The aim of this study was to investigate the effect of human bone marrow mesenchymal stem cells on human T-cell proliferation resulted from stimulation with PHA and possible immunomodulating mechanism. T cells were positively selected by CD3(+) magnetic beads, and were then co-cultured with irradiated MSCs overnight before the addition of PHA. T-cell proliferation was measured by BrdU assay and the degree of apoptosis was assessed by flow cytometry with Annexin V/PI. T cells co-cultured with or without MSCs were treated with PHA for 72 hours, then harvested. They were labeled with anti-CD4, anti-CD8, anti-CD25 antibodies and analyzed by flow cytometry. The results showed that MSCs inhibited T-cell proliferation, but did not induce T cell apoptosis. There were no significant changes in the ratio of CD4(+) and CD8(+) T cells of MSC-treated group, as compared with the control group. After stimulation with PHA, there was an increase in CD4(+) T cells and decrease of CD4(+)CD25(+) cells in MSC co-cultured group. It is concluded that the MSCs inhibit T-cell proliferation after stimulation with PHA, and show more inhibitive effects on CD8(+) and CD4(+) T cells, but CD25(+) regulatory T cells may not be involved in this process.
Bone Marrow Cells ; cytology ; CD4-CD8 Ratio ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Humans ; Mesenchymal Stromal Cells ; cytology ; physiology ; T-Lymphocyte Subsets ; cytology

OBJECTIVETo explore the mechanism of electroacupuncture of "Shuanggu Yitong" prescription on postponing aging.

METHODSForty 3-month SD rats, male only, 30 rats were made sub-acute aging model by D-galactose s.c. injection continuously for 42 d, and rest of the rats, 10, were divided into a normal control group. After the modeling, the sub-acute aging model rats were randomly into a Shuanggu Yitong group [electroacupuncture at "Guanyuan" (CV 4) and "Zusanli" (ST 36), hand needle at "Baihui" (GV 20)], an acupuncture control group [electroacupuncture at "Weizhong" (BL 40) and "Shuifen" (CV 9), hand needle at "Yintang" (GV 29)] and an aging model group, ten in each one. The treatment was given once in a day, six of which made a course. The rats in the normal control group and aging model group were not received any treatment. After the treatment for three weeks, the rats were put to death and their spleen index, thymus index and the T lymphocytes subgroups (CD8(+) T/T cell and CD8(+) CD28(-) T/CD8(+) T cell) were tested.

RESULTSThe spleen index (1.74 +/- 0.059) and thymus index (0.64 +/- 0.039) in the aging model group was obviously lower than those in the normal control group (1.93 +/- 0.061), (0.81 +/- 0.053) respectively (both P < 0.05); the CD8(+) CD28(-) T/CD8(+) T cell percentages (26.28 +/- 4.69)% and CD8(+) T/T cell percentages (43.33 +/- 2.84)% in the aging model group were both significantly higher than those (15.08 +/- 5.58)% (P < 0.01), (34.70 +/- 4.24)% (P < 0.01) in the normal control group. Compared with the aging model group, the spleen index (1.91 +/- 0.081) and thymus index (0.79 +/- 0.080) in the Shuanggu Yitong group were significantly higher (both P < 0.05), but obviously decreased with the percentage of CD8(+) CD28(-) T/CD8(+) T cell (18.07 +/- 1.73) (P < 0.01); the percentage of CD8(+) CD28(-) T/CD8(+) T cell (18.07 +/- 1.73)% in the acupuncture control group was also lower than the aging model group (P < 0.05), but more obvious reduce for the Shuanggu Yitong group (P < 0.05).

CONCLUSIONThe treatment of Shuanggu Yitong prescription could regulate the proportions of the T lymphocyte subset, and slow down the immunosenescence of subacute aging model rats induced by D-galactose.

Acupuncture Points ; Aging ; physiology ; Animals ; Electroacupuncture ; Flow Cytometry ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Spleen ; cytology ; T-Lymphocyte Subsets ; cytology ; Thymus Gland ; cytology

OBJECTIVE: To explore the effect of suppressive oligodeoxynucleotides (Sup ODN) on the Th1 differentiation of CD4(+)T splenetic lymphocytes in mice. METHODS: The splenetic lymphocytes of BALB/c mice were separated, and then CD4(+) cells were purified with immune magnetic CD4(+) microbeads (positive selection). The purification was examined by fluorescence-activated cell sorter. CD4(+) cells, anti-CD3epsilon, anti-CD28, IL-12 and Sup ODN or control oligodeoxynucleotides (Con ODN) were co-incubated for 72 h. IFN-gamma and IL-4 in the supernatant were detected using enzyme-linked immunosorbent assay. The expression of T-bet mRNA in CD4(+) cells was tested by reverse transcription polymerase chain reaction. RESULTS: Sup ODN could significantly inhibit the release of INF-gamma and increase IL-4 production respectively (P<0.01). T-bet mRNA of CD4(+) lymphocytes was remarkably inhibited by Sup ODN as well (P<0.01). CONCLUSION In the presence of pro-Th1-cytokines, Sup ODN may affect the differentiation of CD4(+) T lymphocytes in vitro. Sup ODN can promote CD4(+) T cells to differentiate into Th2, and suppress them into Th1.
Animals ; CD4-CD8 Ratio ; CD4-Positive T-Lymphocytes ; cytology ; immunology ; Cell Differentiation ; drug effects ; Female ; Interferon-gamma ; antagonists & inhibitors ; physiology ; Interleukin-12 ; antagonists & inhibitors ; physiology ; Mice ; Mice, Inbred BALB C ; Oligodeoxyribonucleotides ; pharmacology ; T-Lymphocyte Subsets ; Th1 Cells ; cytology ; immunology

Th17(T helper 17 cell), a newly discovered subset of T cells is associated with IL-23 and characterized by production of IL-17, the functions of which are distinct from those of Th1, Th2 and Treg subsets. The development of Th17 cells can be promoted by TGF-beta1, IL-6, and IL-23; but inhibited by IFN-gamma, IL-4 and Socs3. It is clear that Th17 cells have protective effects on body by facilitating the pro-inflammatory responses. On the other hand, the role of Th17 cells in the pathophysiology of autoimmune diseases has been described.
Autoimmune Diseases ; immunology ; physiopathology ; Interleukin-17 ; biosynthesis ; immunology ; Interleukin-23 ; biosynthesis ; genetics ; T-Lymphocyte Subsets ; immunology ; physiology ; T-Lymphocytes, Helper-Inducer ; classification ; cytology ; immunology ; Transforming Growth Factor beta1 ; biosynthesis ; genetics

The protective efficacy of DNA plasmids encoding avian infectious bronchitis virus (IBV) S1, N, or M protein was investigated in chickens. Chickens were inoculated monovalently (with plasmid pVAX1-16S1, pVAX1-16M, or pVAX1-16N alone) or multivalently (combination of the three different plasmids, pVAX1-16S1/M/N). A prime-boost immunization protocol against IBV was developed. Chickens were immunized with the multivalent DNA vaccine twice and then boosted with an inactivated vaccine once. Antibody titers of the chickens immunized with pVAX1-16S1/M/N were much higher than those of the monovalent groups (p < 0.01). A protective rate up to 90% was observed in the pVAX1-16S1/M/N group. The serum antibody titers in the prime-boost birds were significantly higher than those of the multivalent DNA vaccine group (p < 0.01) but not significantly different compared to the inactivated vaccine group at 49 days of age. Additionally, the prime-boost group also showed the highest level of IBV-specific cellular proliferation compared to the monovalent groups (p < 0.01) but no significant difference was found compared to the multivalent DNA vaccine group, and the prime-boost group completely protected from followed viral challenge.
Aging ; Animals ; Antibodies, Viral/blood ; Cell Proliferation ; Chickens ; Coronavirus Infections/prevention & control/*veterinary/virology ; Immunization, Secondary/veterinary ; Infectious bronchitis virus/*immunology ; Poultry Diseases/*prevention & control/virology ; T-Lymphocyte Subsets/cytology/physiology ; Vaccines, DNA/immunology ; Vaccines, Inactivated/immunology ; Viral Vaccines/*immunology

This study was purposed to investigate the changes in quantum and function of gammadelta T cell subsets, and to explore its significance in pathogenesis of acquired pure red cell aplastic anemia (A-PRCA). Eleven patients were diagnosed as A-PRCA based on bone marrow smear and biopsy, and were treated with cyclosporine A and glucosidorum tripterygll totorum. The flow cytometry technique was used for analyses of T cells subsets and gammadelta T cells. Furthermore, peripheral mononuclear cells (MNC) isolated from A-PRCA patients were cultured in RPMI 1640 medium (10(5) cells/ml) containing 10% FCS, phytohemagglutinin (PHA, 10 microg/ml), and recombinant human interleukin-2 (rIL-2, 50 U/ml) for two weeks, then gammadelta T cells were isolated with the TCRgammadelta Microbead Kit from cultured cells. The collected gammadelta T cells were incubated with normal control bone marrow MNC in RPMI 1640 medium (37 degrees C, 5% CO2 atmosphere) for CFU-E, CFU-GM, and BFU-E colony assay. The result showed that compared with the control group, CD3(+), CD8(+) cells increased significantly in the patient group (P < 0.05), the CD4(+)/CD8(+) ratio decreased and reversed, and gammadelta T cells were significantly increased in patient group (P < 0.05). After treatment with cyclosporine A, 9 out of 11 patients got good response, and CD3(+), CD8(+) cells in the responding patient decreased, the ratio of CD4(+)/CD8(+) returned to normal, and gammadelta T cells also decreased to normal range. Moreover, in vitro culture, the gammadelta T cells isolated from A-PRCA patients showed an inhibiting action to CFU-E and BFU-E but not to CFU-GM in a dose-dependent manner. It is concluded that gammadelta T cells increase in A-PRCA patients, and decrease in parallel to normal range with significant improvement of anemia symptoms after immune suppressive therapy. The gammadelta T cells isolated from A-PRCA patients showed an inhibiting action to CFU-E and BFU-E but not to CFU-GM in vitro culture, suggesting that gammadelta T cells may bring an impact on the research of A-PRCA pathogenesis. Cyclosporine A demonstrated better therapeutic effect on A-PRCA patients.
Adult ; Aged ; CD4-CD8 Ratio ; Cells, Cultured ; Cyclosporine ; therapeutic use ; Female ; Flow Cytometry ; Humans ; Immunosuppressive Agents ; therapeutic use ; Male ; Middle Aged ; Receptors, Antigen, T-Cell, gamma-delta ; physiology ; Red-Cell Aplasia, Pure ; drug therapy ; etiology ; immunology ; T-Lymphocyte Subsets ; cytology ; T-Lymphocytes ; cytology ; T-Lymphocytes, Cytotoxic ; immunology