禽呼肠病毒感染对SPF鸡外周血T细胞亚型变化和细胞因子转录的影响

为探讨禽呼肠病毒(ARV)对SPF鸡外周血淋巴细胞中CD4+、CD8+T细胞数量变化和细胞因子mRNA转录水平的影响,利用流式细胞术和实时荧光定量PCR方法分别测定了ARV感染后1、7、14、21、28、35d感染组和对照组SPF鸡外周血淋巴细胞中CD4+、CD8+T细胞含量和细胞因子IL-1β、IL-6、IL-17、IL-18、IFN-γ、TNF-α基因mRNA相对转录时相。流式细胞术检测结果表明,SPF鸡感染ARV后7d和14dCD4+、CD8+T细胞比值高于对照组,其中感染7d,CD8+T细胞含量差异显著(P0.05);感染后1、21、28、35d感染组CD4+、CD8+T细胞比值均低于对照组,感染1d后CD4+、CD8+T细胞含量均差异显著(P0.05),说明外周血T细胞亚型变化是ARV感染的重要表现之一。实时荧光定量PCR结果表明,与对照组相比,感染组外周血淋巴细胞中IL-1β、IL-6(除7d外)、IL-18(除14d外)和TNF-α在整个感染过程中表达上调,IL-17和IFN-γ除感染1d外,均表达下调,说明IL-1β、IL-6、IL-17、IL-18、IFN-γ和TNF-α均参与了ARV的感染进程。     

IL-6、FGF21和ANGPTL8与能量负平衡奶牛卵巢静止的关系

为了确定组织细胞因子在奶牛产后能量负平衡(NEB)诱导的卵巢静止(IO)发生中的作用。本试验根据产后21 d血浆β-羟丁酸(BHBA)和葡萄糖(Glu)的水平，从100头奶牛中选取能量正平衡(PEB)组和NEB组各30头。随后，根据产后50～60 d是否发情及卵泡发育状况，选取NEB组中IO奶牛(NEB-IO)和PEB组中发情奶牛(PEB-E)各6头进行采样。利用生化和ELISA方法分别检测血浆中代谢指标和细胞因子的水平；利用实时荧光定量PCR(qRT-PCR)和ELISA方法检测组织中细胞因子的水平。结果显示：与PEB组奶牛相比，NEB组奶牛产后21 d血浆中白细胞介素6(IL-6)极显著升高(P<0.01),成纤维细胞生长因子21(FGF21)和血管生成素样蛋白8(ANGPTL8)水平显著降低(P<0.05),并持续到产后50 d; 3个细胞因子与BHBA、Glu和游离脂肪酸(NEFA)之间显著相关(P<0.05);与PEB-E组奶牛相比，NEB-IO组奶牛IL-6 mRNA表达量和蛋白含量在肌肉和卵巢中显著降低(P<0.05);FGF21和ANGPTL8 ...     

蛋氨酸缺乏对雏鸡盲肠扁桃体T淋巴细胞亚群和细胞因子的影响

为了研究蛋氨酸缺乏对雏鸡盲肠扁桃体T淋巴细胞亚群和细胞因子的影响,试验选取100只1日龄健康Cobb(科宝)雏鸡分为对照组和蛋氨酸缺乏组,分别采食对照日粮和蛋氨酸缺乏日粮,试验期为42 d,分别于14、28和42日龄对雏鸡进行剖检,取盲肠扁桃体样品进行酶联免疫吸附试验(ELISA)和实时荧光定量PCR(qRT-PCR)检测T淋巴细胞亚群和细胞因子。结果表明:与对照组相比,蛋氨酸缺乏组中盲肠扁桃体CD3+、CD3++CD4+、CD3++CD8+以及CD4+/CD8+比例均显著或极显著降低(P0.05或P0.01)。细胞因子白介素2(IL-2)、IL-6、IL-10、IL-17、IFN-γ、TNF-α(LITAF)的含量及mRNA表达水平也呈现不同程度的下降(P0.05或P0.01)。说明蛋氨酸缺乏会导致盲肠扁桃体T淋巴细胞亚群和细胞因子下降,进而影响雏鸡的特异性免疫功能。     

茶皂素对奶牛免疫功能的影响

本试验旨在研究茶皂素对荷斯坦奶牛的血清免疫相关细胞因子含量及其mRNA表达量的影响。试验采用4×4拉丁方设计,选取4头瘤胃瘘管奶牛,每头奶牛分别灌注0(对照)、15、30、45 g/d的茶皂素,预试期14 d,正试期21 d,共4期。采用酶联免疫吸附剂测定(ELISA)法试剂盒测定血清中免疫球蛋白(Ig)、白细胞介素(IL)和肿瘤坏死因子-α(TNF-α)的含量,采用实时荧光定量PCR方法测定淋巴细胞IL-1、IL-6、IL-10和TNF-αmRNA表达量。结果显示:1)30 g/d茶皂素组奶牛血清Ig M含量显著高于对照组(P0.05)。2)15 g/d茶皂素组奶牛血清IL-1含量显著高于对照组和45 g/d茶皂素组(P0.05)。3)与对照组相比,15、30、45 g/d茶皂素组奶牛血清IL-1、IL-6、IL-10、TNF-αmRNA表达量均无显著差异(P0.05),但45 g/d茶皂素组可提高以上细胞因子mRNA表达量。由此可见,茶皂素可提高奶牛血清中Ig及免疫相关细胞因子含量,并提高细胞因子IL-6 mRNA表达量,表明茶皂素可提高奶牛免疫功能。     

干扰素α对小鼠Tfh细胞分化的调控作用

通过体外试验观察干扰素-α(IFN-α)对滤泡辅助性T细胞(Tfh)分化及功能的影响。无菌分离小鼠脾细胞,磁珠分选CD4^+T细胞,加入抗CD3单克隆抗体、抗CD28单克隆抗体,重组IL-21、IL-6进行刺激,同时加入重组IFN-α(设不加IFN-α的阴性对照)进行培养,分别在培养后的1、3、5d,流式细胞术检测Tfh细胞(CD4^+CXCR5^+)和CD4^+CXCR5^+PD-1^+细胞的比例;ELISA检测细胞上清中Tfh细胞功能性细胞因子IL-4、IL-21的水平;real-time PCR检测培养3d后Tfh细胞相关转录因子STAT1、Bcl-6以及细胞因子IL-21mRNA的表达水平。结果表明,与阴性组相比,IFN-α能明显促进Tfh细胞的分化及Tfh细胞中PD-1的表达(P<0.05),能提高Tfh细胞功能性细胞因子IL-4、IL-21的分泌水平(P<0.05);IFN-α能正向调控Tfh细胞相关转录因子STAT1、Bcl-6以及细胞因子IL-21mRNA的表达水平(P<0.05)。说明IFN-α可通过正向调控STAT1、Bcl-6的表达,促进Tfh细胞的分化和细胞因子的分泌。     

归芪益母口服液对产后奶牛免疫机能和生产性能的影响

为阐明归芪益母口服液对产后奶牛免疫机能和生产性能的影响,选择40头2～4胎次产后荷斯坦奶牛,随机分为4组,每组10头。对照组灌服温开水800 mL,低、中、高剂量组分别灌服归芪益母口服液300,500和800 mL,连续灌服5 d。采集产后0,5和12 d的血样和乳样,酶联免疫法测定血清IgA、IgM和IgG含量,流式细胞仪检测外周血T淋巴细胞CD4~+、CD8~+,全自动乳体细胞分析仪检测乳成分和体细胞。结果显示,奶牛产后0,5和12 d血清IgA含量变化不显著(P0.05),IgM和IgG逐渐升高(P0.05),外周血液淋巴细胞亚群CD8~+呈上升趋势(P0.05),CD4~+与CD4~+/CD8~+的比值降低(P0.05);中、高剂量归芪益母口服液能显著提高产后奶牛IgM、IgG、CD4~+、CD4~+/CD8~+含量(P0.05),降低CD8~+含量(P0.05),并且能显著提高产奶量(P0.05),降低体细胞数(P0.05)。结果表明,中、高剂量归芪益母口服液具有增强奶牛产后免疫机能和提高生产性能作用。     

脂多糖对奶牛乳腺上皮细胞IL-6、IL-8和TNF-α mRNA表达的影响

本试验以体外培养的奶牛乳腺上皮细胞为模型,研究不同浓度脂多糖刺激奶牛乳腺上皮细胞时IL-6、IL-8及TNF-α的mRNA表达情况。结果显示,LPS浓度为10μg/mL时刺激效果最好;10μg/mL LPS刺激后24h,IL-6和IL-8的表达量最高(P0.05),TNF-α的mRNA表达在刺激后的4h达最高(P0.05)。以上结果说明,本实验室体外培养的奶牛乳腺上皮细胞具有表达与免疫有关的细胞因子功能,可以作为一种模型来研究奶牛乳腺炎的发病机制。     

中药对围产期奶牛血浆IL-2、IL-6含量的影响

为探讨中药对围产期奶牛血浆IL-2、IL-6含量的影响,本试验选择90头年龄、胎次和预产期相近,经临床检查健康的围产期荷斯坦奶牛,随机分成A、B、C三组,每组30头。A组,于预产期15天饲喂时添加中药加味四君子散(400 g/头),每天1次,连用5 d,分娩后当天灌服益母生化散2剂(400 g/头);B组,产前不添加中药,分娩后当天灌服益母生化散2剂(400 g/头);C组为对照组,产前不添加中药,产后灌服常用水2次,每次400 mL/头。在预产期15 d、7 d,产后0 d(分娩当天灌服益母生化散前计为0 d)、7 d和15 d时,在各组随机抽取10头奶牛颈静脉采血,测定血浆IL-2、IL-6的含量。试验结果显示,A组奶牛血浆IL-2含量明显高于B组、C组(P0.05或P0.01);血浆IL-6含量A组在产前7 d时低于B组、C组,差异显著(P0.05),而产后0 d、7 d、15 d,A组高于B组、C组,差异显著(P0.05)。     

不同能量摄入水平对围产期奶牛肝载脂蛋白E mRNA丰度的影响

将年龄、胎次相同，年产奶量大于5000kg的围产期健康奶牛30头随机分为3组，每组10头。低能组饲喂中国奶牛饲养标准（2000）减少20％日粮（能量摄入80％组）、对照组饲喂中国奶牛饲养标准（2000）日粮（能量摄入100％组）和高能组饲喂中国奶牛饲养标准（2000）增加20％日粮（能量摄入120％组），试验从产前28d开始至产后56d结束，产后各组奶牛均饲喂标准日粮。采用内对照RT-PCR方法检测了不同能量摄入围产期奶牛肝组织载脂蛋白E（apolipoprotein E，apoE）mRNA丰度。结果表明：apoE mRNA相对表达量从产前28d至产后56d各组均呈现先升高后降低的趋势，各组apoE mRNA水平均在产后一天达到最大值（P〈0．01），且产后均高于产前（产后56d除外）；产前14d至产后28d低能组apoE mRNA相对表达量均高于其他两组（P〈0．05或P〈0．01），高能组在产后28d和56d高于对照组（P〈0．01）。可见，围产期奶牛能量摄入水平对肝组织载脂蛋白E mRNA丰度有显著影响。     

双翅目昆虫对荷斯坦奶牛护身行为及外周血液中淋巴细胞亚群的影响

为了研究荷斯坦奶牛受到双翅目昆虫侵袭时奶牛个体所表现的护身行为、外周血液中淋巴细胞亚群的变化规律,试验期间每日跟踪观察奶牛护身行为1 h,记录每5 min内侵袭奶牛的双翅目昆虫数目和奶牛的护身行为出现次数,定时采血用流式细胞仪检测外周血液中CD3+、CD4+、CD8+ T淋巴细胞和CD21+B淋巴细胞等的变化规律.结果表明:奶牛护身行为随飞来昆虫的数量而变化;外周血液中的CD3+、CD4+、CD21+水平随双翅目昆虫数量的增多而降低;CD8+水平随双翅目昆虫数量的增多而升高.     

Primary and anamnestic responses of bovine bronchoalveolar and peripheral blood lymphocyte subsets to aerosolized Pasteurella haemolytica A1

Site-specific responses of bronchoalveolar and peripheral blood lymphocyte subsets were compared during primary and anamnestic immune responses against live Pasteurella haemolytica A1 (Ph1). Eight 1-year old calves were sequentially exposed intrabronchially with aerosolized Ph1 on days 0, 14, and 21, and two calves were sham exposed. Bronchoalveolar and peripheral blood lymphocytes were analyzed before each Ph1 exposure, and on days 3 and 7 post exposure using single and two-color flow cytometry to identify CD2+, CD4+, CD8+, CD21+, CD45R+, CD25+ and gammadelta lymphocyte subsets. Significant differences (p < 0.05) in bronchoalveolar and peripheral blood lymphocyte subsets were observed before Ph1 exposure. Subsequent aerosol exposures, resulted in significant (p < 0.05) changes in bronchoalveolar lymphocyte subsets and the CD4:CD8 bronchoalveolar lymphocyte ratio, but concomitant changes were not observed in peripheral blood lymphocytes. Expression of CD2, CD4 and CD8 lymphocyte differentiation antigens was consistently lower and more heterogeneous on bronchoalveolar lymphocytes. Differential analysis of bronchoalveolar leukocytes revealed a significant increase in bronchoalveolar lymphocytes and neutrophils during anamnestic responses.     

Leukocyte subsets and specific antibodies in pigs vaccinated with a classical swine fever subunit (E2) vaccine and the attenuated ORF virus strain D1701

Total white blood cell (WBC) counts and percentages of CD4a+, CD8a+, CD5a+, CD45RA+, CD45RC+, wCD21+ and SWC3a+ cells in the peripheral blood of pigs were analysed in this study. Blood samples were collected before and on days 4, 10, 21 and 28 after vaccination. Group 1 pigs were vaccinated with a subunit E2 vaccine (gp E2 32 microg/dose), and Group 2 received a subunit vaccine combined with an attenuated ORF virus strain D1701 10(6.45) TCID50/dose. Control pigs received a placebo. The total WBC count and percentage of particular cell types were within the normal range in vaccinated and control pigs. Although the mechanism of attenuated ORF virus activity is not clear, changes were observed in CD4a+, CD5a+, CD8a+, CD45RA+ and CD45RC+ cells in pigs that received the combination of a subunit vaccine and ORF virus. However, the percentage of wCD21+ and SWC3a+ did not differ significantly from that recorded in pigs given only the subunit vaccine. At days 4 and 10 the number of pigs positive to E2 antibodies was higher in the group that received the subunit vaccine and ORF virus than in pigs vaccinated with the subunit vaccine only. A higher percentage of memory cells (CD45RC+) as well as Th and Tc lymphocytes in pigs that received the ORF virus and the subunit vaccine could be ascribed to a nonspecific influence of the ORF virus on the development (through cognate interactions between T and B cells) and the duration (presumed according to the finding of the clonal expression of memory cells) of humoral immunity (assessed by a higher number of seropositive pigs in this group). This seems likely since the proportion of these cells was found to be lower in the pigs that received E2 vaccine only.     

The effect of experimental infectious mastitis on leukocyte subpopulations and cytokine production in non-lactating ewes.

The interactions between leukocytes and cytokines during the acute response to intramammary infections in the dry mammary gland of sheep were studied. Dry ewes were experimentally infected in one udder half with either Staphylococcus aureus or Escherichia coli, or infused with saline as control. Udder secretion samples, blood samples and udder tissue samples were collected before and 4, 8 and 24 h after infections/infusions. Total and differential leukocyte counts were calculated in both blood and mammary secretions, and flow cytometry was used to detect the presence of CD4+, CD8+, WC1+, IL-2R+, CD18+ or L-selectin + lymphocytes, CD18+ or L-selectin + neutrophils, and CD14+ leukocytes. Moreover, the concentrations of interleukin-1 beta (IL-1 beta), interleukin-8 (IL-8) and granulocyte-macrophage colony stimulating factor (GM-CSF) in udder secretions were measured using ELISA, and RT-PCR was used to detect the presence of corresponding cytokine mRNA in udder tissue biopsies. The results suggest an association between the concentrations of IL-1 beta, IL-8 and the intensity of neutrophil infiltration of the infected gland. Immunologically relevant changes in proportions of lymphocyte subpopulations might also occur in the acute phase of the inflammatory reaction of the udder. Greater cellular and cytokine responses to E. coli infection may have contributed to the milder clinical picture and more rapid resolution of infection than that seen for S. aureus. Enhancing the production of pro-inflammatory cytokines may improve defence against bacterial mastitis.     

The Effect of Experimental Infectious Mastitis on Leukocyte Subpopulations and Cytokine Production in Non-lactating Ewes

The interactions between leukocytes and cytokines during the acute response to intramammary infections in the dry mammary gland of sheep were studied. Dry ewes were experimentally infected in one udder half with either Staphylococcus aureus or Escherichta coli, or infused with saline as control. Udder secretion samples, blood samples and udder tissue samples were collected before and 4, 8 and 24 h after infections/infusions. Total and differential leukocyte counts were calculated in both blood and mammary secretions, and flow cytometry was used to detect the presence of CD4+, CD8+, WC1+, IL-2R+, CD18+ or L-selectin+ lymphocytes, CD18+ or L-selectin + neutrophils, and CD14+ leukocytes. Moreover, the concentrations of interleukin-1β+ (IL-1β), interleukin-8 (IL-8) and granulocyte-macrophage colony stimulating factor (GM-CSF) in udder secretions were measured using ELISA, and RT-PCR was used to detect the presence of corresponding cytokine mRNA in udder tissue biopsies. The results suggest an association between the concentrations of IL-1β, IL-8 and the intensity of neutrophil infiltration of the infected gland. Immunologically relevant changes in proportions of lymphocyte subpopulations might also occur in the acute phase of the inflammatory reaction of the udder. Greater cellular and cytokine responses to E. coli infection may have contributed to the milder clinical picture and more rapid resolution of infection than that seen for S. aureus. Enhancing the production of pro-inflammatory cytokines may improve defence against bacterial mastitis.     

Cell-mediated immune responses to a killed Salmonella enteritidis vaccine: lymphocyte proliferation, T-cell changes and interleukin-6 (IL-6), IL-1, IL-2, and IFN-gamma production

Two experimental approaches were used to investigate the immunological responses of chickens to a commercial killed Salmonella enteritidis (SE) vaccine. In the first, the effects of host age on antigen-specific proliferative responses and cytokine production were examined. Compared with non-vaccinated controls, 4-wk-old vaccinated chickens showed higher proliferation to SE LPS and flagella. The lymphoproliferation responses to these antigens of 8-mo-old vaccinated chickens were not different compared to the non-vaccinated controls. Increased production of interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) by antigen-stimulated splenocytes following vaccination were, in general, more often observed in 4-wk-old compared with 8-mo-old chickens, whereas serum levels of these cytokines were consistently higher in the vaccinated birds compared with controls regardless of age. The second set of experiments were designed to determine the effects of SE vaccination on mitogen- or antigen-induced splenocyte proliferation and serum nitric oxide (NO) and cytokine levels. Splenocytes from vaccinated chickens stimulated with SE flagella showed significantly increased numbers of TCRgammadelta+ cells at 7 days post-vaccination compared with non-vaccinated birds. In contrast, no differences were noted with CD4+, CD8+, or TCRalphabeta+ cells at any time points examined. Higher levels of NO production were observed following stimulation with SE flagella at 4, 7, 11, and 14 days after SE vaccination while serum levels of IFN-gamma, IL-1, IL-6, and IL-8 were elevated only at day 7 post-vaccination. In conclusion, younger chickens mounted a more robust antigen-specific immune response to the SE vaccine compared with older birds and vaccination induced not only T-cell-mediated responses but also host innate and pro-inflammatory responses.     

Lymphocyte subsets in peripheral blood of dogs--a flow cytometric study

Slight differences in the results of papers describing lymphocyte subsets distribution in the peripheral blood of healthy dogs may be explained by differences in monoclonal antibody clones and sources, breed and age of animals examined, methods of sample treatment, or methods of result analysis. In this paper, we described the effect of sample processing and of sample storage as well as the effect of age, breed, and gender of dogs on lymphocyte subset distribution. No significant differences were found between samples processed following a whole-blood lysis method and samples processed after density gradient separation. Furthermore, no significant differences were found between samples processed within 2h after collection and those stored at 4 degrees C for 12-16 h before processing. Age-related changes were evident in lymphocyte subset distribution in the peripheral blood of 38 Beagles divided according to their age into the six groups: (1) 5-6 days; (2) 2 months; (3) 6 months; (4) 1-2 years; (5) 3-5 years; and (6) >5 years. The percentage of B-lymphocytes (CD21-like positive cells) in the peripheral blood of newborn pups was 39.5+/-5.7 and decreased with advancing age. The percentage of CD8+ lymphocytes was 7.7+/-3.4 after birth and increased with advancing age. No age-related changes were observed in the percentages of CD4+ lymphocytes. The CD4+:CD8+ ratio decreased with advancing age. No significant age-related change was observed for lymphocytes bearing the gammadelta-TCR. Some breed differences were evident. Adult (1-5-year-old) Beagles, German Shepherds, Dalmatians, and Dachshunds were examined. The percentages of lymphocytes were higher in Beagles and Dachshunds than in Dalmatians and German Shepherds. The highest and the lowest absolute lymphocyte counts were found in Beagles and German Shepherds, respectively. As a consequence, German Shepherds showed the lowest absolute counts of the individual lymphocyte subpopulations and the widest neutrophil:lymphocyte ratio. Dalmatians showed the lowest percentage of CD3+ cells, the highest percentage of CD21+ cells, and the lowest CD4+:CD8+ ratio. German Shepherds showed the lowest percentage of CD21+ cells and the highest CD4+:CD8+ ratio. Females in Beagles and Dachshuns had nonsignificantly higher percentages of total lymphocytes, CD3+, CD4+, and nonsignificantly lower percentages of CD21+ lymphocytes. We concluded that there are age-, breed-, and perhaps also gender-related differences in lymphocyte subset distribution in the peripheral blood of dogs. Therefore, there is need to use appropriate control group in the experimental protocols. Among-breed differences could explain, at least partly, breed predisposition for some diseases.     

伯氏疟原虫ANKA株感染小鼠的T细胞、NK细胞及细胞因子变化

旨在系统分析伯氏疟原虫感染引起宿主T细胞、NK细胞、Tim-3表达及细胞因子的变化。选取64只雌性C57BL/6小鼠随机分为8组,每组8只,分别于感染后0、4、7、9、11、13、16和19 d获取小鼠脾及外周血免疫细胞,利用流式细胞术检测小鼠主要免疫细胞亚群及免疫检查点分子Tim-3表达水平的变化;同时检测血清中细胞因子的变化。结果表明,感染疟原虫后,小鼠脾CD3⁺CD4⁺ T细胞、CD3⁺CD8⁺ T细胞及NK细胞的比例均逐渐降低（P<0.01）,且伴随着3种细胞Tim-3表达量的升高（P<0.01）。小鼠外周血CD3⁺CD4⁺ T细胞的比例呈先降低后升高趋势,CD3⁺CD8⁺ T细胞的比例呈先升高后降低趋势（P<0.05）;小鼠外周血CD3^-NK1.1⁺细胞的比例呈先降低后升高趋势,但感染末期,其比例仍低于未感染组（P<0.05）。Tim-3分子在外周血CD3⁺CD4⁺ T细胞、CD3⁺CD8⁺ T细胞及CD3^-NK1.1⁺细胞的表达均显著升高（P<0.05）。感染疟原虫后,小鼠血清中促炎细胞因子IL-2的分泌量均显著高于未感染组（P<0.05）;促炎细胞因子IFN-γ、TNF-α和IL-6在血清中分泌均呈先升高后降低趋势（P<0.05）;具有免疫抑制作用的细胞因子IL-10呈逐渐升高趋势,且感染后期急剧升高（P<0.001）。以上结果表明,感染疟原虫后,小鼠的特异性免疫反应发挥了一定的杀伤作用,但由于Tim-3免疫检查点分子及一些发挥免疫抑制作用的细胞因子（IL-10）的过度表达,有利于疟原虫逃避宿主的免疫捕杀作用。该研究提示了从宿主免疫抑制角度研究疟原虫感染的重要性。     

Proportions and phenotypic expression of peripheral blood leucocytes in pigs vaccinated with an attenuated C strain and a subunit E2 vaccine against classical swine fever

The influence of an attenuated classical swine fever virus C strain vaccine and a subunit E2 vaccine against classical swine fever on the peripheral blood leucocyte proportion and phenotypic expression in 12-week-old pigs was studied. The C strain was amplified in minipig kidney cell culture and final product contained 10(4 +/- 0.15) TCID50/ml, while the subunit vaccine contained 32 microg per dose of gp E2. Haematological findings showed that the vaccines did not cause leucopenia or lymphocytopenia and the number of neutrophils and eosinophils during the observation period was within physiological range. The results of the proportion of CD4a+, CD5a+, CD8a+, wCD21+, CD45RA+, CD45RC+ , non-T non-B, SWC3a+ and CD11b+ cells were gained by single-colour flow cytometry. At the end of the trial a significantly increase of percentage of CD4+, CD5a+, CD8+, wCD21+ cells has been found in pigs that received the subunit vaccine and the percentage of CD4+, CD5a+, CD8+, CD45RA+ and CD45RC+ cells was higher in pigs that received the attenuated vaccine. Twenty-eight days after vaccination the percentage of CD4+, CD45RA+ and CD45RC+ was significantly higher in pigs vaccinated with the C strain than in pigs vaccinated with the subunit vaccine. In contrary, the percentage of the wCD21- cells was higher in pigs that received the subunit vaccine. Statistically higher values of SWC3a+ and lower values of CD11b+ cells was observed in pigs that received the attenuated vaccine than in pigs vaccinated with the subunit vaccine. Taken altogether, our results showed that the subunit vaccine produced a better stimulation of B cells and CD11b+ monocytes/macrophages /granulocytes/NK cells, whereas the attenuated vaccine induced a higher response of Th cells, naive/memory cells and macrophages/neutrophils. Thus, both vaccines were able to influence the porcine immune system, by activating different subsets of the immune effector/accessory cells.     

复方中药秦公散对小鼠脾脏和外周血中T、B淋巴细胞的影响

为探讨治疗奶牛乳房炎的复方中药秦公散对小鼠脾脏和外周血中T淋巴细胞和B淋巴细胞亚群的影响,采用流式细胞仪测定各处理组小鼠脾脏和外周血中用CD3+、CD4+ 、CD8+标记的T淋巴细胞百分率及CD4+/CD8+的比值和CD19+标记的B淋巴细胞百分率。结果表明,与蒸馏水组相比,秦公散高剂量组小鼠脾脏和外周血中CD3+、CD4+细胞百分率和CD4+/CD8+的比值均显著提高,对环孢菌素（CsA）抑制小鼠有显著恢复作用(P<0.05);秦公散高剂量（20 g/kg体重）可显著提高健康小鼠脾脏和外周血中CD19+细胞百分率(P<0.05),对环孢菌素抑制小鼠的CD19+细胞百分率有显著恢复作用(P<0.05)。结果表明,治疗奶牛乳房炎的复方中药秦公散通过提高机体中CD3+细胞、CD19+细胞百分率和调节CD4+/CD8+之间的平衡来显著提高小鼠机体的细胞免疫功能和恢复由环孢菌素（CsA）抑制的小鼠机体免疫力。