Molecular cloning, expression and characterization of an interleukin 27 p28 homolog in pig (Sus scrofa)

IL-27 is the newest member of the IL-12 cytokine family and plays an important role in the immune regulation. It is composed of two subunits, p28 and EBV-induced gene 3(EBI3). Although human and mouse IL-27 p28 genes have been cloned, pig IL-27 p28 gene has not ever been reported. In the present study, we have cloned and characterized the full-length cDNA of IL-27 p28 from pig. The open reading frame of pig IL-27 p28 gene is 720 bp, which encodes a protein of 239 amino acids with a predicted molecular mass of 26.6 kDa. The deduced amino acid sequence of pig IL-27 p28 showed a high degree of homology to human (63%) and mouse (58%). It was a 4-helix cytokine and belonged to 4-helix cytokine superfamily. Pig IL-27 p28 has one transmembrane region, one signal peptide, and one N-glycosylation site, two Protein kinase C phosphorylation sites, three Casein kinase II phosphorylation sites and one N-myristoylation site. For the expression of pig IL-27 p28 protein in a eukaryotic expression system the recombinant plasmid was constructed. The expression of pig IL-27 p28 in mammalian cells were confirmed by flow cytometry analysis, immunofluorescence and Western blot. The analysis also confirmed a cross reactivity with anti-mouse IL-27 p28 antibody. This is the first report of cloning and characterization of IL-27 p28 in pig.     

Development and validation of cytokine quantitative, real time RT-PCR assays for characterization of Asian elephant immune responses

Infectious disease is an important factor in Asian elephant health and long-term species survival. In studying disease pathogenesis, it is important to consider not only the pathogen, but also the effectiveness of the host immune response. Currently, there is a paucity of information available on elephant immune function. Measurement of cytokine levels within clinical samples can provide valuable information regarding immune function during health and disease that may elucidate disease susceptibility. To develop tools for assessment of elephant immune function, Asian elephant partial mRNA sequences for interleukin (IL)-2, IL-4, IL-10, IL-12, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and β-actin were determined. Sequence information was then utilized to design elephant-specific primers and probes for quantitative, real time, RT-PCR assays for the measurement of cytokine mRNA. Greater than 300 bps of Asian elephant mRNA sequence were determined for each cytokine of interest. Consistent and reproducible, real time, RT-PCR assays with efficiencies of greater than 93% were also developed. Assay sensitivities ranged from less than 1 to 5000 DNA copies with the exception of IL-12, which had a sensitivity of 42,200 copies. Employment of molecular techniques utilizing mRNA-based detection systems, such as real time, RT-PCR, facilitate sensitive and specific cytokine detection and measurement in samples from species for which commercial reagents are not available. Future studies utilizing these techniques to compare elephant immune function during health and in the face of infection will be useful for characterizing the contribution of the elephant immune system to disease.     

SOCS2真核表达载体的构建及在3T3 L1前体脂肪细胞中的表达

细胞因子信号抑制蛋白(SOCS)是细胞因子信号通路的重要负调控因子.构建SOCS2真核表达载体并转染3T3-L1前体脂肪细胞,为研究SOCS2对不同细胞因子调控脂代谢奠定基础.取小鼠皮下脂肪提总RNA进行RT-PCR,克隆获得pMDl8-SOCS2质粒载体,双酶切目的基因,并克隆至质粒载体pcDNA3.1,成功构建了重组质粒载体pcDNA3.1-SOCS2;重组质粒瞬时转染3T3-L1前体脂肪细胞系,以未转染和空载体转染作为对照,RT-PCR和Westem blotting检测重组质粒转染组SOCS2核酸和蛋白表达,表达水平均显著高于对照组,为进一步研究SOCS2基因对不同细胞因子调控脂肪代谢提供基础.     

复方人参皂苷纳米乳对OVA接种小鼠免疫功能的影响

旨在研究口服复方人参皂苷纳米乳对抗原免疫效果的影响。将小鼠分为正常对照组、卵清白蛋白(OVA)对照组、空白纳米乳组、盐酸左旋咪唑纳米乳组、人参皂苷纳米乳组、人参皂苷-盐酸左旋咪唑水溶液组和复方人参皂苷纳米乳组(高、中、低剂量),皮下注射OVA抗原2次,在首次和第2次注射前3d连续灌胃给药,第2次注射后14d,测定血清(IgG、IgG1和IgG2a)抗体水平、脾细胞因子(IFN-γ和IL-4)水平以及脾淋巴细胞增殖刺激指数。结果表明,复方人参皂苷纳米乳中剂量(50mg·kg-1)组IL-4、IgG和IgG1水平显著或极显著高于复方人参皂苷纳米乳低剂量组和盐酸左旋咪唑纳米乳组,IFN-γ、IgG2a及OVA诱导脾淋巴细胞增殖刺激指数显著或极显著高于人参皂苷纳米乳组、人参皂苷-盐酸左旋咪唑水溶液组、盐酸左旋咪唑纳米乳组、复方人参皂苷纳米乳高剂量组和低剂量组。口服复方人参皂苷纳米乳能诱导OVA免疫小鼠产生Th1/Th2混合型免疫反应,表明人参皂苷和盐酸左旋咪唑能协同增强小鼠的细胞免疫和体液免疫功能。     

尖海龙提取物对淋巴细胞的作用

[目的]研究尖海龙提取物的免疫调节活性。[方法]制备尖海龙提取物,用密度梯度离心法分离人外周血淋巴细胞（PBMC）,将海龙提取物作用于PBMC,观察PBMC的形态、细胞增殖率,同时检测经尖海龙提取物诱导后细胞因子的生成。[结果]结果表明,尖海龙提取物作用后PBMC圆形、发亮,正常生长并有增殖,随着时间的延长,细胞出现多形的活化状态;经尖海龙提取物诱导的效应细胞中可检测出IL-2和TNF-τ细胞因子。[结论]尖海龙提取物具有免疫调节活性,在癌症的治疗与预防方面具有应用价值。     

Infection with parasitic nematodes confounds vaccination efficacy

T helper (Th) cells produce signature cytokine patterns, induced largely by intracellular versus extracellular pathogens that provide the cellular and molecular basis for counter regulatory expression of protective immunity during concurrent infections. The production of IL-12 and IFN-γ, for example, resulting from exposure to many bacterial, viral, and protozoan pathogens is responsible for Th1-derived protective responses that also can inhibit development of Th2-cells expressing IL-4-dependent immunity to extracellular helminth parasites and vice versa. In a similar manner, concurrent helminth infection alters optimal vaccine-induced responses in humans and livestock; however, the consequences of this condition have not been adequately studied especially in the context of a challenge infection following vaccination. Demands for new and effective vaccines to control chronic and emerging diseases, and the need for rapid deployment of vaccines for bio security concerns requires a systematic evaluation of confounding factors that limit vaccine efficacy. One common albeit overlooked confounder is the presence of gastrointestinal nematode parasites in populations of humans and livestock targeted for vaccination. This is particularly important in areas of the world were helminth infections are prevalent, but the interplay between parasites and emerging diseases that can be transmitted worldwide make this a global issue. In addition, it is not clear if the epidemic in allergic disease in industrialized countries substitutes for geohelminth infection to interfere with effective vaccination regimens. This presentation will focus on recent vaccination studies in mice experimentally infected with Heligmosomoides polygyrus to model the condition of gastrointestinal parasite infestation in mammalian populations targeted for vaccination. In addition, a large animal vaccination and challenge model against Mycoplasma hyopneumonia in swine exposed to Ascaris suum will provide a specific example of the need for further work in this area, and for controlled field studies to assess the impact of other similar scenarios.     

Therapeutic strategies for COVID-19 based on its pathophysiological mechanisms

An outbreak of coronavirus disease 2019 (COVID-19) began in Wuhan, China in early December 2019, which dominated by pneumonia. As of March 11, 2020, the overall confirmed cases in China had reached 80 955. Part of them became seriously ill, such as acute respiratory distress syndrome, and was associated with ICU admission and high mortality. Therefore, how to prevent the patients from becoming serious is a key point in the treatment and control of COVID-19. This review provides an overview of pathophysiology of cytokine storm which was associated with COVID-19 severity, and proposes that anti-inflammation, antioxidation and oxygen-therapy at the early stage of disease are better therapeutic option to save COVID-19 patients.     

三种生殖激素和细胞因子在绵羊妊娠期的变化

研究胚胎移植受体绵羊妊娠期血液激素和细胞因子的动态变化与妊娠的关系.用放免试剂盒测定绵羊妊娠期（0、30 d、45 d、60 d、75 d、90 d、105 d、120 d、137 d）FSH、E2、P4三种激素和IL-6、IGF-Ⅱ、EGF三种细胞因子的含量.结果表明,生殖激素和细胞因子在绵羊妊娠期发生着动态变化,妊娠45 d,所测定激素和细胞因子水平在妊娠羊组（处理组和对照Ⅰ组）和早期流产组（对照Ⅱ）间存在差异.对照Ⅱ血液中FSH、E2、IL-6、EGF的水平显著高于处理组和对照Ⅰ组的妊娠成功绵羊（P＜0.05）,而P4、IGF-Ⅱ水平却低于妊娠成功各组.