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Isolation of Precursor B-cell Subsets from Umbilical Cord Blood
Published on: April 16, 2013
Selective loss of BoLA class I determinants from the lymphocyte surface after acid treatment
P Horín1, P Cermák, P Vojtísek
1Institute of Animal Breeding and Genetics, Faculty of Veterinary Medicine, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic.
Acid treatment of bovine lymphocytes by a buffered solution of 0.263 m citric acid and 0.123 M Na2HPO4 at pH = 3.0, originally described for human and murine lymphocytes, selectively eliminated the antigenicity of MHC (BoLA) class I determinants also from bovine lymphocytes. The viability of acid-treated cell suspension was higher than 90%. The reactivity of acid-treated lymphocytes with BoLA class I typing alloantisera was lost in microcytotoxicity test, while their reactions with cross-reactive anti-HLA class II, anti-BoCD2 and BoCD5 monoclonal antibodies, and with antisera detecting two non-MHC lymphocyte alloantigenic specificities (BoLY w1 and R') remained unchanged in the microcytotoxicity and/or indirect immunofluorescence tests. The results thus show that this approach of modulating cell surface expression of MHC class I determinants may be used in cattle.
Acid treatment of bovine lymphocytes by a buffered solution of 0.263 m citric acid and 0.123 M Na2HPO4 at pH = 3.0, originally described for human and murine lymphocytes, selectively eliminated the antigenicity of MHC (BoLA) class I determinants also from bovine lymphocytes. The viability of acid-treated cell suspension was higher than 90%. The reactivity of acid-treated lymphocytes with BoLA class I typing alloantisera was lost in microcytotoxicity test, while their reactions with cross-reactive anti-HLA class II, anti-BoCD2 and BoCD5 monoclonal antibodies, and with antisera detecting two non-MHC lymphocyte alloantigenic specificities (BoLY w1 and R') remained unchanged in the microcytotoxicity and/or indirect immunofluorescence tests. The results thus show that this approach of modulating cell surface expression of MHC class I determinants may be used in cattle.

