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* Experimental Hematology and Hematopoiesis Section,
Bone Marrow Transplantation Program,
Division of Pathology and Laboratory Medicine, Cleveland Clinic Foundation, Cleveland, OH 44195; and
Case Comprehensive Cancer Center, Cleveland OH 41106
The development of laboratory tests for the diagnosis and monitoring of graft-vs-host disease (GVHD) is hampered by a lack of knowledge of minor histocompatibility Ags triggering alloresponses. We hypothesized that the unique molecular structure of the TCR could be used as a marker for the unidentified Ags and exploited for molecular monitoring of GVHD posttransplant. To identify alloreactive T cell clones, we performed in vitro allostimulation cultures for a cohort of patients undergoing hemopoietic stem cell transplantation and determined the sequence of the CDR3 of immunodominant alloreactive clones; 10 corresponding clonotypes restricted to activated T cells were identified. As an alternative method for the identification of alloreactive clones, molecular TCR analysis was applied to biopsies of GVHD-affected tissues. Culture- and biopsy-derived clonotypes were used to design sequence-specific quantitative PCR assays to monitor the levels of putative allospecific clonotypes in posttransplant blood samples and subsequent biopsies. Because of the rational design of the methods used to identify immunodominant clonotypes, we were able to follow the behavior of potentially GVHD-specific T cells during the transplant course. Based on our results, we conclude that molecular T cell diagnostics can be a powerful tool for monitoring immune responses posttransplantation.
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 C.L.O. designed the research, performed the research, analyzed data, and wrote the paper. L.G. performed the research and analyzed data. R.M.S., Z.M., and R.D. analyzed clinical data. R.M.S. and B.B. provided patient samples for the study. Y.N. performed research. R.T. analyzed biopsies. J.P.M. designed the research and analyzed data.
2 Address correspondence and reprint requests to Dr. Jaroslaw P. Maciejewski, R40 Taussig Cancer Center, The Cleveland Clinic Foundation, 9600 Euclid Avenue, Cleveland, OH 44195.
3 Abbreviations used in this paper: GVHD, graft -vs-host disease; BM, bone marrow; Bx, biopsy sample; CB, 
-chain constant region; HSC, hemopoietic stem cell transplant; mHA, minor histocompatibility Ag; PB, peripheral blood; VB, variable -chain.
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  K. Sugimoto, M. Murata, S. Terakura, and T. Naoe CTL Clones Isolated from an HLA-Cw-Mismatched Bone Marrow Transplant Recipient with Acute Graft-Versus-Host Disease J. Immunol., November 1, 2009; 183(9): 5991 - 5998. [Abstract] [Full Text] [PDF]
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 W. J. Murphy Raising the spectra of T-cell profiling Blood, October 15, 2008; 112(8): 3008 - 3009. [Full Text] [PDF]
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