have indicated that rat NK cells express activating receptors for nonclassical MHC class I allodeterminants from the RT1-C/E/M region. Using an expression cloning system that identifies activating receptors associated with the transmembrane adapter molecule DAP12, we have cloned a novel rat Ly-49 receptor that we have termed Ly-49 stimulatory receptor 3 (Ly-49s3). A newly generated anti-Ly-49s3 Ab, mAb DAR13, identified subpopulations of resting and IL-2-activated NK cells, but not T or B lymphocytes. Depletion of Ly-49s3-expressing NK cells drastically reduced alloreactivity in vitro, indicating that this subpopulation is responsible for a major part of the observed NK alloreactivity. DAR13-mediated blockade of Ly-49s3 inhibited killing of MHC-congenic target cells from the av1, n, lv1, and c haplotypes, but not from the u or b haplotypes. A putative ligand was mapped to the nonclassical MHC class I region (RT1-C/E/M) using intra-MHC recombinant strains. Relative numbers of Ly-49s3(+) NK cells were reduced, and surface levels of Ly-49s3 were lower, in MHC congenic strains expressing the putative Ly-49s3 ligand(s). In conclusion, we have identified a novel Ly-49 receptor that triggers rat NK cell-mediated responses.
Cellular and humoral responses against donor lymphocytes were studied in ten kidney-transplant patients, nine of whom had well functioning grafts. No(T)cell-mediated cytotoxicity against donor cells was demonstrated. Specific anti-donor antibodies were found in two recipients with well-accepted grafts. A single immunological factor responsible for a favourable clinical course was not demonstrated. Neither complete T nor B cell tolerance against donor cells had developed, and a well-tolerated graft could coexist with antibodies directed against donor cells.
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