Expression of A20 in patients with systemic lupus erythematosus / 中国病理生理杂志

[ ABSTRACT] AIM:To investigate the expression and regulation of A20 in healthy individuals and the patients with systemic lupus erythematosus (SLE).METHODS:The expression levels of A20, NF-κB, MALT1, and MALT1V1 in peripheral blood mononuclear cells ( PBMC) of the patients with SLE ( including 2 cases with scleroderma, 1 case with rheumatoid arthritis, and 1 case with lymphoma) were analyzed by real-time PCR.RESULTS:A significantly lower A20 expression level was found in the PBMC from SLE group compared with the healthy controls, while the expression levels of MALT1 and NF-κB were also decreased.In addition, no significant correlation between A20 and NF-κB expression levels in healthy group was observed, but a positive correlation was found in SLE group ( P<0.05) .A significant positive corre-lation between MALT1 and NF-κB expression levels in healthy group ( P<0.05) was observed, and no significant correla-tion was found in SLE group.The expression level of MALT1V1 in SLE group was significantly lower than that in healthy control group, and there was a positive correlation between A20 and MALT1V1 in healthy volunteers (P<0.01), but that did not exist in SLE group.CONCLUSION: The characteristics of the expression pattern of MALT1-A20-NF-κB in the SLE patients were presented.Lower level of A20 expression was found in the SLE patients, in particular with other autoim-mune disease or lymphomas, indicating the lower immune tolerance in SLE.The positive correlation of A20 and NF-κB may relate to positive regulation of MALT1.

Study on the distribution and clonal expansion of TCR Vβ subfamily in peripheral T cells after infusing mesenchymal stem cells in patients with chronic GVHD / 白血病·淋巴瘤

Objective To investigate the distribution of TCR Vβ genealogy and clonal expansion in peripheral blood after infusing mesenchymal stem cells (MSC) in patients with chronic GVHD. Methods The complementarity determining region 3 (CDR3) of 24 TCR Vβ subfamily genes in peripheral blood mononuclear cell from 1 case with cGVHD after allogeneic hematopoietic stem cell transplantation (Allo-HSCT),who were treated with infusing MSC,were amplified using RT-PCR. The blood samples were taken at the first and the fifth day after 1st infusion; and the first day,the 10 th day and the 20 th day after the second infusion of MSC,as well as the MSC infused as control . The products were labelled by fluorescein and then analyzed the CDR3 size with gene scan technique to determine the clonality of T cells. Results There were no expression of TCR Vβ subfamily with the MSC infused and after the 1st day of the first infusion of MSC. Then 3,10,14,10 Vβ subfamilies clones are appeared at the other time points,of which were polyclone and oligoclone predominately. In the same time,the manifestations of cCVHD have been abated. Conclusion MSC played a certain role in reviving the immune function of the patients after Allo-HSCT and mitigating the disease of chronic GVHD. Lineage analysis of TCR Vβ subfamily showed some predominant expression.

Experimental study of prevention effect of andrographolide against radiation exposure in rats / 中华放射医学与防护杂志

Objective To explore the effects of andrographolide(AP),extracted from the traditional Chinese herb Andrographlis paniculata(AP),on injury induced by radiation exposure.Methods Sixty male rats were randomly divided into 4 equal groups and irradiated with 60Co γ-rays at the doses of 1,2,and 4 Gy,respectively:low dose AP group(intragnstrically administered with AP at the dose of 100 ms/kg daily for 10 d before irradiation),and high dose AP group(intragastrically administered with AP at the dose of 200 ms/kg daily for 10 d before irradiation),model group(administered with the same volume of normal saline instead of AP for 10 d before irradiation),and control group(irradiated only at 3 different doses).One day after irradiation all rats were killed with their livers being fixed to make paraffin section.The morphological feature was observed under light microscope after HE staining,and the cell apoptosis was detected by TUNEL technology.Results Compared to the control and model groups,the pathological changes of liver were significantly gentler in the AP treatment groups.The apoptosis rates of the liver cells of all the AP sub-groups were significantly lower than those of the control and model subgroup(t=2.19-4.80.P<0.05).Conclusions AP might have prevention effect against radiation exposure.

Expression of T cell-specific transcriptional factor Elf-1 in mononu-clear cells, CD4~+ T cells and CD8~+ T cells from umbilical cord blood / 中国病理生理杂志

AIM: To investigate the expression level of transcriptional factor Elf-1 in mononuclear cells, CD4~+ and CD8~+ T cells from umbilical cord blood (UCB). METHODS: Real-time PCR with SYBR green I technique was used for detecting the Elf-1 expression in mononuclear cells, sorted CD4~+ and CD8~+ T cells from 12 cases of umbilical cord blood. The relative mRNA expression level of Elf-1 was analyzed by a formula of 2~(-△Ct)×100%. The expression level of β_2-microglobulin gene (β_2M) was used as an endogenous reference. The peripheral blood from 10 cases of healthy adults was served as control. RESULTS: Elf-1 mRNA expressed in all blood samples collected from both UCB and healthy adults. The expression level showed apparent diversity in different individuals. The relative mRNA expression of Elf-1 in both mononuclear cells (18.55%±2.48%) and CD8~+ T cells (3.52%±0.45%) from UCB were significantly higher than those from healthy adults (9.16%±1.92%, 2.02%±0.27%, respectively, P<0.01, P<0.05). CONCLUSION: The results of Elf-1 expression level from umbilical cord blood indicate that the over-expression of Elf-1 gene in mononuclear cells and CD8~+ T cells might be one of the features of T cell immune state in umbilical cord blood.

The feature of TCR ζ chain gene expression in peripheral blood T cells from patients with multiple myeloma / 白血病·淋巴瘤

Objective To investigate T-cell receptor(TCR)ζchain gene expression level in peripheral blood T cells from patients with multiple myeloma(MM),thereby to estimate the feature of T cells activation status.Methods Real-time PCR with SYBR Green I technique Was used for detecting TCR ζchain expression level in peripheral blood mononuelear cells(PBMC)of 24 cages with MM and 24 normal individuals.β2-microglobulin(β2M)gene expression was used as an endogenous reference.Relative changes in TCRζchain expression level were analyzed by the 2-△α×100%method between patients with MM and normal individuals.Results Compared with normal individuals,TCRζchain gene expression was obviously down regulated in PBMC from patients with MM(P=0.019).The expression level of TCR ζchain gene is not significantly age-associated in MM patients[(1.83±1.72)%,(3.46±2.75)%](P=0.525).Conclusion This is the first description in the expression feature of TCRζchain gene in MM patients.TCRζchain expression Was decreased in most of MM patients,which might be related to celhar immunodeficiency.

Effect of cimetidine on clonal expansion of TCR V? subfamily of T cells in cord blood stimulated by K562 cells / 第三军医大学学报

Objective To investigate the effect of cimetidine on the clonal expansion of TCR V? subfamily of T cells in cord blood after the stimulation by K562 cells in vitro.Methods Cimetidine(1?10-5 mol/L) or K562 cells(1?106/ml) or both of them were respectively cultured with mononuclear cells(MNC) isolated from normal human cord blood for 2 weeks.After the induction,specific cytotoxicity of the proliferated T cells were detected with K562 cells as the target cells.The selective usages and clonal expansion of TCR V? subfa-mily of T cells were analyzed by RT-PCR and genescan technique.Results After induction for 2 weeks,the 3 groups showed the increased cell proliferation,in which specific cytotoxicity of T cells induced by both cimetidine and K562 cells against K562 cells was enhanced significantly compared with the other 2 groups(P

Changes in pattern of TCR V? repertoire in peripheral blood from CML patients / 第三军医大学学报

Objective To investigate the distribution and clonality of TCR V? repertoire T cells in peripheral blood mononuclear cells with chronic myeloid leukemia(CML)at chronic phase(CP)and complete remission(CR).Methods The CDR3 of TCR V? 8 subfamily genes were amplified in mononuclear cells of peripheral blood from CML patients in CP(8 cases)and CR(8 cases)phases for observation of the usage of TCR V? repertoire by RT-PCR.The positive PCR products were further labeled with fluorescence and analyzed by genescan technique for the CDR3 size for evaluation of the clonality of the detectable TCR V? T cells.A total of 10 cases of healthy adults served as controls.Results The mean values of detectable TCR V? subfamilies were(3.25?0.89)in patients with CML-CP,predominantly in V?1,2 and V?3,whereas(3.5?0.76)of 8 V? subfamily T cells were expressed in patients with CML-CR,predominantly in V?1,2,3 and V?8.There was no significant difference between the above groups and the healthy adults(3.5?0.52).In addition,a higher frequency expression of V?8 could be found from peripheral blood in CR phase(5/8)rather than from that in CP phase(2/8)and from that of the healthy adults(3/10).Genescan analysis showed that the majority of PCR products from both CML and healthy adults displayed clonal expansion,predominantly in V?1,V?2 and V?3.Clonal expanded V?7 subfamily T cells,however,could be identified from two patients in CR phase but not in all of 10 healthy adults.Conclusion The similar distribution and clonal pattern of TCR V? repertoire T cells can be found in peripheral blood from the three groups,suggesting that the cellular immunosuppression of CML patients might not be mainly involved in the alteration of TCR V? repertoire.In CML-CR groups,the expression and clonal pattern of some V? repertoires are different from those in other groups,which needs further studies.

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>

Analysis of T cell clonality by using T-cell receptor varible β gene repertoire in T-cell acute lymphoblastic leukemia / 中国病理生理杂志

AIM: To analyze T cell clonality in patients with T cell acute lymphoblastic leukemia (T-ALL). METHODS: The complementarity determining region 3 (CDR3) size of 24 T cell antigen receptor variable β (TCR Vβ) region gene was analyzed in peripheral blood mononuclear cell (PBMC) samples from 6 T-ALL cases and 10 normal individuals by using reverse transcriptase-polymerase chain reaction (RT-PCR). The PCR products were further studied by genescan and sequencing analysis. RESULTS: Some TCR Vβ subfamily T cells display mono- or oligoclonal expansions in 3 cases of T-ALL, predominantly in Vβ2, Vβ3, Vβ6, Vβ9, Vβ21 or Vβ24, respectively. Polyclonal expansions of T cells were found in the other three cases, which could also be found in normal samples. CONCLUSION: A part of T cell acute lymphoblastic leukemia cells may arise from a clonal expansion of TCR Vβ subfamily T cell. This method may be useful for the detection of minimal residual disease in clinical study of the disease.

Analysis of T cell clonality by using T-cell receptor varible ? gene repertoire in T-cell acute lymphoblastic leukemia / 中国病理生理杂志

AIM: To analyze T cell clonality in patients with T cell acute lymphoblastic leukemia (T-ALL). METHODS: The complementarity determining region 3 (CDR3) size of 24 T cell antigen receptor variable ? (TCR V?) region gene was analyzed in peripheral blood mononuclear cell (PBMC) samples from 6 T-ALL cases and 10 normal individuals by using reverse transcriptase-polymerase chain reaction (RT-PCR). The PCR products were further studied by genescan and sequencing analysis. RESULTS: Some TCR V? subfamily T cells display mono- or oligoclonal expansions in 3 cases of T-ALL, predominantly in V?2, V?3, V?6, V?9, V?21 or V?24, respectively. Polyclonal expansions of T cells were found in the other three cases, which could also be found in normal samples. CONCLUSION: A part of T cell acute lymphoblastic leukemia cells may arise from a clonal expansion of TCR V? subfamily T cell. This method may be useful for the detection of minimal residual disease in clinical study of the disease.

Clonal expansion and specific cytotoxicity of autologous or allogeneic T cells induced by M_ 2a cells / 中国病理生理杂志

AIM: To investigate clonal expansion and specific cytotoxicity of TCR V? subfamily T cells from acute myelogenous leukemia M_ 2a subtype (AML-M_ 2a)patients and normal individuals induced by AML-M_ 2a cells, respectively. METHODS: Complementarity determining region 3(CDR3) of TCR ? with variable region genes was amplified in autologous or allogeneic T cells from mixed lymphocyte and tumor culture (MLTC) using RT-PCR. The positive products were further analyzed to identify the clonality of T cells by genescan. The specific cytotoxicity of T cells was analyzed by MTT. RESULTS: T cells from both M_ 2a patients and normal individuals after MLTC showed high response to M_ 2a cells with 4-17 TCR V? subfamily dominant utilization, one or two clonal expansion of T cells were identified in some predominant TCR V? subfamilies. Difference of distribution and clonal expansion of TCR V? subfamily T cells were related to source of T cells and the phase during MLTC. Compared with LAK cell, most of T cells from MLTC were CD3+CD8+T cells with higher and more specific cytoxicity to the induced cells, M_ 2a cells, but not HL60 or K562 cell line. CONCLUSION: Clonal expansion of TCR V? subfamily T cells stimulated selectively by M_ 2a cells may be a specific immune response of autologous and allogeneic T cells to M_ 2a cells associated antigen. The T cells induced by M_ 2a cells have the ability of specific cytoxicity to the AML-M_ 2a cells.

The feature of clonal expansion of TCR V? T cells from peripheral blood in patients with acute monoblastic leukemia / 中国病理生理杂志

AIM: To investigate the distribution and clonal expansion of TCR V? subfamily T cells in patients with acute monoblastic leukemia (M5). METHODS:The CDR3 of TCR V? 24 subfamily genes were amplified in peripheral blood mononuclear cells from 9 cases with M5 using RT-PCR and the PCR products were further labeled with fluorescent and analyzed by genescan technique for the CDR3 size, to evaluate clonality of the detectable TCR V? T cells. RESULTS:Only 1-10 V? subfamily T cells were identified in M5 cases. Genescan analysis showed that oligoclonal (clonal expansion) T cells were found in some V? subfamilies from 8 cases with M5, V? 2 oligoclonal T cells were identified in six cases, whereas V? 7- or V? 9 clonal expansion T cells were detected in the other two cases, respectively. In addition, except V? 2, V? 7 or V? 21 oligoclonal T cells could be detected in two cases, respectively. CONCLUSION:The skew distribution and clonal expansion of TCR V? subfamily T cells could be found in patients with M5. It may be a specific anti-leukemia immune response with which the host T cells were activated by the leukemia-associated-antigen. The clonal expansion T cells were tendentious in V? 2, which may be related to the M5 leukemia cells associated antigen.

The Insertional Mutation of Transcription Factor GATA-2 Gene in Acute Non-Lymphocytic Leukemia / 中国实验血液学杂志

To investigate the expression and mutation of transcription factor GATA-2 gene in acute non-lymphocytic leukemia (ANLL), the GATA-2 gene were amplified in peripheral blood mononuclear cells from 85 cases with ANLL using RT-PCR, and the PCR products were further analyzed by single strand conformation polymorphism (SSCP) and sequencing. The results showed that GATA-2 gene was expressed in the most cases with ANLL (89.4%). One larger band was observed by RT-PCR in a case with M(1) subtype. Nucleotide sequence analysis revealed a insertion of 18 nucleotides at position 1 288 bp of the GATA-2 gene cDNA, thereby leading to 6 additional amino acids in second zinc finger complex of GATA-2, and it includes a cysteine which can bind to zinc ion. The insertional mutation may change the function of GATA-2 transcription factor. To our knowledge this is the first description of insertional mutation in the GATA-2 gene.

Detection of Human Herpesvirus-6 DNA in Peripheral Blood Cells of Patients with Myelogenous Leukemia by Nested PCR / 中国实验血液学杂志

Human herpesvirus-6 (HHV-6) is largly disseminated in human population, which may relate to many diseases. Due to different methods previously used, the results of HHV-6 DNA detections in patients with leukemia are not identical. With a control-study, HHV-6 DNA in the blood cells from 38 cases, including 30 cases with acute myeloid leukemia and 8 cases with chronic myelogenous leukemia was detected by nested PCR. The results showed that HHV-6 DNA positive rate (63%) in patients with AML was lower than that in controls, whereas no significant difference between patients with CML (75%) and controls (92%) was shown. The clinical importance of HHV-6 infections in myelogenous leukemia should continue to be investigated later.

Expression of the Cholinesterase-Related Cell Division Controller Gene in Peripheral Blood Cell from Patients with Myelodysplastic Syndrome / 中国实验血液学杂志

To investigate the expression of the cholinesterase-related cell division controller (CHED) gene in the patients with myelodysplastic syndrome (MDS), CHED gene expression was assayed by RT-PCR and its relative expression rate (RER) was determined by the semi-quantitative RT-PCR analysis in peripheral blood mononuclear cells from 21 patients with MDS, 12 normal individuals served as controls. Results showed that RER of CHED in the patients (2.69 +/- 0.76) was significantly higher than that in controls (1.12 +/- 0.51, P < 0.01), the RER out of 85.7% of the patients was higher than the mean value of the controls, in which three patients developed into acute leukemia; the RER out of 61.9% of the patients was higher than the upper limit of the mean value of the controls; three patients whose RER was lower than the mean value of the controls did not developed into leukemia. These findings suggested that the expression of CHED gene in patients with MDS was significantly higher than in controls.

The Feature of Skewing Distribution of TCR Vbeta T Cells from Peripheral Blood and Bone Marrow in Patients with AML-M(5) / 中国实验血液学杂志

The TCR Vbeta 24 subfamily genes were amplified in peripheral blood and bone marrow mononuclear cells from 5 cases with acute monocytic leukemia (AML-M(5)) using RT-PCR, to observe the distribution of TCR Vbeta subfamilies. The results indicated that 1 - 19 Vbeta subfamily T cells could be identified in different samples from AML-M(5) cases. The variation of distribution of TCR Vbeta subfamily T cells could be found in different individual samples. The results provided the feature of cell immune function change in skewed distribution of TCR Vbeta subfamily T cells from peripheral blood and bone marrow of patients with AML-M(5).

The thymic recent output function in patients with acute promyelocytic leukemia / 肿瘤研究与临床

Objective To analyze the T- cell receptor excision DNA circles (TRECs) level and evaluate the thymic recent output nave T cells function in patients with acute promyelocytic leukemia (APL). Methods Quantitative detection of TRECs in DNA of peripheral blood mononuclear cells (PBMC) from 9 cases with APL were preformed by real- time PCR (TaqMan) analysis. The TRECs- number was related to the number of T- cells by determination of the number of CD-3 positive cells. 14 normal individuals served as controls. Results In comparison with normal individuals [ (4.10?3.65) copies/1000 PBMC and (6.36?5.28) copies/1000 CD+3 cells], a dramatic reduction of TRECs values in patients with APL [ (0.13?0.22) copies/1000 PBMC and (0.77?1.60) copies/1000 CD+3 cells] could be found (P =0.0004, P =0.0005). Conclusions This is, to our knowledge, the first description of TRECs level which was markedly reduced in APL patients.

A preliminary study of the mechanism and therapeutic effect in four types of immunic diseases with plasmapheresis / 中国病理生理杂志

This paper presents plasma exchange(P.E) therapy in 8 cases of four types of immunic diseaes. The colony forming unit-T lymphocyte (TL-CFU), the symbol T-cell subsets with monoclone antibody (McAb), and the cisrculating immune complex (CIC)were examined before and after the therapy of P.E. and compared with the interelationship of each other. The experiments indicated that plasmapheresis may remove the CIC and noxious autoantibodies and regulation of the action of cellular immunity. All these functions are synthetic therapeutic effects. The indices of checkup on therapeutics were: TL-CFU, CIC and the ratio of T_H and T_S. After plasmaphersis, the ratio of CIC was lowered but the TL was higer than before (P

The feature of TCR-zeta chain expression in patients with CML by real-time quantitative PCR / 中国病理生理杂志

AIM:To establish a real-time PCR technique for detection and quantification of TCR ? chain expression and to investigate TCR ? chain expression level in patients with chronic myeloid leukemia(CML).METHODS:Real-time PCR with SYBR GreenⅠ technology was used for detecting TCR ? chain expression level in peripheral blood mononuclear cells from 30 patients with CML and 30 normal individuals.?2-microglobulin gene(?2M) was used as an endogenous reference.Relative changes in TCR ? chain expression level were used by the 2-Ct method between patients with CML and normal individuals.RESULTS:The SYBR GreenⅠ real-time technique for quantitative detection of TCR ? chain expression levels was established successfully.The expression level of TCR ? chain in 18 patients with CML was reduced.However,the TCR ? chain expressed increased in 12 patients with CML.CONCLUSION:The TCR ? chain expression level is divided into down expression(60%) and over expression(40%) groups,and the down expression of TCR ? chain might related to cellular immunodeficiency in most of CML patients.

Changes in TCR V? subfamily nave T cells in peripheral blood of patients with multiple myeloma / 中国病理生理杂志

AIM:To detect the existence of signal joint T-cell receptor excision DNA circles(sjTRECs)of 23 TCR V? subfamilies in mononuclear cells of patients with multiple myeloma(MM),and to evaluate the recent thymic emigrants of corresponding V? subfamily nave T cells in MM patients.METHODS:23 TCR V? subfamily sjTRECs were amplified in genomic DNA from 5?104 PBMCs of 12 cases in MM patients by using semi-nest PCR.10 normal individuals served as controls.RESULTS:The number of detectable V? subfamily sjTRECs was 5.00?2.45 from MM patients,as compared with 9.60?5.48 from normal individuals,the difference was significant(P