Flouxetine Improved Intravaginal Ejaculatory Latency TIME Through Decreased Levels of Interferon-gamma and Increased Levels of Serotonin in Patient with Premature Ejaculation

Pathophysiology of premature ejaculation (PE) is very complex because it is associated with many factors, which can be grouped into biological factors and psychological factors. Various diseases have been found correlate between psychological factors and biological factors through cytokines, one of which is IFN-g (IFN-g). IFN-g affect indolamine dioxygenase enzyme (IDO) and decrease levels of serotonin. Low levels of serotonin leads to PE. The purpose of this study was to prove the relationship of serotonin and IFN-g in pathophysiology of PE. This study was designed as a pretest-posttest double-blind cross-over control group design. Patients with PE were divided into 2 groups: control group and treatment group. Treatment group received flouxetine 20 mg for 30 days. Then the control and treatment groups were crossed after passing a 14-days washout period. Previously as a control group to treatment group and received flouxetine 20 mg per day for 30 days. Before and after treatment in each group was examined the levels of serotonin and IFN-g. Of the 26 subjects, each group there was 13 subjects. Flouxetine 20 mg per day for 30 days increased serotonin levels were significantly (p < 0.05), and decreased levels of IFN-g were significantly (p < 0.05). Increased levels of serotonin and decreased levels of IFN-g was significantly associated with improvements (intravaginal ejaculatory latency time) ejaculation in PE patient. From these results it can be concluded that PE occurs because decreased levels of serotonin. Decreased levels of serotonin are associated with increased levels of IFN-g

Interleukin (IL)–12 and IL-23 Are Key Cytokines for Immunity against Salmonella in Humans

Patients with inherited deficiency of the interleukin (IL)–12/IL-23–interferon (IFN)–g axis show increased susceptibility to invasive disease caused by the intramacrophage pathogens salmonellae and mycobacteria. We analyzed data on 154 patients with such deficiency. Significantly more patients with IL-12/IL-23–component deficiency had a history of salmonella disease than did those with IFN-g–component deficiency. Salmonella disease was typically severe, extraintestinal, and caused by nontyphoidal serovars. These findings strongly suggest that IL-12/IL-23 is a key cytokine for immunity against salmonella in humans and that IL-12/IL-23 mediates this protective effect partly through IFN-g–independent pathways. Investigation of the IL-12/IL-23–IFN-g axis should be considered in patients with invasive salmonella disease

Studies on ovine interferon-gamma

Interferons have been recognized as important mediators of cellular communication for many years. There are two types of interferon: Type I interferons have antiviral functions, but Type II interferon (IFN-g) is more important as an immunomodulating molecule. Type II interferon has effects on cellular MHC class II expression, immunoglobulin class-switching, macrophage activation, cellular proliferation and a number of other functions. The role of IFN-g during in vivo immune responses has not been studied in great detail, but the sheep is an ideal species in which to study these phenomena by using the efferent lymphatic vessel cannulation model. This allows access to cells and tissue fluid for cytokine analysis using antibody and genetic probes for the detection of IFN-g.Bovine IFN-g peptides (amino-terminus, carboxy-terminus and central) were used to generate antibodies in rabbits. None of the antipeptide sera reacted with denatured ovine or bovine IFN-g, nor neutralized their antiviral effect. Rabbit antibodies to bovine recombinant IFN-g neutralized ovine IFN-g and detected IFN-g in a sandwich ELISA when used in combination with a monoclonal antibody against a human IFN-g carboxy-terminal peptide. The sensitivity of detection was only 125ng/ml, insufficient for use with efferent lymph fluid samples.The expression of MHC class II molecules on cell surfaces is increased by IFN-g on many cell types. This has been used previously to measure biologically active IFN-g concentrations in fluids. Measurement of ovine class II by slot blot was assessed as a method of adapting this to ovine IFN-g measurement, but the technique proved to be too problematic for regular use. The expression of class II on T lymphocytes is influenced by IFN-g in the surrounding fluid. Analysis by FACS of resting ovine T lymphocytes shows them to express class II, a situation different to that in the human. Incubation of efferent lymph cells with IFN-g enhances the expression of ovine DR-like class II molecules especially on CD8⁺ cells, but also on CD4⁺ cells. The expression of ovine DQ-like class II molecules was much less influenced by IFN-g. This differential expression has been seen previously in human cells. It is likely that such differences are due to variation in transcriptional control between the two types of molecule.The IFN-g genes of many species have been cloned, including bovine IFN-g. Nucleotide primers for the cloning of ovine IFN-g by polymerase chain reaction were chosen from the bovine sequence. Cloning of the central 300bp of the gene revealed 98% identity between ovine and bovine IFN-g at the amino acid level. Subsequent cloning of ovine IFN-g by other groups showed that allelic variation of the gene occurs with no alteration in the amino acid structure. This feature is not found in human IFN-g, but is described in other ruminant cytokines. Attempts to isolate a lambda clone expressing an IFN-g fusion protein using the rabbit anti-bovine rIFN-g sera were unsuccessful.The immune response may be analysed by studying cells and fluid delivered by the cannulation of an efferent lymphatic vessel. A secondary response to ovalbumin was induced in a sheep by inoculation of a dependent area. The lymphocytes collected were isolated into CD4⁺ and CD8⁺ cells by a magnetic separation technique (MACS). Their mRNA was isolated, and cDNA generated from it was subjected to IFN-g-specific PCR. This revealed that both CD4⁺ and CD8⁺ cells contribute to the synthesis of IFN-g during the secondary immune response to a protein antigen in the sheep

Alveolar macrophage priming by intravenous administration of chitin particles, polymers of N-acetyl-D-glucosamine, in mice

Intravenous (i.v.) administration of phagocytosable chitin particles (1 to 10 mm) in C57BL/6 mice and SCID mice primed alveolar macrophages (Mf) within 3 days to yield up to a 50-fold increase in their oxidative burst when elicited in vitro with phorbol myristate acetate (PMA). C57BL/6 mice pretreated with monoclonal antibodies (MAbs) against mouse gamma interferon (IFN-g) or NK1.1 showed a markedly decreased level of alveolar Mf priming following injection of chitin particles. To confirm IFN-g production in vitro, spleen cells isolated from normal C57BL/6 mice and SCID mice were cultured with chitin particles. Significant IFN-g production was observed following stimulation with chitin but not with chitosan or latex beads. When spleen cells were treated with anti-NK1.1 MAb, IFN-g production was significantly inhibited. Another set of experiments showed that when C57BL/6 mice were pretreated i.v. with a small dose IFN-g, a higher level of priming was induced with not only phagocytosable chitin particles but also phagocytosable chitosan and even latex beads. Likewise, the spleen cell cultures preconditioned with IFN-g provided an up-regulation of IFN-g production by these phagocytosable particles. Taken together, the in vivo and in vitro results suggest that (i) the alveolar Mf priming mechanism is due, at least in part, to direct activation of Mf by IFN-g, which is produced by NK1.11 CD42 cells; (ii) IFN-g would have an autocrine-like effect on Mf and make them more responsive to particle priming; and (iii) phagocytosis of particulates, probably by a postmembrane event such as interiorization, appears to be important for the up-regulation of alveolar Mf priming and IFN-g production. Originally published Infection and Immunity, Vol. 65, No. 5, May 199

RHODOCOCCUS EQUI INFECTION AND INTERFERON-GAMMA REGULATION IN FOALS

Rhodococcus equi (R. equi) is one of the most serious causes of pneumonia in young foals. The clinical disease is of great concern to breeding farms worldwide due to the impact of mortality on economic losses. While adult horses are resistant to R. equi, foals exhibit a distinct age-associated susceptibility. The mechanism underlying this susceptibility in foals is not well understood. Interferon-gamma (IFNg) plays an important role in the clearance of R. equi, but its expression is impaired in neonatal foals. Moreover, the regulation of this age-related IFNg expression in foals remains unknown. In humans, IFNg expression has been shown to be regulated by DNA methylation, lymphoproliferation, and influenced by environmental exposure. Therefore, we hypothesized that environmental exposure promotes IFNg expression through regulation of DNA methylation and lymphoproliferation. The objectives were: (1) to estimate the relevance of IFN-g production and R. equi infection in foals; (2) investigate the role of lymphoproliferation and DNA methylation in the regulation of IFN-g expression in foals; (3) to evaluate the effect of environmental exposure on IFN-g expression by housing foals in a barn environment verses pasture.; (4) to investigate the effect of environment exposure on antigen-presenting cells (APC), which sensor the environmental antigens and modulate IFN-g production by T cells. The results demonstrated that the IFN-g expression was inversely correlated with the age-related susceptibility to R. equi infection. lymphoproliferation promoted IFN-g expression in foals, whereas, DNA methylation repressed IFN-g expression. The IFN-g expression was augmented in foals exposed to the barn air which contained higher numbers of aerosol miroorganisms. DNA on the IFN-g promoter was demethylated and the lymphoproliferative activity was elevated in foals with barn-air exposure. The barn-air exposure also promoted the maturation and activation of APC to prime IFN-g expression by T cells in foals. Overall, this body of work demenstrated a relationship between IFN-g expression and R. equi infection, provided novel information on mechanisms that regulate IFN-g expression, and identified the effect of environment on mechanisms responsible for IFN-g expression

Environmental stimuli shape microglial plasticity in glioma

In glioma, microglia and infiltrating macrophages are exposed to factors that force them to produce cytokines and chemokines, contributing to tumor growth and maintaining a pro-tumorigenic, immunosuppressed microenvironment. We demonstrate that housing glioma-bearing mice in enriched environment (EE) reverts the immunosuppressive phenotype of infiltrating myeloid cells, by modulating inflammatory gene expression. Under these conditions, branching and patrolling activity of myeloid cells is increased, and their phagocytic activity is promoted. Modulation of gene expression depends on interferon-(IFN) g produced by natural killer (NK) cells, disappearing in mice depleted of NK cells or lacking IFN-g, and was mimicked by exogenous interleukin-15 (IL-15). Further, we describe a key role for BDNF produced in the brain of mice housed in EE in mediating the expression of IL-15 in CD11b+ cells. These data define novel mechanisms linking environmental cues to the acquisition of a pro-inflammatory, anti-tumor microenvironment in mouse brain

Neutrophil and lymphocyte responses to oral Streptococcus in Adamantiades-Behcet's disease

Immune reactions against microorganisms play an important pathogenic role in Adamantiades-Behçet’s disease (ABD). We had previously obtained Streptococcus sanguinis (strain BD113-20) isolated from the oral cavity of patients with ABD. To investigate the pathogenesis of this isolate, we examined neutrophil 5 reactions and level of cytokine production by lymphocytes after stimulation with the strain. The reactions of neutrophils were examined by chemiluminescence assay using whole blood. The amounts of interferon gamma (IFN-g) and interleukin (IL)-4, IL-8, IL-10, and IL-12 produced by peripheral blood mononuclear cells (PBMCs) were measured by ELISA. 10 Strain BD113-20 activated neutrophils from patients with ABD and healthy volunteers, and, in addition it increased IFN-g production by lymphocytes. Lymphocyte from the patients with ABD showed a dominant T helper 1 (Th-1) immune response. Results indicated that both bacterial stimulation and host hypersensitivity might be involved in the symptoms and pathogenesis of ABD

IL-10 in Human Newborns : Ontogeny of IL-10 secretion and relation to the secretion of IFN-g, immunoglobulin M, G, and A, and mononuclear cell composition in newborns

The purpose of this work was to elucidate the ontogeny of interleukin-10 (IL-10) secretion from newborn mononuclear cells (MCs), and to examine its relation to the secretion of interferon-g (IFN-g) and immunoglobulins (Igs). The initial hypothesis was that the decreased immunoglobulin (Ig) synthesis of newborn babies was the result of immature cytokine synthesis regulation, which would lead to excessive IL-10 production, leading in turn to suppressed IFN-g secretion. Altogether 57 full-term newborns and 34 adult volunteers were enrolled. Additionally, surface marker compositions of 29 premature babies were included. Enzyme-linked immunoassays were used to determine the amount of secreted IL-10, IFN-g, and Igs, and the surface marker composition of MC were analyzed with a FACScan flow cytometer. The three most important findings were: 1. Cord blood MC, including CD5+ B cells, are able to secrete IL-10. However, when compared with adults, the secretion of IL-10 was decreased. This indicates that reasons other than excessive IL-10 secretion are responsible of reduced IFN-g secretion in newborns. 2. As illustrated by the IL-10 and IFN-g secretion pattern, newborn cytokine profile was skewed towards the Th2 type. However, approximately 25% of newborns had an adult like cytokine profile with both good IL10 and IFN-g secretion, demonstrating that fullterm newborns are not an immunologically homogenous group at the time of birth. 3. There were significant differences in the surface marker composition of MCs between individual neonates. While gestational age correlated with the proportion of some MC types, it is evident that there are many other maternal and fetal factors that influence the maturity and nature of lymphocyte subpopulations in individual neonates. In conclusion, the reduced ability of neonates to secrete Ig and IFN-g is not a consequence of high IL-10 secretion. However, individual newborns differ significantly in their ability to secrete cytokines as well as Igs.Tutkimuksen tarkoituksena oli selvittää välittäjä-aine IL-10 tuotto vastasyntyneiden napaverestä eristetyistä mononukleaarisista soluista. On yleisesti tiedettyä, että vastasyntyneet ovat herkkiä saamaan vakavia infektioita. Vastasyntyneiden vasta-ainetuotanto on aikuisiin verrattuna selvästi vähäisempää ja mm myös tärkeän välittäjä-aineen, gamma-interferonin (IFN-g) tuotto, on vähäistä verrattuna aikuisiin. Tutkimushypoteesimme oli, että vastasyntyneiden välittäjä-aine tuotanto on kehittymätöntä johtaen suhteettoman suureen IL-10 tuotantoon joka estäisi IFN-g-tuotantoa. Tutkimuksessa oli mukana kaiken kaikkiaan 57 täysiaikaista vastasyntynyttä, 35 aikuista ja lisäksi 29:den keskosen lymfosyyttien alatyyppien prosentuaaliset määrät tutkittiin. Soluviljelmien tuottama IL-10, IFN-g, IgM, IgG ja IgA tuotanto tutkittiin ELISA-testillä ja lymfosyyttityyppien prosentuaaliset määrät mitattiin virtaus-sytometrillä. Tulosten perusteella vastasyntyneet, myös heidän ns CD5+ B solut, tuottivat IL-10:tä. Aikuisiin verrattuna vastasyntyneet tuottivat kuitenkin vähemmän IL-10:tä. Muut syyt kuin IL-10 ylimäärä vaikuttaa siis vähäiseen IFN-g tuotantoon. IL-10 ja IFN-g tuotantoja vertailemalla voitiin todeta, että vastasyntyneiden välittäjä-aine-tuotanto oli IL-10 voittoista. Kuitenkin noin 25% vastasyntyneistä tuotti molempia välittäjä-aineita aikuisten lailla. Vastasyntyneet eivät ole siis homogeeninen joukko välittäjä-aine-tuotannon suhteen. Vastasyntyneet erosivat huomattavasti myös eri lymfosyyttien prosentuaalisten osuuksien suhteen. Tuloksiin vaikutti selvästi raskauden kesto syntymähetkellä, mutta myös monet muut sekä äitiin että lapseen liittyvät seikat vaikuttivat lymfosyyttien alatyyppien määriin ja kypsyyteen. Edellä mainittujen välittäjä-aineiden ja vasta-aineiden tuotannossa oli siis selviä yksilöllisiä eroja. Yleisesti ottaen vastasyntyneiden vähäinen vasta-ainetuotanto ja IFN-g tuotanto ei kuitenkaan johtunut lisääntyneestä IL-10 tuotannosta

The Role of Interferon Gamma in the Regulation of IL-18 Binding Protein and the Development of Autoimmune Arthritis in a Genetically Non-Susceptible Mouse Strain

The etiology of the autoimmune disease rheumatoid arthritis (RA) is unknown, but the role of cytokines, including IFN-g, as effectors of immune cell function has been established by the examination of cytokine production in RA patients and through the use of animal models. C57BL/6 (B6) mice that express MHC class II molecules of the b haplotype (I-Ab) are not typically susceptible to collagen-induced arthritis (CIA), the most widely studied animal model of RA. When the gene encoding IFN-g is removed by genetic deletion, however, susceptibility to CIA is conferred. In addition, T cell responses against the immunogen that stimulates CIA, type II collagen (CII), are reduced in the B6 strain when compared to susceptible mouse strains that express differing MHC haplotypes such as the I-Aq-expressing DBA mouse. These observations led to the development of the following hypothesis. IFN-g functions as a regulator of T cell responses to weak determinants, and I-Ab MHC class II molecules have low affinity for collagen autoantigen determinants. In the IFN-g-/- B6 mouse, the absence of IFN-g alters T cell function and cytokine production following immunization with CII, disrupting normal immune function and allowing the development of CIA. To reveal the pathogenic mechanisms that mediate susceptibility to arthritis in the B6 IFN-g-/- mouse we examined several aspects of the immune response that follows immunization with CII. Firstly, alterations in cellular immune responses between wild type and B6 IFN-g-/-mice were examined following immunization with CII. Secondly, an antigenic determinant present in bovine CII that stimulates T cell response in mice expressing I-Ab was identified. This determinant was identified by the use of direct binding assays between I-Ab and CII derived peptides as well as by the generation of T hybridomas generated in I-Ab-expressing mice that respond to CII. In a second series of experiments, the role of IFN-g in modulating disease progression was examined by the use of microarray analysis to identify genes that are altered between wild type and IFN‑g‑/‑ B6 mice immunized with CII in CFA emulsion. This allowed the identification of IL-17 and IL-18 Binding Protein (IL-18 BP) as differentially expressed between the two strains. When IL-18 BP was given exogenously to IFN-g-/- B6 mice immunized with CII, the mice were protected from the development of CIA. These results indicate that a CII determinant is present in bovine CII and the low immunogenic properties of this determinant may be responsible for the lack of arthritis development in wild type B6 mice. In the absence of IFN-g, there is disregulation of immune function exhibited as decreased expression of IL-18 BP and increased expression of IL-17. This disregulation allows T cell responses to weakly antigenic CII determinants to progress, thus promoting the development of autoimmune arthritis

Increased porcine circovirus type 2 replication in porcine leukocytes in vitro and in vivo by concanavalin A stimulation

Veterinary Microbiology 2008, Vol. 132: 74–86Previously, it was shown that modulation of the immune system enhances porcine circovirus type 2 (PCV2) replication in pigs. In the present study, the effect of the mitogen concanavalin A (ConA) on PCV2 replication was investigated. Since ConA induces T-lymphocyte activation and initiates the production of interferon-gamma (IFN-g), a cytokine that enhances PCV2 replication in porcine epithelial and monocytic cell lines in vitro, it was examined if the effects observed with ConA were mediated by IFN-g. In an in vitro study, ConA but not IFN-g enhanced PCV2 replication in peripheral blood mononuclear cells (PBMC). Up to 2.08% and 0.96% of PBMC were antigen positive for PCV2 strains 1121 and Stoon-1010, respectively, and a low virus production was observed. PCV2-infected PBMC were identified as CD4+ (40%), CD8+ (54%) and IgM+ (11%). In a subsequent in vivo study, caesarean-derived colostrum-deprived piglets were injected with ConA or IFN-g 12 h before inoculation and every 3 days for 9 days after inoculation with strain 1121. PCV2 was isolated from inguinal lymph node biopsies from 10 days post-inoculation (dpi) in ConA-treated pigs and from 15 dpi in non-treated and IFN-g-treated pigs. ConA increased PCV2 replication levels, but disease was not observed. Half of the ConA-treated and IFN-g-treated pigs showed a delayed humoral immune response, but this delay did not result in increased PCV2 replication in these pigs. These experiments demonstrated that ConA enhances PCV2 replication in PBMC in vitro and in lymphoid tissues in vivo. # 2008 Elsevier B.V. All rights reserved