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Journal of Virology, August 2000, p. 6922-6934, Vol. 74, No. 15
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Quantitation of CD8+ T-Lymphocyte Responses to Multiple Epitopes from Simian Virus 40 (SV40) Large T Antigen in C57BL/6 Mice Immunized with SV40, SV40 T-Antigen-Transformed Cells, or Vaccinia Virus Recombinants Expressing Full-Length T Antigen or Epitope Minigenes

Lawrence M. Mylin,1,dagger Todd D. Schell,1 Debra Roberts,1 Melanie Epler,1 Alina Boesteanu,1,Dagger Edward J. Collins,2,3 Jeffrey A. Frelinger,2 Sebastian Joyce,1,§ and Satvir S. Tevethia1,*

Department of Microbiology and Immunology, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033,1 and Department of Microbiology and Immunology2 and Department of Biochemistry and Biophysics,3 School of Medicine, University of North Carolina, Chapel Hill, North Carolina 27599

Received 1 March 2000/Accepted 3 May 2000

The cytotoxic T-lymphocyte response to wild-type simian virus 40 large tumor antigen (Tag) in C57BL/6 (H2b) mice is directed against three H2-Db-restricted epitopes, I, II/III, and V, and one H2-Kb-restricted epitope, IV. Epitopes I, II/III, and IV are immunodominant, while epitope V is immunorecessive. We investigated whether this hierarchical response was established in vivo or was due to differential expansion in vitro by using direct enumeration of CD8+ T lymphocytes with Tag epitope/major histocompatibility complex class I tetramers and intracellular gamma interferon staining. The results demonstrate that epitope IV-specific CD8+ T cells dominated the Tag-specific response in vivo following immunization with full-length Tag while CD8+ T cells specific for epitopes I and II/III were detected at less than one-third of this level. The immunorecessive nature of epitope V was apparent in vivo, since epitope V-specific CD8+ T cells were undetectable following immunization with full-length Tag. In contrast, high levels of epitope V-specific CD8+ T lymphocytes were recruited in vivo following immunization and boosting with a Tag variant in which epitopes I, II/III, and IV had been inactivated. In addition, analysis of the T-cell receptor beta  (TCRbeta ) repertoire of Tag epitope-specific CD8+ cells revealed that multiple TCRbeta variable regions were utilized for each epitope except Tag epitope II/III, which was limited to TCRbeta 10 usage. These results indicate that the hierarchy of Tag epitope-specific CD8+ T-cell responses is established in vivo.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, H107, The Pennsylvania State University College of Medicine, 500 University Dr., Hershey, PA 17033. Phone: (717) 531-8872. Fax: (717) 531-5578. E-mail: sst1{at}psu.edu.

dagger Present address: Messiah College, Grantham, PA 17027.

Dagger Present address: Wistar Institute, Philadelphia, PA 19104.

§ Present address: Department of Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, TN 37232.

Journal of Virology, August 2000, p. 6922-6934, Vol. 74, No. 15
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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Copyright © 2000 by the American Society for Microbiology. All rights reserved.