[Prediction of T-cell immune reconstitution by investigating T cell receptor excision circle and T-cell receptor beta-chain variable region clonal repertoire in leukemia patients after allogeneic hematopoietic stem cell transplantation].
OBJECTIVE
To predict T-cell immune reconstitution by investigating T cell receptor excision circles (TREC) and T-cell receptor beta-chain variable region (TCRBV) clonal repertoire in leukemia patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT).


METHODS
Real-time quantitative PCR was used to detect the TREC in 43 leukemia patients undergoing matched sibling donor bone marrow transplantation (MSD BMT), matched unrelated donor (MUD) BMT, or haploidentical-stem cell transplantation (HID-SCT), and in 70 normal individuals. RT-PCR was used to amplify 24 subfamily genes of T-cell receptor beta-chain variable region (TCRBV) in 24 of the 43 patients and 5 normal donors as control. The PCR products were further analyzed by genescane to evaluate the clonality of BV subfamily, characteristics of complementarity determining region 3 (CDR3), and the usage rate in BV subfamily.


RESULTS
There were (335.1 +/- 782.5) copies/10(5) cells in the 43 patients before transplantation, far lower than the normal value. The TREC values of the patients of the 3 groups all decreased obviously 3 months after transplantation. The TREC value of the MSD-BMT group recovered faster than the other two groups and reached the value before transplantation in 24 months. The recovery of TREC value in the HID-BMT group was delayed. 3 - 19 months after transplantation, the usage of TCRBV subfamilies was still restricted. There were 6 - 16 BV subfamilies expressed and 33% - 48% of them were polyclonals, the others were monoclones and oligoclones and existed in 24 BV subfamilies, no common monoclone BV subfamilies was expressed.


CONCLUSIONS
Investigation of the TREC and TCRBV clonal repertoire showed that the number of naive T cell is lower and the usage of TCRBV subfamilies skewed 3 - 24 months after allo-HSCT. The immune deficiency of the patients undergoing HID-BMT is more prominent and consistent with the clinical process.
Fu
Yue-Wen
YW
Department of Hematology, First Hospital Affiliated to Suzhou University, Suzhou 215006, China.
Wu
De-Pei
DP
Cen
Jian-Nong
JN
Feng
Yu-Feng
YF
Chang
Wei-Rong
WR
Qiu
Qiao-Cheng
QC
Zhu
Zi-Ling
ZL
Zhu
Ping
P
D000293:Adolescent; D000328:Adult; D002648:Child; D017637:Clonal Deletion / Q000235:genetics / Q000276:immunology; D005260:Female; D015973:Gene Expression Regulation, Leukemic; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006801:Humans; D007938:Leukemia / Q000235:genetics / Q000276:immunology* / Q000601:surgery; D008297:Male; D008875:Middle Aged; D011184:Postoperative Period; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics / Q000276:immunology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism
D011948:Receptors, Antigen, T-Cell; D016693:Receptors, Antigen, T-Cell, alpha-beta