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[Use of flow cytometry in the determination of HLA antigens].

The authors compare the classical method of typing of HLA antigens by the microlymphocytotoxic test (CTX) and the results obtained by flow cytometry (FCTX). They investigated agreement of results as regards estimation of 14 HLA antigens, loci A and B. in 42 sera. They compared possibilities of flow cytometry using microquantities of material and investigated changes of results obtained by cytotoxic reactions after 24 hours. The results evaluated by linear regression analysis proved the comparability of flow cytometry and the classical microlymphocytotoxic test.

Cell Separation↗

Transplantation of cultured pancreatic islets to BB rats.

Pancreatic islets held in tissue culture before transplantation into artificially induced diabetics are not rejected. In animals and human identical twin transplants, the autoimmunity of naturally occurring diabetes may destroy islets, even if rejection is avoided. Therefore we studied whether autoimmune damage of islets can be avoided by pretransplant culture. Recipients were BB rats, which spontaneously developed diabetes. Donors were either Wistar Furth (WF) (major histocompatibility [MHC] identical to BB rats) or Lewis (MHC nonidentical to BB rats). Islets were inoculated into the portal vein either immediately after isolation or after 14 days in tissue culture (95% air, 5% CO2, 24 degrees C). Recipients of cultured islets received a single injection of 1 ml of antilymphocyte serum at the time of transplant. Recurrence of diabetes after transplantation of freshly isolated MHC incompatible Lewis islets occurred rapidly on the basis of rejection or autoimmune damage (or both). Precultured Lewis islets had prolonged or permanent survival. Freshly isolated MHC compatible WF islets were destroyed, and no improvement was seen with culture. We conclude that autoimmune destruction of transplanted islets can be avoided by tissue culture, as can rejection. This is important because this strategy is effective only if recipient and donor differ at the MHC locus. Islet donors may need to be selected on the basis of disparity of histocompatibility factors.

Animals↗

Extra reactivities detected in flow-cytometry-positive, CDC-negative crossmatches are definable HLA specificities.

In this preliminary study, additional reactions were detected in sera that were not found by T-AHG-CDC. The reactions had definable HLA specificities. In our laboratory, the procedures described in this article had the following relative sensitivities for detecting class I HLA alloantibody specificities: FC = B-AHG-CDC greater than T-AHG-CDC greater than B-CDC greater than T-CDC. This study supports the concept that some FC-positive crossmatches, negative by T-AHG-CDC, can be associated with reduced renal allograft survival, since many of the additional reactions detected by FC appear to be due to HLA Class I antibodies.

B-Lymphocytes↗

Patterns of immunological anti-donor reactivity in renal allotransplanted patients--a prospective study.

The immunological reactivity of 20 consecutive renal allotransplanted patients against donor lymphocytes were followed from the day of transplantation and subsequently twice weekly until dismissal or graftectomy. The investigations concerned Complement Dependent Cytotoxic (CDC) antibodies, Antibody Dependent Cell-Mediated Cytotoxicity (ADCC) and Cell-Mediated Lymphocytotoxicity (CML). Five patterns of immunological reactivity were observed parallelling the clinical courses. A specifically positive CML test was highly but not absolutely correlated to clinical graftrejection. Graftrejection was in general not preceeded by positive CML. The patterns of CDC and ADCC reactivity in relation to graftrejection were not evident.

Antibody-Dependent Cell Cytotoxicity↗