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1.
For optimal use of the mixed lymphocyte reaction (MLR) for genetical and transplantation research in dogs, insight into the kinetics of the test is required. Therefore, cell-dose response curves were determined for various genetically defined cell mixtures. The optimal cell concentration was variable. No relation between genetic disparity and optimal cell concentration or responder/stimulator cell ratios was found. In time-course kinetics studies, earlier peak reactivity was observed with combinations from unrelated dogs than with those from related dogs. No difference was found when comparing related dogs with one haplotype difference with those with two haplotype differences. These results can be explained by the assumption that, in those combinations which differ for the major histocompatibility complex (MHC), non-MHC gene products can influence the outcome of the MLR. For a semiquantitative evaluation, a shorter culture period than that used for the determination of MLR negativity is preferred. Apparently, the combined use of different modifications of the test procedure for a single cell combination may increase the value of the test as a genetic tool. It remains to be seen whether such modifications will be sufficient for a complete evaluation of the complex genetic control of the MLR and its relevance to homotransplantation.  相似文献   

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In the course of the culture of an autologous mixed lymphocyte reaction (AMLR), T cells proliferated in response to autologous non-T cells, and differentiated to cytotoxic T cells (AMLR killers). DNA synthesis was necessary to generate AMLR killers, as the elimination of autoreactive proliferating cells with BUdR and UV light completely abrogated AMLR killer cytolysis. Amlr killers lysed various lymphoid cell lines, including autologous B cell lines, autologous or allogeneic mitogen blasts stimulated by Con A, PHA, or pokeweed mitogen, variious nonlymphoid cell lines derived from human, mouse, or rat, and weakly normal autologous or allogeneic non-T cells. KMT-17, methylcholanthrene-induced rat fibrosarcoma, was the only resistant cell line to have been tested. AMLR killers had characteristics similar to NK cells, Major histocompatibility antigens were not the target antigens for AMLR killers. AMLR killers distinguished the blasts stimulated by alloantigens as self from the blasts stimulated by mitogens as non-self.  相似文献   

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Urine dialysate from rats treated orally with 25 mg/Kg 3H-labeled niridazole was fractionated by DEAE-Sepharose column chromatography and was found to contain three radioactive metabolites and no parent compound. When human niridazole urine dialysate (NUD) was fractionated under identical conditions, fractions corresponding to the three rat NUD metabolites were found to inhibit the human one-way MLR. No inhibition was obtained with fractionated control urine dialysate. It was concluded that nonimmunosuppressive niridazole is metabolized by rats and man to produce three active compounds with the ability to suppress the in vitro response to alloantigens.  相似文献   

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Effects of toxic fungal metabolites on mixed lymphocyte reaction (MLR) using mouse splenocytes and on growth of mouse myeloma cells were examined. Among 25 toxins assayed, the IC50 values of emodin, luteoskyrin, sterlgmatocystin, deoxynivalenoi, 4-acetylnivalenol, T-2 toxin and fusaric acid for the MLR were lower than those for the cytotoxicity toward the myeloma cells, suggesting that these toxins possess suppressive activity to the cellular immune system.  相似文献   

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Procedures are described for the quantitation of SV40 virus infectivity by plaque formation within 7 days and T antigen assay by the sensitive and economical indirect immunoperoxidase technique.  相似文献   

8.
Secondary mixed lymphocyte reaction (MLR-II) was studied in A.TH anti A.TL and A.TL anti-A.TH combinations in which stimulation was mainly due toH-2I-region differences. In both cases the MLR-II was specifically inhibited by the responder anti-stimulator Ia serum. The level of inhibition was dependent on the ratio of the amount of immune serum to the number of stimulating cells. The inhibitory activity and Ia antibodies were specifically absorbed and eluted together. The results confirm that the lymphocyte-activating determinants of the MLR-II (1) are carried by the Ia molecules and (2) are identical to the serologically defined Ia determinants. —Anti-Ia sera directed against private and public specificities of the stimulating cell induced a higher level of inhibition than anti-Ia sera directed only against public specificities, indicating that both private and public Ia specificities are involved in restimulation during MLR-II. — These results, in connection with others, suggest that the receptor of the proliferating T cell recognizes the same Ia determinant as the combining site of the Ia-recognizing antibody.Abbreviations used in this paper Lad lymphocyte-activating determinant - MLR mixed lymphocyte reaction - MLR-I, MLR-II primary, secondary MLR - PRC primed responder cells - LCT dye exclusion lymphocytotoxicity microtechnique - RR relative response  相似文献   

9.
Secondary mixed lymphocyte reaction (MLR-II) was studied in A.TH anti A.TL and A.TL anti-A.TH combinations in which stimulation was mainly due to H-21-region differences. In both cases of MLR-II was specifically inhibited by the responder anti-stimulator Ia serum. The level of inhibition was dependent on the ratio of the amount of immune serum to the number of stimulating cells. The inhibitory activity and Ia antibodies were specifically absorbed and eluted together. The results confirm that the lymphocyte-activating determinants of the MLR-II (1) are carried by the Ia molecules and (2) are identical to the serologically defined Ia determinants. - Anti-Ia sera directed against private and public specificities of the stimulating cell induced a higher level of inhibition than anti-Ia sera directed only against public specificities, indicating that both private and public Ia specificities are involved in re-stimulation during MLR-II. - These results, in connection with others, suggest that the receptor of the proliferating T cell recognizes the same Ia determinant as the combining site of the Ia-recognizing antibody.  相似文献   

10.
Splenocyte plaque assay for the detection of murine leukemia virus.   总被引:10,自引:0,他引:10  
A modified XC assay for murine leukemia virus (MuLV) employing splenocytes taken directly from the animal is described. This modification can be more than 1000 times more sensitive than XC plaque assays employing tissue extracts. This technique should lend itself readily to the quantitation of infectious MuLV in defined populations of lymphoid cells.  相似文献   

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Whereas the mixed lymphocyte reaction was essentially normal in inactive lepromatous leprosy and tuberculoid leprosy, it was severely impaired in active lepromatous leprosy. The impairment was found to be contributed by certain unknown factors in their plasma and subnormal reactivity of their T lymphocytes. The plasma derived from active lepromatous leprosy patients depressed the reaction of normal cells and normal plasma enhanced the reaction of active lepromatous lymphocytes. The cellular factor was studied by using a one-way reaction in which one of the two lymphocyte preparations was inactivated with mitomycin C. The impairment of blastogenesis of active lepromatous lymphocytes was partially reversed by substituting inactivated normal cells for similarly treated leprous cells, and conversely the response of normal allogeneic lymphocytes was depressed by substituting inactivated leprous lymphocytes as the stimulator cells.  相似文献   

14.
The development of congenic mouse strains identical at the H-2K and H-2D loci but differing by I-region associated (Ia) determinants has permitted an association to be established between Ia determinants and stimulation in mixed lymphocyte culture reactions (MLR). The present experiments were undertaken to establish whether the Ir-coded control of MLR operated at the level of recognition or of stimulation. Reciprocal MLR were established between A.TH and A.TL mouse spleen cells in the presence or absence of anti-Ia sera directed either at determinants of the stimulating or responding cells. The number of T cells responding was assessed by the virus plaque assay. Anti-Ia sera directed against the responding cells were no more inhibitory of the MLR than normal mouse serum. In contrast, anti-Ia sera directed against determinants of the mitomycin-treated stimulating cells markedly inhibited activation of T cells in the MLR.  相似文献   

15.
The autologous mixed lymphocyte reaction (AMLR) is a proliferative response of T cells to signals from autologous non-T cells. The AMLR has been an enigma to immunologists because spontaneous proliferation of cells removed from the body is usually substantially less than that observed with a strong AMLR. However, the AMLR is thought to represent an important in vitro function, since it has the attributes of other immune responses, and it is abnormal in a variety of disease states thought to have an immune basis. We reasoned that if the AMLR represented a fundamental immune phenomenon, it should be subject to regulation. In the present study, we present evidence for suppression of the AMLR by macrophages and by T cells. Macrophages inhibited the T cell proliferation to (B + null) cells in a dose-dependent fashion and throughout the time course of the AMLR. Elimination of suppressor T cells by a specific antiserum led to an increase in the AMLR, which was again suppressed in a dose-dependent way by addition of the suppressive T cells. It may be concluded that the AMLR itself is subject to immune regulation and that the suppressive influences observed probably strongly inhibit the AMLR in vivo. Removal of the suppressive principles allows the maximal expression of the AMLR in vitro. We believe that our demonstration of regulation of the AMLR should remove the enigma associated with it and lead to a better understanding of normal cell-cell interactions as well as the basis for abnormalities in a variety of immune-mediated diseases.  相似文献   

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Previous work from this laboratory has suggested that a CD8+ T suppressor (Ts) cell network regulated the murine syngeneic mixed lymphocyte reaction (SMLR). We have attempted to disrupt this network by the inoculation of anti-CD8 monoclonal antibodies (mAb) in vivo. Intraperitoneal inoculation of three mAbs resulted in a marked increase in the proliferation of CD4+, self-Ia-reactive splenic T cells in vitro to syngeneic, but not to allogeneic, spleen cells. Suppression was not limited to a specific mouse strain as the enhanced SMLR was reproducible following anti-CD8 treatment of three strains of mice. In vivo depletion of CD8+ T cells was not a prerequisite for enhancement of the SMLR as several mAb to CD8 augmented the SMLR independent of their capacity to cause CD8 T cell depletion. Moreover, enhancement of the SMLR could be mimicked in vitro by inclusion of anti-CD8 mAb in in vitro cultures of responder T cells and irradiated Ia+ syngeneic stimulators. Since the in vitro SMLR was enhanced following mAb treatment, it was expected that the in vivo SMLR would also be increased. However, no evidence of increased in vivo autoreactivity could be detected following in vivo treatment with anti-CD8 mAb, indicating that other mechanisms in addition to CD8+ regulatory T cells acted to regulate the in vivo activity of autoreactive T cells.  相似文献   

19.
Human leucocyte antigens (HLA) and mixed lymphocyte reactions (MLR) were studied in 38 women with severe pre-eclampsia and their husbands. Thirty-nine women with normal pregnancies and their husbands served as controls. Thirty-three of the control women were matched for age and parity with members of the study group. Infants were studied when possible. HLA compatibility was increased in the pre-eclamptic group compared with matched controls and with theoretical estimates for possible matings. The one-way MLR at delivery showed diminished response of maternal to paternal and cord cells in pre-eclamptic women. This reduced maternal reactivity in women with pre-eclampsia may have a role in the illness, and paternal/maternal histocompatibility may be a feature of the severe form.  相似文献   

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