DR alpha: E beta heterodimers in DRA transgenic mice hinder expression of E alpha: E beta molecules and are more efficient in antigen presentation.
HLA-DRA transgenic (tg) mice on H-2d background were constructed to study assembly, expression and function of DR alpha: E beta class II heterodimers when an alternate E alpha chain is available. Cytofluorimetric analysis and immunoprecipitation studies demonstrate that the majority (90%) of E beta d molecules on class II-positive splenocytes from DRA-tg mice are associated with DR alpha rather than E alpha chains. To characterize the functional role of the interspecies as compared with the wild-type I-E molecules, MHC restriction and T cell epitope immunodominance of synthetic peptides spanning the entire sequence of 65 kDa heat shock protein (hsp) from Mycobacterium tuberculosis were determined in hsp-primed DRA-tg and DBA/2 mice. A similar pattern of responsiveness was observed in both strains, but hsp epitopes recalled a higher response in DRA-tg as compared with DBA/2 mice. A panel of T cell hybridomas specific for two hsp peptides or a hen egg white lysozyme peptide presented by both DR alpha: E beta d and E alpha d: E beta d was studied in detail. Surprisingly, DR alpha: E beta d dimers present these peptides more efficiently than E alpha d: E beta d, even when the TCR was selected in mice expressing only E alpha d: E beta d molecules. The higher efficiency of antigen presentation by DR alpha: E beta d dimers does not appear to depend on increased binding affinity for peptides, as demonstrated by competition for antigen presentation, nor on increased efficiency in the interaction with CD4 molecules. Rather, the higher efficiency of antigen presentation could be explained by a more effective ligand-TCR interaction. This is consistent with molecular modeling based on the class II structure, indicating that 16 out of 17 substitutions between the first domain of E alpha d and DR alpha chains ile outside the peptide binding groove and are potentially available for interaction with the TCR.
Trembleau
S
S
Roche Milano Ricerche, Milano, Italy.
Giacomini
P
P
Guéry
J C
JC
Setini
A
A
Hammer
J
J
Sette
A
A
Appella
E
E
Adorini
L
L
D000595:Amino Acid Sequence; D000818:Animals; D017951:Antigen Presentation; D015704:CD4 Antigens / Q000378:metabolism; D018984:Epitopes, T-Lymphocyte / Q000235:genetics; D005260:Female; D006183:H-2 Antigens / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006684:HLA-DR Antigens / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D059805:HLA-DR alpha-Chains; D000949:Histocompatibility Antigens Class II / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008822:Mice, Transgenic; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation
D015704:CD4 Antigens; D018984:Epitopes, T-Lymphocyte; D006183:H-2 Antigens; D006684:HLA-DR Antigens; D059805:HLA-DR alpha-Chains; D000949:Histocompatibility Antigens Class II; C024138:I-E-antigen