Alloreactive T‐cell recognition of bovine major histocompatibility complex class II products defined by one‐dimensional isoelectric focusing

Abstract

T-cell recognition of bovine MHC (BoLA) class I1 antigens was investigated in relation to BoLA class I1 polymorphisms defined by one-dimensional isoelectric focusing (1D-IEF). One-way mixed lymphocyte reactions (MLRs), and allospecific cell lines and clones were used. In general, T-cell responses correlated with the 1D-IEF defined haplotypes (EDF types). However, with MLRs some responses appeared to be associated with BoLA class I differences. All combinations of responder-stimulator pairs produced alloreactive T-cell responses both in MLR and in generation of allolines/clones. Thus allospecific lines and clones were generated to all EDF types tested. Splits in the IEF typing were observed with EDF6 and EDF3, indicating that distinct BoLA class I1 haplotypes are not necessarily distinguished by 1D-IEF alone. Furthermore, the patterns of reactivity with EDF3 expressing cells were complex with the T-cell specificities splitting EDF3 into several distinct types. Also, in some cases it was clear that more than one T-cell specificity per EDF type was detectable. Thus, allospecific lines and clones provide complementary and additional information to the 1D-IEF typing for polymorphism of the BoLA class I1 complex. This extra information is particularly important in terms of the functional significance of the BoLA complex for antigen presentation and immune response gene effects.

Characterization of BoLA-DRB3.2 Alleles in Hanwoo (Korean cattle) by Sequence Based Typing (SBT)

“…In cattle, a number of serological, biochemical and molecular methods have been developed for the efficient typing of DRB3 polymorphisms (Watkins et al, 1989;Davies and Antczak, 1991;Davies et al, 1992;Glass et al, 1992;van Eijk et al, 1992;Paswan et al, 2005). Later on, more efficient methods have been discovered including heteroduplex analysis (Sitte et al, 1995), sequence-specific oligonucleotide typing (Sitte et al, 1996), denaturing gradient gel electrophoresis (Aldridge et al, 1998), and sequencing of the PCR products (Aida et al, 1995).…”