Distinct TCRAV and TCRBV repertoire and CDR3 sequence of T lymphocytes clonally expanded in blood and GVHD lesions after human allogeneic bone marrow transplantation

Abstract

Summary:Acute graft-versus-host disease (GVHD) is a disorder involving the skin, gut and liver that is caused by mismatches of major and/or minor histocompatibility antigens between the HLA-identical donor and recipient. If T lymphocytes infiltrating GVHD lesions recognize antigens expressed in these organs, T cell clones should expand in inflammatory tissues. We previously reported that recipients of allogeneic bone marrow grafts have clonally expanded TCR␣␤ + T lymphocytes soon after transplantation, which leads to a skew of TCR repertoires. To establish whether or not the same antigens cause clonal expansion of T lymphocytes in both blood and GVHD tissues, we examined the usage of TCR ␣ and ␤ chain variable regions (TCRAV and TCRBV) and determined the complementarity-determining region 3 (CDR3) of T lymphocytes clonally expanded in circulating blood and GVHD lesions. We found that the repertoires and CDR3 diversity of TCRAV and TCRBV differed between the GVHD lesions and circulating blood, suggesting the selective recruitment of antigenspecific T cells into GVHD tissues. We also found that the usage of TCRAV and TCRBV by the clonally expanded T lymphocytes and their CDR3 sequences differed between the GVHD tissues and blood. These results suggest that the antigen specificity of TCR␣␤ + T lymphocytes clonally expanded in blood and GVHD lesions is different. Bone Marrow Transplantation (2002) 30, 915-923. doi:10.1038/sj.bmt.1703730 Keywords: human; graft versus-host disease; T lymphocytes; T cell receptors transplantation Acute graft-versus-host disease (GVHD) is a disorder involving the skin, gut and liver that is caused by mismatches of major and/or minor histocompatibility antigens between the donor and recipient. 1 Although several candidates for minor histocompatibility antigens have been reported, including the male-specific H-Y antigen, cellularly defined minor HA antigens and myxovirus-related protein, the mechanism of cellular injury in acute GVHD has not been fully elucidated. [2][3][4] We previously reported a skewing of repertoires of the T cell receptor-␤ chain variable region (TCRBV) and the TCR-␣ chain variable region (TCRAV) in recipients of allogeneic hematopoietic stem cell grafts from HLAmatched donors. 5 This results from the extensive clonal expansion of a limited number of T cells in the graft. 6 Skewed TCR repertoires accompany accumulation of the CD28 Ϫ fraction in both CD4 + and CD8 + T cells. 7,8 Since accumulating evidence indicates an association between the loss of CD28 expression and T lymphocyte aging, 9,10 posttransplant skewing of the TCR repertoires probably results from chronic antigen stimulation in vivo.The above results raise the issue of whether or not clonally expanded T cells in circulating blood recognize the discrepant histocompatibility antigens between donors and recipients, which may be responsible for the induction of acute GVHD. Moreover, crucial questions include whether or not T lymphocytes infiltrating acute GVHD lesions recognize antigens commonly expressed in...

Molecular Analysis of Alloreactive CTL Post-Hemopoietic Stem Cell Transplantation

“…However, we were able to identify immunodominant T cell clones for most of the coculture experiments molecularly analyzed. Several of the clonotypes were present at a very high frequency, approaching levels more reminiscent of clonal expansions seen in other immune conditions (i.e., CMV and EBV) (17)(18)(19) or in large granular lymphocytic leukemia (13,14) and similar to those previously reported in blood posttransplantation (5,6,8,20). No VB family was over-represented among the clonotypes and no sharing was identified between the patients.…”

“…Clonotypic diagnostics have been applied to studying the GVHD-related T cell repertoire posttransplant in both blood (4,5) and tissues affected by GVHD (6,7). Previously, we characterized the VB utilization spectrum and immunodominant clonotypes among CD8 ϩ T cells in 29 HSCT patients and in biopsies from nine patients posttransplant (8,9).…”

“…Recipient PBMC were labeled with PKH26 (Sigma-Aldrich) as per the manufacturer's instructions and cultured with or without donor BM mononuclear cells at a 1:10 ratio in RPMI 1640 medium (Invitrogen Life Technologies) supplemented with 10% heat-inactivated FCS, penicillin/streptomycin (1ϫ), and IL-2 (50 U/ml) for 72 h. Positive controls consisted of cultures of donor cells stimulated with PHA (nos. [1][2][3][4][5][6][7][8] or PMA (nos.9 -17).…”

Accumulation of autoreactive effector T cells and allo-specific regulatory T cells in the pancreas allograft of a type 1 diabetic recipient

“…This has been proven to be suitable for detecting oligoclonality in transplantation [17]. Under 'normal' circumstances a Gaussian distribution within each Vbeta family is expected in polyclonal cell populations, whereas mono-or oligoclonality would lead to over-representation of a certain CDR3-length.…”

Interstitial Pneumonia Associated with Connective Tissue Disease: An Overview and an Insight

“…We then analyzed T cell receptor α-chain variable region (TCR Vα) and TCR β-chain variable region (TCR Vβ) repertoires of T cells infiltrating the lung tissues using an adaptor ligation polymerase chain reaction (PCR)-based microplate hybridization assay [159]. Quantitative assay has been used in many previous studies, and accuracy and reproducibility of this assay have been validated [160,161]. Briefly, total RNA was extracted with TRIzol Reagent (Invitrogen, USA) from lung tissues obtained by VATS biopsy.…”