Domain interactions of H–2 class I antigens alter cytotoxic T-cell recognition sites

Abstract

H-2 class I antigens appear to direct the recognition of virus-infected and neoplastic transformed cells by cytotoxic T lymphocytes (CTLs). Here, to identify the regions of class I antigens involved in CTL recognition, four hybrid class I genes were constructed in which exons were exchanged between the H-2Kb and H-2Db genes. These class I genes were expressed in mouse L cells and recognition of the hybrid Kb/Db antigens by CTLs and monoclonal antibodies specific for either Kb or Db was investigated. The pattern of CTL and monoclonal antibody recognition obtained indicates three correlations between structure and function of class I antigens. First, most CTL recognition sites and alloantigenic determinants are located on domains 1 and 2 of the antigen molecule. Second, these CTL recognition sites and alloantigenic determinants are not influenced by interaction of domains 1 and 2 with polymorphic regions of domain 3. Third, in contrast, interaction between domains 1 and 2 alters these CTL recognition sites and alloantigenic determinants. The alteration of CTL recognition sites by interaction between domains 1 and 2 suggests that a CTL site may be formed by amino acids from both domains 1 and 2, or that the conformation of amino acids at a CTL site may be altered by interactions between domains 1 and 2. Through these two features, the conformation of CTL recognition sites on H-2 class I antigens may be sensitive to alteration by interaction of either domain 1 or 2 with viral antigens.

The murine cytomegalovirus immunoevasin gp40 binds MHC class I molecules to retain them in the early secretory pathway

“…Our data in Fig. 3C support this hypothesis: both the monoclonal antibodies Y3 and B22.249, which bind to the α 1 /α 2 superdomain of class I molecules, preclude gp40 co-precipitation, whereas the antibody 28-14-8S, which binds to the α 3 domain, co-precipitates gp40 (Allen et al, 1984;Nathenson et al, 1989). In the structural model of Wang et al (2012), there is space for class-I-bound peptide, which agrees with the observation that gp40 does not inhibit peptide binding ( Fig.…”

Restoration of MHC Class I Surface Expression and Endogenous Antigen Presentation by a Molecular Chaperone

“…Monoclonal antibody M1/42 (ATCC TIB126) [13] recognizes a monotypic determinant of the murine H-2 (MHC class I) complex of antigens. Monoclonal antibody B22.249 (kindly provided by Dr Alain Townsend) recognizes a conformation-dependent determinant of the a2 domain of peptide-bound D b [14], while monoclonal antibody 28-14-8s (ATCC HB27) recognizes a conformation-independent determinant of the a1 domain of D b [15]. The biotin-conjugated monoclonal antibody 28-8-6 (Pharmingen, San Diego, CA, USA) recognizes H-2 K b and D b independently of conformation and AF6-88.5 (Pharmingen, San Diego, CA, USA) is specific for K b .…”

Identification of cytotoxic T cell epitopes on the VP3 and VP6 rotavirus proteins

“…P815 (H-2 a) cells transfected with the K b and D ~ genes (obtained from P. Romero, Ludwig Institute for Cancer Research, Epalinges, Switzerland) were grown in the same medium but supplemented with G418 (Gibco BRL) at 1 mg/ml to ensure expression of the transfected molecule. The presence of the protein product of transfected genes in these cells was analysed on a FACScan flow cytometer (Becton Dickinson) using the K10 65 1 (anti-K h) and 28 14 8 (anti-D b) (Allen et al, 1984) monoclonal antibodies (MAbs) (a gift of M. Pla, H6pital St Louis, Paris, France). The CV1 and 143B thymidine kinase-negative (TK-) cells used to construct the rVV (a gift from Y. Rivi6re, Institut Pasteur, Paris, France) were grown in DMEM containing 10% heat-inactivated FCS, 100 lag/ml streptomycin sulphate and 100 units/ml penicillin.…”