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Journal of Virology, July 2008, p. 7120-7134, Vol. 82, No. 14
0022-538X/08/$08.00+0     doi:10.1128/JVI.00453-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

An Extremely Diverse CD4 Response to Vaccinia Virus in Humans Is Revealed by Proteome-Wide T-Cell Profiling{triangledown}

Lichen Jing,1 D. Huw Davies,2 Tiana M. Chong,3 Sookhee Chun,2 Christopher L. McClurkan,3 Jay Huang,3 Brian T. Story,3 Douglas M. Molina,4 Siddiqua Hirst,2 Philip L. Felgner,2 and David M. Koelle1,2,5,6,7*

Department of Medicine, University of Washington, Seattle, Washington 98102,1 Department of Medicine, University of California Irvine, Irvine, California 92697,2 Department of Laboratory Medicine, University of Washington, Seattle, Washington,3 ImmPORT Therapeutics Inc., Irvine, California 92618,4 Benaroya Research Institute, Seattle, Washington,5 Fred Hutchinson Cancer Research Center, Seattle, Washington,6 Program in Pathobiology, University of Washington, Seattle, Washington7

Received 29 February 2008/ Accepted 2 May 2008

CD4 T cells are required for the maintenance and recall of antiviral CD8 T cells and for antibody responses. Little is known concerning the overall architecture of the CD4 response to complex microbial pathogens. 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.

* Corresponding author. Mailing address: 1616 Eastlake Ave. E., Suite 500, Box 358117, Seattle, WA 98102. Phone: (206) 667-6491. Fax: (206) 667-7711. E-mail: viralimm{at}u.washington.edu

{triangledown} Published ahead of print on 14 May 2008.

Journal of Virology, July 2008, p. 7120-7134, Vol. 82, No. 14
0022-538X/08/$08.00+0     doi:10.1128/JVI.00453-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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