Evaluation of Soluble Toll-Like Receptors 2, 4, 9 and Their Damps Signaling Molecules (HMGB1 & HSP70) in Breast Cancer Patients of Basrah Province

Pregnancy is a unique symbiotic union between mother and fetus which contradicts many of the general rules of immunology. Clarifying the cellular mechanisms involved in the maternal-fetal cross talk will enable us to establish therapies for gestational problems. Recurrent spontaneous abortion (RSA) one of the most important problems occurs during the first trimester of human pregnancy. RSA with unknown causes is may correlated with immunological alterations. The study aimed to investigate the immune disorders regarded with natural killer cells (NK) and their activation through using the inhibitory and activating receptors associated with human leukocyte antigen-C molecule. The study had been applied to women with recurrent abortion that attending the department of obstetrics and gynecology of al-Fayha teaching hospital and Basrah Maternity and Children hospital from January 2021 until the end of May 2021. A total of 80 samples were included, 55 patients with recurrent miscarriage in the first trimester of pregnancy, their ages ranged from 20-35 years, and 25 healthy pregnant women similar to patients in terms of age and pregnancy period recruited as a control group. The samples were divided into two age groups: group 1 (G1) ranged (2027) years and group 2 (G2) ranged (28-35) years, the results showed no significant difference (p ≤ 0.928) according to age. II Complete blood count (CBC) was performed to determine total lymphocytes which showed elevated levels in women with RSA in the two age groups. Peripheral natural killer (p NK) cells were measured by flow cytometry and determined as CD3CD16+ which showed a significant difference (p ≤ 0.030) in patients in comparison with control. CD16 CD56 a subset that covers 90% of p NK cells had been evaluated and showed significant rising in aborted women than healthy pregnant women. Flow cytometric evaluation of killer-cell immunoglobulinlike receptor (KIR2DL1) that is expressed on NK cells surface had been determinate and showed high significant elevation in healthy pregnant women in comparison with RSA women. Furthermore, soluble human leukocyte antigen-C (HLA-C) showed high significant levels in the control group in comparison with aborted women. Statistically, a positive correlation between KIR2DL1 (CD158a) and HLA-C molecules cells (p ≤ 0.01) had been found.

HLA-G is one of the non-classical HLA class I molecules and it is known to be implicated in a tumor-driven immune escape mechanism in malignancy. The purpose of this study was to investigate soluble HLA-G (sHLA-G) and HLA-G expression in breast cancer. sHLA-G was determined by enzyme-linked Immunosorbent assay (ELISA) from plasma samples of 70 breast cancer patients and 35 healthy controls. HLA-G expression in breast cancer lesions was also analyzed by immunohistochemistry staining. The presence of HLA-G expression were analyzed and found to be involved in breast carcinogenesis. Levels of sHLA-G were higher in the breast cancer group compared to the control group and sHLA-G concentration could be used as a diagnostic marker for detecting breast cancer.

The aim of the present study was to determine the effect of two prepared antigens fromMethicillin-resistantStaphylococcus aureus (MRSA), whichincluded crude alpha toxin (Hla) and heat inactivate bacterial suspension (HIBS) in rats through the examination of the inflammatory response of both antigens on primary and secondary lymphoid organs (thymus gland and spleen), respectively. Examination of sections from spleen of rats treated with HIBS antigen induced high immune response compared with rats treated with Hla antigen which induce mild immune response by established changes. The histological observations on sections from thymus gland of rats treated with HIBS had the ability to induce different changes than Hla antigen compared with control group.

Cathelicidin is important components of the innate defense in the urinary tract. The aim of this study was to characterize whether these anti -microbial peptide are important for developing urinary tract infections (UTIs). This aim was investigated by comparing blood urinary peptide levels of UTI patients’ infection to those of controls. A casecontrol study was conducted at Basrah province (Basrah general hospital and Al-Sadr Educational hospital) during the period from 18 November 2018 to 15 April 2019. 60 patients with confirmed UTI and 30 healthy controls without UTI. Plasma and urine levels of cathelicidin were determined using an enzyme linked immunesorbent assay (ELISA) kit. The mean concentration of anti-microbial peptide cathelicidin (AMPccl37ng/ml) was highly significant P≤0.001, P≤0.000 in urine and sera respectively, with no significant difference correlation between the type of bacterial infection and concentration of ccl37 in urine and sera. Conclusion Urinary cathelicidin is microbial markers that may assist the diagnosis of UTI in woman.

Current study included Seroepidemiological Screening of 450 sera from unmarried women, within age ranged between (15 – 45) year by using IgG and IgM Enzyme Linked Immunosorbent Assay (ELISA) test. Sera collected from areas of Basrah province and from different classes ( students , officials and housewives ) , and according to the primary health care department program the province was divided into eight sections . Results documented that the prevalence rate of the disease in unmarred women in the whole province was 58% while the prevalence rates of sections was as follows : Sec I 63.2% , Sec II 51.8% , Al – Midayna 75% , AI – Qurna 64% , Al – Hartha 70% , Shatt – al – Arab 55% , Abu – al – Khaseeb 67.5% and Al – Zubair 36.3% . The higher rate recorded in Al – Midayna and lower rate in Al – Zubair . According to age group there is a significant difference ( P<0.025 ) and the results showed that (15 19) year with the rate of infection 64.54% , (30-34) year with rate 60.7% , (35-39) year with rate 60%, while the results upon sociodemographic data recorded that the housewives (all of them with low level of learning) represents rate 62.4% in comparison with students 45% and officials 44% , with a significant differences ( P < 0.05 ) .

Corresponding author: Hadeel S. Touma Email: fatimaalzahraa134@yahoo.com Department of Biology College of Science University of Basrah Iraq. ABSTRACT Objective: FOXP3 is a forkhead box transcription factor, a defining hall mark of Treg cells, and its functions as the master regulator in the development and function of regulatory T cell. Treg cells are sub set of T lymphocytes that the main functions regulate the immune response through suppress the proliferation and cytokines production of effector T lymphocyte. The infiltration of FOXP3+ regulatory T cells into invasive tumors has been reported to be associated with survival in a variety of cancers. The prognostic significance of FOXP3+ tumor infiltrating lymphocytes (TILs) in breast cancer, however, remains controversial. We investigated whether there were significant numbers of FOXP3-positive Tregs in breast cancer using immunohistochemistry, and whether the presence of FOXP3-positive Tregs was associated with other prognostic factors, such as stage or histologic grade, receptor status, lymph node involve and metastasis. Methods: A tissue microarray (TMA) including 24 ductal and 11 lobular breast cancers was stained with antibodies recognizing FOXP3 by immunohistochemical techniques. Foxp3+Lymphocyte counts were correlated with clinicopathological parameters. Results: The Foxp3-positive was more prevalent whereas Foxp3-negative was rare. Lymph node displayed significantly (p < 0.01) higher FOXP3+ lymphocyte infiltration and no metastasis was also significant (P˂0.02) associated with higher FOXP3+ TIL counts. In contrast, FOXP3+ lymphocyte infiltration was not linked with other clinic pathological parameters. Conclusion: Our results showed a significant increase in the proportion of these cells in the tissue of breast cancer patients. The prognostic value of FOXP3+ TILs in breast cancer differs depending on lymph node and metastasis.

Background: Currently, cancer as major problem around the world threatens human health and has a high incidence in developing countries. Many reports have showed that there is a dysfunction in the immune system of cancer patients. Interleukin-17 (IL-17) and transforming growth factor-β (TGF-β) are increased in the generated through cancers, and IL-17 is considered as a pro-inflammatory cytokine; IL-17 is an angiogenic factor and promotes tumor growth. Materials and Methods: In this study TGF-β and IL-17, mRNA gene expression has been measured in peripheral blood of 35 breast cancer patients and 35 normal age-matched women using quantitative real-time polymerase chain reaction method with master mix reaction containing SYBER Green. β-actin gene was used as housekeeping gene. Results: Our data demonstrated a significant upregulation of TGF-β and IL-17 gene expression (P < 0.05) in patient's peripheral blood compared to normal healthy control. Levels of genes expression were higher in the breast cancer group compared to the control group. Conclusion: TGF-β and IL-17 expression could be used as a diagnostic marker for detecting breast cancer.

Corresponding author: Nada Fadil Abass Email: dewdropaa90@gmail.com Department of Biology College of Science University of Basrah Basrah Iraq. ABSTRACT Objective: The present study aimed to isolation and identification of C. albicans from women infected with vaginal candidiasis, then Preparation of whole cell fraction (WCF) antigen and testing immunization efficiency by evaluating the humoral immune response (by measuring IgG level in serum) and cellular immune response (by measuring IFN-γ and IL-4 concentration). Methods: Seventy five clinical specimens were collected from women suspected to infect with vaginal candidiasis were cultured on Sabouraud Dextrose Agar (SDA) and CHROMagar Candida media and identified by biochemical test. Three of them identified by molecular diagnosis by using ITS1 and ITS4 primers. BalB/c mice were immunized subcutaneously and intradermaly with C. albicans strain M366B whole cell fraction antigen for 21 days to evaluate humoral and cellular immune response represented by IgG, Th1 and Th2 cytokine level by microiterplate ELISA. Results: Among total positive cases of vaginal infections only 43 (57.3%) was caused by yeast infections, whereas Candida albicans was recorded more dominance (30/43) 69.77% among other yeast infections. Humoral and cellular immune response represented by IgG, Th1 (IFN-γ) and Th2 (IL4) cytokine level were measured respectively. IgG level appeared in high concentration in immunized mice by comparison with control groups in a high significant differences (P<0.01), and immune response represented by (IL-4) concentration in immunized group was higher than those in control mice in a significant differences (P<0.01); while there was no significant differences in (IFN-γ) concentration between immunized and control groups. So WCF antigen can efficiently stimulate both humoral and cellular immune response by elevation both IgG and IL-4 concentration in sera of immunized mice. Conclusion: Different species of yeast are causing vaginal candidiasis, C. albicans was most prevalence than others in infection in V. candidiasis, . WCF antigen was immunogenic in regarding in inducing IgG and IL-4 secretion.

Objective: This study aimed to assess the role of Fas/Fas ligand (FasL) system in the progression of type 2 diabetes mellitus (T2DM). Materials and Methods: This study included 100 participants – 30 persons as a control group and 70 patients with T2DM (35 males and 35 females) and their ages were ranged from 40 to 70 years. The patients were distributed into two groups according to gender and duration of the disease: newly diagnosed group for short duration ≤5 years and chronic diagnosed group for long duration >5 years. Serum sFas and sFasL levels were measured by enzyme-linked immunosorbent assay technique, and also, lipid and glucose profile were measured by COBAS analyzer. Results: The results revealed a significant (P ≤ 0.05) decrement in the levels of FasL in T2DM than controls while the levels of Fas were increasing significantly (P ≤ 0.05) in T2DM than controls. Hemoglobin A1c (HbA1c) and fasting blood glucose were negatively correlated with FasL, while high-density lipoprotein was positively correlated with it, and whereas HbA1c positively correlated with Fas, the gender and duration of disease did not show any correlation with the disease. Conclusion: Our findings suggest that hyperglycemia causes increase in Fas levels which lead to dysfunction of pancreatic β-cell in T2DM.

The study was aimed to investigate the association between IL-10 (-592A/C) gene polymorphism with the progression of type 2 diabeteis mellitus in Basrah Province. This study included (100) subjects (30) person as a control group and (70) patients with T2DM. The patients were distributed as two groups according to Their gender and duration of the disease: group for short duration (≤ 5 years)  and group for long duration( u003e5 years) . Lipid profile and glucose concentrations were measured by COBAS analyzer while IL-10 (-592A/C) gene polymorphism was genotyped by using (RFLP-PCR) technique. CC genotype frequency showed a significant decrease while CA genotype  revealed significant increase ( p≤0.05) in T2DM compared with controls .No significant differences were observed in the allelic frequencies between both groups. Gender and duration of diabetes didn’t show any significant differences. There is significant association between CA genotype with the risk of T2DM  (OR=1.50, 95% CI = 1.035-2.173). We concluded that IL-10 (-592A/C) gene polymorphism contribute in development of T2DM.

This study was included the preparation of vaccine against fascioliasis from three Fasciola gigantica antigens, somatic antigen, excretory-secretory antigen and partially purified cathepsin antigen. Their vaccine efficacy was tested in rabbits, by using 12 Lepus lepus arabica rabbit, aged 2 months, similar weight. The rabbits were subdivided to three groups each of four rabbits and was vaccinated with one antigen emulsified in freund adjuvant. In compare to control groups Which involved 4 rabbits vaccinated with PBS and other four left without vaccination, challenge infection was done by vaccination the animals with 90 metacercaria obtained from experimentally infected intermediate host. All animals were killed 12 weeks after challenge. Protein concentration of all prepared antigens and the molecular weight of P.P.CatA was also measured, which reached (1.5, 1.2 and 0.04) mg/ml in SA, ESA and P.P.CatA respectively, while P.P.CatA 26.7 KD. Protection estimated by followed number of aspects, the first aspect of this study, was the determination the hepatic damage which was evaluated using gross morphometric observations depending upon the severity and intensity of lesions. Significant differences (p<0.05) was noted between vaccinated and control groups. The second aspect was concerned with estimation of fluke number and reduction rate, the high rate (81.3%) recorded in animals vaccinated with P.P.CatA while (39.53%) of those vaccinated with SA. Significant differences (p<0.05) was documented in the length of fluke in animals vaccinated with P.P.CatA. No significant differences were noted in fluke burden (width, length, weight) and fecal egg counts, Significant differences (p<0.01) were noted in egg viability and reduction rate in the vaccinated animals in comparison with control group. ELISA was used for monitoring of IgG levels in all vaccinated and control animals, before and after infection. The IgG increased in the sera of animals vaccinated with SA, ESA and P.P.CatA within 1 week after the first injection. A boosting of the antibody responses was observed in all groups following both the second and third immunization. An increasing in the antibody titers in all vaccinated animals was observed within 2 weeks following challenge. IgG in all vaccinated groups remained high throughout the infection but decreased at approximately 6 weeks after infection, and then remained constant throughout the subsequent weeks. Antibodies to antigens in the sera of the control group increased within the first 2 to 4 weeks after infection but decreased at approximately 8 weeks and then remained constant throughout the subsequent weeks. Accordingly, the results of these aspects could be consider a P.P.CatA as a first best vaccine then secondly ESA.

Forty cases of women sever from recurrent pregnancy loss (RPL) compared with 40 normal women for determining the cytokine gene polymorphism in the promoter region of : tumor necrosis factor-alpha TNF-α (-308 G/A), interleukin IL-6 (-634 G/C) and IL10 (-592 C/A) in aborted women of Basra /Iraq. The results indicated that there was an association between RPL and the polymorphisms in inflammatory cytokines (IL-6, TNF-α), while IL-10 gene did not showed any effect on both cases of RPL and normal cases.

The objective of this study was to compare the concentrations of Th1 (IFN-γ , TNF-α) and Th2 (IL-4,IL-10) cytokines in the serum of normal pregnant and recurrent spontaneous abortion (RSA) women in 1st trimester as well as in the normal fertile non pregnant women. In RSA women there was a significant increasing in the mean concentration of Th1 cytokines (TNF- α and IFN- γ) as well as in Th1/TH2 than normal pregnant and non pregnant women. In normal pregnant women there was significant increasing in the mean concentration of Th2 cytokines (IL-4 and IL-10). RSA was associated with increasing concentration of Th1 cytokines while successful pregnancy was associated with increase concentration of Th2 cytokines in the first trimester of pregnancy, indicating Th2 bias in normal pregnancy and a Th1 –bias in unexplained RSA.

This study designed to analyzed humoral response related to measuring the serum levels of immunoglobulines (IgG , IgM and IgA) and complements corporeats ( C3 and C4) in (20) thalassemic patient and (10) control subject. IgG only recorded a high significant differences between two studied group - whereas no significant differences were found in levels of remaining humoral components ( IgM , IgA , C3 and C4 ) between thalassemic patients and control group .

HLA-G is a nonclassical, class I major histocompatibility complex (MHC) gene that exhibits immunomodulatory properties and likely plays a role in the maintenance of successful pregnancy. In this study, we investigated the role of HLA-G polymorphisms on risk for recurrent pregnancy loss (RPL) and on circulating levels of soluble (s)HLA-G in Iraqi women. DNA and plasma were obtained from blood samples collected at 9-12 weeks gestation from 50 women with RPL and 50 healthy pregnant women in Basrah province, Iraq. As measured by ELISA, median sHLA-G levels were significantly lower in the RPL cases compared to healthy controls (21.4 vs. 38.8 U/ml, respectively; P=0.025), and decreased with increasing maternal age (P=0.0051). However, HLA-G allele and haplotype frequencies did not differ significantly between cases and controls (P values ≥0.12 for all tests). In contrast, homozygosity for the C allele (CC) at a tri-allelic promoter polymorphism, -725C/G/T, was associated with lower concentrations of sHLA-G compared to the CG or CT genotypes (median levels 21.1 vs. 40.1 vs. 42.6 U/ml, respectively; P=0.0089). These results demonstrate that HLA-G genotype influences circulating sHLA-G levels during pregnancy but is not significantly associated with risk of RPL.

Objectives: This study was performed to determine the local (tissue) and systemic immune responses in rats infected with Giardia lamblia. Materials and methods: Rattus noruigicus rats aged 2 months were inoculated with 103 Giardia lablia cyst. After 7 and 14 days of infection samples (blood and small intestine) collected from both control and infected rats. Local immune response was assessed by counting the inflammatory cells in small intestine, mast cell number and lymph node diameter. Whearas systemic immune response was carried out by measurement of total and differential WBC count, phagocytic activity of polymphonuclear cells by NBT stain and histamine levels in blood plasma by ELISA kit. Results: Results showed an infiltration of inflammatory cells in all parts of small intestine and an increasing in lymph nodules diameter. Also, there is an infiltration of mast cells. Results of systemic immune response revealed a high significant differences in total and differential WBC count between two studied groups. Moreover, there was an increasing in phogocytic activity of polymorphonuclear cells and histamine plasma levels. Conclusion: Recent data indicated that there is a local and systemic immune response during acute phase of Giardia lamblia infection in experimental animals.

Present work included the study of cellular and humoral immune response in athletes before and after running on treadmill. Results of cellular immune response related with differential white blood cell count indicated an increasing in mean number of neutrophils after exercise in comparison with their number before exercise but it is not significant, whearas the eosinophils and basophils did not show any significant differences in their numbers between the two intervals of study (before and after exercise). Also there is a significant elevation in numbers of monocytes after an exercise in comparison with athletes before an exercise and control group. The study also indicated that there is non significant increasing in lymphocytes after an exercise when compared with their number before an exercise. Moreover results of total white blood cell count indicated that there is an increasing in their numbers in athletes after an exercise. Other indicator of cellular immune response is the presence of an elevation in phagocytic activity of PMNs after an exercise in comparison with their activity before an exercise. In relation with results of humoral immune response, present results demonstrated the significant increasing in IgM and IgA in athletes after an exercise while IgG donot revealed any significant differences in their level among the studied groups. Introduction : In recent years considerable interest has been directed to the effects of exercise on immune function. As demonstrated in animal experiments ( 1&2), moderate exercise appears to stimulate the immune system. However, several studies indicated that intense training increases susceptibility to illness. These illness range from persistant colds, sore throats to flu-like illnesses and post-viral fatigue syndrome (3). Also, there is a rather evidence indicated that exercise is a life style that offers some protection against malignancy (4). It has become clear that moderate exercise stimulates the immune system and may be somewhat responsible for exercise related reduction in illness. However, strenuous exercise induces AL-Qadisiya Journal For Science Vol .17 No.3 Year2012 9 immunosuppression in the recovery period in athletes (5 &6). Moreover, Regular exercise has been reported to have a several favorable effects on physiological, psychological and immunological functions (7 ; 8 and 9). In present work it has been tried to through a sight on the effect of an exercise related with running on treadmill on immunoglobulines level and phagocytic activity of polymorphonuclear cells before and after exercise in comparison with control group. Materials and Methods: Twenty student from first class in Basrah college of sports were performed an exercise related with running on treadmill for 15 minute with a speed of 12 Km/hr and 20 healthy age-matched sedentary or control group were enrolled in this study. Sample collections: Blood samples (5 ml) were taken once from sports student (before and after exercise) and from control group, then the samples separated as a follows: 1. (1 ml) put in EDTA (ethylin diamin tetra-acetic acid) tube for total and differential white blood cells count. 2. (2 ml) put in heparin tube for measuring the phagocytic activity of poly morphnuclear cells by chemiluminescence. 3. The remaining (2 ml) of blood were centrifuged for serum collection and measuring immunoglobulines (IgG, IgM, IgA) concentration. Methods of cellular immune response determination

Forty sera of patients infected with Leishmania donovani and (10) sera from apparently healthy subject were tested by single radial immunodiffusion to visualize the concentration of IgG, IgM, IgA, C3 and C4. Recent results showed that there was a highly significant increase in the level of IgG, IgM, C3 and C4 but there were no significant differences in the level of IgA between the two—studied groups.