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The Journal of Immunology, 1999, 163: 6867-6875.
Copyright © 1999 by The American Association of Immunologists

Functional Analysis of Antigen-Specific T Lymphocytes by Serial Measurement of Gene Expression in Peripheral Blood Mononuclear Cells and Tumor Specimens

Udai S. Kammula*, Kang-Hun Lee{dagger}, Adam I Riker*, Ena Wang*, Galen A. Ohnmacht*, Steven A. Rosenberg* and Francesco M. Marincola1,{dagger}

* Surgery Branch and {dagger} Department of Transfusion Medicine, Clinical Center, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892

The cloning of cancer Ags recognized by T cells has provided potentially new tools to enhance immunity against metastatic cancer. The biological monitoring of effective immunization has, however, remained a dilemma. We describe here a sensitive molecular quantitation methodology that allows analysis of in vivo immune response to vaccination. Metastatic melanoma patients were immunized with a synthetically modified peptide epitope (209-2M) from the melanoma self-Ag gp100. Using serial gene expression analysis, we report functional evidence of vaccine-induced CTL reactivity in fresh cells obtained directly from the peripheral blood of postimmunized patients. Further, we demonstrate in vivo localization of vaccine-induced immune response within the tumor microenvironment. The results of these molecular assays provide direct evidence that peptide immunization in humans can result in tumor-specific CTL that localize to metastatic sites.




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