小花棘豆生物碱中毒对小白鼠免疫器官脏器生长指数的影响

试验将80只小白鼠随机分组,分别将苦马豆素(SW)、黄花碱(Ts)和两类生物碱混合物的稀释液胃内灌服小白鼠,并在第7、14、21、28和35天,每组随机取4只小白鼠脱颈致死,检测胸腺(脾脏)生长指数,研究小花棘豆生物碱SW、Ts及两类生物碱的混合物对小白鼠免疫器官脏器生长指数的影响,探讨小花棘豆的中毒机理。试验结果显示:短时间(7 d)内,SW能够提高小白鼠胸腺(脾脏)生长指数,但差异不显著;长时间(28 d),SW能极显著降低胸腺(脾脏)生长指数;整个试验周期,Ts和两类生物碱的混合物均能降低胸腺(脾脏)生长指数,但Ts在第35天差异极显著,两类生物碱的混合物在第28天差异极显著。说明小花棘豆生物碱SW和Ts均能不同程度地降低小白鼠的免疫器官脏器生长指数,从而损害动物机体的免疫器官,对动物机体产生毒害作用。     

小花棘豆生物碱对α—甘露糖苷酶活性的影响

为探讨小花棘豆的中毒机理,采用苦马豆素、黄花碱以及这2种生物碱的混合物分别对小白鼠进行传统灌胃,并于试验开始后第7、14、21、28和35天分别从各组中随机取4只小白鼠,眼球采血,肝素钠抗凝,利用紫外可见分光光度计检测血清中α—甘露糖苷酶的活性。结果表明:低浓度的SW具有提高血清中α—甘露糖苷酶活性的作用,中等浓度和高浓度的SW在短时间内也可以提高血清中α—甘露糖苷酶的活性,但时间过长会严重降低血清中α—甘露糖苷酶的活性;Ts在整个试验过程中不影响血清中α—甘露糖苷酶的活性;混合生物碱组表现出与SW组类似的结果。     

营养限制及补偿对羔羊体重及外周血液CD4+和CD8+T淋巴细胞的影响

本试验旨在研究羔羊营养限制期(第1～60天)和营养补偿期(第61～150天)体重和外周血液中CD4+和CD8+T淋巴细胞的变化规律.选用80只体况中等、平均体重为(14.72±1.10)kg的3月龄乌珠穆沁羔羊,随机分为对照组(CG)、限制组Ⅰ(RG Ⅰ)、限制组Ⅱ(RGⅡ)和限制组Ⅲ(RG Ⅲ)4个组.营养限制期4组饲粮能氮水平分别为代谢能(ME):10.88、10.88、9.41和8.62 MJ/kg;粗蛋白质(CP):15%、10%、10%和5.7%.营养补偿期各组饲喂同一能氮水平饲粮(ME:9.75 MJ/kg;CP:12%).在试验期每周称重并在第1天、第30天、第60天、第90天和第150天饲喂前从各组羊颈静脉采血,采用流式细胞仪检测羔羊血液中的CD4+和CD8+T淋巴细胞比例.结果表明:1)营养限制结束,RG Ⅰ组、RG Ⅱ组和RG Ⅲ组平均体重均显著低于CG组(P＜0.05).补偿期RGⅢ组平均日增重显著高于CG组(P＜0.05).2)营养限制前期(第1～30天)各组CD4+和CD8+T淋巴细胞浓度均呈上升趋势.补偿期结束时CD4+T淋巴细胞浓度RG Ⅰ组最低RGⅡ组最高,CD8+T淋巴浓度RGⅡ组最低RG Ⅰ组最高,差异均不显著(P＞0.05).结果提示,乌珠穆沁羔羊受不同能氮营养水平限制后体重和免疫机能有补偿效应,而且低能高氮比高能低氮限饲后得到的补偿效果较好,表明限饲蛋白质比限饲能量对补偿生长的负影响更大.蛋白质和能量同时限饲补偿生长最差.     

高温下清凉冲剂对鸡免疫器官中CD4+、CD8+ T细胞数量的影响

采用免疫组化方法检测鸡免疫器官中CD4^＋、CD8^＋T细胞数量，探讨高温下清凉冲剂对机体细胞免疫机能的影响及其药理作用。108只农大3号35目龄公雏，随机分为3组，每纽36只鸡，即常温对照组、高温对照组和高温用药组。用药组饲喂中药煎荆（浓度为1g／ml），用药量与饲料量比为1：1000。在试验的第1、4、8、10天各组捕杀5只。分别采取胸腺、脾脏、法氏囊检测。结果高温下。用药组免疫器官中CD4^＋、CD8^＋T细胞数量均高于高温对照组。第8、10天差异极显著（P〈0．01）。表明高温下清凉冲剂可以增加鸡免疫器官中CD4^＋、CD8^＋T细胞数量，有效地提高鸡细胞免疫水平。     

益生菌联合新城疫疫苗应用雏鸡消化道局部黏膜组织CD4~+CD8~+T细胞数量变化

100只艾维因肉雏鸡随机分为益生菌组、新城疫(ND)疫苗组、益生菌联合ND疫苗组和对照组。采用组织石蜡切片及免疫组织化学法分别测定ND疫苗免疫后第0天、7天、14天、28天和第42天盲肠扁桃体、十二指肠和回肠派伊尔结CD4+、CD8+T细胞数量的变化。结果表明,益生菌联合ND免疫雏鸡、ND疫苗单独免疫雏鸡上述免疫组织的CD4+T和CD8+T细胞数量分别于免疫后第14天和第7~14天明显高于对照雏鸡和益生菌雏鸡(P0.05或P0.01),表明益生菌作为免疫增强剂,与疫苗联合应用可显著增强机体消化道局部黏膜免疫组织对ND疫苗的细胞免疫应答。     

外周血中CD4+、CD8+T、CD4+CD25+Treg在绵羊接种布鲁氏菌M5-90弱毒疫苗后的动态变化

为研究绵羊接种布鲁氏菌弱毒M5-90株后外周血中CD4+、CD8+T、CD4+CD25+Treg细胞的动态变化规律,本研究选择11只健康绵羊,每10 d免疫一次,共免疫3次,分别在免疫前、免疫后10d、20 d、30 d利用流式细胞术检测外周血中CD4+、CD8+T、CD4+CD25+Treg淋巴细胞亚群.在免疫后的第20 d,CD4+T、CD8+T细胞百分含量达到最高水平(P＜0.05)后均缓慢下降;在第10d,CD4+CD25+Treg细胞缓慢升高,至20 d、30 d均显著升高(p＜0.05);在布鲁氏菌M5-90疫苗免疫应答过程中CD4+CD25+Treg细胞参与了机体的免疫反应调控,对CD4+T、CD8+T淋巴细胞的比例进行调节,并且维持CD4+/CD8+比值稳定,起到平衡Th1/Th2细胞间反应的作用.     

刺五加多糖对雏鸡脾脏中CD4+ 和CD8+T淋巴细胞定位分布的影响

为了研究刺五加多糖（ASPS）对雏鸡脾脏中CD4+和CD8+ T淋巴细胞定位分布的影响,从组织学角度评价ASPS对脾脏的免疫调节作用,试验将1日龄海兰褐公雏饲养至7日龄时选取150只,随机分为3组：空白对照组、ASPS低剂量组（ASPSL）和高剂量组（ASPSH）,每组50只,所有组每天注射1次,连续注射3天。免疫后的第7、14、21和28天分别取其脾脏制作冰冻切片,采用免疫组织化学方法检测CD4+和CD8+ T淋巴细胞的定位分布。结果显示,与空白对照组比较,免疫注射后21天和28天时ASPSL组和ASPSH组CD4+ T淋巴细胞的数量均显著增加（P<0.05）,而且ASPS能够促进红髓中CD4+ T淋巴细胞向动脉周围淋巴鞘迁移,从而使单个动脉周围淋巴鞘面积较对照组明显增加,而ASPS对脾脏中CD8+ T淋巴细胞的数量和分布无明显影响。由此可知,ASPS能够通过影响脾脏中CD4+ T淋巴细胞的定位分布发挥免疫调节作用,这对于进一步揭示ASPS的免疫调节机制具有重要意义。     

抗菌肽Sublancin与黄芪多糖对免疫抑制小鼠免疫功能调节作用的比较分析

旨在比较研究抗菌肽sublancin与黄芪多糖对环磷酰胺(cyclophosphamide,CTX)诱导的免疫抑制小鼠的免疫功能的调节作用。试验选取60只4～6周龄健康雌性BALB/c小鼠,随机分为6个处理组。正常对照组:第1—3天,腹腔注射生理盐水;第4—10天,灌胃生理盐水。其余五个处理组:第1—3天,腹腔注射CTX(80mg·kg-1);第4—10天,阴性对照组:灌胃生理盐水;低浓度抗菌肽组:灌胃4.0mg·kg-1抗菌肽sublancin;高浓度抗菌肽组:灌胃8.0mg·kg-1抗菌肽sublancin;黄芪多糖组:灌胃200.0mg·kg-1黄芪多糖;阳性对照组:灌胃10.0mg·kg-1左旋咪唑。所有处理均为每天一次,每次0.2mL。于试验第1天小鼠腹腔注射CTX前、第4天灌胃前和第11天对小鼠进行称重。第11天称重结束后,采集全部小鼠的外周血及脾,检测外周血生理生化指标、CD4+与CD8+数量和脾细胞的细胞因子mRNA表达等指标。结果表明,与正常对照组相比,阴性对照组中小鼠体重、外周血的红细胞、血红蛋白和白细胞含量显著降低(P<0.05),脾IL-2、IL-4和IL-6的基因表达显著降低(P<0.05)。灌胃后,与阴性对照组相比,黄芪多糖组和高浓度sublancin组体重无差异但有升高趋势,说明高浓度sublancin与黄芪多糖对小鼠体重有积极影响;低、高浓度sublancin、黄芪多糖组和阳性对照组中小鼠外周血的白细胞含量显著升高(P<0.05);高剂量sublancin组中小鼠外周血的红细胞和血红蛋白显著升高;低、高浓度sublancin和黄芪多糖组中小鼠外周血的CD4+显著升高(P<0.05);低、高浓度sublancin、黄芪多糖组和阳性对照组中小鼠外周血的CD8+显著降低(P<0.05);低、高浓度sublancin组中小鼠外周血的CD4+/CD8+细胞显著升高(P<0.05);低、高浓度sublan-cin、黄芪多糖组和阳性对照组中小鼠脾IL-4的表达均显著升高(P<0.05);高浓度sublancin、黄芪多糖组和阳性对照组中小鼠脾IL-2、IL-6的表达显著升高(P<0.05)。综合上述结果,适宜剂量的抗菌肽sublancin和黄芪多糖均可缓解环磷酰胺造成的免疫抑制。与200.0mg·kg-1黄芪多糖相比,8.0mg·kg-1抗菌肽sublancin对缓解环磷酰胺免疫抑制造成的细胞因子降低的效果更好。     

猪瘟兔化弱毒免疫猪淋巴细胞亚类及转化水平、IFN-γ的变化与分析

为研究免疫猪瘟兔化弱毒(HCLV)对猪细胞免疫应答的影响,本研究分别采用HCLV脾淋毒单独(A组)、HCLV细胞毒与兔脾淋组织液联合(B组)、HCLV细胞毒单独(C组)及生理盐水(对照组)免疫仔猪,应用流式细胞术、淋巴细胞增殖试验及ELISA方法测定免疫前后外周血淋巴细胞(PBL)亚类比值、PBL非特异性(NSI)及特异性(SSI)刺激指数、IFN-γ浓度。结果显示免疫后各组CD3+/PBL与CD3+CD8+/PBL差异不显著;CD3+CD4+/PBL于第7 d时A、B和对照组显著高于C组(p0.05);CD3+CD4+/CD3+CD8+于第7 d、14 d时A、B和对照组显著高于C组(p0.05),并且C组CD3+CD4+/CD3+CD8+显著降低(p0.05);NSI于第3 d、7 d时A与B组显著高于C组(p0.05),而C组显著高于对照组(p0.05);SSI于第3 d、7 d时A组显著高于B与C组(p0.05),而B与C组显著高于对照组(p0.05);IFN-γ含量由高至低依次为:A、B、C和对照组,并且于第3 d、7 d时差异显著(p0.05)。研究结果表明,HCLV细胞毒引起机体CD3+CD8+相对更快速增殖,而HCLV脾淋毒引起机体CD3+CD4+相对更快速增殖及NSI、SSI、IFN-γ浓度更高,脾淋毒与细胞毒可能侧重不同的免疫分子途径发挥免疫保护效力;兔脾淋组织液对NSI和IFN-γ浓度具有增强作用。     

肝片吸虫对奶牛免疫水平的影响

本试验选择济南地区3个奶牛场中5头感染肝片吸虫的奶牛和5头健康奶牛,在患牛出现症状并确诊后第5天、第10天、第15天分别进行血常规检测、C-反应蛋白检测、免疫球蛋白含量检测、T细胞亚群检测和淋巴细胞转化率的测定。结果表明,感染肝片吸虫的奶牛其淋巴细胞比率和单核细胞比率明显低于对照组,差异极显著;C-反应蛋白含量与对照组相比显著升高;蛋白含量中免疫球蛋白的含量与对照组相比差异显著,T细胞亚群中CD4~+、CD8~+和CD4~+/CD8~+与对照组相比显著降低;感染20d奶牛淋巴细胞转化率与对照组相比差异显著。这说明肝片吸虫对奶牛的免疫水平有明显的抑制作用,且随着时间的变化,机体免疫水平逐渐降低。     

In vitro effects of dexamethasone on bovine CD25+CD4+ and CD25−CD4+ cells

This paper investigates the in vitro effect of dexamethasone on bovine CD25^highCD4⁺, CD25^lowCD4⁺ and CD25⁻CD4⁺ T cells. Only a small percentage of bovine CD25^highCD4⁺ (2–4%) and CD25^lowCD4⁺ (1–2%) cells expressed Foxp3. Dexamethasone caused considerable loss of CD25⁻CD4⁺ cells, but it increased the relative and absolute numbers of CD25^highCD4⁺ and CD25^lowCD4⁺ lymphocytes, while at the same time reducing the percentage of Foxp3⁺ cells within the latter subpopulations. Considering all these, as well as the intrinsically poor Foxp3 expression in bovine CD25⁺CD4⁺, it can be concluded that the drug most probably increased the number of activated non-regulatory CD4⁺ lymphocytes. It has been found that changes in cell number were at least partly caused by proapoptotic effect of the drug on CD25⁻CD4⁺ cells and antiapoptotic effect on CD25^highCD4⁺ and CD25^lowCD4⁺ cells. The results obtained from this study indicate that the involvement of CD4⁺ lymphocytes in producing the anti-inflammatory and immunosuppressive effect of dexamethasone in cattle results from the fact that the drug had a depressive effect on the production of IFN-γ by CD25⁻CD4⁺ cells. Secretion of TGF-β and IL-10 by CD4⁺ lymphocytes was not involved in producing these pharmacological effects, because the drug did not affect production of TGF-β and, paradoxically, it reduced the percentage of IL-10⁺CD4⁺ cells.     

The role of CD4+CD25+ regulatory T cells in viral infections

Many virus infections result in the suppression of one or more functions of the immune system. Multiple mechanisms have been proposed to explain viral-induced immunosuppression, including an imbalance in the cellular Th1/Th2 or cytokine profile, induction of anergy, depletion of effector cells and most recently the activation of CD4+CD25+ regulatory T (T reg) cells. CD4+CD25+ T reg cells are a subset of circulating CD4+ T cells with suppressive properties. CD4+CD25+ T reg cells were first identified in mice as cells capable of maintaining self-tolerance by suppressing autoreactive T cells. This review focuses on interactions between CD4+CD25+ T reg cells and viral pathogens. Most cases in which CD4+CD25+ T reg cells participate in response to infection reported so far involve chronic or persistent viral infections. Examples have been growing recently and include members of different viral families including retroviridae, herpesviridae and picornaviridae. It is currently not known how microbes are recognized by CD4+CD25+ T reg cells and whether exoantigen-specific T reg cells are of the same lineage as self-reacting natural T reg cells or represent peripherally induced counterparts derived from CD4+CD25- T cells. The findings that T reg cells influence the functional immunity during viral infections, however, might indicate that, in some cases, virus-specific T reg cells not only influence immune pathology or prevent pathogen elimination but also can promote a generalized state of immunosuppression in vivo such that the host is more susceptible to secondary infections with other pathogens or has reduced resistance to tumors. Conceivably, the activities of T reg cells might be one of the contributing reasons why it has been difficult so far to produce effective vaccines against some persisting viral infections.     

In vivo lipopolysaccharide injection alters CD4+CD25+ cell properties in chickens

In chickens, thymic CD4(+)CD25(+) cells are characterized as regulatory T cells. The objectives of this experiment were to study the effects of an in vivo lipopolysaccharide (LPS) injection on the percentage of CD4(+)CD25(+) cells in peripheral organs and the suppressive properties of splenic CD4(+)CD25(+) cells in chickens. Chickens were injected with LPS and CD4(+)CD25(+) cells were analyzed at 1, 2, 3, 5, and 10 d post LPS injection. The LPS injection increased CD4(+)CD25(+) cell percentage approximately 5-fold in the blood at 1 d post LPS injection (P < 0.001), 3-fold in the thymus at 3 d post LPS injection (P = 0.001), and 2.5-fold in the spleen at 2 d post LPS injection (P = 0.001) compared with the no-LPS-injected group. The LPS injection did not alter the CD4(+)CD25(+) cell percentage in the cecal tonsil (P = 0.162), lung (P = 0.098), or bone marrow (P = 0.071) at any time point measured. At 2 d post LPS injection, splenic CD4(+)CD25(+) cells lost their suppressive ability (P < 0.001). At 5 d post LPS injection, splenic CD4(+)CD25(+) cells not only regained their suppressive ability, but also became supersuppressive (P < 0.001). Splenic CD4(+)CD25(+) cells at 5 d post LPS injection produced 5.5-fold more (P = 0.005) IL-10 mRNA than splenic CD4(+)CD25(+) cells at 0 and 2 d post LPS injection. In conclusion, chicken regulatory T cells are differentially activated to facilitate immune response during the early stage of inflammation and to facilitate immune suppression at a later stage of inflammation.     

AA肉鸡血液CD3+、CD4+和CD8+T淋巴细胞的变化规律

采用流式细胞仪检测1、3、5、7、14、21、28、35、42、49日龄AA肉鸡血液中的CD3、CD4、CD8阳性T细胞比例。研究结果表明:1～5日龄T细胞逐渐进入血液参与细胞免疫,7、21日龄注射疫苗起免疫应答作用,28日龄后基本形成稳固的细胞免疫水平。     

CD4+ and CD8+ T- lymphocytopenia in a filly with Pneumocystis carinii pneumonia

Decreased proportion of CD4+ and CD8+ T lymphocytes in peripheral blood likely contributed to susceptibility to Pneumocystis carinii in a foal. Cytological evaluation of bronchoalveolar lavage was required for identification of the pathogen and serial flow-cytometric analysis of peripheral blood lymphocytes documented transient low expression of CD4+ and CD8+ T lymphocytes. Although immunodeficiency is uncommon, it must be included in the differential diagnosis for patients suffering from chronic or opportunistic infections and may provide an indication for immunostimulant therapy.     

Differential cytokine gene expression in CD4+ and CD8+ T cell subsets of calves

The distinct patterns of cytokine expression in CD4+ and CD8+ T cells are well understood in mice and humans. However, little information is available about cytokine expression in bovine CD4+ and CD8+ T cells. In this study, mRNA expression of 19 different cytokines was analyzed in CD4+ and CD8+ T cells of calves with or without Concanavalin A (Con A) stimulation. CD4+ and CD8+ T cell populations were enriched to 98% purity by positive selection using magnetic cell sorting (MACS). CD4+ T cells spontaneously expressed the mRNAs of interleukin-1alpha (IL-1alpha), IL-1beta, IL-2, IL-6, IL-7, IL-8, IL-10, IL-18, IFN-gamma, TNF-alpha, TNF-beta and TGF-beta, and augmented the mRNA expression of IL-10, IFN-gamma and TNF-beta after Con A stimulation. The mRNAs of IL-3, IL-4, IL-5, IL-13 and GM-CSF were newly expressed in Con A-stimulated CD4+ T cells. CD8+ T cells displayed spontaneous mRNA expression of IL-6, IL-18, TNF-alpha, TNF-beta and TGF-beta, and newly expressed the mRNA of IL-2, IL-7, interferon-gamma (IFN-gamma) and GM-CSF after Con A stimulation. It was found that CD4+ T cells expressed the mRNA of 17 cytokines except for IL-12 and IL-15, while CD8+ T cells expressed only the mRNA of 9 cytokines after Con A stimulation. The profile of cytokine mRNA expression was substantially different in the CD4+ and CD8+ T cells of calves, indicating that CD4+ T cells can be distinguished from CD8+ T cells by the cytokine gene expression of IL-1alpha, IL-1beta, IL-3, IL-4, IL-5, IL-8, IL-10 and IL-13. Differential cytokine expression between CD4+ and CD8+ T cells serve to interpret an individual function of T cell subsets in the immune system of calves.     

CD4+CD25+ regulatory T cells are infected and activated during acute FIV infection

HIV-induced AIDS may be mediated by the activation of immunosuppressive CD4+CD25+ T regulatory cells (Treg cells). Treg cells have been shown to regulate CD4+ and CD8+ immune responses to HIV and FIV antigens in vitro. We tested the hypothesis that Treg cells become infected and activated during the acute infection with FIV leading to the suppression of CD4+ T helper cell responses. Cats were experimentally infected with FIV-NCSU1 and blood and lymph node cells were collected at weekly intervals following inoculation. Real-time RT-PCR was used to determine plasma viremia and the relative expression of FIV, FoxP3, TGF-beta, and GAPDH mRNA copies in CD4+CD25+ and CD4+CD25- T cell subsets. Flow cytometry was used to assess the absolute numbers of each cell type and the expression of surface TGF-beta and intracellular FoxP3 in CD4+CD25+ and CD4+CD25- T cells at each time-point. Treg suppression of IL-2 production in CD4+ T helper cells was assessed by ELISPOT assays. Our results showed that peak viremia occurred at 2 weeks post infection and correlated with maximal infectivity in CD4+CD25+ T cell populations. FIV-gag-mRNA levels were higher in CD4+CD25+ T cells than CD4+CD25- T cells throughout the acute phase of infection. Induction of FoxP3 and TGF-beta indicated activation of Treg cells during the acute stage infection, which was confirmed by Treg cell suppression of IL-2 production by CD4+ Th cells in an ELISPOT assay. Our findings support the hypothesis that early activation of Treg immunosuppressor function may limit an effective anti-FIV response, contributing to the establishment of chronic infection and the immunodeficiency caused by this virus.