This study was aimed to investigate the effect of fetal bone marrow-derived mesenchymal stem cells (FBM-MSC) on the development of human Th1 cells. FBM-MSC were isolated, cultured and expanded in vitro. The cells were identified by their phenotype profiles and differential capacity. Human CD4(+) T cells from healthy donors were cultured alone or co-cultured with FBM-MSC (FBM-MSC/CD4). In these two cultures, the quantities of Th1 cells (interferon-γ(+)) were analyzed by flow cytometry. The results indicated that the immunophenotype and multilineage differentiation of FBM-MSC satisfied the generally accepted criteria. FBM-MSC played an inhibitory role in the development of Th1 cells. Flow cytometry analysis showed that the percentage of Th1 cells in FBM-MSC/CD4 was significantly lower than that in CD4(+) T cells cultured alone. The protein level of IFN-γ in FBM-MSC/CD4 detected by ELISA was also lower than that in CD4(+) T cells cultured alone. It was also demonstrated that the expression level of IL-6 in FBM-MSC/CD4 was much higher than that in CD4(+) T cells cultured alone or FBM-MSC. The neutralizing antibody of IL-6 could increase the quantities of Th1 cells and the expression levels of IFN-γ. It is concluded that FBM-MSC may play an inhibitory role in the development of human Th1 cells, and the IL-6 pathway may be one of mechanisms involved in the inhibitory role.
Bone Marrow Cells ; cytology ; Cell Differentiation ; Flow Cytometry ; Humans ; Immunophenotyping ; Interleukin-6 ; metabolism ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Th1 Cells ; cytology

OBJECTIVETo investigate the influence of maternal atopy on cord blood effector T cells and to identify these biologic markers as predictors of atopic dermatitis (AD).

METHODSSeventy mother-infant pairs were recruited in this prospective birth cohort study. Suspected factors for allergy, including maternal allergic history, total serum IgE, and maternal age at birth, were collected. Mother peripheral blood samples and cord blood were obtained and assayed for the percentage of interferon-γ (IFN-γ) and interleukin 4 (IL-4) producing T cells(Th1 and Th2 respectively) using flow cytometry. Their offspring at the age of 2 years old were evaluated by their dermatologist whether they had AD. Statistical analysis was performed using multiple logistic regression models and receiver-operating characteristic curve was employed to predict atopic dermatitis.

RESULTSTwenty-one allergic and 49 nonallergic mothers were recruited in this study. During the first two years of life, 15.7% children (n = 11) developed a physician-diagnosed AD (all children were the only child in the family). In group with maternal allergic rhinitis, a significantly increased percentage of Th2 was observed in peripheral blood of mother (7.10[1.18;16.1]% vs. 0.37[0.25;0.72]%, U = 10.0, P < 0.05) and cord blood of newborns (1.02[0.57;1.34]% vs. 0.21[0.15;0.42]%, U = 127.5, P < 0.05), respectively. Maternal atopic history did not affect the percentage of Th1 cells in cord blood (0.69[0.40;1.12]% vs.0.50[0.31;0.66]%, U = 361.0, P > 0.05). Children with reduced Th1/Th2 ratio in cord blood had a higher risk to develop AD (OR = 1.72, P = 0.001) . The model including Th1/Th2, maternal allergy, maternal age at birth and maternal total IgE showed high ability to discriminate children with and without AD. AUC was 0.907 (95% CI: 0.804-1.011, P < 0.001).

CONCLUSIONSElevated IL-4⁺CD4⁺ T cells in cord blood were of relevance with maternal allergic history. Imbalance between Th1 cell and Th2 cell at birth are associated with maternal allergy and promoted subsequent AD development.

Adult ; Child, Preschool ; Dermatitis, Atopic ; immunology ; Female ; Fetal Blood ; cytology ; Flow Cytometry ; Humans ; Immunoglobulin E ; blood ; Infant ; Infant, Newborn ; Mothers ; Prospective Studies ; Rhinitis, Allergic ; blood ; immunology ; Th1 Cells ; cytology ; Th1-Th2 Balance ; Th2 Cells ; cytology

Animals ; Male ; NF-kappa B ; metabolism ; Otitis Media with Effusion ; immunology ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Th1 Cells ; cytology ; Th2 Cells ; cytology

OBJECTIVETo explore the role of Th17 cells in early onset of acute graft-versus-host disease (aGVHD) and its mechanism.

METHODSMice aGVHD model was established by irradiated BABL/c mice inoculated with mixed suspension of C57BL/6 bone marrow cells and splenocytes. The mice were divided into 4 groups: (1) normal control, (2) irradiated group, (3) allo-BMT + DMSO group, (4) allo-BMT + halofuginone (HF) group. HF was given intraperitoneally at 5 µg per mouse from -1 d to +10 d after allogeneic bone marrow transplantation(allo-BMT).Mice aGVHD symptoms and survival were observed. Th1/Th17 cells ratio was evaluated by flow cytometry.

RESULTSAll the experimental groups (3) and (4) developed aGVHD after transplantation. More severe aGVHD was observed in group (4) than in group (3). HF prevented cutaneous aGVHD in all the mice, but augmented hepatic and small intestine GVHD. The percentage of Th17 cells and the ratio of Th1/Th17 were significantly higher while the percentage of Th1 cells was significantly lower in group (4) at day +6 (P < 0.05).

CONCLUSIONEarly blockage of Th17 cell results in increase of Th1 cell percentage, which exacerbates aGVHD.

Animals ; Bone Marrow Transplantation ; Female ; Graft vs Host Disease ; etiology ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Th1 Cells ; cytology ; Th17 Cells ; cytology ; Transplantation, Homologous

BACKGROUNDSurgical stress causes a helper T-cell type 2 (Th2)-dominant status and disturbs the Th1/Th2 cytokine balance. Anesthesia can suppress the stress response to surgery, therefore it may inhibit the imbalance in the Th1/Th2 ratio. In this study, we assessed if propofol anesthesia and sevoflurane anesthesia influence the Th1/Th2 cytokine balance, and which anesthesia method better attenuates this ratio.

METHODSTwenty-eight patients with an American Society of Anesthesiologists (ASA) physical status of I undergoing laparoscopic cholecystectomy were selected. They were randomly allocated into two groups of 14. Group 1 received propofol anesthesia by a target-controlled-infusion (TCI) pump and group 2 received sevoflurane anesthesia. Non-invasive blood pressure, heart rate, and end-expiration CO2 partial pressure were monitored during anesthesia. The depth of anesthesia was measured using the bispectral index (BIS), and maintained between 50 and 60. During surgery we adjusted the doses of propofol and sevoflurane according to the BIS. Samples of peripheral blood were taken before the induction of anesthesia (T1), after the induction of anesthesia (T2), at the beginning of surgery (T3), at the end of surgery (T4) and on the first day after surgery (D1). Blood samples were analyzed to give the Th1/Th2 ratio and plasma level of cortisol.

RESULTSNon-invasive blood pressure, heart rate and end-expiration CO2 partial pressure were not notably different in the two groups. At T4, the percentage of T1 cells was higher in group 1 and had statistical significance (P < 0.05). The percentage of T2 cells was not significantly different in the two groups. At T4, the difference in the Th1/Th2 ratio was significantly different. At T3, T4, and D1, the plasma level of cortisol was lower in group 1 (P < 0.05).

CONCLUSIONCompared with sevoflurane, propofol can preferably promote Th cells to differentiate into Th1 cells and inhibit surgical stress. Propofol may therefore be immunoprotective for such patients.

Adult ; Cell Differentiation ; drug effects ; Female ; Flow Cytometry ; Humans ; Hydrocortisone ; blood ; Male ; Methyl Ethers ; pharmacology ; therapeutic use ; Middle Aged ; Propofol ; pharmacology ; therapeutic use ; T-Lymphocytes, Helper-Inducer ; cytology ; drug effects ; Th1 Cells ; cytology ; drug effects ; Th2 Cells ; cytology ; drug effects

Increasing importance is being given to the stimulation of Th1 response in cancer immunotherapy because its presence can shift the direction of adaptive immune responses toward protective immunity. Based on chemokine receptor expression, CXCR3+CCR4-CD4+ T cells as Th1-type cells were investigated its capacity in monocyte-derived dendritic cell (DC) maturation and polarization, and induction of antigen specific cytotoxic T lymphocytes (CTL) in vitro. The levels of IL-4, IL-5 and IL-10 were decreased to the basal level compared with high production of IFN-gamma, TNF-alpha, and IL-2 in CXCR3+CCR4-CD4+ T cells stimulated with anti-CD3 and anti-CD28 antibodies. Co-incubation of activated CD4+ or CXCR3+CCR4-CD4+ T cells with DC (CD4+/DC or CXCR3+CD4+/DC, respectively) particularly up-regulated IL-12 and CD80 expression compared with DC matured with TNF-alpha and LPS (mDC). Although there was no significant difference between the effects of the CXCR3+CCR4-CD4+ and CD4+ T cells on DC phenotype expression, CXCR3+CD4+/DC in CTL culture were able to expand number of CD8+ T cells and increased frequencies of IFN-gamma secreting cells and overall cytolytic activity against tumor antigen WT-1. These results demonstrated that the selective addition of CXCR3+CCR4-CD4+ T cells to CTL cultures could enhance the induction of CTLs by DC in vitro, and implicated on a novel strategy for adoptive T cell therapy.
Antigens, CD4/*immunology ; Cell Line ; Cells, Cultured ; Cytokines/immunology ; Cytotoxicity, Immunologic ; Dendritic Cells/cytology/*immunology ; Humans ; Interferon-gamma/biosynthesis ; Receptors, CCR4/*immunology ; Receptors, CXCR3/*immunology ; T-Lymphocytes, Cytotoxic/*cytology/immunology ; Th1 Cells/*immunology

This study was aimed to detect the level of the peripheral blood Breg and CD4(+) T cell subgroups in patients with chronic idiopathic thrombocytopenic purpura (CITP) before and after therapy, and to analyse the charge of related cytokines and their correlation, to explore their roles in the pathogenesis of CITP. A total of 35 CITP cases were taken as the research group and 35 healthy persons were served as the control group. The peripheral blood mononuclear cells (PBMNC) were separated, the percentages of Th1, Th17, Th22 and Breg cells were detected by flow cytometry before and after treatment of glucocorticoid, and the IFN-γ, IL-17, IL-22 and IL-10 levels from PBMNC culture supernatant also were determined by ELISA. The results showed that there was significant difference as compared with the healthy controls, the proportion of peripheral blood Th1, Th17, Th22 cell subgroups all increased in CITP patients before treatment with glucocorticoid, the regulatory B cells (Breg) ratio was reduced, the differences had statistical significance (P < 0.05), but the differences were no statistically significant after treatment with glucocorticoid (P > 0.05). The levels of IFN-γ, IL-17, IL-22 from culture supernatant all increased in CITP patients before treatment, the level of IL-10 was lower than that of the healthy control, the difference was statistically significant (P < 0.05), but the there were no statistically significant differences after treatment (P > 0.05). There were positive correlation between the Breg cells and IL-10 expression in CITP patients (P < 0.05), the Breg cells and Th1, Th17, Th22 cells showed a negative correlation, IL-10 and IFN-γ, IL-17, IL-22 levels also showed a negative correlation. It is concluded that the down-regulation of regulatory B cells proportion and the IL-10 level may participate in the mechanism of CD4(+) T cell immunity disorder in CITP, which can provide new targets and ideas for the clinical immune regulation therapy.
Adolescent ; Adult ; Aged ; B-Lymphocytes, Regulatory ; cytology ; immunology ; CD4-Positive T-Lymphocytes ; cytology ; immunology ; Case-Control Studies ; Cells, Cultured ; Female ; Humans ; Interleukin-10 ; immunology ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; blood ; immunology ; Th1 Cells ; Th17 Cells ; Young Adult

Animals ; Antigens, Neoplasm ; immunology ; Cancer Vaccines ; immunology ; Graft vs Tumor Effect ; Humans ; Immunotherapy ; methods ; Neoplasms ; immunology ; therapy ; Th1 Cells ; cytology ; immunology ; transplantation ; Transplantation, Homologous ; immunology

OBJECTIVETo detect the quantity, proportion and function of producing cytokines of Th1 and Th2 cells in aplastic anemia (AA) patients and their contribution to the hematopoietic failure.

METHODS(1) Eleven patients with severe aplastic anemia (SAA) at diagnosis were observed by Marsh's method for the CFU-E, BFU-E and CFU-GM before and after depletion of CD(4)(+) T lymphocytes from bone marrow mononuclear cells (BMMNC); (2) Th1 (CD(4)(+) IFN-gamma(+)) and Th2 (CD(4)(+) IL-4(+)) cells in peripheral blood mononuclear cells (PBMNC) of 21 SAA patients and 17 normal controls were counted by FACS. (3) mRNA expression of IFN-gamma and IL-4 gene in unstimulated BMMNC from 16 SAA patients, 11 chronic aplastic anemia (CAA) patients, 26 other hematological diseases patients and 11 normal controls were measured by reverse transcriptase polymerase chain reaction (RT-PCR).

RESULT(1) CFU-E, CFU-GM and BFU-E increased significantly after depletion of CD(4)(+) T lymphocytes from BMMNC of SAA patients. (2) The percentage of IFN-gamma producing CD(4)(+) T cell (Th1) of SAA patients was significantly higher than that of controls, the percentages of IL-4 producing CD(4)(+) T cells (Th2) had no difference between SAA patients and normal controls. (3) IFN-gamma mRNA was detected in unstimulated BMMNC in 13 of 16 SAA patients, 6 of 11 CAA patients and one of 6 paroxysmal nocturnal hemoglobinuria (PNH) patients. The IFN-gamma mRNA was not detected in unstimulated BMMNC of 11 normal controls and other hematological diseases patients.

CONCLUSIONSDisbalance of CD(4)(+) T lymphocytes subsets and increases in quantity and IFN-gamma producing function of Th1 cells might be important for the development of bone marrow failure in AA and in distinguishing AA from other kinds of pancytopenic diseases.

Adolescent ; Adult ; Anemia, Aplastic ; blood ; etiology ; Colony-Forming Units Assay ; Erythroid Precursor Cells ; cytology ; Female ; Granulocytes ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Interferon-gamma ; genetics ; Interleukin-4 ; genetics ; Macrophages ; cytology ; Male ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Th1 Cells ; cytology ; metabolism ; physiology ; Th2 Cells ; cytology ; metabolism

BACKGROUND: Interleukin-17 (IL-17)-producing T helper (Th) 17 cells are considered as a new subset of cells critical to the development of rheumatoid arthritis (RA). We aimed to investigate the distribution of Th1 and Th17 cells and their association with disease activity, and determine the Th17-related cytokine levels in the peripheral blood of RA patients. METHODS: Peripheral blood mononuclear cells from 55 RA and 20 osteoarthritis (OA) patients were stimulated with mitogen, and the distributions of CD4+Interferon (INF)+IL-17- (Th1 cells) and CD4+INF-IL-17+ (Th17 cells) were examined by flow cytometry. Serum levels of IL-6, IL-17, IL-21, IL-23, and tumor necrosis factor (TNF)-alpha were measured by ELISA. Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were recorded. The 28-joint disease activity score (DAS28) was also assessed. RESULTS: The median percentage of Th17 cells was higher in RA patients than in OA patients (P=0.04), and in active than in inactive RA (P=0.03), whereas that of Th1 cells was similar in both groups. Similarly, the levels of IL-17, IL-21, and IL-23 were detected in a significantly higher proportion of RA patients than OA patients and the frequencies of detectable IL-6, IL-17, and IL-21 were higher in active RA than in inactive RA group. The percentage of Th17 cells positively correlated with the DAS28, ESR, and CRP levels. CONCLUSIONS: These observations suggest that Th17 cells and Th17-related cytokines play an important role in RA pathogenesis and that the level of Th17 cells in peripheral blood is associated with disease activity in RA.
Adult ; Aged ; Arthritis, Rheumatoid/blood/metabolism/*pathology ; Blood Sedimentation ; C-Reactive Protein/analysis ; Cytokines/blood ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis/blood/metabolism/pathology ; Severity of Illness Index ; Th1 Cells/cytology/immunology/metabolism ; Th17 Cells/*cytology/immunology/metabolism