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This Article Full Text Full Text (PDF) Data Supplement All Versions of this Article: jimmunol.0900903v1 183/7/4800    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Google Scholar Articles by Tsuji, T. Articles by Gnjatic, S. PubMed PubMed Citation Articles by Tsuji, T. Articles by Gnjatic, S.

Characterization of Preexisting MAGE-A3-Specific CD4⁺ T Cells in Cancer Patients and Healthy Individuals and Their Activation by Protein Vaccination¹

Takemasa Tsuji^*, Nasser K. Altorki, Gerd Ritter^*, Lloyd J. Old^* and Sacha Gnjatic^2,*

^* Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, NY 10065; and Presbyterian Hospital, Division of Thoracic Surgery, Weill Medical College of Cornell University, New York, NY 10065

Vaccination with cancer/testis Ag MAGE-A3 in the form of recombinant protein often induces specific humoral and cellular immune responses. Although Ag-specific CD4⁺ T cells following vaccination are detectable by cytokine production after a single in vitro stimulation, their detection before vaccination is difficult because of low frequency. In this study, we have applied a sensitive method using CD154 (CD40L) staining to detect MAGE-A3-specific CD4⁺ T cells. MAGE-A3-specific T cell responses were analyzed in four healthy donors, two lung cancer patients with spontaneous serum Abs to MAGE-A3, and two baseline seronegative lung cancer patients throughout vaccination with MAGE-A3 protein. MAGE-A3-specific CD4⁺ T cells were detected in all individuals tested, at low frequency in healthy donors and seronegative cancer patients and higher frequency in patients seropositive for MAGE-A3. Polyclonal expansion of CD154-expressing CD4⁺ T cells after cell sorting generated a large number of MAGE-A3-specific CD4⁺ T cell lines from all individuals tested, enabling full characterization of peptide specificity, HLA-restriction, and avidity. Application of this method to cancer patients vaccinated with MAGE-A3 protein with or without adjuvant revealed that protein vaccination induced oligoclonal activation of MAGE-A3-specific CD4⁺ T cells. It appeared that MAGE-A3 protein vaccination in the presence of adjuvant selectively expanded high avidity CD4⁺ T cells, whereas high avidity T cells disappeared after multiple vaccinations with MAGE-A3 protein alone.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ This work was supported in part by the Cancer Vaccine Collaborative, funded by Cancer Research Institute and Ludwig Institute for Cancer Research.

² Address correspondence and reprint requests to Dr. Sacha Gnjatic, Ludwig Institute for Cancer Research, New York Branch, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, Box 32, New York, NY 10065. E-mail address: gnjatics{at}mskcc.org

³ Abbreviation used in this paper: DC, dendritic cells; EBV-B cells, EBV-transformed B cells; ICCS, intracellular cytokine staining; T-APC, PHA-activated CD4⁺ T antigen presenting cells.

⁴ The online version of this article contains supplemental material.