弥漫大B型非霍奇金淋巴瘤患者外周血TCR Vβ亚家族T细胞分布和克隆性增生特点

目的：了解弥漫大B淋巴瘤（DLBL）患者外周血T细胞的TCRVβ基因谱系及克隆性增殖情况。方法：利用RT-PCR方法扩增6例DLBL患者外周血单个核细胞中24个TCRVβ基因的互补决定区3（CDR3）,PCR产物进一步经荧光标记和基因扫描分析CDR3长度而确定T细胞的克隆性。结果：6例DLBL患者分别表达9-20个TCRVβ亚家族。患者均存在1个或多个Vβ亚家族的克隆增殖T细胞,增殖形式包括寡克隆、寡克隆增殖趋势及双克隆。Vβ13和Vβ15亚家族出现克隆增殖性T细胞的频率较高（66.7%）。结论：DLBL患者外周血TCRVβ亚家族T细胞的分布具有特点,表现为TCRVβ亚家族T细胞呈限制性选用和克隆性增殖,这可能是淋巴瘤患者机体对淋巴瘤相关抗原产生的特异性免疫反应。     

大剂量化疗并自体外周血干细胞移植治疗原发性干燥综合征二例患者的免疫重建

目的 通过观察大剂量化疗（HDC）并自体外周血干细胞移植（APBSCT）治疗重症原发性干燥综合征（pSS）患者治疗前后T细胞亚群和T细胞受体（TCR）克隆谱型的变化,探讨移植的免疫重建过程。方法 2例长期糖皮质激素和免疫抑制剂治疗病情不能缓解的重症pSS患者,给予HDC及APBSCT治疗。流式细胞仪检测T细胞亚群,观察治疗前后T细胞亚群的变化。RT-PCR扩增患者外周血TCRV区24个β亚家族序列,阳性的亚家族行荧光标记的二次PCR,用分析TCRVβ亚家族CDR3长度的方法检测T细胞克隆性,以及移植前后T细胞克隆谱的变化。结果 2例重症pSS患者顺利完成APBSCT全过程,T细胞亚群有不同程度的恢复,移植后TCRVβ有由寡克隆向多克隆转化的趋势。结论 本组2例重症pSS患者HDC并APBSCT治疗后病情得以控制,免疫功能在3个月时已有恢复的趋势,但其可行性与安全性有待大样本及随诊证实。     

用RT-PCR和Genescan分析CML急变期病人TCRVβT细胞的表达和克隆性

目的：了解CML急变期的TCRVβ亚家族T细胞的表达及其克隆性。方法：采用RT－PCR扩增4例CML急变期病人的外周血单个核细胞的TCRVβ24个亚家族的互补决定区3（CDR3），产物进一步经基因扫描分析确定T细胞的克隆性。结果：病人外周血仅表达4～9个Vβ亚家族T细胞，主要为Vβ1，Vβ2，Vβ3，Vβ5，Vβ15和Vβ17；基因扫描分析显示2例CML急粒变的部分产物为寡克隆性，而2例CML急淋变者的产物均为多克隆性。结论：CML急变期外周血仅选择性表达部分Vβ亚家族T细胞；CML急粒变存在克隆性增殖T细胞，这可能是机体对白血病细胞相关抗原的一种直接反应。该方法可用于检测微小残留病变和判断疾病复发。     

基因扫描分析维持性血液透析患者外周血T细胞克隆性分布的特点

目的:了解慢性肾功能衰竭进行维持性血液透析(MHD)患者的外周血T细胞克隆性增殖的特性和意义. 方法:通过RT-PCR方法扩增11例膜性肾病(MGN)3例、膜增殖性肾炎(MPGN)3例、IgA肾病(IgAN)4例、局灶节段性肾小球硬化症(FSGS)1例.MHD患者外周血单核细胞T细胞受体(TCR)Vβ24个亚家族基因的CDR3,阳性的PCR产物进一步经荧光素标记和基因扫描的方法分析产物CDR3的长度及序列,了解T细胞的克隆性.结果:11例MHD患者中仅表达1～10个TCRVβ的亚家族,其中2例只表达Vβ3一个亚家族,分别为寡克隆及双克隆性.最常见的TCR Vβ亚家族是Vβ3,其次Vβ8、Vβ19、Vβ22,再次为Vβ10、Vβ23.11例患者的透析时间与TCR Vβ表达呈显著负相关(r=-0.646,ρ=0.044);用协方差分析MGN、MPGN、IgAN三个病种的TCR Vβ表达差别:不同透析时间的TCR Vβ表达的数量不同(F=10.193,P=0.019),而MGN、MPGN、IgAN三个病种之间的表达量没有显著的差异(F=1.772,P=0.249).结论:MHD患者外周血存在着TCR Vβ亚家族T细胞倾斜分布的现象,并有克隆性增殖的特征,提示大部分MHD患者存在某些细胞免疫功能低下和有相同的免疫刺激原.透析时间愈长,TCR Vβ表达的数量愈少,提示长期血液透析可能是MHD患者的细胞免疫功能低下的一个原因;而 MGN、MPGN、IgAN三个病种之间TCRVβ的表达量没有显著差异.     

基因扫描分析人T细胞受体CDR3谱型检测T细胞克隆技术的建立

目的建立人T细胞受体（TCR）互补决定区3（CDR3）基因谱型的基因扫描分析术,为分析T细胞克隆提供研究方法。方法分别以Jurkat和Raji细胞作为阳性和阴性对照,应用RT-PCR方法扩增5例健康志愿者外周血T细胞的34个TCR Vα亚家族和26个TCR Vβ亚家族的CDR3,并用基因扫描分析扩增产物以确定T细胞的克隆性。结果T细胞株Jurkat细胞仅表达TCR Vα1．1和Vβ8基因,且为单克隆,而B细胞株Raji细胞则不表达所有的TCR Vα和Vβ基因。5例健康人表达所有的TCR Vα和Vβ基因,均为多克隆T细胞。结论基因扫描分析人TCRCDR3基因谱型是检测T细胞克隆性的精确敏感方法。     

自身免疫性溶血性贫血病人TCRVβ亚家族T细胞表达特点

目的：了解自身免疫性溶血性贫血病ＴＣＲ Ｖβ亚家族Ｔ细胞的表达情况。方法：采用ＲＴ－ＰＣＲ扩增１ 例ＡＩＨＡ 和１ 例Ｅｖａｎ′ｓ 综合征病人的外周血单个核细胞的ＴＣＲ Ｖβ２４ 个亚家族基因，９ 例正常人作为对照。结果：正常人表达除Ｖβ２０ 以外的其它亚家族Ｔ细胞，而病人外周血仅表达３～６ 个Ｖβ亚家族Ｔ细胞，为Ｖβ２、Ｖβ３、Ｖβ５、Ｖβ１３、Ｖβ１６ 和Ｖβ２４。结论：２ 例病人外周血仅选择性表达部分β亚家族Ｔ细胞，这可能是病人细胞免疫功能异常的特征。     

慢性乙型肝炎患者外周血CD8~+T淋巴细胞亚群T淋巴细胞受体β链互补决定区3谱型研究

目的 研究慢性乙型肝炎(CHB)患者外周血CD8+T淋巴细胞亚群T淋巴细胞受体(TCR)β链Ⅴ区(BV)中互补决定区3(CDR3)谱型特点.方法 采集8例CHB患者(ALT>2倍正常值上限)的抗凝血,分离外周血单个核细胞,磁珠分选CD8~+T淋巴细胞亚群,提取RNA并应用逆转录-聚合酶链反应扩增TCR BV中CDR3基因的24个家族,采用免疫指纹技术进行TCRBV各家族基因扫描和谱型分析.不同细胞亚群间数据比较采用配对t检验.结果 8例患者外周血淋巴细胞TCR BV家族CDR3谱型发生明显偏移,表现为单克隆性,寡克隆性及偏峰性克隆增生.8例患者CD8~+T淋巴细胞亚群发生TCR BV CDR3谱型偏移家族总个数高于去除CD8~+T淋巴细胞群的外周血单个核细胞[(10.6±4.7)个比(4.1±3.1)个,t=6.619,P<0.01)];比较单、寡克隆增生2种形式偏移的家族,CD8+T淋巴细胞亚群也高于代表CD4+T淋巴细胞的去除CD8~+T淋巴细胞群的外周血单个核细胞[(8.8±4.5)个比(3.9±2.8)个,t=5.706,P<0.01].对其中3例患者磁珠分选前后TCR BV谱型的比较发现,分选后CD8+T淋巴细胞亚群发生TCR BV谱型偏移家族数均高于分选前.结论 通过淋巴细胞亚群分选方式分析TCR BV家族CDR3谱型变化可以减少不同淋巴细胞亚群之间的峰型重叠干扰;应用这一方法分析外周血CD8~+T淋巴细胞亚群克隆增生程度有助于了解CHB患者炎症的发生机制.     

慢性乙型肝炎患者外周血T淋巴细胞受体谱系分析及互补决定区3序列测定

目的从新的角度探讨慢性乙型肝炎（CHB）炎症活动期外周血T淋巴细胞克隆性增生情况，了解T淋巴细胞免疫应答在CHB发病机制中的状态及作用。方法利用逆转录聚合酶链反应（RT-PCR）扩增T淋巴细胞受体（TCR）β链V区基因各家族（BV），并采用免疫指纹技术（immunoscope）对4名健康献血员及8例处于炎症活动期CHB患者外周血TCR BV家族基因的优势利用情况及克隆性增生T淋巴细胞β链互补决定区3（CDR3）序列进行分析。结果4名健康献血员外周血T淋巴细胞TCR BV家族CDR3谱型均呈正态分布，而8例CHB患者均出现一个或多个TCR BV家族单克隆或寡克隆性增生。对克隆性增生T淋巴细胞β链CDR3区序列测定证实存在不同的CDR3序列。结论CHB活动期外周血T淋巴细胞存在明显克隆性增生，进一步提示T淋巴细胞免疫应答可能参与CHB的发病过程，且可能有多个病毒抗原表位参与了细胞免疫应答。     

T细胞受体V β链基因限制性取用与哮喘患者的关系

目的：探讨T细胞受体β链可变区（TCR Vβ）受体谱限制性取用与哮喘的相关性。方法：采用荧光标记半定量逆转录聚合酶链反应（RT－PCR），借助377 DNA自动测序仪分析36例哮喘患外周血TCR Vβ基因取用格局的变化，通过变性聚丙酰胺凝胶电泳及单链构象多态性分析（SSCP），对哮喘患优势取用的TCR Vβ基因家族进行基因片段重组长度及碱基差异分析。结果：TCRVβ8在哮喘患以及其中对屋尘螨过敏外周血中被优势取用的次数（13／36，10／24）与正常对照组（1／16）比较，差异有显性（P＝0．0230，0．0147），TCR Vβ5．1 在屋尘螨过敏哮喘外周血中被优势取用的次数（10／24）与正常对照组（1／16）比较，差异有显性（P＝0．0186），值得注意的是，36例哮喘患中有2例豚草过敏均优势取用了Vβ1，同时发现被哮喘患优势取用的TCRVβ基因家族在正常人中均为多克隆扩增，同时哮喘患的外周血中大多数亦为多克隆扩增，但有5例哮喘患Vβ8为寡克隆扩增，出现限制性取用，结论：TCR Vβ8，5．1基因片段可能和哮喘患屋尘螨过敏有关，哮喘患的外周血TCR Vβ基因家族呈寡克隆扩增趋势。     

强直性脊柱炎患者外周血T细胞受体β链可变区基因谱系多态性的初步研究

目的 探讨强直性脊柱炎(AS)患者外周血T细胞受体β链可变区互补决定区3(TCRBV CDR3)谱系多态性,为AS免疫发病机制研究提供实验依据.方法 采用反转录-聚合酶链反应(RT-PCR)扩增AS患者外周血单个核细胞(PBMC)中T细胞TCR BV的26个亚家族CDR3,经免疫扫描谱型技术分析TCR BV CDR,3的谱系多态性情况.结果 20例活动期AS患者TCR BV CDR3扫描谱型均出现非正态的异常峰型,包括单峰、寡峰/寡峰趋势、偏峰和不规则异常峰型.其中有18例患者部分亚家族出现寡克隆/寡克隆趋势增生,有1例患者BV16和BV18的2个亚家族出现单克隆增生.5名健康对照PBMC TCR BV CDR3谱型绝大多数呈高斯分布.结论 AS患者外周血TCR BV CDR3谱系具有显著多态性和谱系漂移特点,进一步表明T细胞在AS免疫发病机制中扮演重要角色.单/寡克隆增生的T细胞有可能是AS发病中的自身反应性T细胞.     

Evaluation of TCR Vβ subfamily T cell expansion in NOD/SCID mice transplanted with human cord blood hematopoietic stem cells

Examination of the T cell receptor (TCR) gene repertoire is important in the analysis of the immune status of models, because clonal expansion of T cells permits the identification of specific antigen responses of T cells. Little is known about T-cell immunity in the humanized NOD/SCID mouse model. TCR Vβ repertoire usage and clonality were analyzed to investigate the distribution and clonal expansion of TCR Vβ subfamily T cells in NOD/SCID mice transplanted with human cord blood (CB) hematopoietic stem cells. The NOD/SCID mice were sublethally irradiated (⁶⁰Co, 300cGy) to eliminate residual innate immunity in the host. The experimental mice were transplanted intravenously with CB CD34⁺ cells sorted by MACS. After 6 weeks, RNA was obtained from peripheral blood, bone marrow and thymus of the study animals. The gene expression and clonality of the TCR Vβ repertoire were determined by RT-PCR and GeneScan techniques. A restricted range of TCR Vβ usage was exhibited in the bone marrow of mice, which included TCR Vβ 1, 2, 9, 13 and 19. Further, oligoclonal expression of some TCR Vβ subfamilies (Vβ9, 13, 19) was identified by GeneScan technique. To investigate the reason for oligoclonal expansion of the TCR Vβ subfamily T cells from CB in mouse models, the T-cell culture with tissue-antigen of NOD/SCID mouse was performed in vitro. The cells from peripheral blood mononuclear cells and bone marrow, spleen, thymus in NOD/SCID mice were frozen and thawed, and used as tissue-antigen. CB mononuclear cells were separately cultured with the component from those murine cells for 15–20 days. Oligoclonal expression or oligoclonal trend of some TCR Vβ subfamilies (Vβ10, 11 and Vβ2, 15, 16, 19) was detected in T cells after stimulation with tissue-antigen of NOD/SCID mouse. Interestingly, a similar clonal expansion of the TCR Vβ11 subfamily was found in T cells cultured with peripheral blood, bone marrow and spleen respectively. The TCR Vβ subfamily T cells could be reconstituted in humanized NOD/SCID mouse transplanted with CD34⁺ cells from CB. The restricted expression and clonal expansion of some CB T cell clones may be induced by tissue-antigens of NOD/SCID mice.     

The TCR Vβ repertoire usage of T-cells from cord blood induced by chronic myelogenous leukemia associated antigen

Understanding the clonality and restricted usage of the TCR Vβ repertoire of expanded T-cells induced by the chronic myelogenous leukemia (CML) associated antigen may be useful in helping design new immunotherapeutic strategies specifically for CML. T-cells from cord blood that had been stimulated by different stimulators (IL-2, PHA, CML cells, K562 cells and bcr-abl peptide) were amplified in vitro by liquid T-cell culture and the mixed lymphocyte and tumor cell culture (MLTC) method. By using the RT-PCR, the CDR3 segments of 24 variable region genes of TCR β was analyzed in T-cells from 22 cases of cord blood before and after T-cell culture, to observe the usage of TCR Vβ repertoire. The PCR products were further labeled with fluorescence and analyzed by the Genescan technique for the CDR3 size, to evaluating clonality of the detectable TCR Vβ T-cells. Only a part of 24 Vβ subfamily T-cells (3–15 subfamilies) could be detected, however, all of the 24 TCR Vβ subfamily of T-cells were detected after in vitro culture with PHA or IL-2+anti-CD3 antibody. The number of expressed TCR Vβ subfamilies was gradually reduced by prolonging the time of culture with CML-associated antigens. The restricted expression of TCR Vβ subfamilies and oligoclonal expansion of Vβ21 T-cells were found in cultured T-cells induced by CML cells, K562 cells or bcr-abl peptide. In conclusion, T-cells from cord blood may have the potential capability of proliferation in different TCR Vβ subfamily T-cells, and the ability for restricted expression and clonal expansion, when T-cells were induced by CML associated antigen.     

Random suppression of T cells that bear specific T cell receptor Vβ sequences in adult T cell leukemia/lymphoma (ATLL) patients at each clinical stage: Carrier,smoldering, chronic,and acute

Human T cell leukemia virus type I (HTLV-I) is associated with adult T cell leukemia/lymphoma (ATLL), which is well known as a T cell malignancy. In order to clarify whether HTLV-I plays a role as a virus-encoded superantigen in the neoplastic process, we examined the TCR Vβ families in the peripheral blood at four different clinical stages: carrier, smoldering leukemia, chronic leukemia, and acute leukemia. An increased number of CD4 T cells was found in each of the four clinical stages. However, we found neither uniform specific losses nor uniform clonal expansion of particular TCR Vβ gene families in any case from the four clinical stages. However, a suppression of the random TCR Vβ families was found. Our data did not therefore directly suggest the existence of a common superantigen model of HTLV-I which induces an increase in CD4 T cells. The random suppression in the TCR Vβ repertoire is most likely caused by the influence of HTLV-I neoplastic pathogenesis rather than by virus-encoded superantigens. In the patients with acute leukemia, one or two families of the Vβ repertoires were very strongly expressed, while in chronic leukemia, no such repertoire of strong expression was observed. The immunological reaction of the hosts might thus be different between the above described groups. © 1996 Wiley-Liss, Inc.     

Emergence of oligoclonal t cell populations following therapeutic t cell depletion in rheumatoid arthritis

Objective. To examine the compartment of CD4+ T cells in patients with rheumatoid arthritis (RA) who have developed persistent lymphopenia following antibody-mediated T cell depletion and to investigate why T cell depletion is of limited therapeutic efficacy. Methods. Circulating T lymphocytes from 10 patients with seropositive RA treated with the monoclonal antibody (MAb) CAMPATH-1H were longitudinally monitored by fluorescence-activated cell sorter analysis with MAb. To assess the molecular diversity of repopulating T cells, random samples of T cell clones from the peripheral blood of 3 patients were analyzed by sequencing the T cell receptor (TCR) β chains. At the time of recurring disease, the synovial tissue was examined by immunohistochemistry, and the repertoires of peripheral and synovial tissue T cells were compared by TCR β-chain sequencing and by semiquantitative hybridization with oligonucleotides specific for the V–D–Jβ junctional region of selected clones. Results. The reconstitution of the peripheral T cell compartment was very slow. A mean CD4+ T cell count of 105/μl was reached 34 weeks following MAb treatment. After treatment, the percentage of CD4+ T cells with the CD45RO+ phenotype was significantly increased (P = 0.001), indicating the expansion of antigen-primed memory T cells. TCR β-chain sequences revealed a marked restriction in the diversity of repopulating T cells with the emergence of dominant clonotypes. Despite the low counts of peripheral CD4+ T cells, the synovial tissue was infiltrated by CD4+ T cells to a similar extent as that in RA patients not treated with MAb. Selected clonotypes that had emerged in the peripheral blood compartment dominated the repertoire of tissue-infiltrating T cells in the synovium. Conclusion. In patients with RA, T cell depletion induces a long-term imbalance in T cell homeostasis. Clonal proliferation of CD4+ T cells severely restricts the diversity of available T cell specificities and results in the emergence of dominant clonotypes, which accumulate in the synovial tissue despite peripheral lymphopenia.     

Leukemia Associated Clonal Expansion of TCR Vβ Subfamily T Cells

The analysis of the T cell receptor (TCR) Vβ repertoire is one of the most sensitive methods to identify the clonal expansion T cells which respond to tumor associated antigens. Recently, studies have focused on clonally expanded T cells from patients or normal donors induced by leukemia associated antigens in vivo or in vitro. Understanding such clonality and restricted usage of TCR Vβ repertoire of leukemia-associated expanded T cells may be useful for the design of new immunotherapeutic strategy for leukemia.     

T cell receptor V beta repertoire of the antigen specific CD8 T lymphocyte subset of HIV infected children

OBJECTIVE: Analysis of the T cell receptor V beta repertoire during HIV infection reveals expansions in multiple V beta families of CD8 T cells, but their antigenic specificity is ill-defined. We sought to determine the TCR V beta repertoire of HIV specific CD8 T lymphocytes in infected children. DESIGN/METHODS: We performed flow cytometry to examine TCR V beta families as identified by specific monoclonal antibodies and binding of HIV peptide loaded tetrameric MHC complexes in peripheral blood samples from a group of HIV infected children. RESULTS: Simultaneous assessment of 12 selected expanded V beta families amongst nine HIV infected patients for tetramer binding revealed only one child in whom the expanded V beta population bound HIV Gag or Pol tetramers. In four HIV infected children, percentage tetramer binding cells was determined in 21 TCR V beta families. The tetramer binding cells of three children exhibited a widely distributed TCR V beta repertoire while in the fourth patient they were preferentially localized within two TCR V beta families. Repeat analysis revealed that the TCR V beta repertoire of tetramer binding cells was stable. CONCLUSIONS: These data provide evidence that the HIV-specific CD8 T cell response in children is usually distributed widely among many different TCR V beta families. The heterogeneity of the TCR V beta repertoire usage by the antigen specific CD8 T cells may reflect the dynamic interaction between host and pathogen during the course of HIV infection and may be influenced by the rate of viral mutation, CD4 T cell helper activity, or other factors.     

Immune Reconstitution and Tolerance after Allogeneic Hematopoietic Stem Cell Transplantation

We have evaluated recovery of CD56 positive and other cell types following allogeneic stem cell transplantation and have found that the recovery of CD56 positive cells was faster than other lymphoid cells after allogeneic stem cell transplantation, while the recovery of CD4 positive cells was markedly delayed. Chimerism analysis showed that mixed chimerism was often observed in younger (<30 years old) patients. Mixed chimerism in older (≥30 years old) patients was associated with rejection and relapse, while this was not found in younger patients. Among the chimerism of various cell populations, donor-derived CD56-positive cells are important in early engraftment when determined in allogeneic nonmyeloablative stem cell transplantation (allo-NST), regardless of the proportion of donor-derived CD3-positive cells. Complementarity-determining region three (CDR3) size spectratyping in T-cell receptor (TCR) chain subfamilies (Vβ) showed that high level of diversity in TCR Vβ repertoire is important for a late rejection and skewed TCR Vβ repertoire is correlated with the occurrence of graft-versus-host disease (GVHD) especially chronic GVHD. Expression of inhibitory natural killer (NK) cell receptors such as CD158b and CD94/NKG2A on peripheral CD3-negative and -positive cells were increased in parallel with GVHD. Interestingly, these cells appeared to control GVHD, while preserving graft-versus-leukemia (GVL) effect. Analysis of cytokine gene expression in peripheral blood mononuclear cells showed that type 1 helper T cells (Th1)-derived cytokines increased in severe GVHD, while Th2-derived cytokines such as IL-4, IL-10 and IL-13 increased in mild GVHD. These results indicate that Th2 cells suppress GVHD, although Th1 cells augment GVHD. Taken together, evaluation of immune reconstitution and tolerance in patients receiving allogeneic stem cell transplantation from the various viewpoints is essential and useful to obtain better clinical outcome.     

Evaluation of TCR Vbeta subfamily T cell expansion in NOD/SCID mice transplanted with human cord blood hematopoietic stem cells

Examination of the T cell receptor (TCR) gene repertoire is important in the analysis of the immune status of models, because clonal expansion of T cells permits the identification of specific antigen responses of T cells. Little is known about T-cell immunity in the humanized NOD/SCID mouse model. TCR Vbeta repertoire usage and clonality were analyzed to investigate the distribution and clonal expansion of TCR Vbeta subfamily T cells in NOD/SCID mice transplanted with human cord blood (CB) hematopoietic stem cells. The NOD/SCID mice were sublethally irradiated ((60)Co, 300cGy) to eliminate residual innate immunity in the host. The experimental mice were transplanted intravenously with CB CD34(+) cells sorted by MACS. After 6 weeks, RNA was obtained from peripheral blood, bone marrow and thymus of the study animals. The gene expression and clonality of the TCR Vbeta repertoire were determined by RT-PCR and GeneScan techniques. A restricted range of TCR Vbeta usage was exhibited in the bone marrow of mice, which included TCR Vbeta 1, 2, 9, 13 and 19. Further, oligoclonal expression of some TCR Vbeta subfamilies (Vbeta9, 13, 19) was identified by GeneScan technique. To investigate the reason for oligoclonal expansion of the TCR Vbeta subfamily T cells from CB in mouse models, the T-cell culture with tissue-antigen of NOD/SCID mouse was performed in vitro. The cells from peripheral blood mononuclear cells and bone marrow, spleen, thymus in NOD/SCID mice were frozen and thawed, and used as tissue-antigen. CB mononuclear cells were separately cultured with the component from those murine cells for 15-20 days. Oligoclonal expression or oligoclonal trend of some TCR Vbeta subfamilies (Vbeta10, 11 and Vbeta2, 15, 16, 19) was detected in T cells after stimulation with tissue-antigen of NOD/SCID mouse. Interestingly, a similar clonal expansion of the TCR Vbeta11 subfamily was found in T cells cultured with peripheral blood, bone marrow and spleen respectively. The TCR Vbeta subfamily T cells could be reconstituted in humanized NOD/SCID mouse transplanted with CD34(+) cells from CB. The restricted expression and clonal expansion of some CB T cell clones may be induced by tissue-antigens of NOD/SCID mice.