Phase I Trial of Adoptive Immunotherapy With Cytolytic T Lymphocytes Immunized Against a Tyrosinase Epitope
Authors: Jedd D. Wolchok, Paul B. Chapman, Malcolm S. Mitchell
Journal: Journal of Clinical Oncology (2002)

Article Tools SPECIAL DEPARTMENTS Article Tools OPTIONS & TOOLS Export Citation Track Citation Add To Favorites Rights & Permissions COMPANION ARTICLES No companion articles ARTICLE CITATION DOI: 10.1200/JCO.2002.20.14.3176 Journal of Clinical Oncology - published online before print November 1, 2016 PMID: 12118032 Phase I Trial of Adoptive Immunotherapy With Cytolytic T Lymphocytes Immunized Against a Tyrosinase Epitope Jedd D. WolchokxJedd D. WolchokSearch for articles by this author , Paul B. ChapmanxPaul B. ChapmanSearch for articles by this author Malcolm S. MitchellxMalcolm S. MitchellSearch for articles by this author Show More Memorial-Sloan Kettering Cancer Center, New York, NYKarmanos Cancer Institute, Detroit, MI https://doi.org/10.1200/JCO.2002.20.14.3176 First Page Full Text PDF Figures and Tables © 2002 by American Society of Clinical OncologyjcoJ Clin OncolJournal of Clinical OncologyJCO0732-183X1527-7755American Society of Clinical OncologyResponse15072002In Reply:We are grateful for the opportunity to reply to the comments of Drs Wolchok and Chapman about the relevance and importance of the encoded epitope sequence and their interpretation of our published data.1 Also, although it was not called into question, we would like to clarify our numbering of the tyrosinase epitope as 369 to 377 rather than as 368 to 376, as it is often presented.The first important point is that we conducted preliminary experiments to determine whether immunization with the unmodified epitope led to cross-reactive cytotoxic T lymphocytes (CTLs) capable of lysing melanoma cells. As shown in Fig 1, which was typical of several different experiments, immunization in vitro with YMNGTMSQV led to CTLs cross-reactive against the other epitope but not vice versa. This is what prompted us to test YMNGTMSQV in the study, with full knowledge of the article by Skipper et al,2 especially because in their report only a single CTL clone was evaluated, whereas our cell product was a polyclonal “bulk” culture. Fig 1. YMN effectors recognize both Tyr peptides.Lysis of melanoma cells in our hands, and probably in most others’, is always less than that of heavily peptide-loaded T2 cells. However, that lysis is specific, as shown by far lower lysis of control HLA-matched target cells lacking the epitope (MALME 3) (Figs 2 and 3). Fig 2. Lysis of autologous melanoma cells with effectors generated to tyrosinase peptide 369-377 (YMNGTMSQV). Fig 3. Antipeptide and antimelanoma response (patient no. 05-RE).Perhaps most relevant to this discussion, Visseren et al3 obtained tyrosinase-specific CTL bulk cultures from an HLA-A2 donor against YMNGTMSQV-sensitized target cells. The bulk cultures had strong CTL activity against JY cells loaded with YMNGTMSQV peptide and melanoma cells SK-23 and 453 at low effector-to-target ratios without the addition of exogenous tyrosinase peptide. Ten CTL clones were isolated from the bulk culture that recognized YMNGTMSQV peptide, and eight of the 10 clones cross-reacted with melanoma cell lines SK-23, 453, and Mi 3046/2, in a tumor necrosis factor release assay. Lytic activity of three clones was tested further on the melanoma tumor panel and shown to be identical with that of the bulk culture. The melanoma cell panel was also lysed efficiently by CTL from five further bulk cultures generated from normal donors. The authors concluded that the YMNGTMSQV peptide was naturally processed and presented on melanoma cells, but the reactivity could equally well have been explained by cross-reactivity to YMDGTMSQV on the melanomas.Reduction of the size of the computed tomography scans in the final publication, which was a decision made by the Journal, undoubtedly made it difficult for Wolchok and Chapman to fully appreciate the change in the lesions, which was also observed on physical examination. The mediastinal nodes disappeared after treatment as we noted in the text, and the inguinal and iliac nodes dramatically shrank and became more discrete masses. The scans were read officially by disinterested university radiologists, not by the investigators. We would be pleased to provide either the full-size photographs or a copy of the original computed tomography scans to be certain Wolchok and Chapman see the changes we described.With regard to the numbering of the peptide, the original article by Brichard et al4 describing the cloning of the tyrosinase cDNA (clone 123.B2) presents a cDNA whose amino acid residues 369 to 377 are given as YMNGTMSQV in the direct submission sequence provided to Genbank. The authors note that in the sequence reported by Kwon et al5 in 1987, there is a three-base pair deletion beginning at base 124, which accounts for the numbering of the peptide beginning at 368 rather than 369 in some publications.Finally, we thank Wolchok and Chapman for correcting the typographical error substituting T for Y in the sequence of the YMDGTMSQV peptide in the Introduction.We believe that further refinement of our approach and the addition of other elements such as cytokines to the regimen may well yield more convincing results than our phase I study. Nevertheless, if the article stimulated in other readers the same degree of enthusiasm as we generated in Wolchok and Chapman, we may have achieved one of our aims.1. Mitchell MS, Darrah D, Yeung D, et al: Phase I trial of adoptive immunotherapy with cytolytic T lymphocytes immunized against a tyrosinase epitope. J Clin Oncol 20:: 1075,2002-1086, Link, Google Scholar2. Skipper JC, Hendrickson RC, Gulden PH, et al: An HLA-A2-restricted tyrosinase antigen on melanoma cells results from posttranslational modification and suggests a novel pathway for processing of membrane proteins. J Exp Med 183:: 527,1996-534, Crossref, Medline, Google Scholar3. Visseren MJ, van Elsas A, van der Voort EI, et al: CTL specific for the tyrosinase autoantigen can be induced from healthy donor blood to lyse melanoma cells. J Immunol 154:: 3991,1995-3998, Medline, Google Scholar4. Brichard V, Van Pel A, Wolfel T, et al: The tyrosinase gene codes for an antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas. J Exp Med 178:: 489,1993-495, Crossref, Medline, Google Scholar5. Kwon BS, Haq AK, Pomerantz SH, et al: Isolation and sequence of a cDNA clone for human tyrosinase that maps at the mouse c-albino locus. Proc Natl Acad Sci U S A 84:: 7473,1987-7477, (erratum published in Proc Natl Acad Sci U S A 85:6352, 1988) Crossref, Medline, Google Scholar