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In a whole-proteome approach, 180 predicted open reading frames (ORFs) in the vaccinia virus genome were expressed and tested using responder cells from 20 blood samples from 11 vaccinees. Validation assays established the sensitivity and specificity of the system. Overall, CD4 responses were detected for 122 ORFs (68%). A mean of 39 ORFs were recognized per person (range, 13 to 63). The most frequently recognized ORFS were present in virions, including A3L and A10L (core proteins), WR148 (a fragmented homolog of an orthopoxvirus protein that forms inclusions in cells), H3L (a membrane protein), D13L (a membrane scaffold protein), and L4R (a nucleic acid binding protein). Serum immunoglobulin G profiling by proteome microarray detected responses to 45 (25%) of the ORFs and confirmed recent studies showing a diverse response directed to membrane and nonmembrane antigens. Our results provide the first empirical whole-proteome data set regarding the global CD4 response to full-length proteins in a complex virus and are consistent with the theory that abundant structural proteins are immunodominant.<\/jats:p>","DOI":"10.1128\/jvi.00453-08","type":"journal-article","created":{"date-parts":[[2008,5,14]],"date-time":"2008-05-14T21:39:34Z","timestamp":1210801174000},"page":"7120-7134","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":54,"title":["An Extremely Diverse CD4 Response to Vaccinia Virus in Humans Is Revealed by Proteome-Wide T-Cell Profiling"],"prefix":"10.1128","volume":"82","author":[{"given":"Lichen","family":"Jing","sequence":"first","affiliation":[{"name":"Department of Medicine, University of Washington, Seattle, Washington 98102"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"D. 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