Search

Objective: To investigate the expression and localization of vitamin D3 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. [Copyright &y& Elsevier]

Background: 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., Objective: To evaluate the efficacy of heterologous antigen preparations as a way of mouse immunization in the production of anti-CA 125 mAb., Methods: 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., Results: 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., Conclusion: 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.

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.