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Objective@#To investigate the expressions of CD4+CD45RA+ T cells and CD4+CD45RO+ T cells in peripheral blood of patients with acute coronary syndrome (ACS) and their significance.@*Methods@#A case-control study was conducted. Ninety-four patients receiving coronary angiography (CAG) admitted to Tianjin Chest Hospital from March 5th to April 27th in 2018 were enrolled. They were divided into non-coronary heart disease (CHD) group (n = 12), unstable angina pectoris (UAP) group (n = 27), acute non-ST elevation myocardial infarction (NSTEMI) group (n = 27) and acute ST elevation myocardial infarction (STEMI) group (n = 28) according to the patients' symptoms, electrocardiogram, troponin test and angiographic results. General data, blood routine parameters, and biochemical indicators were collected. The ratios of CD4+CD45RA+ T cells and CD4+CD45RO+ T cells were determined by flow cytometry. Multivariate Logistic regression was used to evaluate whether CD4+CD45RA+ T cells and CD4+CD45RO+ T cells were associated with STEMI.@*Results@#Ninety-four patients were included initially. After excluding the patients who died during the intervention, 93 patients were enrolled in the data analysis finally, with 12 patients in the non-CHD group, 27 patients in the UAP group, and the same as the NSTEMI group and the STEMI group. Compared with the non-CHD group, white blood cell count (WBC) was decreased (×109/L: 6.03±1.30 vs. 6.60±1.30, P > 0.05), and lymphocyte ratio was increased (0.273±0.059 vs. 0.269±0.070, P > 0.05) in patients of the UAP group; however, in the NSTEMI group and STEMI group, WBC was increased (×109/L: 8.29±2.28, 9.86±2.76 vs. 6.60±1.30, both P < 0.05), and lymphocyte ratio was decreased (0.236±0.076, 0.173±0.094 vs. 0.269±0.070, P > 0.05 and P < 0.05), especially in the STEMI group [WBC (×109/L): 9.86±2.76 vs. 6.60±1.30, lymphocyte ratio: 0.173±0.094 vs. 0.269±0.070, both P < 0.05]. There was no significant difference in biochemical indicators among all of the groups. Flow cytometry results showed that the ratios of CD4+CD45RO+ T cells in the UAP group and NSTEMI group were higher than those in the non-CHD group (0.323±0.074, 0.319±0.078 vs. 0.314±0.058, both P > 0.05); however, the ratio of CD4+CD45RO+ T cells in the STEMI group showed a decreased tendency (0.270±0.057 vs. 0.314±0.058, P > 0.05), and it was significantly lower than that in the UAP group and the NSTEMI group (0.270±0.057 vs. 0.323±0.074, 0.319±0.078, both P < 0.05). There was no significant difference in the ratio of CD4+CD45RA+ T cells among all of the groups. Multivariate Logistic regression analysis showed that CD4+CD45RA+ T cells ratio was not significantly correlated with the occurrence of STEMI [odds ratio (OR) = 0.976, 95% confidence interval (95%CI) was 0.907-1.050, P = 0.518], but CD4+CD45RO+ T cells ratio was significantly correlated with the occurrence of STEMI (OR = 0.888, 95%CI was 0.821-0.961, P = 0.003).@*Conclusions@#There was no significant difference in the ratio of CD4+CD45RA+ T cells among UAP, NSTEMI and STEMI patients, and CD4+CD45RO+ T cells ratio in the STEMI group was significantly lower than that in the UAP group and NSTEMI group. CD4+CD45RO+ T cells ratio may be risk factor of STEMI.
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Objective To investigate the expressions of CD4+CD45RA+ T cells and CD4+CD45RO+ T cells in peripheral blood of patients with acute coronary syndrome (ACS) and their significance. Methods A case-control study was conducted. Ninety-four patients receiving coronary angiography (CAG) admitted to Tianjin Chest Hospital from March 5th to April 27th in 2018 were enrolled. They were divided into non-coronary heart disease (CHD) group (n = 12), unstable angina pectoris (UAP) group (n = 27), acute non-ST elevation myocardial infarction (NSTEMI) group (n = 27) and acute ST elevation myocardial infarction (STEMI) group (n = 28) according to the patients' symptoms, electrocardiogram, troponin test and angiographic results. General data, blood routine parameters, and biochemical indicators were collected. The ratios of CD4+CD45RA+ T cells and CD4+CD45RO+ T cells were determined by flow cytometry. Multivariate Logistic regression was used to evaluate whether CD4+CD45RA+ T cells and CD4+CD45RO+ T cells were associated with STEMI. Results Ninety-four patients were included initially. After excluding the patients who died during the intervention, 93 patients were enrolled in the data analysis finally, with 12 patients in the non-CHD group, 27 patients in the UAP group, and the same as the NSTEMI group and the STEMI group. Compared with the non-CHD group, white blood cell count (WBC) was decreased (×109/L: 6.03±1.30 vs. 6.60±1.30, P > 0.05), and lymphocyte ratio was increased (0.273±0.059 vs. 0.269±0.070, P > 0.05) in patients of the UAP group; however, in the NSTEMI group and STEMI group, WBC was increased (×109/L: 8.29±2.28, 9.86±2.76 vs. 6.60±1.30, both P < 0.05), and lymphocyte ratio was decreased (0.236±0.076, 0.173±0.094 vs. 0.269±0.070, P > 0.05 and P < 0.05), especially in the STEMI group [WBC (×109/L): 9.86±2.76 vs. 6.60±1.30, lymphocyte ratio: 0.173±0.094 vs. 0.269±0.070, both P < 0.05]. There was no significant difference in biochemical indicators among all of the groups. Flow cytometry results showed that the ratios of CD4+CD45RO+ T cells in the UAP group and NSTEMI group were higher than those in the non-CHD group (0.323±0.074, 0.319±0.078 vs. 0.314±0.058, both P > 0.05); however, the ratio of CD4+CD45RO+ T cells in the STEMI group showed a decreased tendency (0.270±0.057 vs. 0.314±0.058, P > 0.05), and it was significantly lower than that in the UAP group and the NSTEMI group (0.270±0.057 vs. 0.323±0.074, 0.319±0.078, both P < 0.05). There was no significant difference in the ratio of CD4+CD45RA+ T cells among all of the groups. Multivariate Logistic regression analysis showed that CD4+CD45RA+ T cells ratio was not significantly correlated with the occurrence of STEMI [odds ratio (OR) = 0.976, 95% confidence interval (95%CI) was 0.907-1.050, P = 0.518], but CD4+CD45RO+ T cells ratio was significantly correlated with the occurrence of STEMI (OR = 0.888, 95%CI was 0.821-0.961, P = 0.003). Conclusions There was no significant difference in the ratio of CD4+CD45RA+ T cells among UAP, NSTEMI and STEMI patients, and CD4+CD45RO+ T cells ratio in the STEMI group was significantly lower than that in the UAP group and NSTEMI group. CD4+CD45RO+ T cells ratio may be risk factor of STEMI.
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Objective: To study the expression of transcription factor BTB and CNC homology 2 (Bach2) in CD4+CD25+CD45RA- T cells from patients with systemic lupus erythematosus (SLE) and its effect on cell function. Methods: The CD4+CD25+CD45RA- T cells from active SLE patients and healthy volunteers were sorted by flow cytometry. The expression of Bach2 in CD4+CD25+CD45RA- T cells was detected by fluorescence quantitative PCR and Western blotting. The correlation between the median flourscence indensity (MFI) of Bach2 in CD4+CD25+CD45RA- T cells and the disease activity index of SLE (SLEDAI) was analyzed. The MFI of Bach2 in IL-17+CD4+CD25+CD45RA- T cells was compared with that in IL-17-CD4+CD25+CD45RA- T cells by flow cytometry. In Bach2 overexpression system, the expression of IL-17 in CD4+CD25+CD45RA- T cells was detected by flow cytometry and the concentration of IL-17 in the culture supernants was detected by ELISA. Results: The mRNA and protein expressions of Bach2 in CD4+CD25+CD45RA- T cells from SLE patients were significantly lower than those in healthy controls (P<0.01). There was a significant negative correlation between the MFI of Bach2 and SLEDAI (R2=0.433, P=0.001) in patients with SLE. The expression of Bach2 in IL-17+CD4+CD25+CD45RA- T cells was significantly lower than that in IL-17-CD4+CD25+CD45RA- T cells (P=0.013). When Bach2 was overexpressed, the percentage of CD4+CD25+CD45RA- T cells from SLE patients expressing inflammatory factor IL-17 decreased significantly (P=0.032) and the IL-17 concentration in cell culture supernatants markedly decreased (P=0.008). Conclusion: The expression of Bach2 in CD4+CD25+CD45RA- T cells from SLE patients decreases, and overexpression of Bach2 in the cells leads to the falling expression of IL-17.
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Objective·To study the expression of transcription factor BTB and CNC homology 2 (Bach2) in CD4+CD25+CD45RA-T ceils from patients with systemic lupus erythematosus (SLE) and its effect on cell function.Methods·The CD4+CD25+CD45RA T cells from active SLE patients and healthy volunteers were sorted by flow cytometry.The expression of Bach2 in CD4+CD25+CD45RA T cells was detected by fluorescence quantitative PCR and Western blotting.The correlation between the median flourscence indensity (MFI) of Bach2 in CD4+CD25+CD45RA-T cells and the disease activity index of SLE (SLEDAI) was analyzed.The MFI of Bach2 in IL-17+CD4+CD25+CD45RA-T ceils was compared with that in IL-17-CD4+CD25+CD45RA-T cells by flow cytometry.In Bach2 overexpression system,the expression of IL-17 in CD4+CD25+CD45RA T ceils was detected by flow cytometry and the concentration of IL-17 in the culture supernants was detected by ELISA.Results·The mRNA and protein expressions of Bach2 in CD4+CD25+CD45RA-T cells from SLE patients were significantly lower than those in healthy controls (P＜0.01).There was a significant negative correlation between the MFI of Bach2 and SLEDAI (R2=0.433,P=-0.001) in patients with SLE.The expression of Bach2 in IL-17+CD4+CD25+CD45RA-T cells was significantly lower than that in IL-17-CD4+CD25+CD45RA-T cells (P=-0.013).When Bach2 was overexpressed,the percentage of CD4+CD25+CD45RA-T cells from SLE patients expressing inflammatory factor IL-17 decreased significantly (P=0.032) and the IL-17 concentration in cell culture supernatants markedly decreased (P=0.008).Conclusion·The expression of Bach2 in CD4+CD25+CD45RA-T cells from SLE patients decreases,and overexpression of Bach2 in the cells leads to the falling expression of IL-17.
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To characterize the CD45RA+and CD45RO+T lymphocyte subsets in the peripheral blood of patients with cGVHD induced by allogeneic haematopoietic stem cell transplantation ( allo-HSCT ) and to explore their relations with the disease.Methods:The peripheral blood was collected from 64 patients after allo-HSCT,including 21 non-cGVHD patients,15 light grade cGVHD patients,18 mild grade cGVHD patients and 10 severe grade cGVHD patients,then CD4+CD45RA+,CD4+CD45RO+, CD8+CD45RA+and CD8+CD45RO+T lymphocyte subsets were detected by flow cytometry ( FCM).Results: Compared with the control,the percent of CD4+CD45RA+T lymphocyte in patients with light,mild and severe grade cGVHD decreased markedly (P<0.05),the percent of CD4+CD45RO+T lymphocyte increased markedly (P<0.05).But there were not obviously change in the patient with different grade cGVHD.The percent of CD8+CD45RA+,CD8+CD45RO+T lymphocyte did not change obviously.Conclusion:CD4+CD45RA+and CD4+CD45RO+may play an important role in the pathogenesis of cGVHD.
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BACKGROUND: The leukocyte common antigen (CD45) is a transmembrane-type protein tyrosine phosphatase that has five isoforms. METHODS: We generated seven murine mAbs against human CD45 by injecting cells from different origins, such as human thymocytes, PBMCs, and leukemic cell lines. By using various immunological methods including flow cytometry, immunohistochemistry, and immunoprecipitation, we evaluated the reactivity of those mAbs to CD45 of thymus as well as tonsil lysates. Furthermore, we transiently transfected COS-7 cells with each of gene constructs that express five human CD45 isoforms respectively, and examined the specificities of the mAbs against the transfected isoforms. RESULTS: In case of thymocytes, lymphocytes, and monocytes, all the seven mAbs demonstrated positive reactivities whereas none was reactive to erythrocytes and platelets. The majority of immune cells in formalin-fixed paraffin-embedded thymus and tonsil tissues displayed strong membranous immunoreactivity, and the main antigen was detected near 220 kDa in all cases. Among the mAbs, four mAbs (AP4, DN11, SHL-1, and P6) recognized a region commonly present in all the five isoforms. One mAb, YG27, recognized four isoforms (ABC, AB, BC, and O). Two mAbs, P1 and P14, recognized the isoforms that contain exon A encoded regions (ABC and AB). CONCLUSION: In this study, we confirmed that AP4, DN11, SHL-1, YG27 and P6, are mAbs reactive with the CD45 antigen whereas P1 and P14 are reactive with the CD45RA antigen.
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Animales , Humanos , Anticuerpos Monoclonales , Antígenos Comunes de Leucocito , Plaquetas , Línea Celular , Células COS , Eritrocitos , Exones , Citometría de Flujo , Inmunohistoquímica , Inmunoprecipitación , Leucocitos , Linfocitos , Monocitos , Tonsila Palatina , Isoformas de Proteínas , Proteínas Tirosina Fosfatasas , Timocitos , TimoRESUMEN
Objective:To observe the effects of mycobacterium tuberculosis components (mannose-capped lipoarabinomannan,ManLAM) on the maturation of dendritic cells and downstream immunity.Methods:The peripheral blood mononuclear cells were obtained from healthy individuals,and then were cultured in medium with GM-CSF and IL-4.Five days later the medium was changed and cytokine was added to.On the sixth day,the cells were divided into three groups.Meanwhile ManLAM and LPS were added to stimulate the DCs according to the experimental design.Seven days later,the cells were collected and the level of DC-SIGN,HLA-DR,CD86 and CD83 were detected by flow cytometry.The levels of IL-12 and IL-10 in the supernatant were measured by enzyme-linked immunosorbent assay (ELISA).Proliferative induction of DCs to CD4+T lymphocytes was measured by mixed lymphocyte reaction(MLR).The naive CD4+CD45RA+ T cells and allogeneic DCs were co-cultured for 48 hours.Subsequently,the supernatant IFN-? level in each group was measured by ELISA.Results:(1)The expression of CD86 and CD83 on DCs in ManLAM group was lower than those of the mature group (P
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Objective:To characterize the CD4+CD45RA+,CD4+CD45RO+,CD8+CD45RA+ and CD8+CD45RO+T lymphocyte subsets in the peripheral blood of the patients with chronic hepatitis B and to explore their relations with the disease state.Methods:The peripheral blood was collected from 104 patients with chronic hepatitis B and 30 healthy individuals,then CD4+CD45RA+,CD4+CD45RO+,CD8+CD45RA+ and CD8+CD45RO+T lymphocyte subsets were detected by flow cytometry with three-color fluorescence technology.Results:To compare with the control,the percent of CD8+CD45RA+T lymphocyte in patients with mild and severe grad CHB decreased markedly,the percent of CD8+CD45RO+T lymphocyte increased markedly,CD4+,CD8+,CD4+CD45RA+ and CD4+CD45RO+T lymphocyte did not change obviously. Compared with the mild grade CHB,the percent of CD8+CD45RA+T lymphocyte in patients with severe grade CHB decreased obviously(P