Showing papers by "Eliane Gluckman published in 1977"

Freezing of canine lymphocytes for use in mixed leukocyte culture and cell-mediated lympholysis tests.

Abstract: A satisfactory and reproducible technique of cryopreservation of canine lymphocytes for use in mixed leukocyte culture (MLC) and cell-mediated lympholysis tests has been developed. Dimethyl sulfoxide was used as the cryopreservation agent. Cells were frozen to -50 C at a controlled rate (-1 C/min) and stored at -169 C. The best preservation was obtained with a concentration of 10 x 10(6) lymphocytes/ml in Waymouth's MB-752/1 medium supplemented with 30% dog serum (MB-30). Before use in MLC or cell-mediated lympholysis tests, lymphocytes were rapidly thawed at 40 C and then rapidly diluted with MB-30 at room temperature. For best responses in MLC, one fresh component (either stimulating or stimulated cells or serum) was needed. The magnitudes of responses of frozen lymphocytes to phytohemagglutinin or in MLC were similar to those seen with fresh cells, but peak responses occurred usually 24 hr after those seen with fresh cells.

Cellular presensitization to alloantigens in haemodialyzed patients.

Abstract: The state of cellular alloimmunity in haemodialyzed patients has been assessed by a microcytotoxicity assay and compared with that in healthy donors Some of the patients were also studied during the weeks following a first kidney transplant rejection For comparison, lymphocytes of multiparous women have been studied Skin fibroblasts from a panel of normal donors were used as targets, and effector cells were cryopreserved peripheral blood lymphocytes Lymphocyte-mediated cytotoxicity (LMC) demonstrated restricted reactivity against target cells, an indication of specificity It detected presensitization more often than a standard test for cytotoxic antibodies This difference is not attributable to differences in sensitivity of the two tests, since some patients were at least once LMC negative and cytotoxic antibody positive, and the antigens detected by both tests were different in patients who had positive LMC and cytotoxic antibodies simultaneously In some experiments, LMC target determinants were not HLA serologically detectable antigens Furthermore, lymphocytes obtained after transplant rejection were cytotoxic for targets which did not share the donors' serological mismatches These results suggest that antigens other than HLA-A and B may be the determinants recognized in the microcytotoxicity assay