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Journal of Virology, January 2001, p. 73-82, Vol. 75, No. 1
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.1.73-82.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Evaluation of Cytotoxic T-Lymphocyte Responses in Human and Nonhuman Primate Subjects Infected with Human Immunodeficiency Virus Type 1 or Simian/Human Immunodeficiency Virus

Tong-Ming Fu,1,* Daniel C. Freed,1 Wendy L. Trigona,1 Liming Guan,1 Lan Zhu,1 Romnie Long,1 Natasha V. Persaud,1 Kelledy Manson,2 Sheri Dubey,1 and John W. Shiver1

Department of Virus and Cell Biology, Merck Research Laboratories, West Point, Pennsylvania 19486,1 and Primedica, Worcester, Massachusetts 016082

Received 29 March 2000/Accepted 2 October 2000

Cytotoxic T-lymphocyte (CTL) responses have been implicated as playing an important role in control of human immunodeficiency virus (HIV) infection. However, it is technically difficult to demonstrate CTL responses consistently in nonhuman primate and human subjects using traditional cytotoxicity assay methods. In this study, we systematically evaluated culture conditions that may affect the proliferation and expansion of CTL effector cells and presented a sensitive method for detection of cytotoxicity responses with bulk CTL cultures. We confirmed the sensitivity and specificity of this method by demonstration of vigorous CTL responses in a simian-HIV (SHIV)-infected rhesus macaque. The expansion of epitope-specific CTL effector cells was also measured quantitatively by CTL epitope-major histocompatibility complex tetramer complex staining. In addition, two new T-cell determinants in the SIV gag region are identified. Last, we showed the utility of this method for studying CTL responses in chimpanzee and human subjects.

* Corresponding author. Mailing address: Merck Research Laboratories, WP26-246, West Point, PA 19486. Phone: (215) 652-8270. Fax: (215) 993-0512. E-mail: tong-ming_fu{at}merck.com.

Journal of Virology, January 2001, p. 73-82, Vol. 75, No. 1
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.1.73-82.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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