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Journal of Virology, September 2000, p. 8541-8549, Vol. 74, No. 18
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Identification of Dominant Optimal HLA-B60- and HLA-B61-Restricted Cytotoxic T-Lymphocyte (CTL) Epitopes: Rapid Characterization of CTL Responses by Enzyme-Linked Immunospot Assay

Marcus A. Altfeld,1 Alicja Trocha,1 Robert L. Eldridge,1 Eric S. Rosenberg,1 Mary N. Phillips,1 Marylyn M. Addo,1 Rafick P. Sekaly,2 Spyros A. Kalams,1 Sandra A. Burchett,3 Kenneth McIntosh,3 Bruce D. Walker,1 and Philip J. R. Goulder1,3,*

Partners AIDS Research Center and Infectious Disease Unit, Massachusetts General Hospital and Harvard Medical School,1 and Division of Infectious Diseases, The Children's Hospital,3 Boston, Massachusetts, and Laboratoire d'Immunologie, Institut de Recherches Cliniques de Montreal, Montreal, Quebec, Canada H2W 1R72

Received 19 April 2000/Accepted 21 June 2000

Human immunodeficiency virus type 1 (HIV-1)-specific cytotoxic T-lymphocyte (CTL) responses play a major role in the antiviral immune response, but the relative contribution of CTL responses restricted by different HLA class I molecules is less well defined. HLA-B60 or the related allele B61 is expressed in 10 to 20% of Caucasoid populations and is even more highly prevalent in Asian populations, but yet no CTL epitopes restricted by these alleles have been defined. Here we report the definition of five novel HLA-B60-restricted HIV-1-specific CTL epitopes, using peripheral blood mononuclear cells in enzyme-linked immunospot (Elispot) assays and using CTL clones and lines in cytolytic assays. The dominant HLA-B60-restricted epitope, Nef peptide KEKGGLEGL, was targeted by all eight subjects with B60 and also by both subjects with B61 studied. This study additionally establishes the utility of the Elispot assay as a more rapid and efficient method of defining novel CTL epitopes. This approach will help to define new CTL epitopes that may play an important role in the immune control of HIV-1.

* Corresponding author. Mailing address: MGH-East, CNY, 5th Floor, 149 13th St., Charlestown, MA 02129. Phone: (617) 726-5758. Fax: (617) 726-5416. E-mail: goulder{at}helix.mgh.harvard.edu.

Journal of Virology, September 2000, p. 8541-8549, Vol. 74, No. 18
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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