Biochemical analysis of class I and class II MHC antigens in cynomolgus macaques by one‐dimensional isoelectric focusing

Abstract

To obtain a better estimate of major histocompatibility complex (MHC) polymorphism in the Old World macaque, Macaca fascicularis, class I and class II MHC proteins from 42 animals were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and one-dimensional isoelectric focusing (1D-IEF). The panel represented both related and unrelated animals with a total of at least 30 serologically distinct haplotypes. Cells were sequentially immunoprecipitated with monoclonal antibody (mAb) W6/32 for class I and with mAb L243 for class II molecules, followed by neuraminidase treatment. Both sets of immunoprecipitates yielded 5-7 major bands on IEF. All bands present in offspring were present in at least 1 parent. Siblings which were serologically identical for class I and which were non-stimulatory in mixed lymphocyte culture (MLC) yielded identical IEF patterns for both class I and class II; in other sibling pairs which were serologically identical for class I antigens (Ag), IEF produced convincing evidence that the siblings were indeed nonidentical, or helped to verify that recombination had occurred within the MHC. Specific bands were found which correlated with class I specificities A8, A24, and B25 previously defined by serology. Comparison of serological and biochemical data will broaden our understanding of the MHC in Macaca fascicularis and will increase the potential use of this species in transplantation research, as a model of disease, and for comparative studies.

Molecular Requirements for T Cell Recognition of N-Myristoylated Peptides Derived from the Simian Immunodeficiency Virus Nef Protein

“…Autologous PBMCs (3 ϫ 10 5 /well) were preincubated with a saturating amount (5 g/ml) of the indicated Abs and cultured with 2N5.1 (A and B) and SN45 (C) T cells (5 ϫ 10 4 /well) in the presence (ϩ) or absence (Ϫ) of C14nef5 (5 g/ml). Note that these Abs were used to block relevant human T cell responses specifically and are known to recognize the corresponding monkey molecules efficiently(9,(23)(24)(25)(26). nega.…”

Unusually High Frequency MHC Class I Alleles in Mauritian Origin Cynomolgus Macaques

“…In the 1980s, Ͼ30 MHC I serotypes were described (23), but these serotypes were never resolved at the molecular level. One-dimensional isoelectric focusing (1D-IEF) of MHC I proteins immunoprecipitated from Cynomolgus macaques suggests that these animals express between five and seven MHC I proteins (24). Recently, 14 MHC I A locus allele sequences (25) and 26 MHC I B locus alleles (26) were described.…”

Immunoprecipitation and isoelectric focusing of sheep MHC class I antigens reveal higher complexity than serology

“…IEF can be used to confirm serological results with the added advantage that serologically undefined antigens can also be identified (Oliver et al ., 1989; this study). It can be used to study the structural heterogeneity of antigens (Neefjes et al ., 1986; Choo et al ., 1993; this study) and to verify MHC identity or to further investigate individuals in which serology suggests that recombination has occurred (Kubens et al ., 1991; Manning & Heise, 1991). Lastly, the identification of products of different class I loci is also possible (Joosten et al ., 1992, 1993; Al‐Murrani et al ., 1993, 1994; Adams et al ., 2001; this study).…”