A randomized clinical trial evaluating the immunomodulatory effect of convalescent plasma on COVID-19-related cytokine storm.

Evaluating the effect of convalescent plasma (CP) on some cytokine storm indices in severe COVID-19 patients. Totally, 62 patients were randomly assigned into two groups for this clinical trial. Patients in the intervention group received one unit (500 mL) plasma on the admission day plus standard drugs while the controls merely received standard treatments. Eventually, primary and secondary outcomes were evaluated. In the CP group, compared with controls, the mean levels of lymphocytes and IL-10 significantly increased while the levels of IL-6, TNF-α, and IFN-γ decreased (p < 0.05). The length of in-hospital stay, and mortality rate did not significantly reduce in the CP group compared with controls (p > 0.05) while WHO severity scores remarkably improved (p = 0.01), despite the higher frequency of underlying diseases among the CP group (66.7%) vs. controls (33.3%). Although CP has a remarkable immunomodulatory and antiviral potential to improve the cytokine storm and disease severity in COVID-19 patients, it did not considerably affect the mortality rate.

Salivary Interferon Gamma and Interleukin-4 Levels in Patients Suffering from Oral Lichen Planus.

OBJECTIVE: Oral lichen planus (OLP) is a chronic inflammatory disease. Immunological factor may act as etiological factor. The cellular immune cells such as T cells are impor- tant in pathogenesis. Interferon gamma (IFN-γ) and interleukin 4 (IL-4) are secreted by T-helper 1 (Th1) and Th2, respectively. The aim of this study was to investigate the cor- relation between salivary levels of IFN-γ and IL-4 with OLP. MATERIALS AND METHODS: This case control study included sixty three Iranian OLP patients who were selected from the Department of Oral Medicine of Ahvaz Jundishapur University of Medical Sciences from January to July 2013. An equal number of healthy volunteers were also selected as a control group. The OLP patients were then divided into two follow- ing sub-groups: reticular (n=30) and erythematous/ulcerative (n=33). All patients had no systemic disease and received no medication. IFN-γ and IL-4 levels in whole unstimulated saliva (WUS) were measured using the enzyme-linked immunosorbent assay (ELISA) test. Data analysis was done using t test, ANOVA, least significant difference (LSD) test, and the Kruskal-Wallis test. RESULTS: Reticular OLP patients showed higher salivary IFN-γ (7.74 ± 0.09 pg/ml ) and IL-4 (3.876 ± 0.05 pg/ml) levels compared with the control group, indicating that difference was significant. Salivary IFN-γ/IL-4 ratio significantly increased compared with control group (P=0.042). Salivary IFN-γ and IL-4 levels between sub-groups (re- ticular and erythematous/ulcerative) were not significantly different (2.6 ± 0.06 and 2.3 ± 0.05, respectively, P<0.05). CONCLUSION: Salivary IFN-γ and IL-4 levels were increased in OLP patients. An increase of salivary IFN-γ/IL-4 ratio in OLP patients showed that Th1 might have a dominant role in the OLP pathogenesis.

Investigation of The Association between Salivary Procalcitonin Concentration and Chronic Periodontitis.

OBJECTIVE: Chronic periodontitis is the most common form of periodontal disease. Chang- es in biomarkers seem to be associated with the disease progression. Procalcitonin (PCT) is one of these biomarkers that are altered during infection. This study was established to investigate the relationship between periodontitis as an infectious disease and salivary PCT. MATERIALS AND METHODS: This case-control study was performed on 30 patients with gen- eralized chronic periodontitis and 30 health individuals as control group who were referred to Dental School, Jundishapur University of Ahvaz, Ahvaz, Iran at Feb to Apr 2014. The saliva samples were collected and analyzed by the enzyme-linked immunosorbent assay (ELISA) method. Data analysis was performed using t test with the SPSS (SPSS Inc., Chicago, IL, USA) version 13. RESULTS: In both groups, age and sex distribution values were not significantly differ- ent. The concentrations of salivary PCT in controls and patients ranged from 0.081 pg/ mL to 0.109 pg/mL and from 0.078 pg/mL to 0.114 pg/mL, respectively. The statistically significant differences between the two groups were not observed (P=0.17). CONCLUSION: It seems that salivary PCT concentration is not affected by disease progres- sion. Therefore, PCT is not a valuable marker for the existence of periodontal disease.

Increased P-35, EBI3 Transcripts and Other Treg Markers in Peripheral Blood Mononuclear Cells of Breast Cancer Patients with Different clinical Stages.

PURPOSE: Currently, cancer as a major problem around the world threatens human health and has a high incidence in developing countries. Many reports have indicated that patients suffering from cancer demonstrate decreased antitumor immune responses as well as a high prevalence of T regulatory population. It has been reported that Foxp3(+)Tregs exert suppression by cell contact-dependent mechanisms which are mediated by soluble factors such as immunosuppressive cytokines like IL-10, TGF-ß and IL-35. Consequently there is a great need to identify prognostic and diagnostic biomarkers of regulatory T cells for vaccine and drug development. METHODS: In this study IL-10, TGF-ß, IL-35 and Foxp3 mRNA gene expression has been measured in peripheral blood of 40 breast cancer patients and 40 normal age-matched women using quantitative real-time PCR (qRT-PCR) method with Master Mix reaction containing SYBER Green. GAPDH gene was used as housekeeping gene. RESULTS: Our data demonstrated a significant up-regulation of IL-10, TGF-ß, P35, EBI3 and Foxp3 gene expression in patients' peripheral blood compared to normal healthy controls (p<0.05). CONCLUSION: The data suggests that the immune system is suppressed in breast cancer patients, which may be due to elevated Treg cells population. These results may be useful for diagnostic or therapeutic purposes. However it may require more investigations.

Augmentation of lymphocytes activation and T cell modulation by the extracts from some Euphorbia species.

CONTEXT: Euphorbia is an important Euphorbiaceae genus that is traditionally being used for various infections, inflammation, and cancer. OBJECTIVE: The present study investigated the possible in vitro immunomodulatory effect of three species of Euphorbia genus including Euphorbia microciadia Boiss, Euphorbia osyridea Boiss, and Euphorbia heteradenia Jaub. & Sp. on lymphocyte activation and cytokine secretion. MATERIALS AND METHODS: Human lymphocytes were cultured in the presence of various concentrations (0.1-200 µg/ml) of the butanol/hexane extracts of the plants in the presence or absence of phytohemmagglutinin (PHA). The activation of lymphocytes after 48 h was determined by a proliferation assay. The release of T cell cytokines was studied to determine the dominant T cell subsets involved in the immune response. RESULTS: All three plant extracts increased the proliferation of PHA-treated lymphocytes (maximum; 132% of control). Extract treatment of lymphocytes in the absence of PHA resulted in an increased proliferation of the cells indicating their lymphocyte mitogenic activity (maximum at 10 µg/ml E. microciadia extract; 494.5 ± 42.2% of control, p < 0.01). The extracts of E. microciadia and E. osyridea could increase IL-4 and IL-10 secretion but not IFN-γ production showing their capacity to deviate immune response toward a Th2 pattern. Euphorbia heteradenia did not change the release of IL-4 and IFN-γ cytokines but increased IL-10 production. The three extracts stimulated lymphocytes to produce IL-17 which showed their possible effects on Th17 cells activation. CONCLUSION: The studied extracts had the ability to modulate T cell responses suggesting their possible beneficial effects on immune host defense.

Concomitant Increase of OX40 and FOXP3 transcripts in peripheral blood of patients with breast cancer.

BACKGROUND: Regulatory T cells (T-regs) have an important role in cancer by suppression of protective antitumor immune responses. Regulatory T cells express the forkhead/winged helix transcription factor (FOXP3) and OX40 molecules which have important regulatory roles in the immune system. OBJECTIVE: To evaluate FOXP3 and OX40 transcripts in the peripheral blood mononuclear cells of women with breast cancer. METHODS: Blood samples from 40 women with histologically-confirmed infiltrating ductal carcinoma of the breast and 40 healthy volunteer women without a history of malignancy or autoimmune disorders were collected. The abundance of FOXP3 and OX40 gene transcripts were determined by quantitative real-time PCR (qRT-PCR). RESULTS: There was a significant positive correlation between FOXP3 and OX40 gene expression in women with breast cancer in a stage dependent manner. CONCLUSION: This finding emphasizes the importance of T-regs as predominant targets for breast cancer immunotherapy.

Prevalence of hepatitis B and C infections and HCV genotypes among haemophilia patients in ahvaz, southwest iran.

BACKGROUND: Transfusion-transmitted hepatitis is the most important cause of transmitted infections by the parenteral route in patients with haemophilia. OBJECTIVES: This study was performed to determine the prevalence of HBV, HCV, and different genotypes of HCV among haemophilia patients in Ahvaz city, southwest Iran. PATIENTS AND METHODS: A cross-sectional study was conducted on 87 haemophilia patients referred to the Hemoglobinopathy and Thalassemia research centre during February 2008 to March 2009. Patients, sera were tested for HBsAg and anti-HCV using ELISA and confirmed by PCR (HBV) and RT-PCR (HCV). HCV genotypes were determined with HCV genotype specific primers using HCV genotyping kit. RESULTS: The overall prevalence rate of HBsAg and anti-HCV were 1.1% (95% CI: 0-3.39) and 54% (95% CI: 43.5-64.4), respectively. Forty two of the anti-HCV patients (89.3%) were also HCV RNA positive. The prevalence of anti-HCV seropositivity was significantly higher (P = 0.0008) among patients who had started to receive transfusions before implementation of blood donor screening. Moreover, the number of transfusion were significantly associated with anti-HCV and HCV RNA positivity (P = 0.0041 and P = 0.023, respectively). The predominant HCV genotype among haemophilia patients in our region was 1a (26/42, 61.9%), although genotypes 1b and 3a were found in 26.1% (11/42) and 11.9% (5/42) of the patients, respectively. CONCLUSIONS: It appears stringent donor selection procedures reduced HCV infection in multi-transfused patients, but it is still serious risk for these subjects.

Relationship between cell compatibility and elastic modulus of silicone rubber/organoclay nanobiocomposites.

BACKGROUND: Substrates in medical science are hydrophilic polymers undergoing volume expansion when exposed to culture medium that influenced on cell attachment. Although crosslinking by chemical agents could reduce water uptake and promote mechanical properties, these networks would release crosslinking agents. In order to overcome this weakness, silicone rubber is used and reinforced by nanoclay. OBJECTIVES: Attempts have been made to prepare nanocomposites based on medical grade HTV silicone rubber (SR) and organo-modified montmorillonite (OMMT) nanoclay with varying amounts of clay compositions. MATERIALS AND METHODS: Incorporation of nanocilica platelets into SR matrix was carried out via melt mixing process taking advantage of a Brabender internal mixer. The tensile elastic modulus of nanocomposites was measured by performing tensile tests on the samples. Produced polydimetylsiloxane (PDMS) composites with different flexibilities and crosslink densities were employed as substrates to investigate biocompatibility, cell compaction, and differential behaviors. RESULTS: The results presented here revealed successful nanocomposite formation with SR and OMMT, resulting in strong PDMS-based materials. The results showed that viability, proliferation, and spreading of cells are governed by elastic modulus and stiffness of samples. Furthermore, adipose derived stem cells (ADSCs) cultured on PDMS and corresponding nanocomposites could retain differentiation potential of osteocytes in response to soluble factors, indicating that inclusion of OMMT would not prevent osteogenic differentiation. Moreover, better spread out and proliferation of cells was observed in nanocomposite samples. CONCLUSIONS: Considering cell behavior and mechanical properties of nanobiocomposites it could be concluded that silicone rubber substrate filled by nanoclay are a good choice for further experiments in tissue engineering and medical regeneration due to its cell compatibility and differentiation capacity.

Immunomodulatory effects of Astragalus gypsicolus hydroalcoholic extract in ovalbumin-induced allergic mice model.

Several studies have demonstrated that herbal extracts possess various biological effects including anti-inflammatory and anti-cancer activities. The present study was aimed to investigate the protective effects of the Astragalus gypsicolus (AG) hydroalcoholic extract in early allergic sensitized mice induced by ovalbumin. Phytochemical assay was used to recognize the main active constituents in the AG hydroalcoholic extract. Mice were immunized with subcutaneous injection of ovalbumin and aluminum hydroxide. Efficiency of sensitization was assessed by serum IgE levels and eosinophil count. After sensitization, two doses of extract (250 mg/kg and 500 mg/kg) were injected intrapritoneally. On day 14, mice were challenged with intrapritoneal injection of ovalbumin. IL-4 and IFNγ levels in broncoalveolar lavage fluid, which had been collected on day 15, were assessed by Enzyme-Linked Immunosorbent Assay (ELISA) kit. Our results indicate two main active constituents including flavonoids and terpenoids are present in the AG hydroalcoholic extract. Intrapritoneal injection of the AG hydroalcoholic extract was able to decrease IL-4 and increase IFNγ. It seems the AG hydroalcoholic extract has the potential to modulate the balance of Th1/Th2 cytokines in allergy.

Detection of IL-4, IL-6 and IL-12 serum levels in generalized aggressive periodontitis.

BACKGROUND: Periodontitis is a multifactorial chronic inflammatory disease characterized by destruction of tooth-supporting tissues. Environmental and genetic factors as well as the immune system participate in this process. Recent studies have attempted to elucidate the role of cytokine networks involved in periodontal diseases. OBJECTIVE: To assess and compare the levels of IL-4, IL-6 and IL-12 in serum samples of patients with generalized aggressive periodontitis (GAgP) and control individuals. METHODS: A total of 50 subjects were included in the study of which 25 patients had generalized aggressive periodontitis and 25 were healthy unrelated age and gender matched patients undergoing extraction and surgical crown lengthening (control group). Local blood samples of patients were collected from surgical sites of pocket reduction and from healthy individuals before tooth extraction or crown lengthening from non inflamed sites. The levels of IL-4, IL-6 and IL-12 were determined by an ELISA assay using serum samples separated from the whole blood of both groups. RESULTS: The level of IL-4 increased significantly in control group in comparison with the test group (p=0.002). The amount of IL-6 in GAgP patients increased strongly compared with control group (p<0.0001). There was no significant difference between the two groups concerning the level of IL-12. CONCLUSION: There is an association between generalized aggressive periodontitis and low level of IL-4 as an anti-inflammatory cytokine, and high level of IL-6 as a proinflammatory cytokine. No correlation between IL-12 and generalized aggressive peridontitis was found.

Expression of VLA1, VLA2 and VLA3 Integrin Molecules in Uterine Endometrium of Infertile Women with Unexplained Aetiology in Ahwaz-Iran.

BACKGROUND: Recent attention has focused on the expression of integrin molecules within the endometrium, and their relation to infertility. OBJECTIVE: The present prospective study was undertaken to determine whether the endometrium of women with unexplained infertility differs in the expression of very late activation antigens (VLA) from the endometrium of normal fertile women. METHODS: Thirty samples of endometrial biopsies from hysterectomies with nonendometrial pathology and 28 endometrial samples by uterine curetting from infertile women in secretary phase at implantation time were collected, stained with three monoclonal antibodies against beta1 integrin subunits including VLA-1 to VLA-3 by immunohistochemical technique and then assessed semiquantitively by microscope. Chi-Square test was used to compare the expression of VLA antigens on epithelial cells, stromal cells, lymphocytes and vessels within endometrial tissues between two groups. RESULTS: The results showed that most VLA integrins were present in fertile and infertile endometrium tissues. There were similarities and differences in the expression of VLA molecules in different compartments. VLA-2, VLA-3 expression on endometrial compartments showed an unaltered pattern of staining during the putative window of implantation in either fertile or infertile women with no significant differences (P-value> 0.5). VLA-1 expression on endometrial compartments changed in fertile and unexplained infertile women, the differences were related to the presence or lack of the molecules on epithelial and stromal cells respectively. CONCLUSION: Differences may explain causes of unexplained infertility, and suggests that certain integrins may participate in the cascade of molecular events leading to successful implantation and early placental development which requires more investigations.