Search

Nestin, an intermediate filament protein is expressed by neuroectodermal stem cells and tumors originating from cells of neuroectodermal and mesenchymal lineages. Nestin expression is prominent in embryos and remains upregulated until 3–6 weeks after birth but is downregulated afterward. Sertoli cells are nucleated somatic cells that are spanned in the seminiferous epithelium and play a critical role in supporting and controlling germ-cell development. In this context, we employed immunocytochemical, Western blot, and Flow cytometric analyses to demonstrate nestin expression in bovine sertoli cells. Immunostaining clearly showed that setoli cells express high levels of nestin, a result which was confirmed by Western blot analysis of purified cells. Intracellular staining of sertoli cells by flow cytometry revealed that around 74% of the cells express this marker. Given the high expression of vimentin by sertoli cells, it is proposed that the expression of nestin in these cells might be required for the formation of stable vimentin/nestin intermediate filament network. In light of these findings, it seems that sertoli cells of mature bull have potentiality of proliferation.

Infection with Leishmania major species is endemic in many provinces of Iran. Isolates from four endemic areas located in north (Damghan), center (Kashan), west (Dehloran), and south (Shiraz) of country which showed major distinctive polymorphism by RAPD-PCR method were evaluated. Isolates were inoculated to different groups of BALB/c mice and their clinical and immunological status was compared. Lesion size, parasite burden and T cell phenotype in lymph node (LN), and cytokine secretion in the culture of LN mononuclear cells were determined. The results showed the lowest and highest lesion sizes in mice infected by Shiraz strain (3.02+/-0.52 mm) and Kashan strain (5.20+/-0.45), respectively, 8 weeks after inoculation. No significant difference was observed between other strains. The parasite burden was significantly lower in lymph node of mice infected with strain of Damghan (1.51 x 10(7)) than Kashan (3.60 x 10(9)) and Shiraz (7.08 x 10(9)) strains, 8 weeks post-infection. However, Dehloran strain showed intermediate load of viable parasites (1.51 x 10(9)) in LN, 8 weeks post-infection. High ratios of IFN-gamma/IL-4 were shown in mice inoculated by strain of Dehloran (3.17) and Damghan (2.66), but not in mice infected by other strains, 8 weeks post-infection. The highest and lowest ratios of CD4(+)/CD8(+) T cells were found in LN cells of mice infected with Kashan (1.82) and Dehloran (1.00) strains, respectively. Results indicate that the lowest and intermediate loads of parasites induced by Damghan and Dehloran strains along with higher ratio of IFN-gamma/IL-4 produced by both strains in LN of inoculated mice suggest that these strains have the capacity to shift the immune responses to a predominant Th1 response after 8 weeks infection in BALB/c mice and might be the ideal strains for vaccine studies and development of candidate vaccine against leishmaniasis.

Objective To investigate the expression and localization of vitamin D 3 receptor (VDR) in reproductive organs of cycling mice. Design Experimental animal study. Setting Academic research center. Animal(s) Mature (8 to 12 weeks old) cycling female Balb/c mice. Intervention(s) Reproductive tissue, including endometrium, ovary, and fallopian tubes, were collected at each phase of estrous cycle to examine VDR expression. Main Outcome Measure(s) Expression of VDR messenger (mRNA) was determined by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR). The presence and localization of VDR was assessed by immunohistochemistry, and the intensity of VDR expression was quantified with U.S. National Institutes of Health image-analysis software. Result(s) The VDR mRNA was expressed in the endometrium throughout the estrous cycle. The relative expression of VDR mRNA at the estrus phase was more prominent compared with the other phases. Immunohistochemical analysis revealed that dendritic cells, macrophages, and luminal and glandular epithelial cells of the endometrium, granulosa, and cumulus oophorus cells of the ovary and fallopian epithelial cells strongly express VDR, particularly during the estrus phase. Conclusion(s) Our findings have demonstrated, for the first time, that VDR is present and differentially expressed in murine reproductive organs throughout the estrous cycle. Further studies are required to evaluate the functional immunologic role of VDR.

Monoclonal antibodies (mAbs) are essential tools for many molecular immunology investigations, epitope mapping and molecular modelling, clinical laboratory diagnostic tests and immunotherapy. Humoral immune response of immunized animals largely depends on the nature of antigen and the immunization technique. Polysaccharides and heavily-glycosylated proteins are very elusive targets incapable of mounting long-lasting, high affinity antibody responses. Carcinoma antigen 125 (CA 125), a well known tumor marker of ovarian cancer, is a mucin type antigen consisting of repetitive units of heavily glycosylated moieties which render production of mAbs very difficult.To evaluate the efficacy of heterologous antigen preparations as a way of mouse immunization in the production of anti-CA 125 mAb.Two different protocols of immunization were used for priming of NMRI mice. In the first method, mice conventionally immunized by three intraperitoneal injections of purified CA 125 and boosted by the antigen three days before fusion. In the second approach, mice were primed by three intraperitoneal injections of living CA 125 positive cells of OVCAR-3 cell line, and boosted by intravenous injection of the purified extracellular domain of CA 125. Production of mAb was performed by standard hybridoma technology and mAbs were characterized by different immunoassays.The first method failed to produce stable clones despite six time fusion. A total of ten stable clones, however, were produced in the second approach. Some of the clones were characterized and found to have excellent immunoreactivity when tested by ELISA assay, western blotting, intracellular and surface immunofluorescent staining of OVCAR-3 cell line and immunohistochemical staining of ovarian cancer tissues.Altogether the results of the present study clearly showed that heterologous antigen preparation is the method of choice for immunization when production of monoclonal antibody against highly glycosylated poorly immunogenic antigens is concerned.

Summary In spite of reports on cytokines induction by the Brucella DNA in murine model, there is no comparison between pathogenic and appropriate vaccine strains in human. We investigated the cytokines profile of human peripheral blood mononuclear cells (PBMCs) induced by DNA extracted from pathogenic isolates of Brucella melitensis and B. abortus as well as Rev1 and S19; the appropriate vaccine strains. It was observed that despite differential induction of Interleukin(IL)-12 and IL-10 production, identical IL-12/IL-10 concentration ratio was obtained by all Brucella strains DNAs that was 2 after 24 h and 4 after 5 days of incubation. In addition, IL-2 and Interferon(IFN)-γ production were profoundly increased compared to the medium at day 3 and 5 respectively but IFN-α was not induced. Therefore, Brucella strains DNAs are Th1 inducing component with similar pattern in human PBMCs.

Angiocardiography is an X-ray examination of the blood vessels or chambers of the heart. Cardiologists and staff members applying this procedure are exposed to high levels of scattered radiation. In our previous study the incidence of unstable chromosomal aberrations and cytokinesis-blocked micronuclei were found to be significantly higher in exposed individuals than the age and sex matched controls. In the present study we assessed cytokine production by peripheral blood mononuclear cells of the above cases and the percentage of Treg cells. According to film dosimeter analysis, personnels received 0.25-15 mSv during the previous year (average of 3 mSv/y). Isolated PBMCs from the test and control groups were stimulated with Phorbol Myristate Acetate/ Ionomycin (PMA/I). Cytokine production was measured in the supernatants of cultured lymphocytes. The percentage of Treg cells was studied by flow cytometry. The production of IL-10 and IL-5 was significantly down-regulated in the test group compared to the control group. In contrast, IL-12 was up-regulated. Yet, no statistically significant difference was found for IFN- gamma between two groups. In addition, we found higher percentage of CD4+CD25+(bright) Treg cells in the study group compared to the controls. Taken together, it was shown that low doses of scattered X-rays could skew cytokine profile of peripheral blood mononuclear cells in favour of inflammatory response causing the increase of Treg cells.