Effects of Jianpi Jiedu Recipe on TCRVβCDR3 Spectratyping of Liver Cancer Rats with Pi Deficiency Syndrome / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe anti-cancer effects of Jianpi Jiedu Recipe (JJR) on liver cancer (LC) rats with Pi deficiency syndrome (PDS) and its relation with the third complementary-determining region gene spectratyping of TCRVβ-chain (TCRVβCDR3).</p><p><b>METHODS</b>Rats were divided into 8 groups according to random digit table, i.e., the blank control group (normal), the PDS group, the LC model group, the LC-PDS group, high, middle, and low dose JJR groups (75.00, 37.50, 18.75 g/kg, respectively by gastrogavage, once per day), the thymus pentapeptide group (5 mg/kg, intramuscular injection, twice per week), 8 in each group. Rats in the normal group were administered with physiological saline by gastrogavage once per day. PDS rat model was prepared by bitter-cold purgation. LC model was prepared by orthotopic transplantation method. Twenty gene subfamilies of TCRβCDR3 in the thymus, liver, and LC tissues were detected by Gene Scan.</p><p><b>RESULTS</b>High and middle dose JJR could postpone the growth of LC volume (P < 0.05), with equivalent liver index and thymus index to those of the normal group (P > 0.05). In thymus and liver tissue of the normal group, the number of clones (20 and 19), gene fragment number (220 and 113), Quasi-Gaussian distribution ratio of TCRVβCDR3 gene repertoire (100.0% and 42.1%), and fragment fluorescence peak area (6,539 ± 2,325 and 1,238 ± 439) were at the highest level among the 8 groups. TCRVβCDR3 expressions in thymus and liver tissue of high and middle dose JJR groups were approximate to those of the normal group. They were in the middle of the thymus pentapeptide group, the PDS group, the LC model group, and poorest in the LC-PDS group. TCRVβCDR3 in liver tissue expressed the best in the thymus pentapeptide group.</p><p><b>CONCLUSION</b>JJR might inhibit the growth of LC cells, and its mechanism might be related to enhancing TCRVβCDR3 spectratype expression.</p>

Characterization of human αβTCR repertoire and discovery of D-D fusion in TCRβ chains

The characterization of the human T-cell receptor (TCR) repertoire has made remarkable progress, with most of the work focusing on the TCRβ chains. Here, we analyzed the diversity and complexity of both the TCRα and TCRβ repertoires of three healthy donors. We found that the diversity of the TCRα repertoire is higher than that of the TCRβ repertoire, whereas the usages of the V and J genes tended to be preferential with similar TRAV and TRAJ patterns in all three donors. The V-J pairings, like the V and J gene usages, were slightly preferential. We also found that the TRDV1 gene rearranges with the majority of TRAJ genes, suggesting that TRDV1 is a shared TRAV/DV gene (TRAV42/DV1). Moreover, we uncovered the presence of tandem TRBD (TRB D gene) usage in ~2% of the productive human TCRβ CDR3 sequences.

Apoptose e perfil do repertório da cadeia variável beta do receptor de células T em linfócitos T CD8+ de pacientes com leishmaniose cutânea / Apoptosis and profile of the repertoire of beta chain variable T cell receptor on CD8 + T lymphocytes of patients with cutaneous leishmaniasis

A resposta imune envolve a participação de subpopulações de células naïve, efetoras e de memória, as quais são funcionalmente distintas, e a atividade destas células depende do reconhecimento antigênico, que ocorre no contexto MHC-peptídeo-TCR. A manutenção destas subpopulações e a homeostasia do sistema imune são controlados, entre outros fatores, pela apoptose. No entanto, este processo de morte também tem sido relacionado à diversas patologias, sendo responsável pela modulação de eventos imunológicos, interferindo no curso da infecção. Em pacientes com leishmaniose cutânea (LC), apesar da indução preferencial de linfócitos T CD4(positivo), os linfócitos T CD8(positivo) estão envolvidos nos processos de progressão e controle da infecção. Neste contexto, a citometria de fluxo foi utilizada definir simultaneamente as subpopulações de linfócitos T CD8(positivo) naïve, efetora, de memória central e de memória efetora; a diversidade do repertório VBeta destes linfócitos; e a ocorrência de apoptose, na resposta imune da LC. Neste estudo foram analisadas amostras de sangue periférico, obtidas ex vivo de pacientes durante e após o tratamento, e de indivíduos sadios; e após cultivo in vitro frente a antígenos de Leishmania braziliensis. Os resultados mostram que a apoptose exerce um papel modulados, diminuindo a freqüência de linfócitos T CD8(positivo), o que parece contribuir para a persistência da lesão ativa, no décimo dia de tratamento; enquanto a cura clínica parece ser conseqüência da diminuição de apoptose nestes linfócitos T CD8(positivo) contribuindo para o restabelecimento da freqüência destas células. A análise da distribuição das subpopulações de linfócitos T CD8(positivo) revelou que a proporção de células naïve e efetoras é invertida ao longo do tratamento e do processo de resolução da lesão. Paralelamente, apesar do maior percentual de linfócitos T CD8(positivo) efetores observados em pacientes durante o tratamento, a ocorrência de apoptose nestas células pode comprometer uma resposta imune eficaz, estando associada à presença de lesão ativa ulceradaO caráter modulador que este processo de morte exerce na resposta imune em ambos os grupos de pacientes sugere que a ocorrência de apoptose, mais intensa na fase ativa da doença, esteja relacionada à manutenção da infecção. A reunião das análises de repertório VBeta nas diferentes subpopulações de linfócitos T CD8(positivo) mostraram uma indução oligoclonal na qual as células que expressam VBeta 9, VBeta 13.2 ou VBeta 23 podem ser os clones selecionados a se diferenciar em células de memória central após a cura clínica; enquanto que os clones que expressam a cadeia VBeta 22, parecem ser preferencialmente selecionados a se diferenciar em células de memória efetora, durante a fase ativa da doença. Os únicos clones de linfócitos T CD8(positivo) efetores que apresentaram uma expansão significativa foram aqueles que expressam a cadeia VBeta 12, a qual parece estar associada à resposta imune efetora desencadeada na fase ativa da LC. Sendo assim, a heterogeneidade entre o repertório VBeta das diferentes subpopulações de linfócitos T CD8(positivo) ressalta a importância de avaliar separadamente o repertório das células naïve, efetoras, de memória central e de memória efetora.

Characteristics of CDR3 of TCR beta on CD8+ T cells in chronic hepatitis B patients / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To analyze the characteristics of CDR3 of TCRbeta on CD8+ T cells in chronic hepatitis B patients.</p><p><b>METHODS</b>Eight patients with chronic hepatitis B (ALT more than 2 ULN) were enrolled in this study. CD8+ T cells were isolated from peripheral blood. RT-PCR was proformed to amplify the CDR3 of TCRbeta, and the PCR products were sequenced and analyzed.</p><p><b>RESULTS</b>The chronic hepatitis B patients showed obvious clonal expansion of T cell, and three perturbation patterns of T cell expansion were showed in the CDR3 of TCRbeta, including monoclonicity, oligoclonicity and skewed peak patterns. The number of perturbation families of CD8+ subpopulation was significantly higher than that of CD8- subpopulation (10.6+/-4.7 vs. 4.1+/-3.1, t = 6.619, P less than 0.01). In 3 out of 8 patients, the number of perturbation families of CD8+ subpopulation was also higher than that of PBMCs without depleting CD8+ subpopulation.</p><p><b>CONCLUSIONS</b>The characteristics of CDR3 of TCRbeta may help to understand the inflammatory response in CHB patients.</p>

Changes of T-cell clonality after induction-cultivation of peripheral T lymphocytes in adoptive immunotherapy for leukemias / 中国实验血液学杂志

This study was purposed to analyze the changes of T-cell clonality after induction of peripheral T lymphocytes by autogenous DC and cytokines in the preparation of adoptive immunotherapy for leukemias. The bone marrow and peripheral blood from 21 leukemia patients at remission stage after treatment and subjected to adoptive immunotherapy were collected. Their DCs and T-cells were stimulated with cytokines and then were mixed to activate T-cells. T-cell receptor beta variable region (TCRBV) families were amplified by RT-PCR, and genescan method and sequencing of the PCR products were used to observe the clonality changes of T-cells before and after the induction and cultivation of T-cells. The flow cytometry was used to identify CD3(+), CD4(+), CD8(+), CD3(+)CD56(+) and CD4(+)CD25str(+)FOXP3(+) cells to disclose the ratio change of cytotoxic T lymphocytes (CTL), helper T-cells, regulatory T-cells and NK T-cells before and after induction and cultivation of T-cells. The results showed that in the 21 patients, most of the 24 TCRBV families presented as oligoclonal distribution on genescan, several families were not expressed, and only a few families remained polyclonal. TCRBV24 was found to be oligoclonal in all of the 21 patients. DNA sequence analysis of TCRBV24 revealed a common motif of VAG in CDR3 in 3 cases and a common motif of GGG in CDR3 in 2 cases. In patient 5, both TCRBV 24 and TCRBV8 contained the same motif of GGG in CDR3. The identical motif in these patients may suggest that these T-cells recognize the same antigen. The peripheral lymphocytes demonstrated recovery of clonal profile on genescan from oligoclonal profile and absence of several families before the induction and cultivation to typical polyclonal profile in all TCRBV families after the induction by DC and cytokines for 13 days. After the induction and cultivation, the number of lymphocytes increased to 3.38 +/- 1.20 times. CD3(+), CD4(+), CD8(+), CD3(+)CD56(+) and CD4(+)CD25str(+)FOX P3(+) cells were 71.1 +/- 11.8%, 26.7 +/- 11.4%, 35.7 +/- 12.9%, 3.1 +/- 1.6% and 0.12 +/- 0.1% respectively before the induction and cultivation, and changed to 95.4 +/- 3.2% (p < 0.01), 27.0 +/- 13.1% (p > 0.01), 55.5 +/- 13.8% (p < 0.01), 9.8 +/- 6.1% (p < 0.01) and 0.22 +/- 0.18% (p < 0.01) respectively after the induction and cultivation. It is concluded that the major action of this induction and cultivation method on T-lymphocytes in vitro is the promotion of CTL and NK T-cell proliferation. In leukemic patients at the remission stage, the TCRBV profile is characterized by the oligoclonal proliferation of T-lymphocytes. Several proliferated clones may have the same motif in CDR3, suggesting the recognition of the same antigen by these lymphocyte clones. Cytokine induction and co-culture with autogenous DCs can stimulate the T-lymphocytes to recover their immunocompetence as manifested by the polyclonal profile and the proliferation of CTL and NK-T cells.

Role of T-cell receptor V beta 8.3 peptide vaccine in the prevention of experimental autoimmune uveoretinitis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>T-cell receptor (TCR) plays an important role in the development of autoimmune diseases. Recently, it was reported that immunization of animals with TCR peptide derived from the pathogenic cells could prevent autoimmune diseases. The aim of this study was to investigate whether vaccination with a synthetic peptide from the hypervariable region of TCR V(beta) 8.3, an experimental autoimmune uveoretinitis (EAU)-associated gene, was able to prevent the disease.</p><p><b>METHODS</b>EAU was induced in Lewis rats by immunization with IRBP R16 peptide emulsified in complete Freund's adjuvant (CFA). The clinical and histological appearances were scored. Delayed type hypersensitivity (DTH) and lymphocyte proliferation were detected. Cytokine levels of aqueous humour, supernatants of cells from spleen and draining lymph nodes were measured by enzyme linked immunosorbent assay (ELISA). Gene expression of TCR V(beta) 8.3 on CD(4)(+) T cells was examined by real time quantitative polymerase chain reaction (PCR).</p><p><b>RESULTS</b>After vaccination, the intraocular inflammation was significantly mitigated, antigen specific DTH and lymphocyte proliferation responses were suppressed, interleukin (IL)-2 in aqueous humour, interferon (IFN)-gamma and IL-2 produced by the spleen and draining lymph node cells were significantly decreased, whereas the production of IL-4 and IL-10 were increased. The response of draining lymph node cells to TCR V(beta) 8.3 peptide was enhanced after vaccination. Inoculation with CFA alone did not affect the severity of EAU and the above parameters. The suppression of EAU was much stronger in the group of four fold inoculations than the group of two fold inoculations. The expression of TCR V(beta) 8.3 gene was significantly reduced in the group of fourfold inoculations.</p><p><b>CONCLUSION</b>Vaccination with the synthetic TCR V(beta) 8.3 peptide could remarkably inhibit the development of EAU.</p>

Relationship between Staphylococcal superantigens and the dominant expression of T-cell receptor V beta gene in chronic rhinosinusitis with nasal polyp / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To analyse the relationship between superantigens produced by Staphylococcus aureus and the mRNA expression of T-cell receptor V beta region (TCR Vbeta), and to investigate the possible role of Staphylococcal superantigens in the pathogenesis of nasal polyps.</p><p><b>METHODS</b>Sinonasal mucus and polyp/mucosa tissue were obtained from patients with chronic rhinosinusitis (22 patients with bilateral nasal polyps, 15 without nasal polyps) and 12 normal subjects as comparative negative controls. Mucus specimens were assayed by enzyme-linked immunosorbent assay (ELISA) for Staphylococcal exotoxins,and analyzed for the expression of TCR Vbeta genes using the technique of reverse transcriptase-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The percentages of Staphylococcus exotoxins in nasal polyps were 54.54% (21/22) for chronic rhinosinusitis with nasal polyp (CRSwNP) subjects. There were no positive results in the CRSsNP or control groups. The expressional intensity of Vbeta3 (10.02), Vbeta14 (3.54), Vbeta15 (2.39), Vbeta17 (3.48), and Vbeta20 (2.94) was increased significantly for Staphylococcal exotoxin B (SEB) positive subjects (P < 0.05). Vbeta2 (13.8) and Vbeta6. 1-3 (6.53) were significantly highly expressed for toxic shock syndrome toxin-1 (TSTf-1) positive subjects in CRSwNP group (P < 0.05). There were no dominantly used Vbeta fragments in ELISA- negative specimens. In the group of chronic rhinosinusitis without nasal polyp (CRSsNP), most of TCR Vbeta gene subfamilies demonstrated a trend toward higher expressional levels compared with those of normal controls, although there was no statistical difference (P > 0.05).</p><p><b>CONCLUSIONS</b>There was relationship between Staphylococcal superantigens and the excursion of TCR Vbeta gene spectra in nasal polyp, and superantigens possibly play an important role in the pathogenesis of CRSwNP.</p>

Characteristics of T cell receptors recognizing antiphospholipid syndrome associated antigens / 中国实验血液学杂志

To understand the characteristics of T cell receptors recognizing antiphospholipid syndrome associated antigen, the characteristics of T cells were analyzed using T cell receptor beta variable region (TCRbetaV) gene spectrotyping in a case of antiphospholipid syndrome (APS). The results indicated that in the case of APS there were 2 dominant T cell clones. The TCRbetaVs sequences of the 2 T cell clones showed the TCRbetaVs belonged to 8 and 23 gene families respectively. The peptides of third complementarity-determining regions (CDR3) in the TCRbetaVs were CASSLLVAGGPRAYNEQFFGPG and CASSLAGFGQPQHFGDG. Comparing the motifs in CDR3 with another autoimmune disease, the motif YNEQFFGPG in TCRbetaV8 and motif QHFGDG in TCRbetaV23 were identical with that of idiopathic thrombocytopenic purpura and systemic lupus erythematosus reported before. In conclusion, some T cell clones proliferating in these autoimmune diseases may recognize the same antigens.

Complementarity-determining region 3 analysis of T cell receptor beta chain variable region in peripheral blood mononuclear cells of patients with systemic lupus erythematosus / 南方医科大学学报

<p><b>OBJECTIVE</b>To analyze the drift of the complementarity-determining region 3 (CDR3) of T cell receptor (TCR) beta chain variable region (TCR BV) in peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus.</p><p><b>METHODS</b>Immunoscope spectratyping techniques was used to analyze the distribution of TCRbeta chain CDR3 in 5 normal blood donors and the dominant CDR3 in the PBMCs in 5 SLE patients. Sequence analysis of the CDR3 region in monoclonal or oligoclonal T cells was performed.</p><p><b>RESULTS</b>The spectratypes of TCR BV gene CDR3 region showed Gaussian distribution in the 5 normal blood donors. The 5 SLE patients, however, displayed anomalous proliferation and oligoclonal expansion of the T cells was observed in different TCR BV families with different CDR3 sequences.</p><p><b>CONCLUSION</b>Noticeable drift of TCRbeta chain CDR3 can be seen in active SLE, indicating possible association of selective expression of TCR with immune pathogenesis in SLE. Determination of specific TCR CDR3 sequence provides a new means for studying the pathogenesis and personalized treatment of SLE.</p>

The Effects of Intradermal Vaccination with DNA Encoding for the T-cell Receptor on the Induction of Experimental Autoimmune Encephalomyelitis in B10.PL Mice

Intradermal gene administration was found to induce a more profound immune response than direct intramusclular gene injection. We performed intradermal vaccination of B10.PL mice with DNA encoding for the V 8.2 region of the T-cell receptors (TCR). Three weeks later, these mice were immunized with rat myelin basic protein (MBP). Daily mean clinical scores and mortality rate were lower in this group compared with controls. The proliferative responses of lymph node cells to rat MBP were slightly less in the vaccination groups than in the control groups (p<0.05). However, we detected no differences between the two groups with regard to the production of MBP-specific IgG, IgG1, & IgG2a antibodies. The levels of cytokine mRNA expression in the vaccination groups were observed higher than in the control groups without antigen-specific stimulation, but all of cytokine expressions between the vaccination and control groups after antigen-specific stimulation were identical. These results demonstrate that intradermal DNA vaccines encoding for TCR might prove to be useful in the control of autoimmune disease.

Peripheral blood naive T cell level and its T cell receptor Vbeta repertoire usage profile in patients with chronic myelogenous leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze peripheral blood naive T cell level, its T cell receptor (TCR) Vbeta repertoire usage profile and clonality for evaluating the recent thymic output function and the expansion feature of TCR Vbeta subfamily T cells in patients with chronic myelogenous leukemia (CML).</p><p><b>METHODS</b>Quantitative detection of T-cell receptor excision DNA circles (TRECs) in peripheral blood mononuclear cells (PBMNC) from 20 cases of CML was preformed by real-time PCR (TaqMan) analysis, and TRECs-number in T-cells was calculated from peripheral blood CD3-positive cell rate. The expression and clonality analysis were detected by RT-PCR and Genescan technique in PBMNC from 14 out of the 20 patients. Nine normal individuals served as controls.</p><p><b>RESULTS</b>A dramatic reduction of TRECs value in patients with CML was detected as compared with that in normal controls. The mean value of TRECs was 0.06 +/- 0.16 copy/1000 CD3(+) cells in CML patients while 6.84 +/- 4.71 copies/1000 CD3(+) cells in normal controls (P < 0.01). The 1 - 12 Vbeta subfamilies were variably expressed in samples from 14 patients. Genescan analysis identified clonal expanded T cells of some Vbeta subfamily from 13 cases. Vbeta3, Vbeta10, Vbeta19, Vbeta21 and Vbeta22 subfamilies clonal T cells were more frequently seen.</p><p><b>CONCLUSION</b>There is a prominent reduction of recent thymic output naive T cells function in CML. The predominant usage and clonal expansion of TCR Vbeta subfamily T cells could be identified, indicating that CML patients have specific immune response to leukemia associated antigen, in spite of their T cell immunodeficiency.</p>

Análise do perfil do repertório TCR Vβ e da sua natureza clonal em indivíduos infectados pelo HIV-1, antes e durante a terapia anti-retroviral combinada / TCR Vβ profile and T cell clonality evaluation on HIV-1 infected individuals, before and along highly active antiretroviral therapy

A imunopatogenia da infecção pelo HIV é multifatorial e envolve múltiplos eventos virológicos e inerentes ao hospedeiro, e que de um delicado balanço entre eles resulte a severidade dos diferentes distúrbios imunológicos observados e a conseqüente progressão ou não para a aids. Porém, diferenças qualitativasobservadas para a resposta imune de células T entre os diferentes indivíduos, durante a infecção primária, pode ter uma importante participação nesta determinação do curso clínico da doença. A introdução da terapia anti-retroviral combinada (HAART) no tratamento da infecção pelo HIV-1 foi responsável por um intenso controle da replicação viral, seguido pelo dramático aumento das contagens absolutas de células T CD4+ e pela redução das infecções oportunistas, sugerindo um processo de reconstituição imunológica em curso nesses pacientes. Deste modo, o objetivo deste trabalho consiste na avaliação da resposta imune de células T, através da análise do perfil do repertório Vβ em indivíduos cronicamente infectados pelo HIV-1, antes e durante a HAART.Além disto, a natureza clonal das células T envolvidas na resposta também foi analisada, paralelamente a outros parâmetros imunológicos e às propriedades virais. O comprometimento imunológico foi significativamente revertido durante o primeiro ano de tratamento, assim como o fenótipo viral X4 deu lugar ao R5. Com relação ao repertório, uma utilização Vβ não aleatória foi observada para ambos os grupos controles e dos pacientes, tanto para a subpopulação T CD4+ e CD8+ como para os linfócitos não fracionados, e o mesmo perfil oligoclonal de distribiuição do repertório inicialmente observado para os pacientes foi detectado após 24 semanas de terapia anti-retroviral. Por outro lado, outros segmentos Vβ foram mobilizados e uma natureza mais oligoclonal foi observada para as células T envolvidas contra uma identidade monoclonal inicial. Portanto, um padrão de expressão do repertório foi detectado para a...

Clonal expansion of TCR Vbeta subfamily T cells induced by bcr3-abl2 peptide / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the clonal expansion of T cell receptor (TCR) Vbeta subfamily T cells from cord blood induced by bcr3-abl2 peptide in vitro.</p><p><b>METHODS</b>T cells from 3 units of cord blood were amplified by anti-CD(3) monoclonal antibody (McAb) and IL-2 with or without synthetic b3a2 peptide. T cell specific cytotoxicity was analyzed by lactate dehydrogenase (LDH) assay, TCR Vbeta subfamilies by using reverse transcriptase-polymerase chain reaction (RT-PCR) and genescan technique.</p><p><b>RESULTS</b>bcr3-abl2 peptide specific cytotoxicity T cells were successfully induced from the 3 units of cord blood by synthetic b3a2 peptide. Compared with that in CD(3) McAb induced cells, distribution pattern of TCR Vbeta repertoire was different in T cells induced with b3a2 peptide. Oligoclonal and oligoclonal tendency TCR Vbeta subfamily T cells could be identified in cord blood T cells induced by b3a2 peptide in 1 or 2 weeks, whereas those induced by anti-CD(3) McAb and IL-2 were mostly polyclonal.</p><p><b>CONCLUSION</b>The cytotoxicity T cells with anti-CML specificity could be induced by b3a2 peptide. The specific anti-CML cytotoxicity may be derived from the clonal expansion TCR Vbeta subfamily T cells.</p>

The T-lymphocyte clonal proliferation in a patient with chronic eosinophilic leukemia / 中国实验血液学杂志

The gene fingerprinting of T cell receptor beta varial region (TCRbetaV) was used to analyze the T cell clone variations in peripheral blood lymphocytes from a patient with chronic eosinophilic leukemia (CEL) and to find out the characters of CDR3 amino acid sequence related to CEL. RT-PCR was used to amplify betaV1-24 family genes and the gene fingerprinting was set up by denatured polyacrylamide sequence gel electrophoresis. The expanded band was cut and then sequenced. The results showed that one clonal expansion of T cells could be found in TCRbetaV13.2 family. Its nucleotide sequence indicated a monoclonality. The CDR3 amino acid sequence was SFSYEQY and the modif SFSY was special reserved sequence. The other TCRbetaV families remained unchanged. It is concluded that the monoclonal T cells in TCRbetaV13.2 may be related to CEL and it is conjectured that the monoclonal T cells proliferation is induced by CEL malignant cells.

Analysis of selected usage and clonal expansion of TCR Vbeta repertoire of peripheral blood T cells in patients with non-Hodgkin's lymphoma / 中国实验血液学杂志

To investigate the distribution and clonal expansion of TCR Vbeta subfamily T cells in patients with B-NHL and T-NHL, the CDR3 of TCR Vbeta 24 subfamily genes was amplified in peripheral blood mononuclear cells from 4 cases with B-NHL and 2 cases with T-NHL using RT-PCR, and to observe the usage of TCR Vbeta repertoire, the PCR products were further labeled with fluorescence and analyzed by genescan technique for the CDR3 size, to evaluating clonality of the detectable TCR Vbeta T cells. The results indicated that only selected expression of 6-12 Vbeta subfamily T cells could be identified in the 6 cases with NHL, and Vbeta1, Vbeta8, Vbeta13 and Vbeta19 were expressed in all samples, Vbeta2 and Vbeta16 could be found in 5 samples, whereas Vbeta4-6, Vbeta10-12, Vbeta15, Vbeta17-18, Vbeta20 and Vbeta22-23 were absent in all samples. Genescan analysis showed that clonal expansion of T cells could be found in 1-3 Vbeta subfamilies from 2 cases with B-NHL and 1 case with T-NHL. In conclusions, the similar selected usage of TCR Vbeta subfamily T cells could be found in peripheral blood from patients with B and T NHL, clonal expansion of T cells which were considered to be related to lymphoma cell antigen could be detected in a part of patients with B or T NHL.

The distribution feature of TCR Vbeta repertoire in peripheral blood T cells from patients with Ph(+) and Ph(-) CML / 中国实验血液学杂志

To investigate the T cell distribution characters of TCR Vbeta repertoire in Ph(+) and Ph(-) CML. The 24 subfamilies of the TCR Vbeta genes were amplified in peripheral blood T cells from 13 patients with CML (Ph+ b3a2, 5 cases; Ph+ b2a2, 5 cases; Ph(-), 3 cases) by RT-PCR, to analyze the usage of Vbeta subfamilies in different CML patients. The results showed that the expression pattern of Vbeta repertoire was different in normal individuals and in patients with CML which only have part of Vbeta subfamily T cells. 4 - 16 (mean 10.2) Vbeta subfamily T cells were detected in the Ph+ b3a2 CML, 8 - 11 (mean 8.8) Vbeta subfamily T cells in the Ph(+) b2a2 CML and 5 - 6 (mean 5.7) in Ph(-) CML. Moreover, the expression of Vbeta subfamily T cells was different among these three types CML.Vbeta10 and Vbeta16 were detected in the all cases with Ph(+) b3a2 and Ph(+) b2a2 CML, whereas Vbeta9 and Vbeta22 could be found in the most cases with Ph(+) b3a2 CML or Vbeta24 and Vbeta8 in Ph(+) b2a2 CML. In patients with Ph(-) CML, Vbeta24 were detected in all samples, and Vbeta9, Vbeta10, Vbeta13 and Vbeta22 were found in the most cases. The results suggest that skew distribution of TCR Vbeta subfamily T cells was existed in peripheral blood of Ph(+) and Ph(-) CML patients. The selected usage of TCR Vbeta is different in various types of CML patients. It may relate to difference of CML cells associated antigen and individual special immunity reaction.

Clonal expansion T cells identified in acute monoblastic leukemia by CDR3 size analysis of TCR V beta repertoire using RT-PCR and genescan / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the distribution and clonality of T cell receptor (TCR) V beta repertoire in patients with acute monoblastic leukemia (AML-M5).</p><p><b>METHODS</b>Expression of the TCR V beta repertoire was analyzed using reverse transcription-polymerase chain reaction (RT-PCR), which amplified the complementarity determining region 3 of 24 TCR V beta genes in peripheral blood from 9 cases with acute myclogenous leukemia subtype 5 or acute monoblastic leukemia (AML-M5). PCR products were further studied by genescan analysis to identify T cell clonality.</p><p><b>RESULTS</b>Expression of 1-10 V beta subfamilies was found in samples from 9 patients. Genescan analysis showed that some V beta subfamily products from 8 of 9 cases contained an oligoclonal peak. Oligoclonal T cells of the V beta 2 subfamily could be found in 6 patients with AML-M5.</p><p><b>CONCLUSIONS</b>T cell clonality expansion was found in AML-M5 cases and were tendentious in the V beta 2 subfamily, suggesting a the specific immune response for leukemia cell (M5) associated antigen and may display antileukemia activity.</p>

T cell receptor Vbeta gene bias in rheumatoid arthritis / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To explore the pathogenesis of rheumatoid arthritis (RA) by studying the expression of T cell receptors (TCRs).</p><p><b>METHODS</b>T cell receptor Vbeta (TCR Vbeta) gene usage and expression were analyzed from synovial membrane and peripheral blood of 8 RA patients, 2 osteoarthritis patients and 2 accident amputees. The complementary determining region 3 (CDR3) of 25 TCR Vbeta subfamily genes in unselected T cell populations were amplified semi-quantitatively by reverse transcription-polymerase chain reaction (RT-PCR). The products were further studied by genescan for frequency of Vbeta usage.</p><p><b>RESULTS</b>The numbers of Vbeta subfamilies expressed by T cells from RA peripheral blood and synovial membrane were not significantly restricted. More importantly, biased Vbeta gene expression in RA synovium was observed and Vbeta6, Vbeta17, and Vbeta22 genes were the predominant subfamilies. It was noteworthy that the expression of Vbeta17 in RA synovium was significantly increased.</p><p><b>CONCLUSION</b>Our data were consistent with the hypothesis that several antigen or superantigen-driven processes may be involved in the pathogenesis of RA.</p>

Effect of hejie decoction on T-cell receptor V beta 7 gene expression in patients of chronic hepatitis B / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the therapeutic effect of Hejie Decoction (HJD) in treating chronic hepatitis B and its relationship with T-cell receptor V beta 7 (TCRV beta 7) gene expression.</p><p><b>METHODS</b>Forty-five patients of chronic hepatitis B were randomly divided into two groups. The 30 patients in the treated group were treated by HJD, and the 15 patients in the control group were treated by conventional western medicine. The therapeutic effect and changes of TCRV beta 7 gene expression after treatment were observed.</p><p><b>RESULTS</b>After 6 months treatment, the ALT level in the two groups were obviously decreased (P < 0.01). No significant difference was shown in comparison of the total effective rate between the two groups but it did show in comparison of markedly effective rate between them. TCRV beta 7 expression was detected in 5 patients of the treated group, and HBV-DNA and HBeAg in the 5 patients were all negatively converted. While in the control group, no one had TCRV beta 7 expression detected, either no one with negative conversion of HBV-DNA and HBeAg. TCRV beta 7 could not be detected in all the patients whose HBV-DNA and HBeAg hasn't negatively converted, though their liver function could be normalized.</p><p><b>CONCLUSION</b>HJD might have the effect of regulation on TCRV beta 7 expression, it possibly is the important way for HBV replication inhibition and virucidal action of HJD.</p>

Study on clonal proliferation of TCR Vbeta subfamily T cells induced by AML-M(2a) cells in vivo and in vitro / 中国实验血液学杂志

To observe the expression and clonal expansion of TCR Vbeta subfamily T cells induced by AML-M(2a) cells in vivo and in vitro, complementary determining region 3 (CDR3) of TCR beta with variable region genes was amplified by using RT-PCR in both peripheral blood mononuclear cells (PBMNC) and T cells from mixed lymphocyte and tumor culture (MLTC) from four AML-M(2a) patients. The positive products were further analyzed to identify the clonality of T cells by genescan. The results showed that the similarity distribution of TCR Vbeta subfamily T cells was found Vbeta in PBMNC and MLTC. One or two clonality expansions of T cells could be found in predominant TCR Vbeta subfamily T cells induced by A ML-M2a cells from 3 cases in vivo and in vitro. It was concluded that clonal expansion of TCR Vbeta subfamily T cells stimulated selectively by AML-M(2a) cells may be a specific immune response of patient's T cells to AML-M(2a) cells associated antigen. The clonal proliferation of TCR Vbeta subfamily T cells were affected somewhat by environmental difference in vivo and in vitro.