Effect of sitagliptin on depression in male wistar rats

Background: Growing evidence supports relationship between depression and inflammation. The hypothesis of involvement of inflammatory pathways in depression is supported by the findings of increased levels of proinflammatory cytokines. So, we decided to evaluate the effect of sitagliptin on depression using forced swim test (FST) and possible effects of sitagliptin on serum oxidative stress markers and cytokine gene expression in rat hippocampus.Methods: FST model was used to evaluate antidepressant effect in male wistar rats. Rats in group I (control group) were given normal saline, group II (standard group) were given fluoxetine, group III and IV (test groups) were given sitagliptin 5 mg/kg and sitagliptin 9 mg/kg respectively. All the drugs in all groups were given per orally. At the end, animals were sacrificed and blood was collected. Hippocampus of rat brain was dissected out. Serum oxidative stress markers and hippocampal pro inflammatory cytokine gene expression analysis was carried out.Results: Sitagliptin 5 mg/kg and 9 mg/kg showed reduction in depressive symptoms and hippocampal cytokine gene expression in comparison to control. In case of serum oxidative stress markers, there was statistically significant reduction in nitric oxide levels with stagliptin 9 mg/kg. Although there was a decrease in the levels of catalase and increase in the levels of glutathione with standard and test groups, the results were not statistically significant.Conclusions: The present study showed significant antidepressant effect activity of standard and test groups. Hence, further research should be carried out to substantiate above results.

Subsets of Inflammatory Cytokine Gene Polymorphisms are Associated with Risk of Carcinogenic Liver Fluke Opisthorchis viverrini-Associated Advanced Periductal Fibrosis and Cholangiocarcinoma

Opisthorchis viverrini infection induces chronic inflammation, and a minor proportion of infected individuals develop advanced periductal fibrosis (APF) and cholangiocarcinoma (CCA). Inflammatory cytokines and/or their gene polymorphisms may link to these biliary pathologies. We therefore investigated associations among cytokine gene polymorphisms and cytokine production in 510 Thai cases infected with O. viverrini who presented with APF+ or APF−, as established by abdominal ultrasonography as well as in patients diagnosed with CCA. Levels of pro-inflammatory and anti-inflammatory cytokines were determined in culture supernatants after stimulation of peripheral blood mononuclear cells (PBMCs) with O. viverrini excretory-secretory (ES) products. Pro-inflammatory cytokines, IL-1β, IL-6, IFN-γ, LT-α, and TNF-α were significantly increased in CCA patients compared with non-CCA (APF− and APF+) cases. Polymorphisms in genes encoding IL-1β-511C/T, IL-6-174G/C, IFN-γ +874T/A, LT-α +252A/G, and TNF-α−308G/A were then investigated by using PCR-RFLP or allele specific-PCR (AS-PCR) analyses. In the CCA cases, LT-α +252A/G and TNF-α−308G/A heterozygous and homozygous variants showed significantly higher levels of these cytokines than the wild type. By contrast, levels of cytokines in wild type of IFN-γ +874T/A were significantly higher than the variants in CCA cases. IFN-γ +874T/A polymorphisms were associated with advanced periductal fibrosis, whereas IL-6 −174G/C polymorphisms were associated with CCA. To our knowledge, these findings provide the first demonstration that O. viverrini infected individuals carrying several specific cytokine gene polymorphisms are susceptible to develop fibrosis and CCA.

Correlation between cytokine gene polymorphism and aGVHD in allo-HSCT recipients / 中国免疫学杂志

Objective:To investigate the relationship between gene polymorphisms of disease-relevant multiple cytokines including TNF-α,IL-6,IL-10,TGF-β1,IFN-γand acute graft versus host disease(aGVHD) in allogeneic hematopoietic stem cell trans-plantation ( allo-HSCT ) . Methods:32 cases of recipients received allo-HSCT and 36 cases of normal groups in January 2014 to December 2015 were selected as objects of study. We detected genotypes on specific SNP of target genes by polymerase chain reation ( PCR) combined with gene sequencing and observed the occurrence of aGVHD in postoperative recipients. The influence of cytokine gene polymorphisms on prognosis of allo-HSCT patients was analyzed,and the potential relationship between specific SNP mutation of the disease-relevant cytokine genes and severity of aGVHD was discussed. Results:Distribution of cytokines gene polymorphism including TNF-α-308(G/A),IL-6-174(G/C),IL-10-1082(A/G),TGF-β1+915(G/C),IFN-γ(T/A) had no significant differences with incidence of severe aGVHD(P>0. 05). However,the occurrence of severe aGVHD in allo-HSCT recipients with C/T genotype was significantly higher than C/C and T/T in SNP of TGF-β1+869(P<0. 01). Conclusion:Gene polymorphism of TGF-β1+869(C/T) in allo-HSCT patients was closely related to the occurrence of severe aGVHD. The research show allo-HSCT patients with C/T genotype occurred severe aGVHD more frequently, which is an important potential risk factor to induce the incidence of severe aGVHD. Therefore,detecting gene polymorphism of TGF-β1+869 ( C/T ) in allo-HSCT recipients and developing the appropriate therapeutic regimen may be helpful to reduce the incidence of aGVHD.

Membrane-bound p35 Subunit of IL-12 on Tumor Cells is Functionally Equivalent to Membrane-bound Heterodimeric Single Chain IL-12 for Induction of Anti-tumor Immunity

In this study, we compared two different tumor cell vaccines for their induction of anti-tumor immunity; one was a tumor cell clone expressing a membrane-bound form of IL-12 p35 subunit (mbIL-12 p35 tumor clone), and the other was a tumor clone expressing heterodimeric IL-12 as a single chain (mb-scIL-12 tumor clone). The stimulatory effect of mb-scIL-12 on the proliferation of ConA-activated splenocytes was higher than that of mbIL-12 p35 in vitro. However, the stimulatory effect of mbIL-12 p35 was equivalent to that of recombinant soluble IL-12 (3 ng/ml). Interestingly, both tumor clones (mbIL-12 p35 and mb-scIL-12) showed similar tumorigenicity and induction of systemic anti-tumor immunity in vivo, suggesting that tumor cell expression of the membrane-bound p35 subunit is sufficient to induce anti-tumor immunity in our tumor vaccine model.

Footprints of genetic susceptibility to pulmonary tuberculosis: Cytokine gene variants in north Indians.

Background & objectives: Tuberculosis is (TB) responsible for high morbidity and mortality worldwide. Cytokines play a major role in defense against Mycobacterium tuberculosis infection. Polymorphisms in the genes encoding the various pro- and anti-inflammatory cytokines have been associated with tuberculosis susceptibility. In this study we examined association of 25 sequence polymorphisms in six candidate cytokine genes namely IFNG, TNFB, IL4, IL1RA, IL1B and IL12 and their related haplotypes with risk of developing pulmonary tuberculosis (PTB) among north Indians. Methods: Pulmonary TB (n=110) patients and 215 healthy controls (HC) from north India were genotyped. Purified multiplex PCR products were subjected to mass spectrometry using Sequenom MassARRAY platform to generate the genotypes in a population-based case-control study. Results: Using multiple corrections, significant overall risk against PTB was observed at seven loci which included variants in IFNG at rs1861493 and rs1861494; IL1RA at rs4252019, IL4 variant rs2070874, IL12 variants rs3212220, rs2853694 and TNFB variant rs1041981. Analysis of gene structure revealed two haplotype blocks formed by IFNG variants rs1861493 and rs1861494. The TA haplotype was significantly over-represented (P=0.011) in the cases showing a two-fold risk in the current population (Odds ratio=1.59 CI=1.101 to 2.297) and TNFB variants at rs2229094 and rs1041981 contributed to two haplotypes which were in strong linkage disequilibrium (LD) with AT haplotype showing a three-fold risk (P=0.0011, Odds ratio=3, CI=0.1939 to 0.7445) of developing PTB in north Indians. Interpretation & conclusions: Our study showed six novel associations of cytokine gene variants with susceptibility to PTB in north Indians. Variants of IFNG and TNFB emerged as factors imposing a significant risk of developing PTB in north Indians apart from risk indicated by IL1RA, IL4 and IL12.

Cytokine gene polymorphisms in Irritable bowel syndrome in Indian population - A pilot case control study.

Aim: This study aimed at evaluating the single nucleotide polymorphisms (SNPs) in five cytokine genes regulating inflammation at altogether 8 different loci and compared their frequencies in patients with Irritable bowel syndrome (IBS) versus healthy age and sex matched controls. Methods: Peripheral blood was collected for DNA cytokine analysis from 23 patients with IBS and 20 healthy controls. The cytokine SNPs studied include TNF-á (-308G/A), TGF-â1 (codon10T/C, codon25G/C), IL-10 promoter (-1082A/G; -819T/C; -592A/C), IL-6 promoter (-174G/C), and IFN-ã (+874T/A). Results: There was a significant difference between a SNP in IL-10 (-592A/C) among cases and controls. There was also a trend to significance as regards to IL-6 promoter (-174G/C). Frequencies of other SNPs were not significantly different between the two groups. Conclusion: This pilot study shows that there are polymorphism differences in cytokine genes between patients with IBS and healthy controls from India.

Association of cytokine genetic polymorphism with hepatites B infection evolution in adult patients

The infection by the hepatitis B virus (HBV) has different forms of evolution, ranging from self-limited infection to chronic hepatic disease. The objective of this study was to evaluate the influence of cytokine genetic polymorphisms in the disease evolution. The patients were divided into two groups, one with chronic HBV (n = 30), and the other with self-limited infection (n = 41). The genotyping for TNF (-308), TGFB1 (+869, +915), IL-10 (1082, -819, and -592), IL-6 (-174), and IFNG (+874) was accomplished by the PCR-SSP (polymerase chain reaction with sequence specific primers technique using the One Lambda kit. Although no statistically significant differences were found between the groups, the combination of TNF -308GG and IFNG +874TA was found in a lower frequency in chronic patients than in individuals with self-limited infection (26.7 versus 46.3 percent; P = 0.079; OR = 0.40; IC95 percent = 0.14-1.11). In chronic patients with histological alterations it was not observed the genotype TGFB1+869 C/C, against 24.4 percent in the self limited infection group (100 versus 75.6 percent; P = 0.096; OR = 7.67; IC95 percent = 0.42-141.63). Further studies in other populations, and evaluation of a greater number of individuals could contribute for a better understanding of the cytokine genetic polymorphism influence in HBV infection evolution.

Quantification of bovine cytokine gene expression using real-time RT-PCR methodology

T cells produce cytokines that affect host response to infection. This paper reports real-time RT-PCR conditions and validation steps for accurate quantification of Bos indicus cytokines, interleukin (IL)-2, IL-4, IL-8, IL12p-35, IL-13, tumoral necrosis factor (TNF)-alpha, interferon (IFN)-gamma, monocyte chemoattractant proteins (MCP)-1 and MCP-2, and the glycoprotein mucin (MUC)-1 in two groups of Nelore cattle, one resistant and the other susceptible to gastrointestinal nematode infections. RPL-19 was shown to be an ideal internal control gene, since its expression was constant across treatments and presented lower variation when compared to the GAPDH gene. The optimized conditions established in the present study can be used to determine the immune response of cattle under different experimental conditions, such as viral, bacterial and parasite infections.

Expression and location of IL-2/IFN-? fusion protein in colorectal tumor cell lines / 中国实用内科杂志

Objective If IL-2 and IFN-? are united,perhaps we may see a more significant antitumor effect.A fusion gene of IL-2 and IFN-? was designed and constructed by DNA engineering.Methods CEA promotor、IL-2cDNA and IFN-?cDNA were obtained by PCR and RT-PCR.SDS-PAGE was used to examine the results;pcDNA-IL-2、pcDNA-IFN-?、pCEA-IL-2/ IFN-?、pEGFP-IL-2/ IFN-? were transfected to CEA high expression cell line(Lovo)by use of liposome.The fusion protein expression was examined by fluorescence microscope and Western blotting.Results IL-2/ IFN-? fusion gene can be expressed in tumor cells.Its expression is more obvious in CEA high expressed cells(Lovo),and can be located in cell plasma.Conclusion The fusion gene can be expressed in plasma of CEA high expressed cell lines.It may have a new anti-tumor effects.

Diagnosis of acute rejection by bile cytology and cytokine gene expression in rat liver transplantation model / 中国普通外科杂志

Objective To evaluate a reliable and noninvasive method for the diagnosis of acute rejection after liver transplantation.Methods The allogeneic, syngeneic and immunosuppresive models of orthotopic rat liver transplantation were established, and acute rejection was graded according to histopathological change. Bile was sent for cytology by microscopy,and the sequential detection of bile IL 2,IFN ?,IL 4 and IL 6 gene expression was performed at day 1 through day 7. Results The number of cells in bile was persistently high in allogeneic group,and blast and lymphocytes appeared in bile. IL 2 and IFN ? were detected only in allogeneic group,with a specificity of 70% and 67%(7/10 and 8/12),and a sensitivity of 39% and 44%(7/18 and 8/18)respectively.Conclusion Bile cytology and detection of IL 2 and IFN ?mRNA transcript are useful adjuvant diagnosic methods for acute rejection after liver transplantation.

The gene expression of cytokines in human tumor cell lines and specimens / 中国免疫学杂志

Objective:To study the role of IL-2,IL-4,IL-6,IL-10,IL-13 and IFN? in pathogenesis of human tumor.Methods:The gene expression of 6 cytokines in human tumor cell lines and specimens were detected by RT-PCR or RNA dot hybridization.Results: There were predominant expression of Th2 type cytokines in 183 different human tumor cell lines and specimens.The positive rate of IL-4,IL-6,IL-10,IL-13 .IFN? and IL-2 in 183 tumor cell lines and specimens were 65%,70.5%,83.6%,61.7%, 12.6% and 22.9%,respectively.Conclusion: Predominant expression of Th2 type cytokines play an important role in the pathogenesis and development of tumor, may be related to the evasion of tumor cells from immune surveillance.

Follow-up Study on Cell-mediated Immune Responses In Syphilitic Patients after in vitro Stimulation with the 47 kDa Antigen of Treponema pallidum / 대한면역학회지

Present study was aimed to investigate the immunological activities of the 47 kDa protein antigen from Treponema pallidum and conducted on 12 patients with syphilis (early, late, spontaneously healed, congenital and treated patients) followed by therapy. Peripheral blood mononuclear cells (PBMC) were obtained three times from each patient, on admission before the initiation of therapy, 1 and 6 months later. Eleven (96.7%) of the patients prior to therapy, showed depressed lymphoproliferative responses to the 47 kDa antigen (stimulation index <4) by 3H-thymidine incorporation assay. However, these T cell responses were seemed to be transient because most of the patients (63.6%) exhibited significantly higher lymphoproliferation after therapy. Before therapy, PBMC from spontaneously healed syphilis patients resulted in significantly increased gene expression of IFN- and proinflarnmatory cytokines, such as TNFa, IL-1B and IL-6, in response to the 47 kDa. Patients with late latent and late congenital syphilis exhibited lower IFN-r and proinflammatory cytokine mRNA expression than spontaneously healed syphilis group did. After therapy, IFN-r and proinflammatory cytokine mRNA expressions were gradually reduced in these groups. On the other hand, IFN- and proinflammatory cytokine gene expressions were considerably depressed in early syphilis patients, but these patients went on to express prominent IFN-r and proinflamrnatory cytokine mRNA with treatment. These data suggest that the pattern of cellular immune response in response to the 47 kDa antigen may be involved in the evaluation of the clinical course and outcome of syphilis followed by therapy.

Cell-mediated Immune Responses in Syphilitic Patients after In vitro Stimulation with the 47 kDa Antigen of Treponema pallidum / 대한면역학회지

Present study was aimed to investigate the immunological activities of the 47 kDa glycoprotein antigen from Treponema pallidum and conducted on 24 patients with syphilis, (early, late, spontaneously healed, congenital and treated patients) and on 17 normal healthy controls. Two opposite lymphoproliferative manifestations to the 47 kDa antigen were observed in syphilis patients by H-thymidine incorporation assay. Ten responders (stimulation index [Sl] >4) showed a 3-fold-higher proliferation than the nonresponders, and four of those responders were spontaneously healed patients. Furthermore, analysis by flow cytometry indicated a preferential expansion of CD4' T lymphocytes by the 47 kDa antigen in the spontaneously healed syphilis patients. Stimulation of PBMCs of spontaneously healed syphilis patients with the 47 kDa antigen for greater than 72 hrs resulted in piogressive augmentation of IFN-r, IL-2Ra and IL-2 mRNA measured by RT-PCR, but considerably reduced IL-4 and IL-10 mRNA expression. However, patients with late latent syphilis exhibited more increased IL-4 and IL-10 mRNA expressions in response to the 47 kDa antigen than spontaneously healed syphilis group. In contrast to other groups, when cultured with the 47 kDa antigen very low IFN-#y, IL-2Ra and IL-2 mRNA expressions were shown in early syphilis group. These data suggest that the Th1-predominant cellular responses induced by the 47 kDa antigen may be involved in the clinical outcome of syphilis and provide the immunologic basis for further functional studies regarding the role of the 47 kDa in the immunopathogenesis of syphilis.

Analysis of Cytokine Gene Expression in Thyroid Aspirates and Peripheral Blood Mononuclear Cell and in vitro Production of Interferon / 대한내분비학회지

Cytokine production was studied in thyroid fine needle aspirates and peripheral blood and the production of interferon-gamma by peripheral blood mononuclear cell(PBMC) culture in response to interleukin-2(IL-2) stimulation was also studied from patients with hyperthyroidism, non toxic goiter, thyroid nodule. The expression of glycer aldehyde 3-phosphate dehydrogenase(GAPDH), interleukin-1beta(IL-1beta), IL-2, interleukin-8(IL-8), platelet- derived growth factor-A(PDGF-A) and interferon-gamma(IFN-gamma) chain was assessed by RT-PCR(reverse transcriptase polymerase chain reaction) in fine needle aspirates of thyroid and peripheral blood mononuclear cell : the samples were obtained from 7 patients with hyperthyroidism, 6 patients with non toxic goiter, 7 patients with thyroid nodule. A dose of IL-2(25 U/ml) was utilized to induce IFN-gamma production by PBMC from all patients.The results were as follows:1) In case of cytokine expression of fine-needle aspirates, GAPDH and IL-1beta, IL-8 were expressed highly but IFN-gamma, IL-2 were not expressed in hyperthyroidism and non-toxic goiter, thyroid nodule. PDGF-A was expressed in hyperthyroidism and thyroid nodule but not in non toxic goiter. 2) In case of cytokine expression of PBMC, GAPDH, IL-1beta were expressed in hyperthyroidism and non toxic goiter, thyroid nodule and highly expressed after IL-2 stimulation than before. But PDGF-A was more expressed in non toxic goiter and thyroid nodule than hyperthyroidism. Also, IFN-gamma was less expressed in thyroid nodule than hyperthyroidism and non toxic goiter. 3) The incremental increase in IFN-gamma value in supernatants of PBMC culture was significantly higher in patients with non toxic goiter than that in PBMC from hyperthyroidism and thyroid nodule(p<0.05).Therefore it seems that the cytokine production was found in hyperthyroidism and non toxic goiter and thyroid nodule. There were variability in their distribution each other, in general, higher expressed in hyperthyroidism than non toxic goiter. And RT-PCR Method that employed should be sufficiently sensitive to permit the analysis of cytokine gene expression in fine needle aspiration biopsies from patients with thyroid disease.

Anergy and Characteristics of Human T Cells Induced by Combination with B7-1 mAb and CsA / 中国肿瘤生物治疗杂志

To study the induced condition and characteristics of T cell anergy in vitro. Methods: Anergic Tcell was induced by combination of B7-1 mAb and cyclosporin A (CsA) in vitro, cytokine gene of anergic T cells was detected by RT-PCR. Results: T cell anergy was antigen-specific. The state of T cell anergy can be reversed by PHA, CD3 mAb and PMA plus A23187. IL-2 can prevent the induction of T cell anergy, but it can not reverse the state of un-responsiveness. IL-2 and IFN mRNA can not express in anergic T cells. In contrast, IL-4 and IL-10 mRNA were detectable. Conclusion: T cell anergy can be induced in vitro , cytokine profile of anergic T cells deviated to Th2-like phe-norype.

Effects of sinomenine on IL- 1?, LL-8 gene expression of human peripheral blood / 中国免疫学杂志

Objective:To study the anti-inflammatory and anti-rheumatic mechanisms of sinomenine, a pure alkaloid extracted from the Chinese medical plant sinomeniuim acutum, on cytokines-related genes modulation. Methods: Cultured normal human peripheral blood mononuclear cytes(PBMIN) with sinomenine in vitro, isolated total RNA from different groups of treated and controlled cells to investigate the effects on mRNA expression of IL-1?, LL-8 with LPS stimulation by one-step and two-step RT-PCR techniques. Results: The results from the one-step RTPCR showed that the inhibition ratio on IL-l?mRNA expression in the group of mmol/L sinomenine was about( 15.7?5.52) % ; with two-step RT-PCR techneque demonstrated that the inhibition ratio of sinomenine in the group of sinomenine with the concentration of 1 mmol/L, 100 umol/L on IL-1?mRNA expression were ahout( 17.07 ? 7 .69)% ,(25 .99 ? .84)% respectively and the inhitition ratio in the group with sinomenine concentration of 1 mmol/L,100umol/L on IL-8mRNA expression were(2l.01 ?7.79)%,(16.04?7.55)% respectively .Conclusion: The therapeutic effects of sinomenine on rheumatoid arthritis(RA) may partially related to downregulating gene expression of IL-1?, IL-8 cytokines of the peripheral blood mononuclear cells.