Dietary mustard seeds (Sinapis alba Linn) suppress 1,2-dimethylhydrazine-induced immuno-imbalance and colonic carcinogenesis in rats

In a Wistar rat model, prolonged supplementation of mustard seed (MS) to the diet significantly ameliorates the induction of colorectal carcinomas by 1,2-dimethylhydrazine (DMH). The expression of the splenocyte major histocompatibility complex class I (MHCI) was found significantly enhanced, whereas that of the major histocompatibility complex class II (MHCII) was significantly decreased. Compared to that of control animals, the proportion of spleenic B- and dendritic cells (DC) was amplified in the MS group. The expressions of MHCI, as well as that of MHCII, were increased in DC cells; whereas in B cells, MHCI expression was augmented but that of MHCII moderately decreased. The percentages of CD8+CD28+ and CD4+CD28+ cells were increased in the MS group, while the CD4+CD25+Foxp3+ subset was depressed. Plasma analysis showed that DMH-exposure induced amplified amounts of interleukin (IL)-4, IL-5, IL-10, and transforming growth factor-beta, whereas MS feeding counteracted this effect but enhanced IL-2, IL12p70, IL21, TNF-alpha, and interferon-gamma. In the SW480 colon adenocarcinoma cell-line, the cytotoxicity of spleenic T-cells from MS-fed animals was significantly increased. In the DMH-exposed rats, the expression of perforin in the spleenic T-cells was dramatically decreased, whereas MS abolished this depression. In summary, dietary MS suppresses DMH-induced immuno-imbalance as well as colon carcinogenesis in rats.     

Interactions of proteoliposomes from serogroup B Neisseria meningitidis with bone marrow-derived dendritic cells and macrophages: adjuvant effects and antigen delivery

Exposure to proteoliposomes from serogroup B Neisseria meningitidis (PL) induced up-regulation of MHC-II, MHC-I, CD40, CD80 and CD86 expression on the surface of murine bone marrow-derived dendritic cells (DC). CD40, CD80 and CD86 were up-regulated on bone marrow-derived macrophages (MPhi) upon stimulation with PL. Both DC and MPhi released TNFalpha, but only DC produced IL12(p70) in response to PL. A small increase in the expression of MHC-II, CD40 and CD86, as well as production of IL12(p70), was observed on the cell surface of DC, but not MPhi from LPS-non-responder C3H/HeJ after exposure to PL. DC, but not MPhi, incubated with PL containing ovalbumin (PL-OVA) presented OVA-specific peptides to CD4+ and CD8+ OVA-specific T-cell hybridomas. These data clearly indicate that PL exert an immunomodulatory effect on DC and MPhi, with some contribution of non-LPS components besides the main role of LPS. The work also shows the potential of PL as a general system to deliver antigens to DC for presentation to CD4+ and CD8+ T-cells.     

Bacterial derived proteoliposome as ideal delivery system and cellular adjuvant

We explored the potential of a proteoliposome (PL) from the outer membrane of N. meningitidis B, as an immunopotentiator and as a vector for antigen delivery to dendritic cells (DC). DC were incubated with PL resulting in up-regulation of MHC-II, CD40, CD80, and CD86 expression and production of TNFalpha and IL12(p70). Ovoalbumin (OVA) was incorporated within PL (PL-OVA). PL-OVA presented OVA-specific peptides to CD4+ and CD8+ OVA-specific T-cell hybridomas. PL exerts an immunomodulatory effect on DC and is a general system to deliver antigens for presentation to CD4+ and CD8+ T-cells possibly implicated in the induction CD8+ cytotoxic T lymphocytes (CTLs) responses.     

Hepatitis B virus-like particles access major histocompatibility class I and II antigen presentation pathways in primary dendritic cells

Virus-like particles (VLPs) represent high density displays of viral proteins that efficiently trigger immunity. VLPs composed of the small hepatitis B virus envelope protein (HBsAgS) are useful vaccine platforms that induce humoral and cellular immune responses. Notably, however, some studies suggest HBsAgS VLPs impair dendritic cell (DC) function. Here we investigated HBsAgS VLP interaction with DC subsets and antigen access to major histocompatibility complex (MHC) class I and II antigen presentation pathways in primary DCs. HBsAgS VLPs impaired plasmacytoid DC (pDC) interferon alpha (IFNα) production in response to CpG in vitro, but did not alter conventional DC (cDC) or pDC phenotype when administered in vivo. To assess cellular immune responses, HBsAgS VLPs were generated containing the ovalbumin (OVA) model epitopes OVA₂₅₇–₂₆₄ and OVA₃₂₃–₃₃₉ to access MHCI and MHCII antigen presentation pathways, respectively; both in vitro and following immunisation in vivo. HBsAgS VLP-OVA₂₅₇–₂₆₄ elicited CTL responses in vivo that were not enhanced by inclusion of an additional MHCII helper epitope. HBsAgS VLP-OVA₂₅₇–₂₆₄ administered in vivo was cross-presented by CD8⁺ DCs, but not CD8⁻ DCs. Therefore, HBsAgS VLPs can deliver antigen to both MHCI and MHCII antigen presentation pathways in primary DCs and promote cytotoxic and helper T cell priming despite their suppressive effect on pDCs.     

Effects of parenteral lipid emulsions with different fatty acid composition on immune cell functions in vitro

BACKGROUND: Numerous studies suggest that immune function may be compromised by lipid emulsions rich in polyunsaturated fatty acids (PUFAs). In our study, we compared the effect of a new olive oil-based lipid emulsion (ClinOleic) containing a moderate level of PUFAs, with emulsions based on soybean oil (Intralipid or Ivelip), on immune functions of human cell in vitro. METHODS: Peripheral white blood cells were collected from healthy volunteers. Lymphocyte proliferation was evaluated by [3H]-thymidine incorporation after stimulation with either phytohemagglutinin (PHA) or antibodies against T-cell specific antigens. Lymphocytes subsets and T-cell activation markers (CD25 and HLA-DR) were measured by flow cytometry. The release of cytokines (interleukin [IL]-2, IL-1beta, and tumor necrosis factor-alpha [TNF-alpha]) was measured by enzyme-linked immunosorbent assay (ELISA), after lymphocytes or monocytes/macrophages stimulation with PHA or lipopolysaccharide (LPS). RESULTS: A significant dose-dependent inhibition of thymidine incorporation was observed with Intralipid and Ivelip (incorporation down to 39.9% of control, p < .001) whereas ClinOleic showed no inhibitory effect. Activation antigen expression on both CD4+ and CD8+ T-cells tended to decrease with Intralipid (CD25: -53.4% on CD4+ and -57.4% on CD8+; HLA-DR: -61.5% on CD4+ and -58.5% on CD8+) but not with ClinOleic (from -2.9% for CD25 on CD4+ to 16.7% for HLA-DR on CD4+). Intralipid decreased significantly IL-2 production (-39.0%, p < .05) whereas ClinOleic had little effect (-13.0%, NS). Intralipid and ClinOleic tended to inhibit to a similar extent the release of pro-inflammatory cytokines (TNF-alpha: -21.5% and -34.8%, IL-1beta: -45.1% and -40.3%; respectively). CONCLUSIONS: Our results suggest that an olive oil-based lipid emulsion could modulate immune response selectively, maintaining protective immunity and reducing inflammatory response. Olive oil may offer an immunologically neutral alternative to soybean oil for use in parenteral lipid emulsions.     

Interferon gamma (IFN-γ) negative CD4+ and CD8+ T-cells can produce immune mediators in response to viral antigens

Evaluation of antigen-specific T-cell responses to viral antigens is frequently performed on IFN-γ secreting cells. However, T-cells are capable of producing many more functions than just IFN-γ, some of which, like Perforin, are associated with immune protection in HIV-1 disease elite controllers. We evaluated the extent of missed T-cell functions when IFN-γ secretion is used as a surrogate marker for further evaluation of T-cell functions. Intracellular cytokine staining assay and flow cytometry were used to assess peripheral blood mononuclear cells (PBMCs) from 31 HIV-infected ART-naive individuals for the extent to which gated CD4+ and CD8+ IFN-γ producing and non-producing T-cells also secreted IL-2, Perforin, and TNF-α functions. Similarly, the extent of missed virus-specific responses in IFN-γ ELISpot assay negative T-cells from 5 HIV-1 uninfected individuals was evaluated. Cells from HIV-infected individuals were stimulated with pooled consensus group M (Con M) peptides; and those from healthy individuals were stimulated with pooled adenovirus (Ad) peptides. Overall, frequencies of virus-specific IFN-γ secreting CD4+ and CD8+ cells were low. Proportions of IFN-γ negative CD4+ expressing IL-2, Perforin, or TNF-α to Con M were significantly higher (5 of 7 functional profiles) than the corresponding IFN-γ positive CD4+ (0 of 7) T-cell phenotype, p?=?0.02; Fisher’s Exact test. Likewise, proportions of CD8+ T-cells expressing other functions were significantly higher in 4 of the 7 IFN-γ negative CD8+ T-cells. Notably, newly stimulated Perforin, identified as Perforin co-expression with IL-2 or TNF-α, was significantly higher in IFN-γ negative CD8+ T-cell than in the positive CD8+ T-cells. Using SEB, lower responses in IFN-γ positive cells were most associated with CD4+ than CD8+ T-cells. These findings suggest that studies evaluating immunogenicity in response to HIV and Adenovirus viral antigens should not only evaluate T-cell responsiveness among IFN-γ producing cells but also among those T-cells that do not express IFN-γ.     

Treg抑制DC活性调控小鼠肺部炎症发生发展

目的 探讨调节性T细胞(Treg)在变态反应性肺泡炎中的作用及Treg对变态反应性肺泡炎发生发展过程中树突状细胞(DC)活性影响。方法 腹腔注射抗CD25单克隆抗体建立小鼠Treg消除模型,通过气管灌注1-3-β葡聚糖建立小鼠变态反应性肺泡炎模型,采用流式细胞术检测小鼠肺门淋巴结中树突状细胞活性;苏木素-伊红染色进行病理观察,比较各组小鼠肺组织切片中炎症反应和肉芽肿程度。结果 与葡聚糖模型组小鼠比较,Treg消除小鼠肺门淋巴结中CD4+CD25+Treg百分比[(6.76±1.0)%]明显降低(P < 0.05);组织病理观察显示,Treg消除小鼠肺组织局部炎症反应程度明显重于葡聚糖模型组小鼠;流式细胞术检测发现,葡聚糖灌注后早期,小鼠肺门淋巴结中CD80+MHCII+DC的比例较对照组明显增加;Treg消除小鼠肺门淋巴结和脾细胞中CD80+MHCII+DC比例[(37.3±5.8)%]较葡聚糖模型组小鼠[(24.3±1.9)%]显著升高,差异具有统计学意义(P < 0.05)。结论 Treg参与抑制变态反应性肺泡炎的发生发展,其机制可能与抑制DC活性发挥免疫抑制作用有关。     

White button mushroom enhances maturation of bone marrow-derived dendritic cells and their antigen presenting function in mice

Mushrooms have been shown to enhance immune response, which contributes to their antitumor property. White button mushrooms (Agaricus bisporus) (WBM) constitute 90% of the total mushrooms consumed in the United States; however, the health benefit of this strain in general is not well studied. Furthermore, little is known about WBM's immunologic effects. Dendritic cells (DC) are the most potent antigen presenting cells and play a pivotal role in immune response by linking innate and adaptive immune responses. In this study, we investigated the effect of in vitro supplementation with WBM on maturation of bone marrow-derived DC (BMDC) of C57BL mice. BMDC were differentiated in the presence of whole mushroom concentrate at 50, 100, or 200 mg/L. Results showed that mushroom supplementation dose dependently increased the expression of maturation markers CD40, CD80, CD86, and major histocompatibility complex-II. Consistent with the changes in the phenotypic markers, functional assay for DC maturation showed that mushroom supplementation decreased DC endocytosis and increased intracellular interleukin (IL)-12 levels. Furthermore, using a syngeneic T cell activation model, we found that WBM-supplemented DC from BALB/c mice presented ovalbumin antigen to T cells from DO11.10 mice more efficiently as demonstrated by increased T cell proliferation and IL-2 production. In conclusion, WBM promote DC maturation and enhance their antigen-presenting function. This effect may have potential in enhancing both innate and T cell-mediated immunity leading to a more efficient surveillance and defense mechanism against microbial invasion and tumor development.     

Investigation of regulatory T cells in anorexia nervosa

OBJECTIVE: The aim of our study was to determine, how severe calorie restriction in anorexia nervosa (AN) may influence regulatory T (Treg) cells and their cellular networks, that is, their main inducers (dendritic cells (DC) and monocytes) and their target cells, CD4+ lymphocytes. DESIGN: We measured the prevalence of Tregs, myeloid and plasmocytoid DC. The prevalence of tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-12-positive monocytes, IL-2, IL-4 and interferon (IFN)-gamma positive CD4+ cells was determined by intracellular staining after activation. SETTING AND SUBJECTS: In total, 21 AN patients and 19 healthy age-matched controls (body mass index values, median (range): 14.9 (11.1-17.4) vs 23.2 (19.5-27.4) kg/m(2)) have been recruited. RESULTS: Prevalence of Tregs, DCs, TNF-alpha and IL-12-positive monocytes, IL-4 and IFN-gamma-producing CD4+ cells were similar in AN and controls. The prevalence of IL-2-positive CD4+ cells was somewhat lower in AN (% value, median (range): 12.05 (7.50-16.70) vs 14.40 (12.00-22.00), P<0.05). None of these parameters correlated with the patients' clinical characteristics. CONCLUSIONS: Our results suggest that the antigen presenting cell - regulatory T cell - CD4+ lymphocyte axis is not affected by calorie and nutritional deficiency.     

Th-1/Th-2 type cytokine profiles of pig T-cells cultured with antigen-treated monocyte-derived dendritic cells

To examine the effects of cytokine environment at the time of antigenic exposure on T-cell cytokine profiles following T-cell-antigen presenting cell (APC) interaction, pig monocyte-derived dendritic cells (mDCs) were treated with hen egg white lysozyme (HEWL) or killed Mycobacterium tuberculosis (Mtb) alone or with a recombinant pig cytokine (TNF-alpha, interleukin (IL)-12, IL-10, interferon (IFN)-gamma or IL-6) and then incubated with autologous T-cell-enriched lymphocytes. Messenger RNA was isolated from the T-cells and used to evaluate the effects of treatment on IL-12p35, IFN-gamma, IL-4, IL-10 and IL-13 expression using RT-PCR. T-cells exposed to HEWL-treated mDCs expressed high IL-13 and moderate IL-10 and IFN-gamma, suggesting T-helper 2 (Th-2) bias. Addition of any cytokine during HEWL treatment of mDCs reduced subsequent expression of IL-10 and IL-13 by T-cells. Added IL-12 increased IFN-gamma mRNA. T-cells exposed to Mtb-treated mDCs expressed increased IFN-gamma and decreased IL-10 suggesting Th-1 bias. Addition of cytokines to mDCs treated with Mtb altered T-cell cytokine mRNA expression such that TNF-alpha, IFN-gamma or IL-12 increased IFN-gamma; IL-12 and IFN-gamma suppressed IL-10, while IL-10 and IL-12 enhanced IL-13. Messenger RNA for IL-4 and IL-12p35 was not detected in the T-cells. Results suggest Th-1/Th-2 type response bias in pigs T-cells as a function of antigen type and that cytokine environment at the time of antigen-mDC interaction alters cytokine profiles of T-cells responding to antigen-pulsed mDCs. Hence, cytokines may allow designed steering of porcine immune response.     

Therapeutic vaccination with dendritic cells pulsed with tumor-derived Hsp70 and a COX-2 inhibitor induces protective immunity against B16 melanoma

Prophylactic immunization of mice with autologous tumor-derived heat shock proteins (Hsp) generates effective anti-tumor immunity. However, this approach is ineffective when used therapeutically, partly due to the immunosuppressive effects of the growing tumor. Here we sought to overcome this problem by therapeutic vaccination with dendritic cells (DC) pulsed with Hsp70 and a COX-2 inhibitor. We found that Hsp70 induces IL-6 and IL-10 production and suppressed expression of CD40 on DC. Incubation of DC with tumor-conditioned medium attenuated Hsp70-induced expression of CD80 and induced expression of COX-2. Inhibition of COX-2 partially reversed the stimulatory effect of Hsp70 on DC IL-6 and IL-10 production and enhanced expression of CD80 and MHC classes I and II. Therapeutic administration of DC pulsed in vitro with Hsp70 in the presence of a COX-2 inhibitor significantly reduced progression of B16 tumors in mice and significantly enhanced survival. This was associated with a reduction in the frequency of IL-10-producing CD4(+) T cells and enhancement of IFN-gamma-producing CD8(+) T cells. Our findings provide a novel immunotherapeutic approach against cancer based on attenuation of COX-2-mediated immunosuppression using in vitro modulated DC.     

苯中毒再生障碍性贫血患者骨髓细胞免疫功能和Fas、CD34抗原变化

目的　探讨细胞免疫功能、Fas和CD3 4抗原与苯中毒再生障碍性贫血发生、发展及转归的关系。方法　用流式细胞仪检测2 4例苯中毒再障(BPAA)和16例无苯制剂接触史的再障(AA)患者治疗前后骨髓单个核细胞Fas、CD3 4的表达率和T细胞亚群变化。结果　苯中毒再生障碍性贫血和再障患者骨髓单个核细胞Fas表达率显著增高(P <0 . 0 0 1) ,CD3 4阳性率明显降低(P <0 . 0 0 1) ;CD3 + CD8+ T淋巴细胞明显升高(P <0 . 0 5 ) ,CD4+ /CD8+ 比值明显降低(P <0 . 0 0 1) ;治疗缓解后Fas、CD3 4阳性率、CD3 + CD8+ T淋巴细胞和CD4+ /CD8+ 比值趋于正常(P >0 . 0 5 )。但在AA组Fas、CD3 4阳性率和CD4+ /CD8+ 比值较正常对照组差异仍有显著性。两组再障患者之间比较差异无显著性。结论　细胞免疫功能紊乱、Fas抗原的异常表达和CD3 4+ 干/祖细胞损伤,可能与苯中毒再生障碍性贫血病理发病机制有关,用环孢霉素A治疗可能有效。     

Dendritic cells stimulated with outer membrane protein A (OmpA) of Salmonella typhimurium generate effective anti-tumor immunity

Gram-negative bacterial outer membrane proteins (Omps) have an important role in pathogenesis and signal reception. We previously reported that Acinetobacter OmpA (AbOmpA) induced maturation of bone marrow-derived dendritic cells (BMDCs) and that AbOmpA-primed DCs produced IL-12 which generated Th1 CD4⁺ T-cells. We analyzed the effects of Salmonella typhimurium OmpA (OmpA-Sal) on dendritic cell (DC) maturation in the present study, and determined that tumor antigen-pulsed DCs stimulated with OmpA-Sal induced anti-tumor responses in a mouse model. OmpA-Sal activated BMDCs by augmenting expression of MHC class II and of the co-stimulatory molecules CD80 and CD86. RT-PCR revealed that IL-12(p40) gene expression is highly augmented in OmpA-Sal-stimulated BMDCs. DNA (CRT/E7) vaccination combined with OmpA-Sal stimulation generated more antigen-specific CD8⁺ T-cells in the present study. Certain antigen-pulsed BMDCs stimulated with OmpA-Sal induced strong PADRE-specific CD4⁺ and E7-specific CD8⁺ T-cell responses. In addition, BMDCs stimulated with OmpA-Sal (OmpA-Sal-BMDCs) and pulsed with both E7 and PADRE peptide generated greater numbers of E7-specific CD8⁺ effector and memory T-cells than those pulsed with E7 peptide alone. E7- and PADRE-expressing OmpA-Sal-BMDC vaccines resulted in significant long-term protective anti-tumor effects in vaccinated mice. Our data suggested that E7- and PADRE-expressing BMDCs that were matured in the presence of OmpA-Sal might enhance anti-tumor immunity and support the therapeutic use of OmpA-Sal in DC-based immunotherapy.     

三叶青提取物对人树突状细胞功能的影响

目的探讨三叶青提取物TH-b对树突状细胞(dendritic cells,DC)功能的影响。 方法分离健康人外周血中单个核细胞,用粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony stimulating factor,GM-CSF)、白介素-4 (interleukin-4,IL-4)、γ-干扰素(interferon-γ,IFN-γ)和脂多糖(lipopolysaccharide,LPS)诱导培养DC。用念珠菌孢子法检测经TH-b作用后的DC吞噬能力;将经TH-b作用后的DC与同种外周血淋巴细胞进行混合培养,以检测淋巴细胞增殖率;采用流式细胞术检测DC表面标记CD195和TH-b作用后DC的表面标记CD80、CD83、CD86、HLA-DR;用酶免疫法检测TH-b作用后DC的培养上清液中的IL-12含量。 结果培养7 d后DC CD195表达水平(85.3%)明显高于培养前(2.4%)。三叶青提取物TH-b部分对DC吞噬能力有作用明显,TH-b浓度为0.62 μg/ml时吞噬能力最强。各种TH-b浓度作用后DC的细胞表面CD80、CD83、CD86和HLA-DR表达均明显升高,以0.62 μg/ml最明显,分别为88.56%±8.16%、86.75%±7.59%、99.34%±8.24%、96.15%±8.27%,显著高于对照组的71.38%±7.31%、43.61%±4.29%、93.41%±8.34%、79.26%±5.12%,差异有统计学意义(P<0.05)。TH-b作用后的DC与淋巴细胞在1:1、1:10、1:100的比例下,对同种外周血淋巴细胞具有强烈的激发和促增殖作用,以1:1最明显。TH-b作用DC 48 h后,DC分泌的IL-12均明显升高,在0.62 μg/ml时IL-12分泌达最高值(526.3 pg/ml),显著高于对照组(260.1 pg/ml)(P<0.05),并有浓度窗现象。 结论低浓度三叶青提取物TH-b部分能够显著促进人DC吞噬功能,促进DC表面标志物CD80和CD86的表达,提高DC成熟标记和IL-12分泌能力。     

Tuberculin skin test reaction is related to memory,but not naive CD4+ T cell responses to mycobacterial stimuli in BCG-vaccinated young adults

Bacillus Calmette-Guérin (BCG) is the only vaccine available against tuberculosis and the tuberculin skin test (TST) is the most widely used method to detect BCG take. However, subjects may remain TST-negative, even after several BCG administrations. To investigate some of the potential reasons underlying this inability of developing tuberculin sensitivity in response to BCG we compared the effect of different mycobacterial stimuli in the groups differently responding to tuberculin. TST was performed on 71 healthy adults aged 25–30 years, who had received BCG in their childhood, and considered TST-positive at ≥10 mm. Dendritic cells (DCs) were incubated with PPD, live BCG or rBCGhIL-18, producing human IL-18. The latter strain was used to investigate whether the production of IL-18 could overcome some of the immune read-out limitations in the TST-negative subjects. CD86, CD80, CD40, and DC-specific intracellular adhesion molecule-3 grabbing nonintegrin (DC-SIGN) expression was analysed by flow cytometry and IL-10, IL-23 and IP-10 secretion in culture supernatants by ELISA. In DCs-T cell co-cultures with naive and memory CD4⁺ T cells, the IFN-γ and IL-10 levels were determined by ELISA. We found no difference in IL-10 and IFN-γ production by naive T cells between the TST-negative and TST-positive subjects. However, IFN-γ was produced in significantly higher amounts by memory T cells incubated with PPD, BCG or rBCGhIL-18-pulsed DCs in TST-positive than in TST-negative subjects, whereas the numbers of the IFN-γ-producing T cells were similar in both groups. This difference may be partially due to a decreased CD40 and enhanced reduction in DC-SIGN expression by DCs of TST-negative versus TST-positive subjects. A strong effect of IL-18 expression by rBCGhIL-18 on IL-23 production by the DC was seen in both groups, which likely was the reason for the increased IFN-γ production by naïve T cells upon incubation with mycobacteria-pulsed DC, regardless of the TST status.     

Regulation of functional and regressing stages of corpus luteum development in mice. Role of reactive oxygen species

The endocrine and immune systems modulate ovarian function. The aim of the present work was to compare the status of various modulating factors in two well-defined stages of corpus luteum (CL) development (the functional stage and the regressing stage) by means of a gonadotropin-synchronised mouse model. At the regressing stage of CL development, we found that ovarian tissue showed increased prostaglandin (PG) F(2alpha) and diminished PGE levels concomitantly with enhanced protein abundance of ovarian cyclooxygenase 2, the inducible isoform of the limiting enzyme of PG synthesis. We also found both enhanced lipid peroxidation and enhanced total superoxide dismutase activity, as well as inhibited catalase activity and inhibited total hydroxyl radical scavenger capacity, when compared with ovaries at the functional stage. In addition, at the regressing stage we observed an increased percentage of CD8+ (cytotoxic/suppressor) T-cells and a decreased percentage of CD4+ (helper) T-cells from ovarian-draining lymph nodes. Also, the serum interleukin (IL)-2, IL-4 and IL-10 were diminished as compared with the functional stage. We conclude that a pro-oxidant status together with a pro-inflammatory response is responsible for the loss of luteal function.     

Decrease in Mucosal IL17A,IFNγ and IL10 Expressions in Active Crohn’s Disease Patients Treated with High-Dose Vitamin D Alone or Combined with Infliximab

Background: Vitamin D treatment may reduce Crohn’s disease (CD) activity by modulating the mucosal immune function. We investigated if high-dose vitamin D +/− infliximab modulated the mucosal cytokine expression in active CD. Methods: Forty CD patients were randomized into: infliximab + vitamin D; infliximab + placebo-vitamin D; placebo-infliximab + vitamin D or placebo-infliximab + placebo-vitamin D. Infliximab (5 mg/kg) and placebo-infliximab were administered at weeks 0, 2 and 6. Oral vitamin D was administered as bolus 200,000 international units (IU) per week 0 followed by 20,000 IU/day for 7 weeks or placebo. Endoscopy with biopsies was performed at weeks 0 and 7 where endoscopic activity was measured and mucosal mRNA cytokine expression was examined. C-reactive protein (CRP), fecal calprotectin and Harvey-Bradshaw Index (HBI) were measured at weeks 0, 2 and 6. Results: High-dose vitamin D treatment alone and combined with infliximab decreased the IL17A, IFNγ and IL10 expression. High-dose vitamin D alone did not significantly decrease the disease activity, CRP or calprotectin. Combined infliximab and vitamin D treatment was not clinically significantly superior to monotherapy with infliximab. Conclusions: High-dose vitamin D as monotherapy and combined with infliximab decreases IL17A, IFNγ and IL-10 expression in mucosa within treatment groups. This did not induce a statistically significant decreased disease activity. EudraCT no.2013-000971-34.     

H9N2 avian influenza virus enhances the immune responses of BMDCs by down-regulating miR29c

Avian influenza virus (AIV) of the subtypes H9 and N2 is well recognised and caused outbreaks-due to its high genetic variability and high rate of recombination with other influenza virus subtypes. The pathogenicity of H9N2 AIV depends on the host immune response. Dendritic cells (DCs) are major antigen presenting cells that can significantly inhibit H9N2 AIV replication. MicroRNAs (miRNAs) influence the ability of DCs to present antigens, as well as the ability of AIVs to infect host cells and replicate. Here, we studied the molecular mechanism underlying the miRNA-mediated regulation of immune function of mouse DCs. We first screened for and verified the induction of miRNAs in DCs after H9N2 AIVstimulation. We also constructed miR29c, miR339 and miR222 over-expression vector and showed that only the induction of miR29c lead to a hugely increased expression of surface marker MHCII and CD40. Whilst the inhibition of miR29c, miR339 and miR222 in mouse DCs would repressed the expression of DCs surface markers. Moreover, we found that miR29c stimulation not only up-regulate MHCII and CD40, but also enhance the ability of DCs to activate lymphocytes and secrete cytokines IL-6 or TNF-a. Furthermore, we found that Tarbp1 and Rfx7 were targeted and repressed by miR29c. Finally, we revealed that the inhibition of miR29c marvelously accelerated virus replication. Together, our data shed new light on the roles and mechanisms of miR29c in regulating DC function and suggest new strategies for combating AIVs.