Recombinant herpesviral proteins produced by cell-free translation provide a novel approach for the mapping of T lymphocyte epitopes.
The availability of viral genes in cloned DNA forms, in addition to the parallel development of an efficient cell-free system capable of producing viral proteins in vitro, has provided new opportunities for the identification of important T cell Ag encoded within viral genomes. We have cloned bovine herpesvirus type 1 surface glycoproteins gI, gIII, and gIV into the RNA transcription vector pE5LVPO. After translation of the resulting viral mRNA in rabbit reticulocyte lysates, each cloned viral protein specifically stimulated bovine T lymphocytes previously primed to BHV-1. In addition, a panel of specifically truncated peptides has allowed the identification of at least two T lymphocyte epitopes encoded within gIII of BHV-1. Our results represent a novel approach for viral Ag synthesis and subsequent detection of lymphocyte recognition. These findings will likely have important implications in the mapping of T lymphocyte epitopes.
Leary
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Department of Veterinary Science, University of Wisconsin-Madison, Madison 53706.
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D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis; D000956:Antigens, Viral / Q000235:genetics*; D018801:CD2 Antigens; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D002417:Cattle; D002460:Cell Line; D002474:Cell-Free System; D000939:Epitopes; D005434:Flow Cytometry; D007242:Herpesvirus 1, Bovine / Q000276:immunology*; D014176:Protein Biosynthesis; D011971:Receptors, Immunologic / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology; D014759:Viral Envelope Proteins / Q000276:immunology*
D000945:Antigens, Differentiation, T-Lymphocyte; D000956:Antigens, Viral; D018801:CD2 Antigens; D000939:Epitopes; D011971:Receptors, Immunologic; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins