Evaluation of PBMC Distribution and TLR9 Expression in Patients with Systemic Lupus Erythematosus.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease which results in damage to various organs. Some animal studies have revealed that activation of Toll-like receptors (TLRs) is important in the pathogenesis of SLE. In the present study, the percentage of different immune cell subsets in 35 SLE patients and 38 control subjects was analyzed by flow cytometry. We also assessed the expression of TLR9 in the population of peripheral blood mononuclear cells (PBMCs) including T lymphocytes (CD4+ and CD8+), B lymphocytes (CD19+), NK cells (CD56+) and monocytes (CD14+) in SLE patients and healthy controls. The results showed that the percentage of CD8+ T lymphocytes and CD14+ monocytes were significantly higher (p˂0.001) in the SLE patients than the healthy control subjects. Moreover, the percentage of CD56+ NK cells were significantly lower in the SLE patients than the healthy control subjects (p=0.001). The findings indicated that the expression of TLR9 was significantly higher in CD4+ and CD8+ T lymphocytes and CD19+ B lymphocytes of SLE patients than in control subjects (all p˂0.05). The difference in TLR9 expression are involved in pathogenesis of the SLE, hence it can be used as an indicator for SLE diagnosis.

Different Doses of Transforming growth factor-ß on In vitro Differentiation of Human Naïve CD4+ T Cells to T Helper 17.

An appropriate differentiation of distinct human CD4+ T cell subset is critical for manipulating these cells for using in immunity related diseases. Despite various attempts to clarify the role of different factors involved in Th17 differentiation, many crucial contradictions yet remained to be optimized. Although it has been shown that the differentiation of in-vitro Th17 cells culture conditions requires the presence of IL-1beta, IL-23, IL-2, IL-21, IL-6 and TGF-ß, the optimum amount of TGF-ß regulating in vitro human Th17 cell differentiation is still unclear. In the current study, a flow cytometric assay was used to evaluate the effect of different concentrations of TGF-ß and a combination of IL-1beta, IL-23, IL-2 without using IL-6 on development of Interleukin (IL)-17-producing T helper (Th17) cells. According to our findings, 0.1 ng/ml of TGF-ß significantly increases the expression of IL-17 in comparison to other concentrations of this cytokine. Results indicated the vital role of TGF-ß cytokine in the polarization of human Th17 cells in vitro.

Arteether exerts antitumor activity and reduces CD4+CD25+FOXP3+ T-reg cells in vivo.

BACKGROUND: Chemo-immunotherapy is one of the new achievements for treatment of cancer, by which the success of anti-cancer therapy can be increased. In vitro studies have been shown that Arteether (ARE) induces apoptosis in tumor cells, but not in normal cells. OBJECTIVE: To investigate the cytotoxic and immunomodulatory properties of Arteether in-vivo and in-vitro. METHODS: In this study, we used MTT assay for evaluation of cytotoxicity of Arteether on tumor cell line and Peripheral Blood Mononuclear Cells (PBMCs) from healthy individuals. Balb/c mice were subcutaneously transplanted with tumor tissue taken from Spontaneous Mouse Mammary Tumor (SMMT) bearing female mice. Arteether was administered to breast tumor-bearing Balb/c mice at a dose of 6mg/kg/day intraperitoneally. Tumor sizes, lymphocyte proliferation, cytokines production, and the percentage of splenic T-reg cells were measured. RESULTS: We observed that ARE could reduce the cell growth of 4T1 cell line in a dose-dependent manner but it had no cytotoxic effect on the growth of peripheral blood lymphocytes. ARE administered intraperitoneally to tumor-bearing Balb/c mice could reduce the tumor growth rate and splenic T-reg cells. No difference in the IFN-γ, IL-10 and IL-4 production was observed between tumor antigen-stimulated splenocytes of mice treated with ARE and control mice. CONCLUSION: These results underscore antitumor properties of Arteether that may aid in development of more effective antitumor agents.

Study of immunomodulatory effects of arteether administrated intratumorally.

Recent studies have indicated the profound anti-tumor activity of artemisinin's compounds, among which; arteether is an oil-soluble derivative of artemisinin with an endoperoxide bridge that can induce apoptosis in tumor cells but not in the normal cells. An experiment was carried out on tumor-bearing Balb/c mice to estimate the effects of Arteether on tumor growth and antitumor immune responses. Briefly, 6mg/kg/day of Arteether and diluents were administered to two groups of mice. Tumor sizes were measured using digital verniercallipers. Mice were sacrificed and splenocytes were harvested for lymphocyte proliferation assay, the level of IL-4 and IFN-γ cytokines, and the percentage of splenic T regulatory cells were measured. According to the findings, there were no statistical differences between the groups with respect to the level of IFN-γ, IL-4 and proliferation assay; while our results showed that Arteether is effective in the reduction of tumor growth rate. In general, intra-tumoral injection of Arteether as an oil-soluble derivative of artemisinin brings to light some antitumor properties that may aid in development of more effective antitumor agents.