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Objective(s): Breast cancer is one of the most common cancers among women, and current treatments are inadequate due to unwarranted side effects and lack of specificity resulting in off target consequences. Artesunate is a synthetic anti-malarial drug that exerts inhibitory effects on cancer cell lines via apoptosis and has been used treating some cancers. This study investigated the anticancer effects of Fe3O4 magnetic nanoparticles conjugated with chitosan, polyethylene glycol, folic acid, and artesunate in vivo and in vitro. Materials and Methods: Nanoparticles were synthesized by co-precipitation; morphology and size were determined by scanning electron microscopy (SEM), and the presence of the components was verified by nanoscale Fourier transform infrared spectroscopy (FTIR). 4T1 murine mammary tumor cells were treated with nanoparticles, and cell viability was determined by MTT assay. 4T1 cells were also subcutaneously injected into BALB/c mice, and magnetic resonance imaging was carried out two weeks later to determine tumor size among the groups. Interferon gamma (IFN-γ) and IL-4 levels (in splenocytes culture supernatant) were measured by ELISA, and tumors, surrounding tissues, and mouse livers were histopathologically studied. Results: The nanoparticle made in this article had good anticancer effects and caused apoptosis in cancer cells in breast cancer, and also strengthened the cellular immune system and further increased interferon gamma and increased the half-life of mice with cancer, while this nanoparticle It did not have the side effects of chemotherapy drugs. Conclusion: Artesunate-containing nanoparticles decreased 4T1 cell viability and increased apoptosis to a greater extent than nanoparticles without the drug. In vivo, artesunate nanoparticles showed no toxicity and were more effective in decreasing tumor size than control. They were also associated with increased survival, increased IFN-γ, and decreased IL-4 levels in the spleen. The findings show the drug targets cancer cells effectively with minimal side effects due to its herbal nature and targeted nano delivery. [ABSTRACT FROM AUTHOR]

Human Wharton's Jelly-derived Mesenchymal Stem Cells (hWJ-MSCs) are easily available cells without transplant rejection problems or ethical concerns compared to bone-marrow-derived MSCs for prospective clinical applications. These cells display immunosuppressive properties and may be able to play an important role in autoimmune disorders. Regulatory T-cells (Treg) are important to prevent autoimmune disease development. Interleukin 35 (IL-35) induces the proliferation of Treg cell populations and reduces the activity of T helper 17 (Th17) and T helper 1 (Th1) cells, which play a central role in initiation of inflammation and autoimmune disease. Recent studies identified IL-35 as a new inhibitory cytokine required for the suppressive function of Treg cells. We created IL-35-producing hWJ-MSCs as a good vehicle for reduction of inflammation and autoimmune diseases. We isolated hWJ-MSCs based on explant culture. HWJ-MSCs were transduced at MOI=50 (Multiplicity of Infection) with lentiviral particles harboring murine Interleukin 35 (mIL-35). Expression of IL-35 in hWJ-MSCs was quantified by an IL-35 ELISA kit. IL-35 bioactivity was analyzed by inhibiting the proliferation of mouse splenocytes using CFSE cell proliferation kit. Frequency of CD4+CD25+CD127 low/neg Foxp3+ Treg cells was measured by flow cytometry. There was an up to 85% GFP positive transduction rate, and the cells successfully released a high level of mIL-35 protein (750 ng/ml). IL-35 managed to inhibit CD4+ T cell proliferation with PHA, and improved the frequency of Treg cells. Our data suggest that transduced hWJ-MSCs overexpressing IL-35 may provide a useful approach for basic research on gene therapy for autoimmune disorders. [ABSTRACT FROM AUTHOR]

Cytokines have been introduced as critical inducers in the development of Th subpopulations.Cytokines like IL-10 are involved in inducing regulatory T cells such as Type 1 regulatory T (Tr1) cells cells. IL-22 is a member of IL-10 family of cytokines, and IL-28A is a member of IFN-γ family. In this study, cord blood mononuclear cells (CBMC) from normal healthy individuals were isolated by Ficoll and then naïve T cells were purified by CD4+CD25+ Regulatory T cell Isolation kit. The effect of these two cytokines on production of IL-5, TGF-β, IL-10, IL-4 and IFN-γ cytokines from cord blood T cells was investigated to identify Tr1 cells as well as Th1 and Th2 polarization. Flow cytometric analysis showed that IL-28A and IL-22 were not effective in expression of IL-5 and TGF-β either alone or in synergy, but in view of IL-10, IL-4 and IFN-γ, the results showed that IL-22 increased IL-10 and IL-4 but had a decreasing effect on IFN-γ. The results showed that IL-28A was not effective in increasing or decreasing the level of IL-10, IL-4 and IFN-γ. Therefore, according to these results, IL-22 and IL-28A were not effective in inducing Tr1 cells. [ABSTRACT FROM AUTHOR]

Objective(s): Multi-epitopic protein vaccines and direction of vaccine delivery to dendritic cells (DCs) are promising approaches for enhancing immune responses against mutable pathogens. Escherichia coli is current host for expression of recombinant proteins, and it is important to optimize expression condition. The aim of this study was the optimization of multi-epitopic HIV-1 tat/pol/gag/env recombinant protein (HIVtop4) expression by E. coli and conjugation of purified protein to anti DEC-205 monoclonal antibody as candidate vaccine. Materials and Methods: In this study, expression was induced in BL21 (DE3) E. coli cells by optimization of induction condition, post induction incubation time, temperature and culture medium formula. Some culture mediums were used for cell culture, and isopropyl-beta-Dthiogalactopyranoside was used for induction of expression. Protein was purified by Ni-NTA column chromatography and confirmed against anti-His antibody in western-blotting. To exploit DCs properties for immunization purposes, recombinant protein chemically coupled to αDEC-205 monoclonal antibody and confirmed against anti-His antibody in western-blotting. Results: The optimum condition for expression was 1 mM IPTG during 4 hr cultures in 2XYT medium, and final protein produced in soluble form. Conjugation of purified protein to αDEC-205 antibody resulted in smears of protein: antibodies conjugate in different molecular weights. . Conclusion: The best cultivation condition for production of HIVtop4 protein is induction by 1 mM IPTG during 4 hr in 2XYT medium. The final concentration of purified protein was 500 μg/ml. [ABSTRACT FROM AUTHOR]