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Copyright of Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu is the property of Chinese Journal of Tissue Engineering Research and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

This study was purposed to investigate the immune reconstitution of T-cells in patients who received haploidentical hematopoietic stem cell transplantation (hiHSCT). The peripheral blood was harvested from 22 patients before transplantation and at month 1, 3, 6 after hiHSCT. The proportions of T lymphocyte subtypes including CD3(+), CD4(+), CD8(+), CD45RO(+), and CD45RA(+)CD62L(+) were analyzed by flow cytometry, followed by the calculation of T cell numbers according to the amounts of peripheral blood leukocytes. Adenosine triphosphate (ATP) value in CD4(+) T cells was measured by ImmuKnow method to evaluate the function of lymphocytes. The results showed that the CD3(+) cell absolute value before transplantation was 833.75 ± 359.84/µl, but those values at month 1, 3, 6 after transplantation were 318.87 ± 266.71/µl, 1006.76 ± 512.32/µl and 1296.38 ± 958.77/µl respectively. The CD4(+) cell absolute value before transplantation was 336.99 ± 211.11/µl, but such values at month 1, 3, 6 after transplantation were 45.89 ± 44.21/µl, 142.97 ± 114.85/µl, and 181.78 ± 120.61/µl respectively. The CD8(+) cell absolute value before transplantation was 430.21 ± 159.48/µl, but those values at month 1, 3, 6 after transplantation were 230.44 ± 195.89/µl, 621.64 ± 318.83/µl, and 823.07 ± 633.55/µl respectively. The CD4(+)CD45RO(+) memory T cell absolute value before transplantation was 227.44 ± 73.34/µl, but such values at month 1, 3, 6 after transplantation were 43.47 ± 43.40/µl, 138.69 ± 110.17/µl, 147.73 ± 82.94/µl respectively. The CD8(+)CD45RO(+) memory T cell absolute value before transplantation was 212.70 ± 98.48/µl, but such values at month 1, 3, 6 after transplantation were 184.76 ± 168.65/µl, 445.90 ± 252.50/µl, 519.80 ± 475.53/µl respectively. CD4(+)CD45RA(+)CD62L(+) naive T cell number before transplantation was 68.94 ± 59.74/µl, but such cell numbers at month 1, 3, 6 after transplantation decreased to 2.44 ± 2.93/µl, 3.14 ± 3.48/µl, 23.22 ± 38.38/µl respectively. The CD8(+)CD45RA(+)CD62L(+) naive T cell absolute value before transplantation was 124.82 ± 60.95/µl, but those values at month 1, 3, 6 decreased to 19.37 ± 17.71/µl, 76.63 ± 50.85/µl, and 114.49 ± 174.29/µl respectively. The ATP value in CD4(+) T cells decreased to 210.19 ± 119.37 ng/ml at month 1 after transplantation and increased to 280.62 ± 110.03 ng/ml at month 3, and 357.28 ± 76.18 ng/ml at month 6 after transplantation. It is concluded that CD8(+) memory T cell reconstruction contributes critically to T cell recovery early after hiHSCT, while the thymic output function remains low. However, T cell function recovers to normal range at month 3 after 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., Conclusion: 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.

Objective: To investigate the dynamic changes of lymphocyte subsets before and after autologous hemopoietic stem cell transplantation (HSCT) in severe/refractory autoimmune disease (AID) and study the post-transplantation immunological reconstitution in AID., Methods: Thirteen patients with severe/refractory AID who registered for HSCT from April 2003 to April 2005 in Peking Union Medical College Hospital, including 8 patients with systemic lupus erythematosus, 4 patients with rheumatoid arthritis, and 1 patient with primary Sjögren's syndrome (pSS) were enrolled in this study. Blood samples were collected before/after mobilization, before conditioning, and 2 weeks, 1 month, 3 months, 6 months, 12 months, and 18 months post-transplantation. Lymphocyte subsets were tested by flow cytometry as follows: T cell (CD3 +), B cell (CD19 +), natural killer (CD3-CD16 + CD56 +), Th (CD3 + CD4 +), Tc (CD3 + CD8 +), naïve T (CD4 + CD45RA), memory T (CD4 + CD45RO), and CD4/CD8 ratio., Results: Lymphocyte subsets for SLE patients were severely abnormal compared to normal or RA patients (both P < 0.01). B cell reconstituted to normal level within 18 months, meanwhile NK and T cell remained low. The repopulations of Th and naive T cell were delayed, which caused the up-side-down of CD4/CD8 ratio and low level of naYve T cell percentage for a relatively long time., Conclusions: Lymphocyte subsets abnormality in SLE patients are more severe than in RA patients. Although most autoimmune T/B cell in the grafts and patients can be effectively removed after transplantation, nonmyeloablative conditioning may be a risk for the relapse of AID. The long-term inhibition of CD4 + T cell may be related with the relief of AID after transplantation.