Anti-BoLA-w8 monoclonal antibody: production of a tissue typing reagent after blocking monomorphic sites on bovine mononuclear cells.

A murine monoclonal antibody, WIM-8 with specificity for BoLA-w8, was generated after using an immunization protocol designed to reduce the response to bovine monomorphic determinants. BoLA-w8/w9 peripheral blood mononuclear (PBM) cells were incubated with an antiserum from mice immunized with BoLA-w10/w20 PBM cells. Splenocytes from a mouse that received three intravenous injections of the treated cells were fused with murine myelomas. A hybridoma supernatant with alloreactivity in ELISA against BoLA-w8 positive but not BoLA-w8 negative PBM cells was tested for lymphocytotoxicity in parallel with parous alloantisera. WIM-8, an IgG2b monoclonal antibody, identified the 44,000 dalton protein of the bovine MHC class I molecule present on Bos taurus and Bos indicus animals.