A one‐step monoclonal antibody typing procedure that simplifies HLA class I and class II typing

Abstract

We have developed monoclonal antibodies to most HLA specificities, making it possible for us to devise a simple, rapid, one-step microcytotoxicity test. The test is performed by adding 1 microliter of cells to 1 microliter of antibody-complement mixture predotted on the microtest tray. The reactions are read following a 1-hour incubation period (30 minutes in some instances). The analysis of reactions seen on testing 105 class I antibodies and 50 class II antibodies is shown. A comparison of typing by the standard NIH method and the new one-step procedure showed a > 96% concordance in the 500 T cells and 200 B cells we examined. Class I and class II typing could be performed using B cells, thus obviating the need to isolate both T and B cells for HLA typing.

Correlation of Thymic Pathology with HLA in Myasthenia Gravis

“…MHC class I typing (HLA-A and -B loci) was performed by means of CD8-positive lymphocytes purified by immunomagnetic beads from heparinized peripheral blood in a standard cytotoxic assay using monoclonal antibodies (OneLambda, Canoga Park, USA) or polyclonal antisera (Biotest, Dreieich, Germany) and complement (7). MHC II alleles (HLA-DRB1, -DRB3, -DRB4, -DRB5, -DQA1, and -DQB1 loci) were determined by DNA analysis using allele-specific hybridization after PCR amplification of each locus, slightly modified from the method described by Erlich and associates (8).…”

Diversity in HLA class I and class II in kidney donors and recipients according to race in KwaZulu-Natal (South Africa) implications for transplantation

“…All tissue typing tests on blood collected by venesection on kidney recipients and donors described above, were performed at The Laboratory of Immunology, South African Blood Bank Services, Durban, South Africa. MHC Class I typing was done with serological methods as previously described [16]. MHC Class II HLA typing was done using serological and molecular methods as previously described [17].…”

PANEL REACTIVE ANTIBODY IN THALASSEMIC SERUM INHIBITS PROLIFERATION AND DIFFERENTIATION OF CORD BLOOD CD34+CELLS IN VITRO

“…The collection of cord blood for this study was approved by the Ethical Committee of our hospital, and informed consent was obtained in all cases. The HLA typing of cord blood was analyzed by the monoclonal antibody typing procedure [9]. Mononuclear cells were separated from the cord blood by Ficoll-Hypaque gradient centrifugation (1.070 g/cm 3 ).…”