Antigens, CD ; metabolism ; Antigens, Differentiation, T-Lymphocyte ; metabolism ; CD3 Complex ; metabolism ; Cells, Cultured ; Humans ; Kinetics ; Lectins, C-Type ; metabolism ; Lymphocyte Activation ; T-Lymphocyte Subsets ; drug effects ; immunology ; metabolism

The pathogenesis of some autoimmune diseases has been considered to be related to abnormal differentiation of T cell subsets. This study was aimed at investigating the change of Th1-like and Th2-like cells balance in ITP children, and analyzing the role of T cell subsets disequilibrium in the pathogenesis of ITP. Peripheral blood T cells were collected from 30 ITP patients, the T-cells were isolated and purified. The ratios of Th/Tc, Th1/Th2 and Tc1/Tc2 in peripheral blood T cells were analyzed by immunofluorescence staining and bicolor flow cytometry (FCM) in vitro. The results showed that as compared with the ratios of Th1/Th2 (48.76% +/- 6.17%) and Tc1/Tc2 (18.90% +/- 4.12%) in healthy children, the ratios of Th1/Th2 (56.21% +/- 5.95%) and Tc1/Tc2 (23.09% +/- 3.31%) in ITP children increased obviously. FCM analysis revealed that average percentages of Th, Th1, Th2, Tc, Tc1 and Tc2 were 22.31% +/- 6.51%, 21.92% +/- 6.42%, 0.39% +/- 0.14%, 31.12% +/- 6.15%, 30.95% +/- 5.45% and 1.34% +/- 0.84% in ITP children versus 39.24% +/- 5.82%, 39.01% +/- 5.47%, 0.80% +/- 0.16%, 30.25% +/- 5.63%, 28.72% +/- 5.20% and 1.52% +/- 0.68% in healthy children. The average percentages of Th, Th1 and Th2 decreased obviously, while the average percentages of Tc, Tc1 and Tc2 did not change. It is concluded that the ratios of Th1/Th2 and Tc1/Tc2 in peripheral blood T cells increase obviously in ITP children and the cellular immunity in ITP children shifts to Th1 type immunity superiority, which suggest that the abnormal differentiation of T cell subsets may play an important role in the pathologic process of ITP.
Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; T-Lymphocyte Subsets ; immunology ; metabolism ; pathology ; T-Lymphocytes, Cytotoxic ; chemistry ; Th1 Cells ; immunology ; Th2 Cells ; immunology

BACKGROUND: We developed a single-color multitarget flow cytometry (SM-FC) assay, a single-tube assay with graded mean fluorescence intensities (MFIs). We evaluated the repeatability of SM-FC, and its correlation with multicolor flow cytometry (MFC), to assess its application as a routine FC assay. METHODS: We selected CD19, CD3, CD4, and CD8 as antigen targets to analyze a lymphocyte subset. MFIs were graded by adjusting monoclonal antibody (mAb) volumes to detect several cell populations. Dimly labeled mAb was prepared by decreasing mAb volume and the optimum diluted volume was determined by serial dilution. SM-FC repeatability was analyzed 10 times in 2 normal controls. The correlation between SM-FC and MFC was evaluated in 20 normal and 23 patient samples. RESULTS: CV values (0.8-5.0% and 1.3-4.1% in samples 1 and 2, respectively) acquired by SM-FC with CD3-fluorescein alpha-isothyocyanate (FITC)dim+CD4-FITCbright and with CD19-FITCdim+CD3-FITCbright showed good repeatability, comparable to that acquired by MFC (1.6-3.7% and 1.0-4.8% in samples 1 and 2, respectively). Excellent correlation was observed between the 2 methods in the 20 normal samples (B cells, T cells, non-Thelper cells, and Thelper cells; r2=0.87, 0.97, 0.97, and 0.98, respectively; P<0.05). There were also linear relationships between SM-FC with CD19-FITCdim+CD3-FITCbright and CD8-PEdim+CD4-PEbright, and MFC, in the 23 patient samples (B cells, T cells, Tcytotoxic cells, and Thelper cells; r2> or =0.98, 0.99, 0.99, and 0.99, respectively; P<0.05). CONCLUSIONS: The multicolor, single-tube SM-FC technique is a potential alternative tool for identifying a lymphocyte subset.
Antibodies, Monoclonal/chemistry/*immunology ; Antigens, CD19/chemistry/metabolism ; Antigens, CD3/chemistry/metabolism ; Antigens, CD4/chemistry/metabolism ; Antigens, CD8/chemistry/metabolism ; B-Lymphocyte Subsets/immunology/metabolism ; Color ; Flow Cytometry/*methods ; Fluorescein-5-isothiocyanate/*chemistry ; Humans ; T-Lymphocyte Subsets/immunology/metabolism

OBJECTIVETo evaluate the clinical value of T lymphocyte subsets in prediction of chemotherapy responses of patients with pulmonary adenocarcinoma.

METHODSFifty-five chemotherapy-naive patients with pathologically or cytologically confirmed pulmonary adenocarcinoma were examined for peripheral blood T lymphocyte subsets using flow cytometry, including CD3(+) T cells, CD3(+)CD4(+) T cells, CD3(+)CD8(+) T cells, CD45RO(+) T cells and CD45RA(+) T cells.

RESULTSPatients who responded favorably to chemotherapy (CR(+)PR) showed a significantly lower percentage of CD45RA(+) T cells than those who failed to respond to chemotherapy (P=0.04). CD45RO(+) T cell percentage were slightly higher in the response group than in the non-response group, but this difference was not statistically significant (P=0.25). The other T cell subsets, namely CD3(+), CD3(+)CD4(+), and CD3(+)CD8(+) T cells showed no significant differences between the two groups.

CONCLUSIONA high percentage of peripheral blood CD45RA(+) T cells is associated with a poor short-term outcome of chemotherapy in patients with advanced pulmonary adenocarcinoma. Peripheral blood CD45RA(+) T cell level can be a reliable index for predicting chemotherapy efficacy in these patients.

Adenocarcinoma ; blood ; immunology ; Adult ; Aged ; Female ; Humans ; Leukocyte Common Antigens ; metabolism ; Lung Neoplasms ; blood ; immunology ; Lymphocyte Count ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology

OBJECTIVETo study of serum level of cortisol and peripheral T lymphocyte subsets state in the hepatitis B virus (HBV) carriers.

METHODSSixty chronic HBV carriers and ten healthy controls were all enrolled in this present study. Serum expression of cortisol was determined by radioimmunoassay, and also flow cytometry was performed to evaluate peripheral blood T lymphocyte subset.

RESULTSCompared with those in normal controls, the serous levels of cortisol in chronic HBV carriers were significantly elevated, while there was no distinct difference in the proportion of CD4+ T lymphocytes ( P > 0.05) with the decreased odds of CD4+/CD8+ lymphocytes( P < 0.05) and obvious higher proportion of CD8+ T lymphocytes( P < 0.05). In comparison between HBeAg positive group and HBeAg negative group, the serous levels of cortisol of the former group were significantly higher ( P < 0.05), and so proportion of CD8+ T was too ( P < 0.05). However, there is no significant differences in the proportion of CD4+ T lymphocyte ( P > 0.05).

CONCLUSIONThe elevated serum cortisol and increased CD8+ T lymphocytes subsets in the chronic HBV carriers, suggested that there was disturbance of endocrine-immune response in the chronicity of HBV infection.

Adult ; Carrier State ; immunology ; pathology ; virology ; Female ; Hepatitis B ; blood ; immunology ; metabolism ; pathology ; Hepatitis B virus ; immunology ; Humans ; Hydrocortisone ; blood ; immunology ; Male ; T-Lymphocyte Subsets ; immunology

Immune mediated suppression of hematopoiesis has been considered as one of the most important mechanisms leading to pancytopenia in myelodysplastic syndromes. This research was aimed at evaluating immune state of the MDS patients, analyzing the peripheral blood T cell subsets and CD3zeta chain expression and searching the possible reasons of hematopoietic disorders in 11 cases of MDS. Peripheral blood mononuclear cells were collected from 11 patients whose diagnosis was confirmed according to the new WHO diagnostic criteria. Flow cytometry was used for the counts of IFNgamma(+)CD4(+) cell (Th1), IL4(+)CD4(+) cell (Th2), IFNgamma(+)CD8(+) cell (Tc1), and IL4(+)CD8(+) cell (Tc2), and for the analysis of expression of CD3zeta chain in T cell subsets. The results showed that CD8(+) cells increased significantly in MDS patients; there was no significant difference between Th1/Th2, Tc1/Tc2 ratios of T cell subsets and normal control; CD3zeta chain, the functional protein in the signal transduction pathway of T cell, was over expressed in the CD8(+) cell. In conclusion, research indicates that abnormal changes of T cell subgroups exist in peripheral blood of MDS patients. Enhancement of CD8(+) cells and over-expression of CD3zeta chain are important features, which suggest that CD8(+) cells play the most critical role in the pathologic process as compared with other T cell subsets. The over active immunity mediated by T cell subset may be one of the major mechanisms resulting in cytopenia in MDS.
Aged ; Aged, 80 and over ; CD3 Complex ; biosynthesis ; CD8-Positive T-Lymphocytes ; immunology ; metabolism ; pathology ; Female ; Flow Cytometry ; Humans ; Lymphocyte Count ; Male ; Myelodysplastic Syndromes ; immunology ; metabolism ; pathology ; T-Lymphocyte Subsets ; immunology ; metabolism ; pathology

BACKGROUNDAmong HIV-infected patients initiating antiretroviral therapy (ART), early changes in CD4+ T-cell subsets are well described. However, HIV-infected late presenters initiating treatment present with a suboptimal CD4+ T-cell reconstitution and remain at a higher risk for AIDS and non-AIDS events. Therefore, factors associated with CD4+ T-cell reconstitution need to be determined in this population, which will allow designing effective immunotherapeutic strategies.

METHODSThirty-one adult patients with baseline CD4+ T-cell count <350 cells/mm3 exhibiting viral suppression after ART initiation were followed in the HIV/AIDS research center of Peking Union Medical College Hospital in Beijing, China, from October 2002 to September 2013. Changes in T-cell subsets and associated determinants were measured.

RESULTSMedian baseline CD4+ T-cell count was 70 cells/mm3. We found a biphasic reconstitution of T-cell subsets and immune activation: a rapid change during the first 6 months followed by a more gradual change over the subsequent 8 years. Baseline CD4+ T-cell count >200 cells/mm3 in comparison to CD4+ T-cell count ≤200 cells/mm3 was associated with more complete immune Reconstitution (77.8% vs. 27.3% respectively; P = 0.017) and normalized CD4/CD8 ratio. We showed that the baseline percentage of naive CD4+ T-cell was a predictive marker for complete immune reconstitution (area under receiver operating characteristic curve 0.907), and 12.4% as cutoff value had a sensitivity of 84.6% and a specificity of 88.2%.

CONCLUSIONSBaseline naive CD4+ T-cell percentage may serve as a predictive marker for optimal immune reconstitution during long-term therapy. Such study findings suggest that increasing thymic output should represent an avenue to improve patients who are diagnosed late in the course of infection.

Adult ; Antiretroviral Therapy, Highly Active ; methods ; CD4 Lymphocyte Count ; CD4-CD8 Ratio ; CD4-Positive T-Lymphocytes ; metabolism ; Female ; HIV Infections ; drug therapy ; immunology ; metabolism ; HIV-1 ; drug effects ; immunology ; pathogenicity ; Humans ; Male ; Prospective Studies ; T-Lymphocyte Subsets ; immunology

The objective of this study was to investigate the effect of human bone marrow mesenchymal stem cells (MSCs) on T lymphocyte killing K562 cells. MSCs were isolated from bone marrow and cultured, T cells were harvested by using nylon column method from peripheral blood. The T cells were co-cultured with MSCs, the phenotype expressions of T cell subsets were detected by flow cytometry. Killing effects of T cells (culture alone and co-culture with MSCs) on K562 cells were detected by LDH, expressions of IFN-gamma and IL-4 were detected by ELISA. The results showed that after T cells were co-cultured with MSCs for three days, the proportion of CD4+ and CD4+CD25+ T cells raised significantly (p<0.05) as compared with group of culture alone, but the proportion of CD8+ T cell were not significantly changed (p>0.05). In group of T cells co-cultured with MSCs, killing effects of T cells on K562 cells weakened, at the same time, expression of IFN-gamma decreased while expression of IL-4 increased. It is concluded that the MSCs weaken killing effects of T cells on K562 cells, which associates with increase of CD4+CD25+ T cell subsets and changes of IFN-gamma and IL-4 levels.
Bone Marrow Cells ; cytology ; CD4 Antigens ; immunology ; Coculture Techniques ; Humans ; Interferon-gamma ; metabolism ; Interleukin-2 Receptor alpha Subunit ; immunology ; Interleukin-4 ; metabolism ; K562 Cells ; Mesenchymal Stromal Cells ; cytology ; immunology ; T-Lymphocyte Subsets ; immunology ; metabolism ; T-Lymphocytes ; cytology ; immunology

OBJECTIVETo investigate CD4+ CD25+ regulatory T cell frequencies in the peripheral blood of poor or non-responsiveness to Hepatitis B vaccine, and try to understand the relationship between CD4+ CD25+ regulatory T cell and poor or non-responsiveness to Hepatitis B vaccine.

METHODSFlow cytometric analysis was employed for CD4+ CD25+ regulatory T cell frequencies in the peripheral blood of 25 cases of non-responsiveness, 30 cases of poor-responsiveness, and collected 20 cases of responsiveness as control.

RESULTSCD4+ CD25+ regulatory T cell frequencies of responsiveness was (4.32 +/- 1.21)%, poor-responsiveness was (7.01 +/- 1.06)% and non-responsiveness was (12.75 +/- 2.01)%. It was found that non and poor-responsiveness showed a high percentage of CD4+ CD25+ regulatory T cell as compared with responsiveness (t = 8.426, t = 3.289, P<0.01).

CONCLUSIONThe poor and non-responsiveness should be related with the increase of CD4+ CD25+ regulatory T cell and this might be considered as an important cause of poor and non-responsiveness.

Adolescent ; Adult ; Blood ; immunology ; CD4 Antigens ; immunology ; Female ; Flow Cytometry ; Hepatitis B Vaccines ; immunology ; Humans ; Interleukin-2 Receptor alpha Subunit ; immunology ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes, Regulatory ; immunology ; metabolism ; Young Adult

Adult ; Case-Control Studies ; Female ; Hepatitis B, Chronic ; immunology ; metabolism ; Humans ; Male ; Middle Aged ; T-Lymphocyte Subsets ; cytology ; T-Lymphocytes, Cytotoxic ; cytology