Effects of SSRIs on peripheral inflammatory cytokines in patients with Generalized Anxiety Disorder.

BACKGROUND: Extensive research into psychoneuroimmunology has led to substantial advances in our understanding of the reciprocal interactions between the central nervous system and the immune system in neuropsychiatric disorders. To date, inflammation has been implicated in the pathogenesis of depression and anxiety. The immunomodulating effects of antidepressants on depression have been reported, however, there is no evidence of the similar effects of antidepressants on anxiety. The aim of the study was to investigate the effects of selective serotonin reuptake inhibitors (SSRIs) on peripheral inflammatory cytokines in patients with first episode generalized anxiety disorder (GAD). METHODS: A prospective cohort design was employed: 42 patients with first episode GAD were treated with either escitalopram or sertraline for 12 weeks. Anxiety was measured by the Generalized Anxiety Disorder Scale and the State Trait Anxiety Inventory, serum pro-inflammatory cytokine levels were measured by the enzyme-linked immunosorbent assay (ELISA), and CRP determined by an immunoturbidimetric method before and after SSRIs treatment RESULTS: Baseline levels of anxiety and pro-inflammatory cytokines including IL-1α, IL-6, IL-8, IL-12, IFN-γ, and CRP were significantly reduced after treatment of SSRIs (p < 0.05 in all cases). In addition, the change of anxiety measures co-vary with the change of peripheral cytokine levels (p < 0.05 in all cases). The regression model revealed that log transformed baseline levels of CRP and IL-6 predicted treatment response (p < 0.05 in both cases). CONCLUSIONS: This study is the first to investigate the effects of SSRIs on pro-inflammatory cytokines in patients with first episode GAD. The findings indicate moderate acute anti-inflammatory effects of SSRIs in GAD, and suggest that these anti-inflammatory effects may underlie anxiolytic effects of SSRIs. The study also indicates that serum levels of CRP and IL-6 may predict treatment response. However, data from randomized controlled trials is warranted to confirm these findings.

Diosgenin attenuates neuropathic pain in a rat model of chronic constriction injury.

Diosgenin is a steroidal saponin extract from numerous plants, including Solanum and Dioscorea species, and has been reported to possess neuroprotective activity. However, the role of diosgenin in neuropathic pain remains unclear. The present study examined the effects of diosgenin on allodynia and the levels of inflammatory mediators in rats following neuropathic pain evoked by chronic constriction injury (CCI). In addition, the underlying molecular mechanisms involved in diosgenin­induced suppression of neuropathic pain were examined. The results of the present study demonstrated diosgenin reversed CCI­decreased mechanical withdrawal threshold and thermal withdrawal latency. Furthermore, diosgenin inhibited CCI­induced upregulated levels of the pro­inflammatory cytokines tumor necrosis factor­α, interleukin (IL)­1ß and IL­2, and suppressed oxidative stress induced by CCI in the spinal cord. Furthermore, diosgenin significantly inhibited the expression of phosphorylated­p38 mitogen activated protein kinase (MAPK) and nuclear factor (NF)­κB in the spinal cord in CCI rats compared with sham­operated rats. In conclusion, the present study demonstrated that diosgenin attenuates neuropathic pain in CCI rats by inhibiting activation of the p38 MAPK and NF­κB signaling pathways. These results implicate diosgenin in the treatment of neuropathic pain, which merits further clinical investigation.

Areca nut extracts suppress the differentiation and functionality of human monocyte-derived dendritic cells.

BACKGROUND AND OBJECTIVE: Areca quid chewing, a major risk factor contributing to the occurrence of oral cancer and precancer, has been reported to be associated with the severity and high prevalence of periodontal diseases in areca quid chewers. As dendritic cells are critically involved in the regulation of innate and adaptive immunity in oral mucosa, the objective of the present study was to investigate the effect of areca nut extracts (ANE) on the differentiation and reactivity of dendritic cells derived from monocytes. MATERIAL AND METHODS: Human peripheral blood monocytes were cultured in the presence of granulocyte-monocyte colony-stimulating factor and interleukin-4 for 7 d to generate dendritic cells. To examine the effect of ANE on the generation of dendritic cells, the monocytes were exposed to ANE throughout the 7 d culture period. In addition, the effect of ANE on the maturation of monocyte-derived dendritic cells induced by lipopolysaccharide (LPS) was examined. RESULTS: Monocytes cultured in granulocyte-monocyte colony-stimulating factor and interleukin-4 exhibited a typical phenotype of dendritic cells, as evidenced by the heightened expression of human leukocyte antigen (HLA)-DR, CD11c and the co-stimulatory molecules CD40, CD80 and CD86. Exposure of the monocytes to ANE did not influence the expression of HLA-DR and CD11c, but markedly attenuated the proportion of CD40-positive cells and the mean fluorescence intensity of CD86. The expression of co-stimulatory molecules in LPS-activated dendritic cells was not affected, whereas the mRNA expression of interleukin-12 induced by LPS was markedly suppressed by ANE treatment in a concentration-dependent manner. CONCLUSION: These results suggest that ANE exposure interfered with the differentiation of dendritic cells from monocytes. Moreover, the functionality of mature monocyte-derived dendritic cells was attenuated in the presence of ANE.

Clinical and immunological responses in ocular demodecosis.

The purpose of this study was to investigate clinical and immunological responses to Demodex on the ocular surface. Thirteen eyes in 10 patients with Demodex blepharitis and chronic ocular surface disorders were included in this study and treated by lid scrubbing with tea tree oil for the eradication of Demodex. We evaluated ocular surface manifestations and Demodex counts, and analyzed IL-1ß, IL-5, IL-7, IL-12, IL-13, IL-17, granulocyte colony-stimulating factor, and macrophage inflammatory protein-1ß in tear samples before and after the treatment. All patients exhibited ocular surface manifestations including corneal nodular opacity, peripheral corneal vascularization, refractory corneal erosion and infiltration, or chronic conjunctival inflammatory signs before treatment. After treatment, Demodex was nearly eradicated, tear concentrations of IL-1ß and IL-17 were significantly reduced and substantial clinical improvement was observed in all patients. In conclusion, we believe that Demodex plays an aggravating role in inflammatory ocular surface disorders.

The extract of Japanese soybean, Kurosengoku activates the production of IL-12 and IFN-γ by DC or NK1.1(+) cells in a TLR4- and TLR2-dependent manner.

During the search for immuno-improving foods, we found that a variety of the Japanese soybean, Glycine max cv. Kurosengoku (Kurosengoku), which activated Type-1 immunity in a Toll-like receptor (TLR)4- and TLR2-dependent manner. Namely, the extract of Kurosengoku first caused production of IL-12 from DC and sequentially induced IFN-γ production by NK1.1(+) NK cells and NKT cells. The IFN-γ production was significantly blocked by neutralizing mAb against IL-12 or TLR4- and TLR2-deficient condition, indicating that TLR4- and TLR2-dependent activation of DC to produce IL-12 was essential for the production of IFN-γ from spleen cells by Kurosengoku. Moreover, the extract of Kurosengoku also enhanced production of IFN-γ from human PBMC by co-stimulation with anti-CD3 mAb in a TLR2- and TLR4-dependent manner. Thus, our findings strongly suggest that Kurosengoku might a novel immuno-improving food, which would be a useful tool for preventing the tip of immune balance in developed countries.

Comparative analysis of a large panel of non-starch polysaccharides reveals structures with selective regulatory properties in dendritic cells.

SCOPE: Structural-based recognition of foreign molecules is essential for activation of dendritic cells (DCs) that play a key role in regulation of gut mucosal immunity. Orally ingested non-starch polysaccharides (NSP) are ascribed many health-promoting properties, but currently we lack insight into the impact of structure and size for their capacity to affect immune responses. METHODS AND RESULTS: This study addresses the importance of chemical structure, size, origin and presence of contaminants for the capacity of both dietary and non-food NSP to modulate DC. Of 28 NSP products, ß-glucans of microbial and plant origin and the galactomannan guar gum were found to modulate the DC cytokine pattern induced by the Toll-like receptor 4-ligand LPS giving rise to reduced IL-12p70 and increased IL-10 levels, whereas IL-6 production was unaffected. A large proportion of the tested NSP were able to down-regulate LPS-induced IL-12p70 production. The most potent NSP induced up-regulation of CD86 on DC independently of LPS stimulation. Cereal-based ß-glucans showed less potency than ß-glucans of microbial origin, but proper molecular weight composition and preparation may improve effectiveness. CONCLUSIONS: Collectively, this comparative study revealed that some plant-derived NSP besides those of microbial origin exert modulation of the DC phenotype, with the exact structure being important for the activity.

Clinical and Immunological Responses in Ocular Demodecosis

The purpose of this study was to investigate clinical and immunological responses to Demodex on the ocular surface. Thirteen eyes in 10 patients with Demodex blepharitis and chronic ocular surface disorders were included in this study and treated by lid scrubbing with tea tree oil for the eradication of Demodex. We evaluated ocular surface manifestations and Demodex counts, and analyzed IL-1beta, IL-5, IL-7, IL-12, IL-13, IL-17, granulocyte colony-stimulating factor, and macrophage inflammatory protein-1beta in tear samples before and after the treatment. All patients exhibited ocular surface manifestations including corneal nodular opacity, peripheral corneal vascularization, refractory corneal erosion and infiltration, or chronic conjunctival inflammatory signs before treatment. After treatment, Demodex was nearly eradicated, tear concentrations of IL-1beta and IL-17 were significantly reduced and substantial clinical improvement was observed in all patients. In conclusion, we believe that Demodex plays an aggravating role in inflammatory ocular surface disorders.

The antiallergic effects and acute toxicity of Lactobacillus crispatus KT-11 cultured in food grade medium.

We synthesized a medium consisting of commercial food supplements (food grade medium) that could be used to cultivate Lactobacillus crispatus KT-11 (KT-11), and investigated the antiallergic effects and acute toxicity of KT-11 cultured in this medium. We found that the growth of KT-11 in the food grade medium was comparable to that in DeMan-Rogosa-Sharpe (MRS) medium. Sneezing event was reduced in ovalbumin (OVA)-sensitized BALB/c mice given a diet supplemented with KT-11 grown in the food grade medium (FG-KT-11 group) when compared to mice given a diet supplemented with KT-11 grown in MRS medium (MRS-KT-11 group). The number of CD80(+)CD11b(+) Peyer's patch cells was significantly lower in the FG-KT-11 group than in the MRS-KT-11 group, while IL-12(+)CD11b(+) Peyer's patch cells were higher in the FG-KT-11 group. Only minimal acute toxicity was observed in ICR mice given 1000 or 2000 mg of FG-KT-11/kg body weight. These results suggest that FG-KT-11 represents a safe antiallergic food material.

Aqueous extract of Phyllanthus niruri (Euphorbiaceae) enhances the phenotypic and functional maturation of bone marrow-derived dendritic cells and their antigen-presentation function.

Decoctions of Phyllanthus niruri (PN) (Fam. Euphorbiaceae) is promoted in traditional medicine of Africa, Asia, and South America as beneficial supplement for different infectious diseases, especially for viral hepatitis, tumor, and for immune compromised patients. This stimulated the interest in understanding the mechanisms by which the whole extract of the plant could stimulate the immune system. Dendritic cells (DCs) are professional antigen-presenting cells and provide a link between the innate and the adaptive immune responses. In the present study, the effects of lyophilized aqueous extract of PN on structural and functional maturation of murine bone marrow-derived DCs (BM-DCs) were investigated. Bone marrow cells were cultured in the presence of granulocyte macrophage-colony stimulating factor and interleukin-4 (IL-4) and the generated immature DCs were stimulated with PN (25, 50, and 100 microg/mL) or lipopolysaccharide (10 microg/mL) for 48 h. Results showed that treatment with PN increased the expression of major histocompatibility complex-II and the various makers for DCs maturation (CD40), activation (CD83), and costimulation (CD86) in a concentration-dependent manner. Consistent with the increase in phenotypic makers, functional maturation assay showed that treatment of BM-DCs with PN caused a decrease in fluorescein isothiocyanate-dextran pinocytosis and an increase in IL-12 in the supernatant. In a transgenic T-cell activation model, PN-treated BM-DCs presented Ova antigen to Ova-specific CD8(+) T cells from OT-1 mice more efficiently as demonstrated by increased T-cells proliferation and IL-2 production. Therefore, PN enhances the structural and functional maturation of BM-DCs and their antigen-presenting function. These effects are relevant in immunodeficient conditions, tumor control, and in infectious diseases.

Natural killer cell activities of synbiotic Lactobacillus casei ssp. casei in conjunction with dextran.

We have reported previously that Lactobacillus casei ssp. casei, together with specific substrate dextran, exhibited an adjuvant effect of stimulating humoral immune responses against bovine serum albumin (BSA) as a model antigen in BALB/c mice. In the present study, among the Lactobacillus species tested, L. casei ssp. casei with dextran significantly elevated the natural killer (NK) cell activities in spleen mononuclear cells from BALB/c mice in comparison to L. casei ssp. casei alone or other Lactobacillus species with or without dextran. Oral administration of L. casei ssp. casei together with dextran also resulted in a significant increase of NK cell activities in healthy human volunteers. Further, L. casei ssp. casei induced significant production of interleukin (IL)-12 in human peripheral blood mononuclear cells and IL-15 mRNA expression in the human intestinal epithelial cell line Caco-2. L. casei ssp. casei with dextran in food also significantly elevated the survival rate of BALB/c mice bearing Meth-A cells. Taken together, these results demonstrate that dietary synbiotic supplementation which is a combination of the L. casei ssp. casei used as a probiotic together with the dextran, a specific substrate as a prebiotic, efficiently elicits murine and human NK cell activities.

Dietary soy phytoestrogens and ERalpha signalling modulate interferon gamma production in response to bacterial infection.

Diets rich in soy phytoestrogens have many potential health benefits but isoflavones such as genistein may suppress cell mediated immune function. The effect of dietary phytoestrogens on the host response to infection has not been extensively examined. Mice were fed a diet containing soy phytoestrogens and infected with Mycobacterium avium to establish a chronic infection and inflammatory response. As phytoestrogens may act through classical oestrogen receptors (ER), mice deficient in ERalpha signalling and wild type mice were evaluated for a panel of Type 1-associated cytokines (IFNgamma, IL-12 and IL-18) in the spleen. IFNgamma production in the spleen was increased approximately 4-fold in ERalpha-deficient mice fed a casein-based diet over wild type mice fed a casein-based diet (P < 0.05), suggesting a role for ERalpha in suppressing IFNgamma production. IL-18 levels in spleens of wild type mice were decreased compared to ERalpha-deficient mice on a casein diet. Splenic IL-12 and IL-18 levels were not affected in wild type and ERalpha-deficient mice on the phytoestrogen containing diets, with the exception that whole soy increased IL-12 levels in the tissues of ERalpha deficient mice. We conclude that ERalpha and dietary phytoestrogens can influence production of key regulatory cytokines in response to chronic bacterial infection.

Immune-enhancement effect of the herbal combination Allergina.

BACKGROUND: The herbal formulation, Allergina, has long been used for various diseases. It is known to have an anti-microbial and anti-virus activity. However, it is still unclear how Allergina has these effects in experimental models. We investigated the effect of Allergina on the proliferation of T cell and production of cytokines in human T-cell line, MOLT-4 cells, and mouse peritoneal macrophages. METHODS: The MOLT-4 cells were cultured for 24 h in the presence or absence of Allergina. Allergina significantly increased the cell viability by 26.9+/-5.4% (P<0.05) and interleukin (IL)-2, IL-4 and interferon (IFN)-gamma production compared with media control (about 4-fold for IL-2, 2.5-fold for IL-4 and 3.4-fold for IFN-gamma, P<0.05). Maximal effective concentration of Allergina was 1 mg/ml for IL-2 and, 0.01 mg/ml for IL-4 and IFN-gamma. Allergina alone or Allergina plus recombinant IFN-gamma (rIFN-gamma) increased the production of tumor necrosis factor (TNF)-alpha, but Allergina decreased the production of TNF-alpha on rIFN-gamma plus LPS-stimulated macrophages. In addition, Allergina increased the production of IL-12 on mouse peritoneal macrophages and peripheral blood mononuclear cells. CONCLUSION: Allergina may have an immune-enhancement effect through the cytokine production.

[Role of cytokines in the diagnosis of chronic prostatitis]. / Rol' tsitokinov v diagnostike khronicheskogo prostatita.

Chronic prostatitis affects 30-60% males and significantly deteriorates quality of their life. Clinical and experimental investigations have revealed changes in immune status in the onset and development of prostatic inflammation. As some other urologists, we made an attempt to determine the role of cellular immunity and immunoglobulins in the diagnosis of chronic prostatitis. The study was made in 30 patients with chronic abacterial prostatitis (mean age 42.5 years, duration of the disease 1.8 years). In addition to standard examination, all the patients have undergone analysis of the immune status and measurement of proinflammatory cytokines (TNF alpha and IL-12b) in biological media: blood serum, urine, ejaculate, prostatic secretion. The patients had moderate symptoms: IPSS--10.4 scores, life quality index--4.3 scores, on the average. Prostamol-uno was given to all the patients in a standard dose 1 capsule (320 mg) a day for 3 to 6 months. The results were processed statistically. A good effect of prostamoluno was registered in 26 patients, a satisfactory one--in 2. Two patients refused to take prostamol-uno because of lack of a prominent effect. The scores of IPSS lowered from 10.4 to 6.3 (by 39%), life quality improved by 42%. Ultrasound monitoring of the size of the prostate showed no significant changes in the size. Tolerance was good in all 30 patients. Side effects were absent. After 3 months of the treatment serum, urine, ejaculate and prostatic secretion cytokines changed. TNF alpha elevated while IL-1 beta level lowered almost to normal value. In 6 months both IL-1 beta and TNF alpha returned to normal values confirming stabilization of cytokine system and the end of inflammation. Cellular immunity did not change much. Thus, as inflammation in prostatic tissue is characterized by elevation of proinflammatory cytokines, in diagnosis of chronic prostatitis it will be valid to use markers TNF alpha and IL-1 beta as criteria of immune prognosis of prostatic exacerbation. Prostamol-uno does not induce changes in lymphocyte populations and impairment of immune status.

Intranasal delivery of a truncated recombinant human SP-D is effective at down-regulating allergic hypersensitivity in mice sensitized to allergens of Aspergillus fumigatus.

C57BL/6 mice were sensitized to Aspergillus fumigatus 1-week culture filtrate, which is rich in the non-glycosylated allergen Asp f1, a major allergen in allergic bronchopulmonary aspergillosis (ABPA). A comparison of the effect of treatment of allergen challenged mice by intranasal administration of a 60-kDa truncated recombinant form of human SP-D (rfhSP-D) or recombinant full length SP-A (rhSP-A) was undertaken. Treatment with rfhSP-D produced significant reduction in IgE, IgG1 and peripheral blood eosinophilia and treatment with rfhSP-D, but not rhSP-A resulted in a significant reduction in airway hyperresponsiveness as measured by whole body plethysmography. Lung histology revealed less peribronchial lymphocytic infiltration in mice treated with rfhSP-D. Intracellular cytokine staining of spleen homogenates showed increases in IL-12 and IFN-gamma and decrease in IL-4. The level of endogenous mouse SP-D was elevated sixfold in the lungs of sensitized mice and was not affected by treatment with rfhSP-D. Taken with our previous studies, with a BALB/c mouse model of ABPA using a 3-week A. fumigatus culture filtrate, the present results show that rfhSP-D can suppress the development of allergic symptoms in sensitized mice independent of genetic background and using a different preparation of A. fumigatus allergens.

Inhibitory effects of whole and parotid saliva on immunomodulators.

Based on the presence of cytokines in whole saliva and their association with resistance and susceptibility to infectious disease, the present study was designed to evaluate the diagnostic potential of a large panel of cytokines and chemokines in saliva. Despite the endogenous presence of Th1/Th2 and pro-inflammatory cytokines and several chemokines in whole and parotid saliva of most individuals tested, the detection of known concentrations of several recombinant cytokines and chemokines was inhibited immediately following their addition to each type of saliva. In contrast, purified immunoglobulins were unaffected by either whole or parotid saliva. Further studies revealed that the inhibition of immunoreactivity involved sequestration of the majority of cytokines affected and degradation of chemokines. These results suggest that absolute concentrations of cytokines/chemokines may not be fully detectable in saliva. Therefore, the diagnostic value of any cytokine/chemokine is questionable and should be evaluated independently as such.

1alpha,25-dihydroxyvitamin D3 shows strong and additive immunomodulatory effects with cyclosporine A in rat renal allotransplants.

BACKGROUND: Vitamin D3 and its metabolites have long been found to exert immunosuppressive effects both in vivo and in vitro. The present study investigated the effect of 1alpha,25-dihydroxycholecalciferol (1,25DHC) on vascularized renal allografts in rats. METHODS: Three days prior to transplantation, two groups of animals were subjected to 1,25DHC (1 microg/kg/day IP) and a low calcium diet, which was continued until the end of the experiments. Recipient organs were removed and single allografts were transplanted in a high responder strain combination (ACI --> Lewis). Following transplantation, low-dose cyclosporine A (3.2 mg/kg/day CsA) administration was started in two experimental groups of recipients (one group receiving 1,25 DHC additionally) whereas the control allograft recipients received no immunosuppression (control III). Graft survival and renal function was monitored until death or the end of experiments and allograft rejection was assessed histologically using the Banff classification. RESULTS: 1,25DHC significantly prolonged allograft survival in comparison to control III (9.6 +/- 1 vs. 5.7 +/- 0.2 days; P=0.009). In addition, a combination of 1,25DHC and low-dose CsA increased allograft survival compared to CsA administration alone (24 +/- 0.9 vs. 13 +/- 0.3 days; P=0.008). 1,25DHC preserved renal creatinine clearance and decreased proteinuria in comparison to control III, and the combination of 1,25DHC and low-dose CsA again showed an additive effect on preservation of renal function. 1,25DHC and low-dose CsA both decreased interleukin (IL)-2 and IL-12 expression levels in serum and allografts, and a combination treatment produced the strongest attenuation of IL-2 and IL-12 expression. In addition, 1,25DHC increased IL-4 and IL-10 expression levels in allografts, whereas CsA alone did not alter IL-4 and IL-10 expression. In contrast, combination of 1,25DHC and low-dose CsA showed a significant increase in IL-10 expression levels whereas IL-4 expression was not elevated. CONCLUSION: Monotherapy with 1,25DHC significantly prolongs survival of renal allografts and preserves graft function in rats. A combination of 1,25DHC and CsA caused an additive effect on graft survival with differential regulation of pro- and anti-inflammatory cytokines, as compared to 1,25DHC administration alone.

Therapeutic effects of FTY720, a new immunosuppressive agent, in a murine model of acute viral myocarditis.

OBJECTIVES: This study examines the efficacy of FTY720 (FTY), a new immunosuppressor, in the treatment of acute viral myocarditis in a murine model. BACKGROUND: Immunosuppressive agents have no proven therapeutic efficacy in experimental or clinical myocarditis. METHODS: Encephalomyocarditis virus was inoculated i.p. in DBA/2 mice on day 0. Postinoculation treatment consisted of FTY 10 mg/kg/day p.o. (FTY group), or cyclosporine A (CsA) 40 mg/kg/day p.o. (CsA group) or distilled water p.o. only (control group). Survival until day 14, as well as cardiac histopathology, virus concentrations, cytokines (interleukin [IL]-2, IL-12, interferon [IFN]-gamma and tumor necrosis factor [TNF]-alpha) and nitric oxide (NO) on day 5 were examined. RESULTS: In the control and CsA groups, all mice died within 10 and 7 days, respectively. However, in the FTY group, 27% of the animals survived up to day 14. Compared with the control group, 1) histological scores were significantly lower in the FTY group but unchanged in the CsA group; 2) virus concentration was significantly higher in the CsA group but not in the FTY group; 3) expressions of IL-2, IL-12 and IFN-gamma in the heart were suppressed in both the FTY and CsA groups, though suppression was weaker in the FTY group; 4) TNF-alpha and NO were significantly increased in the CsA group but not in the FTY group. CONCLUSIONS: FTY720 had a significant therapeutic effect in acute experimental myocarditis without inducing excessive virus replication. This report is the first to describe a beneficial effect by an immunosuppressive agent in the treatment of acute viral myocarditis.

Interleukin-12 in human milk.

The presence of interleukin-12 (IL-12) in 39 samples of human milk was investigated using the enzyme-linked immunosorbent assay. IL-12 (>40 pg/mL) was detected in 24 of the 39 samples collected (1408 +/- 2256 pg/mL, mean +/- SD, n = 24). A range of concentrations of IL-12 was observed in colostrum, transitional, and mature milk, with an apparent decrease in the mean concentration over time postpartum.