OCT-4, an embryonic stem cell marker expressed in breast, brain and thyroid carcinomas compared to testicular carcinoma

Cancer stem cells are a small subpopulation of cells within a tumor which are responsible for maintaining the tumor mass. A number of factors such as OCT-4 that govern the fate of adult stem cells also play a role in malignant cell transformation. OCT-4 is a key regulator of self-renewal in embryonic stem cells; its expression is potentially correlated with tumorigenesis and can affect some aspects of tumor behavior such as tumor recurrence or resistance to therapies. We have investigated the potential expression of OCT-4 on a panel of tumors including breast, brain, thyroid and testicular carcinomas, using immunohistochemistry. The level of expression of OCT-4 was then compared to different tumor types and degree of differentiation. OCT-4 was expressed at the highest levels on nuclear site of seminoma compared with other tumors. The expression of OCT-4 was detectable in both nucleus and the cytoplasm of almost all breast tumors, but it was detectable at much lower level in normal breast tissues. OCT-4 expression was noted on poorly differentiated papillary carcinoma of thyroid compared to normal follicles of thyroid gland adjacent to the tumor. Breast carcinomas and papillary carcinomas of thyroid express elevated levels of embryonic stem cell gene OCT-4, suggesting that these tumors may contain cells indicative of embryonic-like stem cells. Identification of cancer stem cells in different malignant tumors may be useful for prognostic evaluation and administration of a new treatment which target this sub-population of tumor cells

expression profile of indolamine 2,3-dioxygenase in murine endometrium during estrous cycle

Activation of Indolamine 2,3-dioxygenase [IDO], an enzyme responsible for tryptophan catabolism, has been reported to be a necessary requirement to achieve immunological tolerance against the fetus and protection against intracellular and extracellular pathogens. The objective of this study was to evaluate the expression of IDO gene in murine endometrium and its expression rate in different phases of estrous cycle. Noticing the role of this enzyme especially in the survival of a semi-antigenic embryo, the results of this study may be used as a basis for practical studies on the immunologic bases of recurrent abortions. In this experimental study, we studied the expression of IDO in the female BALB/c mice endometrium during four stages of estrous cycle. The phases of estrous cycle were determined by examining vaginal cytology .At each phase, endometrium was pealed away and the relative expression of IDO mRNA was detected by semi-quantitative RT-PCR using specific primers to IDO and mGAPDH as a housekeeping gene. The specificity of reaction was confirmed by enzymatic digestion of amplicon which yielded to 138bp and 259bp fragments. Our results showed, for the first time, that IDO is expressed in the endometrium of cycling mice during all stages of estrous cycle. The expression of IDO was highest at estrus and lowest at diestrus [p<.001]. Expression of IDO in endometrium during all phases of estrous cycle reveals that this enzyme as an effective arm of innate immune system may serve a role in protecting the female reproductive tract against ascending infections. Also regarding the fact that, mating only occurs at estrus phase, the high expression of IDO in this phase, may act as the main mechanism in inducing immunological tolerance to the fetus

Effect of pregnant mouse serum on induction of indolamine 2, 3-dioxygenase in dendritic cells

Several studies have shown that many factors are involved in the maternal tolerance to the fetus. Indolamine 2, 3- dioxygenase [IDO] enzyme which catabolizes tryptophan is one of the factors that have been reported to play an important role leading to a successful pregnancy. The objective of this study was to evaluate the effects of pregnant mouse serum on the induction of indolamine 2, 3 dioxygenase in dendritic cells [DCs] which may be used as a basis for practical studies on the immunological bases of recurrent abortions. Allogenic pregnant mice sera were collected in mid-pregnancy. DCs were isolated from Balb/c mouse spleen through a three-step method, including: Collagenase digestion of spleen tissue, low density cells separation via the Nycodenz density gradient centri-fugation and plastic adherence. T cells were isolated from C57BL/6 mouse lymph nodes through nylon wool method. As stimulator cells, pregnant and non-pregnant mice sera treated DCs were irradiated and co-cultured with purified T cells [allogenic MLR]. 1-methyl-tryptophan [1-MT], as the specific inhibitor of IDO, was added to some wells of MLR assay in different concentrations and T cells proliferation response was measured by [3]H-Thymidine incorporation. The MLR supernatant was also analyzed by HPLC for its tryptophan and kynurenin [Trp metabolite] content. All tests were repeated for 5 times. Man-Whitney's non- parametric test was used to evaluate the differences among groups. Confidence interval was 95% and p-values <0.05 were regarded as significant. The results showed the ability of pregnant mice sera to reduce the dendritic cells ability in T cell proliferation induction compared to non-pregnant mice sera but addition of 1-MT did not have any significant effect on this inhibition. Additionally, IDO metabolites concentration assessment in the presence or absence of 1-MT, through HPLC method, did not show any significant difference. There are many factors in pregnant mice sera such as progesterone, IL-10, Vit D3, etc. Which might cause inhibition of T lymphocyte proliferation response in allogenic MLR through affecting DCs' efficiency. Although it seems that IDO expression by DCs is not responsible for decrease in T cell proliferation after treatment of DCs by pregnant mice sera, thus some other mechanisms might be responsible for this phenomenon which their identification needs more investigation

case report of recurrent hydrops fetalis due to maternal alloimmunization with Kell antigen

During pregnancy, irregular blood group antibodies originating either from earlier pregnancies or from blood transfusions may severely affect child health. In this report, a case of maternal alloimmunization to Kell antigen is described.The mother had a history of partial mole and four repeated intrauterine fetal death due to hydrops fetalis.Screening of irregular blood group antibodies revealed that she has anti-Kell with the titer of 1:4096. Also in genetic analysis, a C677T homozygous mutation of MTHFR gene was found, which could potentially enhance destructive effects of anti-Kell antibody. The described case emphasizes the importance of being informed about the presence of irregular blood group antibodies during pregnancy which may cause recurrent hydrops

Seroepidemiologic HGV in blood donors haemodialysis patients, haemophiliacs and major beta thalassemics with history of liver disease

The prevalence of GBV-C and HGV in blood donor populations in developd countries based on HGV detection and anti-E2 screening ranges from 1 to 5 and 3 to 14% respectively. The aim of this study was to investigate seroepidemiologic hepatitis G virus [HGV] in blood donors, heamodialysis patients, hemophiliacs, and beta thalassemics with a history of liver disease by Elisa technique. In this descriptive study, blood samples of 330 volunteer blood donors, 44 heamodialysis patients, 16 haemophiliacs, and 40 beta major thalassemics with a history of liver disease were studied by Elisa technique for their seroepidemiologic status of hepatitis G virus and their past record HGV infection. For data analysis, Ch-square, Fisher exact test, and SPSS version 11.5 were used. This study showed that out of 330 healthy blood donors 14[4.2%], out of 44 heamodialysis patients 10[22.7%], out of 16 haemophiliacs 5 [30.3%] and out of 40 beta thalassemics 10 [25%] were positive for HGV-anti-E2. These data are significant evidence for HGV to be considered as a transfusion-transmitted infection. The prevalence of anti-HGV and anti-HCV [co-infection] was found to involve 10 [30.3%] of heamodialysis patients, 4 [28.6%] of haemophiliacs and 9 [23.7%] of beta thalassemics. It was also found that 1 [8.3%] of heamodialysis patients, 1 [33.3%] of haemophiliacs, and 1 [50%] of beta thalassemics were infected with anti-HGV and HBsAg co-infection. The prevalence of HGV was high in multitransfused individuals including heamodialysis patients, haemophiliacs, and thalassaemics. Therefore, HGV was a transfusion-transmittable agent. Co-infection of anti-HGV with HCV was observed in viruses. It is recommended that further studies focus on evaluating sexual and vertical transmission routes so as to cast light on relatively high rate of HGV in donor population

[Allostimulatory efficiency of splenic dendritic cells from pregnant and non pregnant mice]

Mammalian reproduction looks like an immunological paradox, because fetal alloantigens encoded by father genes should induce cell mediated immune responses leading to fetal loss. Maternal immune system, in addition to local modulation, undergoes systemic modulations during pregnancy. Dendritic cells [DCs], as professional antigen presenting cells, play a key role in initiation and control of immune response and it seems that functional changes in these cells during pregnancy may contribute to the systemic immune tolerance. To address this issue, in this study we isolated and purified DCs from pregnant mice and evaluated their stimulatory potential to induce proliferative response of allogenic T cells in unidirectional mixed leukocyte reaction [MLR]. Following collagenase digestion of splenic tissue, using density gradient centrifugation [13% Nycodenz] and adherence properties of DCs to the bottom of tissue culture dish, 7x10[5] DCs were isolated from each spleen with more than 95 percent purity. Allogenic T cells were isolated by nylon wool column, using their non-adhesive character to nylon wool. After radiation, isolated dendritic cells from pregnant and non-pregnant Balb/c mice were used in mixed leukocyte culture with C57BL/6 mice T lymphocytes. T lymphocyte proliferation was measured after 72 hours by [3]H- thymidine incorporation. 7x 10[5] dendritic cells with the purity of >95% were isolated from each spleen. Also the yield of T- lymphocyte form Inguinal and Brachial lymph nodes was about 3-5x10[5] with the purity of%85-90. The results showed that there is no statistical difference between stimulatory potential of DCs form pregnant [cpm=33000] and non- pregnant [cpm=35000] mice in induction of allogenic T-Cell proliferation. These findings can result from low concentration of immune suppressor factors in circulatory system of pregnant mice or due to separation of dendritic cells from pregnancy microenvironment and their maturity in vitro in the absence of the immune suppressor factors

[Designing an ELISA method using sperm antigens for detection of antisperm antibody in comparison with SpermMar test

The role of antisperm antibodies with a prevalence of 6-26% is well known in immunological infertility. Thus, there is clinical importance to determine ASA levels in both male and female. Nowadays, one of the most important discussed controversies in the field of immunological infertility is establishing an standard method to determine ASA. It seems that ELISA method will be more sensitive, specific and more diagnostic in determination of ASA if sperm surface antigens could be used as coated antigens, with least contamination to sperm intracellular antigens and nonspermic antigens. So, the aim of this study is designing an ELISA method by using the best method of sperm antigens extraction with at least contamination. In this study we designed an ELISA method with three different extraction methods of sperm antigens including sonication method, using SDS detergent, and application of LIS detergent, then we compared ELISA method based on the three extraction methods as well as two similar commercial ELISA kit [IBL Co, and Bioserv Co] with SpermMar test. Comparing designed method with commercial kit indicated that among 28 sera which had 16 positive sera and 12 negative sera by SpermMar, 14 sera were true positive by LIS method and only 2 cases were false negative without any false positive results, whereas there were 5 true positive results and 11 cases false negative by the sonication method. The SDS method also had 13 true positive results with 3 false negative and 4 false positive results. In addition, two commercial kit had in turn 7 and 4 cases true positive and both of them had 1 case false positive and in turn 9 and 12 cases with false negative result. ELISA method designed by LIS detergent has adequate sensitivity [87.5%] with higher specificity [100%] and efficacy [92.8%] than other extraction methods. There is a significant correlation between this designed method and SpermMar test [r=0.572]. The results of this study indicated that ELISA method by LIS antigens has at least contamination with nonspermic antigens and it is better than other extraction methods and commercial ELISA kits for detection of antisperm antibody

[Optimization of Dendritic cells purification from mouse spleen a recommended method for purification of Dendritic cells from reproductive organs]

Dendritic cells [DC] are the principal antigen-presenting cells [APC] responsible for induction of primary immune responses by T lymphocytes. Although DCs are present in most lymphoid tissues, they occur in very low frequency accounting for 0.5% or less of nucleated cells in peripheral lymphoid organs. In the present study, we report the purification of DCs from mouse spleen with high yield and purity using a three-step purification technique including: collagenase digestion of tissue, selection of low-density cells using Optiprep density gradient medium and plastic adherence. By using techniques outlined above, we obtained 5-7x10[7] DC/spleen with purity >/= of 97%. Such large numbers of purified DCs enables us to further document their different characteristics including morphology, immunophenotype and to evaluation of their role in immune system. Finally, since DCs have been reported to be present in all reproductive organs, we suggest that this protocol be used for isolation and purification of DCs from those organs for further in vitro studies