The Journal of Experimental Medicine
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© The Rockefeller University Press, 0022-1007/1999/9/705/ $5.00
The Journal of Experimental Medicine, Volume 190, Number 5, September 6, 1999 705-716

Original Article

High Frequencies of Naive Melan-a/Mart-1–Specific Cd8+ T Cells in a Large Proportion of Human Histocompatibility Leukocyte Antigen (Hla)-A2 Individuals

Mikaël J. Pitteta, Danila Valmoria, P. Rod Dunbarc, Daniel E. Speisera, Danielle Liénarda,b, Ferdy Lejeuneb, Katharina Fleischhauerd, Vincenzo Cerundoloc, Jean-Charles Cerottinia, and Pedro Romeroa

a From the Division of Clinical Onco-Immunology, Ludwig Institute for Cancer Research, Lausanne Branch,
b Multidisciplinary Oncology Center, Centre Hospitalier Universitaire Vaudois, 1011 Lausanne, Switzerland
c Institute of Molecular Medicine, Nuffield Department of Medicine, John Radcliffe Hospital, Headington, Oxford OX3 9DU, United Kingdom
d Tissue Typing Laboratory, Department of Biology and Biotechnology (DIBIT), Istituto Scientifico H.S. Raffaele, 20132 Milano, Italy
Division of Clinical Onco-Immunology, Ludwig Institute for Cancer Research, Centre Hospitalier Universitaire Vaudois – BH 19-602, 1011 Lausanne, Switzerland.41-21-314-74-7741-21-314-01-78

pedro.romero{at}isrec.unil.ch

Using fluorescent HLA-A*0201 tetramers containing the immunodominant Melan-A/MART-1 (Melan-A) tumor-associated antigen (Ag), we previously observed that metastatic lymph nodes of melanoma patients contain high numbers of Ag-experienced Melan-A–specific cytolytic T lymphocytes (CTLs). In this paper, we enumerated and characterized ex vivo Melan-A–specific cells in peripheral blood samples from both melanoma patients and healthy individuals. High frequencies (≥1 in 2,500 CD8+ T cells) of Melan-A–specific cells were found in 10 out of 13 patients, and, surprisingly, in 6 out of 10 healthy individuals. Virtually all Melan-A–specific cells from 6 out of 6 healthy individuals and from 7 out of 10 patients displayed a naive CD45RAhi/RO– phenotype, whereas variable proportions of Ag-experienced CD45RAlo/RO+ Melan-A–specific cells were observed in the remaining 3 patients. In contrast, ex vivo influenza matrix–specific CTLs from all individuals exhibited a CD45RAlo/RO+ memory phenotype as expected. Ag specificity of tetramer-sorted A2/Melan-A+ cells from healthy individuals was confirmed after mitogen-driven expansion. Likewise, functional limiting dilution analysis and interferon {gamma} ELISPOT assays independently confirmed that most of the Melan-A–specific cells were not Ag experienced. Thus, it appears that high frequencies of naive Melan-A–specific CD8+ T cells can be found in a large proportion of HLA-A*0201+ individuals. Furthermore, as demonstrated for one patient followed over time, dramatic phenotype changes of circulating Melan-A–specific cells can occur in vivo.

Key Words: melanoma • tetramer • influenza matrix • immunotherapy • tumor immunity

1used in this paper: CTL, cytolytic T lymphocyte; CTLp, cytolytic T lymphocytes precursor(s); LDA, limiting dilution analysis; Melan-A, Melan-A/MART-1

© 1999 The Rockefeller University Press


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