Humoral immune responses in koalas (Phascolarctos cinereus) either naturally infected with Chlamydia pecorum or following administration of a recombinant chlamydial major outer membrane protein vaccine

The development of a vaccine is a key strategy to combat the widespread and debilitating effects of chlamydial infection in koalas. One such vaccine in development uses recombinant chlamydial major outer membrane protein (rMOMP) as an antigen and has shown promising results in several koala trials. Previous chlamydial vaccine studies, primarily in the mouse model, suggest that both cell-mediated and antibody responses will be required for adequate protection. Recently, the important protective role of antibodies has been highlighted. In our current study, we conducted a detailed analysis of the antibody-mediated immune response in koalas that are either (a) naturally-infected, and/or (b) had received an rMOMP vaccine. Firstly, we observed that naturally-infected koalas had very low levels of Chlamydia pecorum-specific neutralising antibodies. A strong correlation between low IgG total titers/neutralising antibody levels, and higher C. pecorum infection load was also observed in these naturally-infected animals. In vaccinated koalas, we showed that the vaccine was able to boost the humoral immune response by inducing strong levels of C. pecorum-specific neutralising antibodies. A detailed characterisation of the MOMP epitope response was also performed in naturally-infected and vaccinated koalas using a PepScan epitope approach. This analysis identified unique sets of MOMP epitope antibodies between naturally-infected non-protected and diseased koalas, versus vaccinated koalas, with the latter group of animals producing a unique set of specific epitope-directed antibodies that we demonstrated were responsible for the in vitro neutralisation activity. Together, these results show the importance of antibodies in chlamydial infection and immunity following vaccination in the koala.     

T、B淋巴细胞及细胞因子在三氯乙烯诱发过敏反应中的作用

目的利用三氯乙烯(TCE)染毒的小鼠模型,研究T、B淋巴细胞及细胞因子在TCE诱发过敏反应中的作用。方法以皮肤接触同时结合吸入对12只小鼠进行致敏处理,6周后耳部涂抹TCE进行激发。以耳肿胀系数作为评价小鼠过敏反应的指标;分离脾细胞体外培养,采用噻唑蓝(MTT)法测定脾淋巴细胞抗原特异性增殖反应;酶联免疫吸附测定(ELISA)方法测定细胞培养上清液中IgG和白介素-4(IL-4)、γ型干扰素(IFN-γ)水平;应用流式细胞技术进行淋巴细胞亚群分析。结果TCE致敏组小鼠脾淋巴细胞加入TCE体外培养后,细胞存活率明显高于溶剂对照组[(79±10)%vs(63±11)%,P<0·05];细胞培养上清液中IgG水平(与二甲基亚砜对照孔比较的相对值)在两组间比较差异有显著性[(70±5%)vs(53±6)%,P<0·01]。流式细胞分析结果显示,TCE致敏组脾淋巴细胞与TCE共培养后,CD3 T细胞占总淋巴细胞的比例[(41·6±4·4)%]及CD4 /CD8 (2·1±0·6)与对照组比较[分别为(39·4±4·0)%和(2·3±0·9)],差异无显著性(P>0·05)。TCE致敏组小鼠激发时未见耳肿胀,然而TCE致敏组小鼠IFN-γ/IL-4比值(0·54±0·12)明显低于溶剂对照组(0·90±0·22,P<0·01)。结论TCE能够诱导T淋巴细胞的增殖活化,分泌Th2型细胞因子,并刺激B淋巴细胞分泌抗原特异性的IgG抗体。     

Adjuvanticity and protective immunity of Plasmodium yoelii nigeriensis blood-stage soluble antigens encapsulated in fusogenic liposome

In our previous studies we established fusogenic properties of lipids isolated from edible yeast Saccharomyces cerevisiae (S. cerevisiae). We demonstrated that liposomes prepared from S. cerevisiae membrane lipid (saccharosome) can deliver encapsulated antigen into cytosol of the antigen presenting cells and elicit antigen specific cell mediated as well as humoral immune responses. In this study, we evaluated immunological behavior of saccharosome encapsulated cytosolic proteins (sAg) of Plasmodium yoelii nigeriensis in BALB/c mice. Immunization with antigen (sAg) encapsulated in saccharosome resulted in enhancement of CD4+ and CD8+ T cell populations and also up-regulated the expression of CD80 and CD86 molecules on the surface of antigen presenting cells. Further, immunization with saccharosome-encapsulated sAg-induced elevated levels of both IFN-gamma and IL-4 cytokines in the immunized mice when compared to egg PC liposome encapsulated sAg or its IFA emulsified form. Saccharosome-mediated immunization resulted in induction of high level of total antibody response with preponderance of IgG2a isotype as well. The data of this study suggest that saccharosome-based vehicle can emerge as an effective vaccine in imparting protection against various intracellular pathogens including Plasmodium yoelii nigeriensis.     

北半球季节性甲型流感疫苗株HA抗原表位的分析

摘要:目的　分析近20年北半球甲型流感疫苗株血凝素（Hemagglutinin,HA）抗原的氨基酸和N-糖基化位点变异情况。方法　利用生物信息学软件分析1990-2012年季节性流感疫苗株H1N1的4个抗原决定簇Sa、Sb、Ca、Cb和H3N2的4个抗原决定簇A、B、C、D的氨基酸和N-糖基化位点变异情况。结果　（1）H1N1疫苗株Sb抗原决定簇28、40位氨基酸N-糖基化位点相对保守,而142位氨基酸发生改变。（2）Sb抗原决定簇上无N-糖基化位点。（3）H3N2疫苗株B、D抗原决定簇2、479、181、301位氨基酸N-糖基化位点相对保守,而149位氨基酸发生改变。（4）H3N2疫苗株的N-糖基化位点不在抗原位点上。结论　（1）流感疫苗株HA抗原氨基酸变异集中在HA1蛋白环状部位。（2）流感疫苗株的N-糖基化位点在相对保守的序列,并且相同亚型的流感疫苗株有相似的N-糖基化位点。（3）HA蛋白变异可以推测流感病毒基因的改变,为选择与之匹配的流行株提供参考。     

Seasonal influenza vaccines induced high levels of neutralizing cross-reactive antibody responses against different genetic group influenza A(H1N1)pdm09 viruses

Influenza A(H1N1)pdm09 viruses have been circulating throughout the world since the 2009 pandemic. A/California/07/2009 (H1N1) virus was included in seasonal influenza vaccines for seven years altogether, providing a great opportunity to analyse vaccine-induced immunity in relation to the postpandemic evolution of the A(H1N1)pdm09 virus. Serum antibodies against various epidemic strains of influenza A(H1N1)pdm09 viruses were measured among health care workers (HCWs) by haemagglutination inhibition and microneutralization tests before and after 2010 and 2012 seasonal influenza vaccinations. We detected high responses of vaccine-induced neutralizing antibodies to six distinct genetic groups. Our results indicate antigenic similarity between vaccine and circulating A(H1N1)pdm09 strains, and substantial vaccine-induced immunity against circulating epidemic viruses.     

天然盐藻提取物对小鼠免疫功能影响的研究

目的 研究天然盐藻提取物对小鼠免疫功能有否影响。方法 分别以12.5、25.0、37.5 mg/（kg BW d）3个剂量天然盐藻提取物以玉米油溶解经口灌胃给予小鼠30 d,另设玉米油对照组;于试验的30 d测定小鼠的细胞免疫、体液免疫、单核巨噬细胞、NK细胞活性。结果 天然盐藻提取物能增强小鼠迟发型变态反应和小鼠的淋巴细胞转化能力、能提高小鼠的半数溶血值和小鼠抗体生成细胞数、能提高小鼠单核-巨噬细胞功能和增加NK细胞活性,以上差异均有统计学意义（均P〈0.05）。结论 在一定剂量范围内天然盐藻提取物具有增强免疫力功能作用。     

Evaluation of immune response,microbiota, and blood markers after probiotic bacteria administration in obese mice induced by a high-fat diet

ObjectiveObesity is associated with alterations in intestinal microbiota and immunity. The aim of this study was to determine the effect of probiotic Lactobacillus casei CRL 431 administration on intestinal and humoral immune response, clinical parameters, and gut microbiota was evaluated using a high-fat diet to induce obesity in a mouse model.MethodsAdult mice received a conventional balanced diet or a high-fat diet supplemented with milk, milk fermented by Lactobacillus casei (FM), L. casei as suspension, or water over 60 d. Histology of liver and small intestine (SI), immunoglobulin A-positive cells and macrophages in SI, phagocytic activity of spleen and peritoneal macrophages, and humoral immune response to ovalbumin were studied. Clinical parameters in serum and gut microbiota were also analyzed.ResultsFM was the most effective supplement for decreasing body weight and clinical parameters in serum. The histology of liver and SI was also improved in obese mice given FM. These animals had increased numbers of immunoglobulin A-positive cells and macrophages in SI. The gut microbiota showed that obese mice given probiotics had increased Bacteroides and bifidobacteria. Administration of FM or L. casei as suspension enhanced the phagocytic activity of macrophages. The anti-ovalbumin specific immune response was not increased by any supplement assayed.ConclusionAdministration of probiotics to obese hosts improved the gut microbiota and the mucosal immunity altered by obesity, down-regulated some biochemical parameters in blood associated with metabolic syndrome, and decreased liver steatosis. These results demonstrate the potential use of probiotics in obese individuals to decrease the body weight and to improve the biochemical and immunologic parameters altered by obesity.     

纳豆片对小鼠免疫功能的影响

目的:探讨纳豆片对小鼠免疫功能的影响。方法:用0.25 g/kg.bw、0.50 g/kg.bw、1.50 g/kg.bw剂量的纳豆片给小鼠灌胃30 d,检测各项免疫指标。结果:纳豆片能提高小鼠的脾淋巴细胞增殖能力、血清溶血素、腹腔巨噬细胞吞噬指数以及碳廓清吞噬指数,对小鼠的NK细胞活性的有一定的升高趋势。结论:提示纳豆片具有增强小鼠免疫功能的作用。     

中和性单克隆抗体检测陕西省70株脊髓灰质炎病毒抗原结果分析

我们应用中和性McAb对陕西省1979～1988年从病人分离的70株Polio病毒进行型内抗原分析,将32株Ⅰ型病毒分为PI—2(28株)、P1—3(3株)和P1—5(1株)3个类型。27株Ⅱ型病毒分为P2—1(19株)和P2—2(8株)2个类型,11株Ⅲ型病毒分为P3—1(5株)、P3—2(3株)、P3—3(1株)和具有新抗原特征的陕P3—1(2株)4个类型。结果反映了陕西省近10年Polio病毒型内抗原特征类型的分布、变迁及其与疾病流行的关系。     

Antibody responses and cross protection against lethal influenza A viruses differ between the sexes in C57BL/6 mice

A mouse model was used to determine if protective immunity to influenza A virus infection differs between the sexes. The median lethal dose of H1N1 or H3N2 was lower for naïve females than males. After a sublethal, primary infection with H1N1 or H3N2, females and males showed a similar transient morbidity, but females generated more neutralizing and total anti-influenza A virus antibodies. Immunized males and females showed similar protection against secondary challenge with a homologous virus, but males experienced greater morbidity and had higher lung viral titers after infection with a lethal dose of heterologous virus. Females develop stronger humoral immune responses and greater cross protection against heterosubtypic virus challenge.     

鱼油脂肪乳对胃肠道恶性肿瘤病人术后营养状况和体液免疫的影响

目的:观察鱼油脂肪乳对胃肠道恶性肿瘤病人术后营养状况和体液免疫的影响. 方法: 将30例胃肠道恶性肿瘤病人随机分为研究组和对照组,每组15例.术后两组病人使用等氮、等热量的PN支持,研究组添加鱼油脂肪乳.于术前、术后第1天和第6天分别检测血肝肾功能、血脂、Hb、ALB、TF、淋巴细胞计数(TLC)、B淋巴细胞亚群(B1、B2)、免疫球蛋白(IgG、IgM、IgA)和补体(C3、C4)水平. 结果: 两组病人术后第6天肝肾功能、血脂水平与术前相比均无显著性差异.研究组病人术后第6天TLC、IgG、IgM、C3显著高于对照组(P<0.05).两组病人Hb、ALB、TF和B淋巴细胞亚群无显著性差异. 结论: 胃肠道恶性肿瘤病人术后使用鱼油脂肪乳安全可耐受,能提高体液免疫功能,但在短期内病人的营养状况未见改善.     

Local cytokine and inflammatory responses to candidate vaginal adjuvants in mice

The current study was undertaken to explore the correlation of adjuvanticity and local inflammatory response elicited in the murine vagina and the draining lymph nodes following local administration of two candidate vaginal adjuvants, Toll like receptor (TLR) 9 agonist CpG ODN, and a non-TLR targeting molecule α-galactosylceramide (α-GalCer). Using real-time PCR array analysis, we could show that a group of 13 common cytokine genes are activated in the vagina within 24 h after vaginal administration of these adjuvants, including Ccl2, Ccl7, Ccl12, Ccl19, Ccl20, Ccl22, Cxcl1, Cxcl5, Il10 and the Th1-inducing molecules Ifng, Cxcl9, Cxcl10 and Cxcl11. A high degree of inflammation in and damage to the epithelium was exclusively observed in the vagina of the CpG ODN treated mice, which was reversed within 48 h. These results indicate that there is a group of common genes that correlate with the adjuvanticity of CpG ODN and α-GalCer in the vagina, and that α-GalCer induces less of local inflammatory reactions in the murine vagina compared to CpG ODN.     

Cross-protective immunity against multiple influenza virus subtypes by a novel modified vaccinia Ankara (MVA) vectored vaccine in mice

Development of an influenza vaccine that provides cross-protective immunity remains a challenge. Candidate vaccines based on a recombinant modified vaccinia Ankara (MVA) viral vector expressing antigens from influenza (MVA/Flu) viruses were constructed. A vaccine candidate, designated MVA/HA₁/C13L/NP, that expresses the hemagglutinin from pandemic H1N1 (A/California/04/09) and the nucleoprotein (NP) from highly pathogenic H5N1 (A/Vietnam/1203/04) fused to a secretory signal sequence from vaccinia virus was highly protective. The vaccine elicited strong antibody titers to homologous H1N1 viruses while cross-reactive antibodies to heterologous viruses were not detectable. In mice, this MVA/HA₁/C13L/NP vaccine conferred complete protection against lethal challenge with A/Vietnam/1203/04 (H5N1), A/Norway/3487-2/09 (pandemic H1N1) or A/Influenza/Puerto Rico/8/34 (seasonal H1N1) and partial protection (57.1%) against challenge with seasonal H3N2 virus (A/Aichi/68). The protective efficacy of the vaccine was not affected by pre-existing immunity to vaccinia. Our findings highlight MVA as suitable vector to express multiple influenza antigens that could afford broad cross-protective immunity against multiple subtypes of influenza virus.     

重组日本血吸虫抗原诱导小鼠抗卵免疫的研究

目的 探讨重组日本血吸虫31／32kDa抗原(rSj31／32)诱导小鼠产生抗卵免疫的效果。方法 采用rSj31／32抗原免疫小鼠，攻击感染后42d，定量检测粪卵、肝肠组织内虫卵及雌虫子宫内虫卵数。结果 与对照组比较，rSj31／32抗原免疫小鼠肝、肠组织内总卵数和粪卵数分别减少46．0％，76．0％和83．0％，其中肝、肠组织内成熟虫卵数明显减少(69．0％和91．0％)，而肝组织内死亡虫卵数显增加(242．5％)。此外，雌虫子宫内虫卵数亦明显下降(49．8％)。结论 rSj31／32抗原能诱导小鼠产生抗雌虫生殖和抗卵胚发育的免疫，可望作为抗卵疫苗的候选抗原。     

Repeat tick exposure elicits distinct immune responses in guinea pigs and mice

Ticks deposit salivary proteins into the skin during a bite to mediate acquisition of a blood meal. Acquired resistance to tick bites has been demonstrated to prevent Borrelia burgdorferi sensu lato (s.l.) transmission. However, the mechanism of resistance, as well as the protective antigens, have remained elusive. To address these unknowns, we utilized a guinea pig model of tick resistance and a mouse model of permissiveness. Guinea pigs developed immunity after multiple Ixodes scapularis tick infestations, characterized by rapid tick detachment and impaired feeding. In comparison, mice tolerated at least 6 infestations with no significant impact on feeding. We analyzed the bite sites by RNA-sequencing and histology, identifying several inflammatory pathways in tick immune animals, such as FcεRI signaling and complement activation, and activation of coagulation pathways that could impair local blood flow. Together, these results identify important pathways altered during tick rejection and potential tick proteins that could serve as vaccine candidates.     

Heparanase DNA vaccine delivered by electroporation induces humoral immunity and cytoimmunity in animal models

Electroporation (EP)-assisted DNA vaccination has been proven effective as an approach to the treatment of cancer. Although heparanase (HPA) is a potential target for patients with advanced tumor diseases, the efficacy of immunotherapeutic strategies targeting HPA has never been evaluated. In this study, humoral immunity was elicited using genetic vaccinations between C57BL/6J mice and Macaca fascicularis. The immunized serum neutralized HPA activity and attenuated the invasion of B16 cells in vitro. In addition, T lymphocytes from the splenic cells of the immunized mice induced HPA-specific cytotoxic lymphocytes (CTLs), which verified cytoimmunity. Prophylactic vaccination significantly suppressed tumor growth and metastasis in vivo and prolonged the survival rate in tumor-bearing murine models. In addition, RT-PCR and Western blot analyses of the primary tumors indicated less proliferation and angiogenesis and more apoptosis in the HPA-immunization immunotherapy groups. Simultaneously, the levels of IL-2, IL-4, and IFN-γ were not significantly greater in the HPA-immunized group than in PBS controls. Thus, we conclude that the combination of an anti-HPA antibody and a CTL response in HPA-immunization gene therapy is enough to attenuate tumor growth and metastasis. This is the first time that a DNA vaccine targeting HPA immunization assisted by EP has induced humoral immunity and cytoimmunity in vivo. This provides a basis for the continued development of DNA vaccines targeting HPA and the use of such vaccines in clinical settings.     

葡萄籽多酚在拟衰老模型小鼠中抗氧化功能的研究

为探讨葡萄籽多酚 (PC)在拟衰老小鼠模型中的抗氧化功能 ,将动物按体重随机分组 ,除对照组外 ,每日通过颈背部皮下注射灭菌D -半乳糖 ,同时经口灌胃给予受试物或对照物 ,阳性对照组给与维生素E ,连续 6 0天后检测结果 :模型对照组相对于空白对照组血清、肝、脑中丙二醛 (MDA)水平上升 ;血、肝脏超氧化物歧化酶 (SOD)、谷胱甘肽过氧化物酶 (GSH PX)活性分别下降 ;脑中单胺氧化酶B(MAO B)水平上升 ,肝中MAO B水平无明显变化。经灌胃给予PC剂量分别为 2 5、5 0、10 0mg kgBW的抗氧化效果为使血清、肝脏、脑组织中LPO水平均有不同程度的下降。血、肝中SOD活性水平有不同程度的上升。脑MAO B活性水平下降 ;肝脏MAO B活性水平基本无变化。维生素E组亦有不同程度的抗氧化作用。本次实验结果显示 ,PC具有良好的抗氧化功能 ,能在一定程度上延缓由于注射D 半乳糖所引起的小鼠体内的拟衰老变化     

Immune response to intranasal and intraperitoneal immunization with Kaposi's sarcoma-associated herpesvirus in mice

A vaccine for Kaposi's sarcoma-associated herpesvirus (KSHV) is not currently available. To obtain the fundamental data in animals for vaccine development, KSHV particles were immunized to Balb/c mice through intraperitoneal and intranasal routes in the present study. Intranasal immunization with KSHV induced IgA to KSHV in not only serum, but also nasal wash fluid and saliva. A neutralization assay using recombinant KSHV that expressed green fluorescent protein revealed that nasal wash fluid and saliva from the KSHV-immunized mice neutralized KSHV infection to human embryonic kidney 293 cells in vitro in a dose-dependent manner to KSHV copies immunized. The serum and nasal wash fluid of KSHV-encoded K8.1 protein-immunized mice neutralized KSHV infection to 293 cells in vitro. These data suggest a possibility of mucosal vaccine for prophylaxis of KSHV infection.