Study the effect of recombinant leukemia inhibitory factor on maintenance of pregnancy and frequency of regulatory T cells in abortion-prone mice.

Recurrent spontaneous abortion (RSA) can have a significant impact on a woman's quality of life. Understanding the mechanisms behind abortion is crucial for developing potential treatments. Among various models of abortion, the CBA/J(♀) × DBA/2J(♂) model stands out as the most extensively studied. This model reveals the influence of an altered immune system on resorption during pregnancy. The leukemia inhibitory factor (LIF) holds considerable importance as a secretory glycoprotein essential for successful implantation. Regulatory T cells (Tregs) have been found to produce high levels of LIF in both mice and humans. LIF plays a vital role in the development of Tregs by upregulating the expression of the Foxp3 transcription factor while downregulating the expression of RORγt. To investigate the impact of recombinant LIF (rLIF) on pregnancy maintenance and Treg cell frequency in abortion-prone (AP) mice, a specific recombinant protein was used in this study. The AP group consisted of CBA/J(♀) × DBA/2J(♂) mice, while the control group comprised CBA/J(♀) × BALB/c(♂) mice. Intraperitoneal injections of rLIF were administered to the AP group on the third day of pregnancy, and its effects on Treg cell frequency and pregnancy maintenance were examined during this period. Following rLIF injections on the fourteenth day of pregnancy, the expression of Foxp3 significantly increased in AP mice (p = 0.02,0.008). Additionally, AP mice injected with rLIF demonstrated a significant reduction in resorption rate (p = 0.01) and a notable increase in birth rate (p = 0.01,0.0005). These findings provide new insights into the potential benefits of LIF in treating RSA patients.

The effect of the anti-leukemia inhibitory factor on the immune system in the Balb/c mice bearing breast cancer induced with 4T1 cells.

BACKGROUND: Breast cancer is one of the most common cancers. Leukemia inhibitory factor (LIF) is considered as one of the effective factors in the growth of breast cancer, and anti-leukemia inhibitory factor antibody is considered as one of the treatment options for this type of cancer. METHODS: Mice models of breast cancer were made with 4T1 cell line and were randomly divided into four groups. The first group included the mice that received anti-LIF (Anti LIF group). The mice in the second group received anti-LIF and doxorubicin (Anti LIF & DOX). The mice in the third group received only doxorubicin (DOX). Finally, the mice in the fourth group did not receive any intervention. 22 days after tumor induction, some of the mice were killed, and their tumor tissues, lymph nodes, and spleens were separated for evaluating P53, Caspase-3, TIM-3, LAG-3, CTLA-4, and PD-1 genes expression. The percentage of regulatory T cells and level of interferon gamma (IFN-γ) and transforming growth factor-beta (TGF-ß) were evaluated. The rest of the mice were kept to check the tumor size and their survival rate. RESULTS: The proposed intervention did not have any significant effect on the tumor growth and the survival rate. However, the expression of P53 gene and Caspase-3 in the tumor tissue of the Anti LIF group had a significant enhancement. In tumor tissues and lymph nodes, the expression of T-bet, PD-1, TIM-3, and LAG-3 genes in the Anti LIF group showed a significant increase. There was no significant difference between groups in the percentage of regulatory T cells and level of IFN-γ and TGF-ß. CONCLUSIONS: The proposed interventions were able to have a direct effect on tumors, but no significant effect was observed on the immune system.

HLA study in Iranian desert Yazd province inhabitants.

Yazd City (1,200,000 inhabitants) is placed in the middle of its Iran desert province and it was constructed on a oasis in ancient times.However,it was a central point on the Silk Road and merchants from both Asia and Mediterranean/European areas crossed through Yazd City.We have studied HLA-A,-B,-DRB1 and DQB1 alleles in Yazd population.Analysys of nine most frequent extended class I and class II haplotypes shows that four of them are specific of this population.The other six haplotypes are also found in Asian and Mediterranean populations in significant frequency. This supports that the nowadays relatively isolated in desert Yazd area also contains people that may bear HLA genes probably originated because of long lasting merchants route between Europe and Asia through the European/Asian Silk Road in addition to other HLA genes close to other Iranian populations, including Kurds.

Association of cytotoxic T-lymphocyte-associated protein 4 polymorphisms with recurrent pregnancy loss: A case-control study.

Background: A large proportion of cases of recurrent pregnancy loss (RPL) are associated with immunological factors. Objective: This study investigated the association between single nucleotide polymorphisms of cytotoxic T-lymphocyte-associated protein (CTLA)-4 gene in women with a history of RPL compared to healthy women. Materials and Methods: A case-control study was performed on 2 groups consisting of 120 healthy women with no history of abortion and at least one delivery (control) and 120 women with a history of 2 or more primary RPLs (case). In addition, 5 mL of peripheral blood sample was taken from all subjects. The frequencies of CTLA-4 rs3087243 and rs231775 polymorphisms were assayed by restriction fragment length polymorphism polymerase chain reaction and rs5742909 using the high-resolution melting real-time polymerase chain reaction method. Results: The mean age of the women in the control and RPL groups were 30.03 ± 4.23 (range 21-37), and 28.64 ± 3.61 yr (range 20-35), respectively. Pregnancy loss numbers ranged between 2-6 in women with a history of RPL, and between 1 and 4 in the successful pregnancy group. Statistical analysis showed a significant difference between the genotypes of GG and AG in the 2 groups in rs3087243 polymorphism (OR 1.00 for GG genotype and OR 2.87 for AG genotype, p = 0.0043). No significant difference was observed in the genotype frequencies of rs231775 and rs5742909 polymorphisms, of the 2 groups (p = 0.37, and p = 0.095), respectively. Conclusion: Our findings indicated that CTLA-4 polymorphism, rs3087243, might be associated with a risk of RPL in Iranian women.

Immunologic deviations in recurrent spontaneous abortion mouse model.

BACKGROUND: Recurrent spontaneous abortion (RSA) or recurrent pregnancy loss (RPL) is an abnormality that has a great impact on women's quality of life. RSA is defined as at least three unexplained abortions (without prior live birth) occurring before the 20th week of pregnancy. AIM: The present review attempts to discuss immunologic deviations in mouse models of RSA. CONTENT: The mating of DBA/2J males with CBA/J female mice has provided specialists with a homologous model of RSA. Much of the research using the CBA/J × DBA/2J mouse model has shown immune system alteration results in rejection. The link between RSA and the immune system suggests new approaches to prevent RSA from an immunological perspective. Rejection in this model is linked with the changed immune system during pregnancy, including change in Th1/Th2 ratio and defects in T and NK cells function, and so forth. IMPLICATIONS: The use of animal models prone to RSA can help a lot to solve the remained mysteries. This study reviews the existing knowledge of immune system roles in the RSA mouse models.

Association between interleukin-32 gene polymorphism and susceptibility to preeclampsia.

OBJECTIVE: To determine association between IL-32 gene polymorphism, and serum levels of IL-32 and susceptibility to preeclampsia (PE). METHODS: The frequency of IL-32 rs9927163 and rs4786370 polymorphisms was determined by PCR-RFLP. Also ELISA was used to determine the levels of serum IL-32. RESULTS: Regarding rs4786370 C/T SNPs, the frequencies of CT, TT genotypes, and T allele were shown to be higher in the PE patients. IL-32 serum level significantly increased in the PE patients. CONCLUSION: Variety of allele and genotype IL32 rs4786370 as well as a rise in serum level of IL-32 can be regarded as a risk factor for PE.

Protective immune response against P32 oncogenic peptide-pulsed PBMCs in mouse models of breast cancer.

High expression of p32 in certain tumors makes it a potential target for immunotherapy. In the present study, the first goal was to design multi-epitope peptides from the P32 protein and the second goal was to compare the prophylactic effects of DCs- and PBMCs- based vaccines by pulsing them with designed peptides. For these purposes, 160 BALB/c mice were vaccinated in 5 different subgroups of each 4 peptides using PBS (F1-4a), F peptides alone (F1-4b), F peptides with CpG-ODN (F1-4c), F peptides with CpGODN and DCs (F1-4d), and F peptides with CpG-ODN and PBMCs (F1-4e). We found a significantly higher interferon-γ (IFN-γ) and granzyme B levels in T cells of F4d and F4e subgroups compared to control (p ≤ 0.05). The result of challenging spleen PBMCs of vaccinated mice with 4T1 cells showed significant up- and down- regulation of Fas ligand (FasL) and forkhead box P3 (Foxp3) gene expression between F4d and F4e subgroups with control, respectively. In addition, a significant change was seen in Caspase3 gene expression of F4d subgroup compared to control (p ≤ 0.05). Supernatant levels of IFN-γ and perforin were significantly increased in F4d and F4e subgroups compared to control. Consequently, significantly lower tumor sizes and prolonged survival time were detected in F4d and F4e subgroups compared to control after challenging mice with 4T1 cells. Accordingly, these results demonstrated that PBMCs pulsed F4 peptide-based vaccine could induce a protective immune response while it is a simple and less expensive vaccine.

Preventive cancer vaccination with P5 HER-2/neo-derived peptide-pulsed peripheral blood mononuclear cells in a mouse model of breast cancer.

This study compared the prophylactic effects from vaccines based on dendritic cells (DCs) and peripheral blood mononuclear cells (PBMCs) by pulsing the cells in-vitro with p5 peptide. The different test groups of mice were injected with free peptide or with peptide pulsed with DCs or PBMCs. Two weeks after the last booster dose, immunological tests were performed on splenocyte suspensions from three mice in each group and the remaining mice (5/each group) were evaluated for tumor growth and survival time. The levels of IFN-γ, granzyme B, and IL-10 were detected in T cells. Additionally, IFN-γ and perforin as well as mRNA levels of some genes associated with immune responses were assessed after challenging the splenocytes with TUBO cells. A significant increase was observed in frequency of CD4+ IFN-γ+, CD8+ IFN-γ+, and CD8+ granzyme B+ T cells, and the perforin of supernatants from mice in the DC and PBMC treatment groups. Significant expression levels of Fas ligand (FasL) and forkhead box P3 (Foxp3) were observed in the DC and PBMC groups. These responses led to smaller tumors and longer survival time in our mouse model of breast cancer. The efficacy of the PBMC-based vaccine in improving the protective immune response makes it a simpler and less expensive candidate vaccine compared with DC-based vaccines.

Contraceptive and molecular function of a novel recombinant vaccine based human leukemia inhibitory factor on Balb/c mice: An experimental in vivo study.

The functional competence of leukemia inhibitory factor (LIF), as immunocontraceptive vaccine in mice, was investigated. Balb/c mice were divided into two groups of vaccinated and controls. The recombinant human LIF (rhLIF) protein and phosphate buffer saline was emulsified with Freund's adjuvant and injected into vaccinated and control groups, respectively. Theinhibition of implantation was evaluated in mice uterine. The concentration of secreted interferon-γ (IFN-γ) and interleukin (IL)-4 were measured in cultured splenocyte of mice stimulated by rhLIF. The expressions of immune responsive gene 1 (IRG-1), cochlin (COCH), amphiregulin(Ar), and heparin-binding EGF-like growth factor (HB-EGF) genes were determined. Mice were assessed for inhibition of fertility after delivery, reversibility of immune response against rhLIF, and survival rate. Active immunization of mice with rhLIF resulted in reduction of the implantation and fertility rate up to 80.49% and 75%, respectively. All mice produced a high titer of anti-rhLIF antibodies in serums and vaginal fluids washes after 16 weeks; however, these antibodies were cleared from vaginal fluid washes after six months. A significant down-regulation in mRNA levels of IRG-1, Ar and HB-EGF was observed in vaccinated group compared to controls; however, no significant change in the expression profile of cochlin gene was detected. The results showed that rhLIF prevented pregnancy in a high percentage of female mice. Although the immunization of female Balb/c mice with rhLIF inhibited fertility and expression of genes associated with this molecule, further studies are needed to support this protein as a suitable candidate for contraceptive vaccine in mammals.

Production and characterization of recombinant human leukemia inhibitory factor and evaluation of anti-fertility effects of rabbit anti-rhLIF in Balb/c mice.

Human leukemia inhibitory factor (hLIF) is a cytokine of interleukin-6 family. This study aimed to evaluate the recombinant production rate of active hLIF by different vector-host systems under various conditions. Moreover, a rabbit polyclonal antibody (pAb) against recombinant hLIF (rhLIF) was produced and its anti-fertility effects were explored in Balb/c mice. Four different constructs including pET22b/hLIF, pET28b/hLIF, pET32b/hLIF and pColdI/hLIF were designed and transformed into BL21-(DE3), Rosetta-(DE3), Origami-(DE3) and Shuffle T7-(DE3) host cells. The expression level and proliferative effect of rhLIF were measured by SDS-PAGE and MTT assays, respectively. Rabbit pAb to rhLIF was produced and characterized using enzyme-linked immunosorbent assay and western blot techniques. The Balb/c mice were divided into two intervention and control groups. Then, they were intraperitoneally injected by purified rabbit anti-rhLIF and non-immunized rabbit pAb, respectively. After sacrifice on day 7, the number of implantation sites was counted. The rhLIF was successfully expressed by pET32b/hLIF and pColdI/hLIF vectors in all hosts with no significant difference in the rate of their expression. The rhLIF was purified and checked for activity. The results showed that it is functionally active and the produced anti-rhLIF pAb could specifically bind to commercial rhLIF. Passive immunization results showed that anti-rhLIF antibody completely inhibited fertility in all injected Balb/c mice compared to controls. Although previous studies showed expression of rhLIF using various methods, using different vector-host systems ensures us of successful biological active expression of it. The pAb against rhLIF could be a powerful tool for inducing in vivo infertility.

Paroxetine modulates immune responses by activating a JAK2/STAT3 signaling pathway.

Paroxetine, a representative of serotonin reuptake inhibitors, has recently gained attention due to its anti-inflammatory properties. However, underlying mechanisms responsible for its immunosuppressive effects remain to be unveiled. To understand the responsible signaling mechanisms, we examined paroxetine's effect on the Janus kinase 2-signal transducer and activator of transcription 3 (JAK2-STAT3) signaling pathway on lipopolysaccharide + phytohemagglutinin-induced human peripheral blood mononuclear cells culture. We also evaluate the possible dependency of paroxetine immunomodulation effects on the 5-HT system of immune cells. Our results indicated that paroxetine attenuates proinflammatory cytokine production (interleukin-1ß [IL-1ß], IL-17, and tumor necrosis factor-α) and increases expression of IL-10 and JAK2/STAT3 evidence for macrophages polarization to M2 subset and functional dendritic cells depletion. In conclusion, paroxetine can exert its anti-inflammatory effects via both the 5-HT systems present in immune cells and the JAK2-STAT3 pathway. Our results also suggest that paroxetine exerted its immunosuppressive effects partially via serotonin. Nonetheless, JAK2/STAT3-modulated paroxetine effects were independent of serotonin, hence sufficiently applicable for inflammation repression.

Association of IL-17 and IL-23 follicular fluid concentrations and gene expression profile in cumulus cells from infertile women at risk for ovarian hyperstimulation syndrome.

This study determined the association between the levels of interleukin (IL)-17 and IL-23 in follicular fluid (FF), as well as their mRNA levels in cumulus cells from infertile women at risk for ovarian hyperstimulation syndrome (OHSS). In this case-controlled study, the control group (n = 40) was infertile women whose partners had male factor infertility, whereas the case group (n = 40) was infertile women at risk of OHSS. IL-17 and IL-23 concentrations in FF were measured using an enzyme-linked immunosorbent assay method, whereas the mRNA expression levels of IL-17 and IL-23 of cumulus cells were determined using RT-PCR. Significantly higher levels of IL-17 were seen in the case group (p = 0.04), whereas there was no significant difference in IL-23 concentrations between the two groups (p = 0.3). The mRNA levels of IL-17 and IL-23 showed no significant differences. In the case group, there was a positive significant correlation between the IL-23 concentration in FF and the oocyte maturation rates (p = 0.01). In the case group, the number of follicles, MII oocytes, immature oocytes, fertilized oocytes and number of embryos were significantly higher than the control group (p < 0.05). Our findings showed that the mRNA expressions of IL-17 and IL-23 were similar in the two groups, and IL-17 was increased in the case group.

Vitamin C and E supplementation effects on secretory and molecular aspects of vascular endothelial growth factor derived from peritoneal fluids of patients with endometriosis.

Endometriosis is an extremely heterogeneous disease and affects about ten percent of the female population during their reproductive years. Recent studies showed that endometriosis is an angiogenesis-dependent disease. Peritoneal macrophages are a well-characterised source of vascular endothelial growth factor (VEGF). The aim of this study was to determine the VEGF gene expression and production in peritoneal macrophages of patients with endometriosis under the effects of vitamins C and E in comparison with control. The lab trial study carried out on 50 patients undergoing laparoscopy and peritoneal fluid samples were collected from them. We compared the VEGF gene expression and production in peritoneal macrophages among groups by using real-time polymerase chain reaction and enzyme-linked immunosorbent assay methods, respectively. Our results showed that gene expressions influenced by vitamin C increased in different concentrations and incubation times, except for the incubation time after 48 h. In the case of vitamin E, this was evident with the exception of vitamin E 50 µM after 24 h and vitamin E 100 µM after 48 h. Our findings indicated that vitamin C and E in different concentrations and incubation times altered VEGF gene expression in the peritoneal macrophages but they had not affected on VEGF productions. Impact statement What is already known on this subject? Previous studies showed that antioxidants play a key role in the inhibition of oxidative stress-induced damages and the reduction of pelvic pain in patients with endometriosis. Vitamin E and vitamin C are the main components in neutralising free radicals. Also, antioxidant consumption such as vitamin C and vitamin E in women with endometriosis showed an inverse correlation between antioxidant intake and endometriosis pathology. What do the results of this study add? Vitamin C and E in different concentrations and times of incubation altered vascular endothelial growth factor gene expression and production in peritoneal macrophages. What are the implications of these findings for clinical practice and/or further research? Further studies are needed to determine the effects of C and E vitamins in different concentrations on vascular endothelial growth factor gene expression and production in peritoneal macrophages and the possible roles of these vitamins in treating endometriosis.

Relative Expression of OX40, OX40L mRNA, and OX40L Serum Levels in Women with Recurrent Spontaneous Abortion.

This study determined the roles of OX40 and OX40L in women with recurrent spontaneous abortion (RSA). We compared the expression of OX40 and OX40L genes in peripheral blood mRNA levels and serum levels of OX40L in women with a history of RSA to the control group. In this case-control study, 40 women with a history of RSA (case group), and 40 others with no history of abortion (control group) were investigated. The expressions of OX40 mRNA and OX40L mRNA were determined in the two groups using the quantitative polymerase chain reaction. Also, the enzyme-linked immunosorbent assay was used to determine the levels of serum OX40L in the two groups. There were no significant differences in the maternal age of women in the two groups (30.1 ± 4.28 years in the case vs. 30.03 ± 4.23 years in the control group). There was no difference in terms of the levels of OX40 and OX40L mRNA between the groups (p = 0.08 and p = 0.56, respectively). In addition, there was no significant correlation between the expression of OX40 and OX40L mRNA levels with age or the number of abortions. The correlation between OX40 and OX40L mRNA levels was not significant. RSA history group turned to show a higher level of serum OX40L than the control group (p = 0.03). In conclusion, our findings demonstrated that the expression of OX40 mRNA and OX40L mRNA was similar between women with a history of RSA and the control group. The elevation of serum OX40L level may be considered as a risk factor for RSA.

Human Leukocyte Antigen (HLA) Class I and II Polymorphism in Iranian Healthy Population from Yazd Province.

The major histocompatibility complex (MHC) genes are the most polymorphic loci in the human genome and have been widely studied in various populations and ethnic groups. Investigations into the HLA genes and proteins have been useful tool for anthropological, transplantation and disease association studies. The polymorphism of the HLA class I (A, B, C) and class II (DRB1, DQA1, DQB1) genes were investigated in 90 unrelated Iranian individuals from Yazd province located in the center of Iran using sequence-specific primers (PCR-SSP). Allele and haplotype frequencies, expected/observed heterozygosity, unbiased expected heterozygosity, number of effective alleles, deviations from Hardy-Weinberg (HW) equilibrium and genetic diversity were computed. A total of 23, 48, 23, 24, 13 and 16 alleles for HLA-A, -B,-C, -DRB1, -DQA and -DQB loci were determined, respectively in the population study. The most frequent allele identified in this study were A*02:01 (18.889%), HLA-B* 51:01 (12.778%), HLA-C* 12:03 (17.033%), HLA-DRB* 11 (24.4%), HLA-DQA* 05:05 (20.55%) and HLA-DQB*03:01 (22.8%).Furthermore, the most frequent 3-locus haplotypes were DRB*11-DQA*05:01-DQB*03:01 (21.1%), HLA-A*02:01- B *50:01- DRB*07:01 (4.9%) and A*26:01 -B* 38:01 -C*12:03(5%). The most 4-locus haplotype were A*11:01 -B* 52:01 -C*12:03 -DRB!*15(2.5%) and A*02:01 -B* 50:01 -DRB1*07:01 -DQB1*02:01(4.5%). The heterozygosity of the study population was confirmed the expected value and not deviated from Hardy-Weinberg equilibrium for all loci (p>0.05). Our study shows a close relatedness between Yazd population and other ethnic group of Iran despite some differences, which may be due to admixture of each one of these groups with each other or foreigner subpopulations during centuries. Moreover, the results of this study suggest that the Iranian population from Yazd province is in close vicinity with the Caucasians populations and far from the Korean and Japanese populations.

Tumor Necrosis Factor Alpha Gene Polymorphism and Association With Its Serum Level in Iranian Population with Rheumatoid Arthritis.

OBJECTIVES: This study aims to determine whether promoter -238 G/A polymorphism in tumor necrosis factor-alpha (TNF-α) gene is associated with susceptibility to rheumatoid arthritis (RA) in Iranian population and serum level of TNF-a. PATIENTS AND METHODS: This case-controlled study was performed on two groups including 90 RA patients (20 males, 70 females; mean age 50.3 years; range 26 to 65 years) and 90 healthy controls (21 males, 69 females; mean age 48.6 years; range 27 to 63 years). We determined the frequency of -238 G/A TNF-a gene polymorphism by polymerase chain reaction restriction fragment length polymorphism. We measured the serum level of TNF-a using enzyme-linked immunosorbent assay method. Also, we determined the association of serum TNF-a level with the polymorphism in RA patients. RESULTS: There was no significant difference in terms of sex and age in the two groups. In the RA group, the genotype frequency of -238 G/A polymorphism was GG (76.6%), GA (17.8%), and AA (5.6%). In the control group, the genotype frequency of -238 G/A polymorphism was GG (83.5%), GA (8.8%), and AA (7.7%). Statistical analysis showed no significant difference in the genotype frequency of this polymorphism between two groups (p=0.07). The serum level of TNF-a were 5.21±1.69 Pg/mL (range 0 to 11.6) in the control group and 62.4±27.1 Pg/mL (range 0 to 117.22) in the RA group (p<0.0001). There was no significant difference in terms of serum TNF-a level and different genotypes in the RA group (p=0.5). CONCLUSION: Our findings demonstrate that the TNF-a -238 G/A gene polymorphism may not represent a significant risk factor for RA in Iranian population and there is no association between the polymorphism and serum TNF-a level in RA patients.

The association between single nucleotide polymorphism in interleukin-27 gene and recurrent pregnancy loss in Iranian women.

BACKGROUND: Recurrent pregnancy loss (RPL) has been defined as two or more miscarriages before 20(th) week of gestation. It seems that IL-27 may reduce inflammatory responses and affect the survival of the embryo during human pregnancy. IL-27 polymorphisms may influence RPL by altering the levels or the activity of gene product. OBJECTIVE: We studied for the first time the association of IL-27 -964 A>G single nucleotide polymorphism (SNP) with RPL in Iranian women. MATERIALS AND METHODS: A case-controlled study was performed on two groups consisting of 150 healthy women with at least one delivery (control group) and 150 women with two or more primary RPLs history (RPL group). The -964 A>G SNP in IL-27 gene was determined by PCR-RFLP technique. Genotype and allele frequencies were compared using (2) tests between two groups. RESULTS: There was no difference between the two groups regarding age of women (29±4.4 [control] vs. 30.84±5.2 years [case]). In the RPL group, the genotype frequencies of -964 A>G polymorphism were AG (49.3%), AA (40%), and GG (10.7%), and in the control group, they were AG (43.3%), AA (48.7%), and GG (8%). There was no significant difference between the genotypes of AA, AG, and GG in two groups (p=0.23). As the frequency of allele A was 64.7% in the RPL group and 70.3% in the control group, the difference in frequency of allele A in -964 A>G between two groups was not significant (p=0.19). CONCLUSION: Our findings indicate that SNP of -964 A>G in IL-27 gene is not a risk factor for RPL in Iranian women.

Relative Expression of Toll-Like Receptors 2 and 7 mRNA in Peripheral Blood of Patients With Hepatitis C.

BACKGROUND: Hepatitis C virus (HCV) is an important human pathogen affecting an estimated 120 - 170 million individuals in the world. Toll-Like receptors (TLRs) are pattern-recognition receptors that recognize pathogen-associated molecular patterns, and stimulate immune responses. OBJECTIVES: The aim of this study was to determine the mRNA expression level of TLR2 and TLR7 in HCV-infected patients in comparison with normal controls. PATIENTS AND METHODS: Nineteen consecutive patients with HCV infection and nineteen sex and age-matched healthy controls were studied in a case-controlled research. RESULTS: Our results showed that the expressions of TLR7 in HCV infected samples were significantly increased in comparison those of the controls (P = 0.02), while the expression of TLR2 was similar between the case and the control group (P = 0.8). There were no associations between the expression levels of TLR2 and TLR7 with HCV viral load and HCV genotypes. Also, there was no association between viral load and genotypes of the virus. CONCLUSIONS: Our findings showed that HCV infection could lead to increased expression level of TLR7 mRNA in peripheral blood cells of HCV infected samples. The viral load and genotypes of HCV did not affect the mRNA expression levels of TLR2 and TLR7.

The serum level of transforming growth factor beta1 and its association with Foxp3 gene polymorphism in Iranian women with recurrent spontaneous abortion.

The aim of the study was to determine the role of transforming growth factor-beta1 (TGF-ß1) and Foxp3 (rs3761548) promoter polymorphisms in Iranian women with recurrent spontaneous abortion (RSA). Eighty women with RSA were compared with eighty in a control group. Serum levels of TGF-ß1 were measured using ELISA and Foxp3 (rs3761548) promoter polymorphisms using a PCR-RFLP technique. In addition, serum levels of TGF-ß1 were compared in different genotypes in the two groups. The women's ages in the two groups were similar (30.15 ± 4.42 years [RSA] vs. 29.97 ± 4.51 [control]) as were serum TGF-ß1 concentrations in case and control groups (53.42 ± 2.08 ng/ml in control and 56.31 ± 2.58 ng/ml in the RSA group; p = 0.4). Furthermore, there was no significant difference in the genotype frequencies of the rs3761548 Foxp3 gene between the two groups (p = 0.3) and the levels of TGF-ß1 were similar in different genotypes. In conclusion, the data indicate that serum TGF-ß1 levels and Foxp3 (rs3761548) promoter polymorphism is not a risk factor for RSA and that there is no association between the polymorphism and serum TGF-ß1 levels.

Association of IL-17A and IL-17 F gene polymorphisms with recurrent pregnancy loss in Iranian women.

PURPOSE: Recurrent pregnancy loss (RPL) is defined as the occurrence of two or more miscarriages before the 20th week of pregnancy. T helper17 cells are a novel subset of T cells, which secrete IL (Interleukin)-17 and are known to be involved in inflammation, autoimmunity and rejection of non-self tissues. Herein, we studied the association between IL-17A rs2275913 and IL-17F rs763780 gene polymorphisms with RPL in Iranian women. METHODS: A case-controlled study was performed on two groups consisting of 85 healthy women with at least one delivery and 85 women with the history of two or more RPLs. The frequency of IL-17A rs2275913 and IL-17 F rs763780 polymorphisms were determined by PCR-RFLP. RESULTS: In the RPL group, the genotypes frequencies of rs2275913 polymorphism were GG (8.2 %), AG (30.6 %), and AA (61.2 %) and in the control group, were GG (3.5 %), AG (42.4 %) and AA (54.1 %). Statistical analysis showed no significant difference between the genotypes of AA, AG and GG in the two groups (p = 0.1). The genotypes frequencies of rs763780 polymorphism were TT (43.5 %), TC (49.4 %) and CC (7.1 %) in the RPL group; whereas the frequencies were TT (25.9 %), TC (70.6 %) and CC (3.5 %) in the control group. Statistical analysis revealed a significant difference in the TT, TC, and CC genotypes frequencies between the case and the control groups (p = 0.01). CONCLUSIONS: Our findings indicate that IL-17F polymorphism, rs763780, might be associated with a high risk of RPL in Iranian women.