八珍汤对乳牛产后免疫状态的影响

为了探索提高乳牛产后期免疫状态的新方法，对试验乳牛分4组（产前第30d至产前第1d灌喂组、产前第15d至产后第15d灌喂组、产后第1d至第30d灌喂组、不灌喂中药对照组）灌喂中药八珍汤，检测产后期淋巴细胞及其亚群数量和淋巴细胞增殖活化功能。结果发现，产前组CD3细胞数量在产后第1d升高；产前产后组CD3、CD4和CD8细胞数量在产后第1d和第15d升高；产后组CD3细胞数量在产后第15d和第30d升高，CD4细胞数量也在产后第30d升高。淋巴细胞对ConA的反应能力，产前产后组在产后第1～30d明显提高，产后组在产后第15d和第30d明显提高。各组乳牛产后期CD21细胞数量的变化相近。结果表明，从产前第15d开始到产后第15d每日喂八珍汤，能明显提高乳牛产后期T细胞及其亚群数量和增加淋巴细胞增殖活化功能，而对B细胞数量增加的作用不明显。     

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.     

雏鸡淋巴细胞变化规律的流式细胞仪检测研究

为进一步研究鸡在出孵1个月左右时间内T、B淋巴细胞及其亚群的发育状况，将SPF雏鸡分别在3、5、7、9、11、14、17、22、27、32日龄时取血液、脾脏、法氏囊及胸腺，分离并制成淋巴细胞悬液，流式细胞仪双染色法检测Bu-la^＋B淋巴细胞、CD3^＋、CD4^＋、CD8^＋和TCRγδ^＋CD3^＋T淋巴细胞含量及血液淋巴细胞的凋亡规律。结果：在出孵1个月内T、B细胞处于分化发育阶段的初期，除胸腺外，呈上升趋势，但T细胞亚群的变化无规律，B细胞在法氏囊中和T细胞在脾脏中3～9日龄发育最快；血液淋巴细胞凋亡趋于下降，尤其是在9日龄以后：此间不断有成熟的淋巴细胞产生，不断进入血液和脾脏，使其数量有上升趋势。但是，血液中第7日龄时Bu—la^＋B淋巴细胞和CD3^＋、CD4^＋、CD8^＋T淋巴细胞数量均最低，说明体液免疫和细胞免疫水平第7日龄时都最低，并在17日龄时进入水平较高的又一个低谷。TCRγδ^＋CD3^＋T细胞则变化幅度不大。     

猪繁殖与呼吸综合征病毒ORF5基因疫苗肌肉注射诱导小鼠的细胞免疫

将猪繁殖与呼吸综合征病毒（PRRSV）ORF5基因疫苗（pcDNA—PRRSV—ORF5）以50、100、200μg 3个剂量肌注免疫BALB／c小鼠，以PBS和空载体质粒pcDNA为对照，采用流式细胞仪（FACS）、淋巴细胞增殖试验（MTT法）分别时小鼠外周血中CD4^＋、CD8^＋ T淋巴细胞数和淋巴细胞转化功能进行了检测。结果，3个剂量的pcDNA—PRRSV—ORF5接种小鼠后，其外周血都对ConA有明显的反应性。试验组与对照组比较差异极显著（P〈0．01）或显著（P〈0．05），CD4^＋ T淋巴细胞数在免疫后7d高于对照组（P〈0．01），CD8^＋ T淋巴细胞在免疫后28d高于对照组，大剂量基因疫苗免疫小鼠的细胞免疫应答高于中、小剂量。结果表明，pcDNA—PRRSV—ORF5免疫小鼠能够诱导机体产生良好的细胞免疫应答，并表现一定的剂量依赖性。     

不同剂量猪繁殖与呼吸综合征病毒ORF5基因疫苗肌肉注射诱导仔猪细胞免疫的研究

将猪繁殖与呼吸综合征病毒(PRRSV)ORF5基因疫苗(pcDNA-PRRSV-SC2-ORF5)分别以200 ug,100 ug和50 ug 3种剂量肌肉注射免疫21日龄仔猪,同时设空载体对照和生理盐水空白对照.于免疫后7 d、15d、30d、45d、60d和70d采用流式细胞仪(FACS)和淋巴细胞增殖试验(MTT法)分别对免疫猪外周血中CIM+、CD8+T淋巴细胞数和淋巴细胞的转化功能进行检测.结果表明3个不同剂量的pcDNA-PRRSV-SC2-ORF5基因疫苗免疫仔猪后均能诱导产生良好的细胞免疫应答,实验组与对照组比较差异极显著(p<0.01)或显著(p<0.05).大剂量基因疫苗免疫仔猪的外周血T淋巴细胞的增殖能力及CD4+、CD8+百分含量高于中剂量和小剂量基因疫苗免疫的仔猪,表现出一定的量效关系.     

鸡回肠中T淋巴细胞及其亚群发育的试验

为探寻鸡回肠中T淋巴细胞及其亚群的发育规律,本试验通过免疫组织化学方法,应用CD3、CD4和CD8单克隆抗体研究鸡回肠中T淋巴细胞及其亚群出现、迁移、定位分布及数量变化过程.结果显示,CD3+、CD8+T淋巴细胞最初于18胚龄时出现,CD4+T淋巴细胞于出壳后1日龄时出现.在定位分布上,CD3+细胞在黏膜上皮内以及固有层中均匀分布,CD4+细胞以固有层中的分布为主,黏膜上皮内的分布较少.CD8+细胞最初主要分布在黏膜固有层中;随后,CD8+细胞逐渐向上皮内迁移;最终,黏膜上皮内出现广泛的CD8+细胞浸润.在数量变化上,CD3+、CD4+及CD8+整体呈逐渐增加趋势,第2周时阳性细胞数量稍有下降,21日龄时显著增加到达较高水平后保持稳定.结果表明,鸡出壳后,回肠的细胞免疫功能逐渐增强,并在21日龄时到达成熟水平.     

鸡眼区相关淋巴组织对ND疫苗点眼免疫应答Ⅱ.眼区局部T淋巴细胞亚群的变化

本研究采用流式细胞仪技术对鸡眼区相关淋巴组织(HALT)在鸡新城疫疫苗点眼免疫后的局部T淋巴细胞亚群的变化进行了研究。研究发现,LaSota 系ND疫苗点眼免疫后眼区相关淋巴组织对ND疫苗可产生良好的细胞免疫应答。免疫组结膜相关淋巴组织(CALT)和哈氏腺(HG)中CD4 T及CD8 T淋巴细胞数量于首免后迅速升高。免疫期间其百分比含量明显高于非免疫组,到40 日龄时,有恢复到正常值的趋势。     

新鱼腥草素钠对雏鸡细胞免疫的影响

将采集的健康鸡抗凝血按常规方法分离外周血淋巴细胞并于体外培养24h后，加入不同剂量的新鱼腥草素钠（SNH），然后以MTT法测定SNH对淋巴细胞增殖的影响。结果显示，SNH在体外具有刺激淋巴细胞增殖的能力，而且其刺激淋巴细胞的能力与剂量有关，其中以终浓度为250mg/L效果最明显。为进一步探讨SNH在体内对动物免疫功能的影响，将120只非免疫健康鸡随机分为5组；其中第Ⅰ～Ⅲ组为试验组，分别注射不同剂量（4mg／kg、2mg／kg、1mg／kg体重）的SNH；第Ⅳ组为白细胞介素对照；第Ⅴ组为疫苗对照；所有动物于14日龄以新城疫病毒La Sota株弱毒活疫苗进行第一次免疫，2周后加强免疫1次，并于第一次免疫后的第7d、14d、21d、28d、35d分别测定各组动物的外周血CD4^＋／CD8^＋淋巴细胞比值。结果表明，体内注射SNH具有提高外周血CD4^＋／CD8^＋淋巴细胞比值的作用，并与对照组差异显著（P〈0．05），且这种作用同样与给药剂量相关，其中以2mg／kg体重最为理想。     

葡萄球菌性关节炎肉鸡免疫器官的病理学变化

利用鸡源致病性金黄色葡萄球菌复制鸡葡萄球菌性关节炎的病理模型,研究免疫器官的主要病理学变化及其内CD4 和CD8 T淋巴细胞的动态变化。结果表明:鸡接种细菌后呈典型的关节炎症状。脾脏肿大,表面呈网格状,法氏囊黏膜增厚,盲肠扁桃体和胸腺上散在出血点。光镜下脾脏淋巴小结早期数目增多,后期呈灶状坏死,法氏囊水肿,淋巴小结坏死液化。CD4 和CD8 T淋巴细胞的变化为:脾脏在接种后7 d淋巴小结内的CD4 和CD8 T淋巴细胞已明显多于对照组,与对照组比差异极显著(P<0.01);接种后7 d,法氏囊的淋巴滤泡周围的间隙中检出较多的阳性细胞,在接种后14 d CD4 T淋巴细胞达到高峰,随后下降,与对照组相比在接种后7 d差异显著(P<0.05),在接种后14 d差异极显著(P<0.01);而CD8 T淋巴细胞在接种后7 d就达到高峰,与对照组相比差异极显著(P<0.01);盲肠扁桃体的CD8 T淋巴细胞在接种后7 d上升,21 d达到最高值;胸腺组织中阳性细胞数对照组和试验组都很少,试验组的阳性细胞数的变化没有明显的规律。鸡接种金黄色葡萄球菌后免疫器官内CD4 和CD8 T淋巴细胞数目增多,表明T淋巴细胞参与了金黄色葡萄球菌引起鸡的关节炎的发生发展过程。     

高锌对雏鸡免疫功能影响的研究

1日龄艾维茵肉鸡健雏200只随机分为4组，Ⅰ组(基础日粮，Zn100mg／kg)、Ⅱ组(基础日粮 Zn1400mg／kg)、Ⅲ组(基础日粮 Zn1900mg／k)和Ⅳ组(基础日粮 Zn2400mg／k)，试验期7周。病理组织学观察可见免疫器官的淋巴细胞减少，淋巴滤泡形成较少，其皮质变薄、髓质增宽并可见髓质部淋巴细胞减少。淋巴器官发育和外周血淋巴细胞的分裂增殖受抑，外周血T淋巴细胞的成熟率显著降低并引起其亚群CD4、CD8数量和组成的变化，导致机体细胞免疫和体液免疫功能的降低；雏鸡红细胞C3b受体花环率下降和免疫复合物花环率升高，引起红细胞免疫黏附功能降低。结果表明，日粮高锌可致雏鸡免疫功能受损。     

福建黄兔泌乳性能初探

用本所兔场自繁的１１只福建黄兔健康经产母兔，进行泌乳性能测定。结果表明，本品种的泌乳高峰期出现于产后第９天，第１５天达到高峰，最高日泌乳量１１０．９克，３０天泌乳量２５１１．８克，日平均泌乳量８３．７３克，泌乳力９２５．６克。仔兔适宜补料诱食期应在产后第１５天。     

Postnatal development of leukocyte subset composition and activity in dogs

The aim of the presentation is to summarise our data on the counts and activity of circulating canine leukocytes at birth and on their changes in the first 3 months of life. On day 1, neutrophil counts were almost three times higher than lymphocyte counts. During the first week of life, a decrease of neutrophil and an increase of lymphocyte counts, resulting in a predominance of lymphocytes, were observed. Neutrophil counts reached values comparable with those in adults in 1 month. Lymphocyte counts were higher than those in adults during the first 3 months. From birth to the age of 3 months, the phagocytic activity of neutrophils was nonsignificantly higher than in young adults. When compared with adults, the peripheral blood of new-born pups contained a lower proportion of T lymphocytes (detected by CD3 and CD5 markers), with a very low percentage of CD8(+) cells and a higher proportion of CD21(+) B lymphocytes. The counts of individual subsets levelled out during the first 3 months of life, although the proportion of CD21(+) B cells remained higher all the time. Lymphocytes of new-born pups were able to respond to nonspecific mitogen stimulation. Spontaneous proliferation in vitro was higher during the first week of life. Although in vitro stimulation of lymphocytes with Concanavalin A in some pups was comparable with that of adult dogs, mean activity was weaker. Pups with zero or very low levels of maternal antibodies were able to develop specific immune responses to a parvovirus antigen as early as at 2 weeks of age. On the basis of these data, we assume that pups are born with an immune system that can respond to external stimuli. Nevertheless its development continues in the postnatal period and some parameters differ from adult values for at least 3 months after birth.     

Long‐term impact of puerperal metritis on the profiles of peripheral blood leukocytes in peripartum dairy cows

To determine the effects of puerperal metritis on the immune response, changes in the differential peripheral blood leukocyte counts were analyzed during the peripartum period in cows with or without metritis. Multiparous Holstein cows were examined for uterine health disorders and classified into two groups: healthy (n = 11) or metritis (n = 5) cows. The lymphocyte and monocyte counts and the proportion of CD8⁺ lymphocytes were higher in cows with metritis compared to healthy cows. Moreover, the effects of puerperal metritis on the lymphocyte counts and CD4⁺/CD8⁺ ratio persisted weeks after the uterine inflammation had self‐resolved. Taken together, the findings of the present study indicate the possible long‐term alterations of systemic immune responses in cows with puerperal uterine inflammation. © 2015 Japanese Society of Animal Science     

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.     

Bovine colostral CD8-positive cells are potent IFN-gamma-producing cells

IFN-gamma plays an important role in cellular immunity contributing to microorganism elimination. We have previously reported that bovine colostrum contains high levels of IFN-gamma as well as immunoglobulin. Lymphocytes are potent producers of IFN-gamma, so establishing the lymphocyte population in colostrum will help to identify the source of colostral cytokines. In this study, we used flow cytometric analysis to quantify the population of three types of lymphocytes found in colostrum; namely, CD4 (Th) cells, CD8 (cytotoxic T) cells, and gammadelta-T cells. We also quantified the concentration of colostral IFN-gamma using ELISA. IFN-gamma concentrations were measured in colostrum obtained from 96 healthy Holstein cows; the levels tended to decrease on the first day post-parturition. Flow cytometric analysis showed that many gammadelta-T- and CD8-positive cells were present in the colostrum, and that the CD4/CD8 ratio was low. The ratios of CD8- and gammadelta-T-positive cells to cells of other types decreased during the 5 days after parturition, but that of CD4-positive cells showed no change during the observation period. To identify IFN-gamma-expressing colostral lymphocytes, we used magnetic separation technology to separate the lymphocytes (CD4, CD8 and gammadelta-T) from colostral cells, then examined them for IFN-gamma mRNA expression with real-time PCR (RT-PCR). RT-PCR analysis revealed potent expression of the IFN-gamma gene in CD8-positive cells, reaching higher levels than in CD4- or gammadelta-T-positive cells. These results suggest that the CD8-positive T cells in colostrum play a role as producers of IFN-gamma.     

松针对雏鸡免疫功能和血清胆固醇的影响

试验旨在研究松针对雏鸡免疫功能和血清胆固醇的影响。选用1日龄健康罗曼公鸡240只,随机分为4组,每组3个重复,每个重复20只鸡,对照组直接饲喂基础日粮和正常饮水,Ⅰ、Ⅱ组分别在基础日粮中以0.5、1.0 g/只剂量的松针粉拌料饲喂,Ⅲ组以0.5 g/只剂量的松针水煎液饮服。预试期4 d,正试期70 d。结果表明,Ⅰ、Ⅱ、Ⅲ组CD3⁺CD4⁺T淋巴细胞百分率在首免后第15天时高于对照组,但Ⅰ、Ⅱ组CD3⁺CD4⁺T淋巴细胞百分率在首免后第30、45、60天时则低于对照组,Ⅲ组在首免后第45天时则低于对照组;Ⅰ、Ⅱ、Ⅲ组CD3⁺CD8⁺T淋巴细胞百分率在首免后第15、30、45天时高于对照组,但Ⅰ、Ⅲ组在首免后第60天则低于对照组。Ⅰ、Ⅱ组新城疫抗体水平在首免后第14和21天时低于对照组,但在第28、35天时则高于对照组;Ⅲ组新城疫抗体水平在首免后第7、35天时低于对照组,但在第14、21天时则高于对照组。Ⅰ、Ⅱ、Ⅲ组血清总胆固醇和甘油三酯水平与对照组相比均有降低趋势,其中Ⅰ、Ⅱ组血清甘油三酯水平显著低于对照组和Ⅲ组（P<0.05）;Ⅱ、Ⅲ组血清低密度脂蛋白水平低于对照组,Ⅱ组血清高密度脂蛋白水平高于对照组,但差异均不显著（P>0.05）。综上所述,松针粉和水煎剂可促进T淋巴细胞的增殖与分化,提高CD3⁺CD4⁺和CD3⁺CD8⁺T淋巴细胞含量,协同疫苗增强特异性免疫,不同程度的降低血清胆固醇水平。     

Effect of a dietary n-6 polyunsaturated fatty acid supplement on distinct immune functions of goats

The aim of this study was to investigate whether dietary n-6 polyunsaturated fatty acids (PUFA) alter immune functions as lymphocyte blastogenesis, expression of lymphocyte and monocyte markers, and cell adhesion molecules of the integrin family in goats. Dutch White castrated male goats received a diet either rich in olive oil (control group, n = 4) or rich in corn oil (test group, n = 4) over a period of 3 weeks. The animals in the test group had significantly higher linoleic acid (LA, 18:2n-6) levels in plasma on days 7, 14 and 21 of the diet than control animals. This was also true for washed erythrocyte membranes at day 21. A significant reduction in the percentage of alpha-4 integrin (CD49d) expressing lymphocytes was observed in goats fed LA rich corn oil at day 21. However, no changes were observed in either lymphocyte proliferative responses to ConA and PHA or the expression of other lymphocyte/monocyte markers. In conclusion, feeding corn oil rich in LA caused significant increases in the proportion of LA in the fatty acid composition of plasma and erythrocyte membranes and was accompanied by a significant decrease of the adhesion molecule alpha-4 integrin (CD49d) on lymphocytes but not on monocytes.     

Early-life longitudinal survey of peripheral blood lymphocyte subsets in Beagle dogs

A longitudinal study of peripheral blood lymphocyte subsets from 7 to 15 months of age was performed in Beagle dogs employing a multiparametric flow cytometry. The data were compared with data obtained from adult Beagle dogs that were housed in the same animal facilities and that were subjected to the same controls during the 34 weeks of the study. Absolute counts of total lymphocytes and CD3+ T, CD3+CD4+ Th and CD21+ B lymphocytes decreased during the entire 34 weeks period of the study in the young dogs group. The same was observed with regard to the percentage of CD3+CD4+ Th lymphocytes and the CD4/CD8 ratio, while the percentage of CD3+CD8+ Tc lymphocytes increased from 7 to 15 months of age. These age-related changes found in lymphocyte subsets distribution of young dogs led to level the absolute and relative values of adult dog lymphocytes. The observations of this longitudinal study illustrate the changes related to maturation of lymphocyte subsets that occur during early life in Beagle dogs.     

Phenotypic analysis of hepatic lymphocytes from healthy dogs

Phenotypes of lymphocytes from laparoscopically biopsied liver tissues of eleven healthy beagle dogs were analyzed. The proportion of CD3(+) lymphocytes (T cells), CD3 (-)CD21(+) lymphocytes (B cells) and CD3 (-)CD21(-) lymphocytes (non-T non-B lymphocytes), and the CD4(+)/CD8(+) ratio in the canine hepatic lymphocytes were 54.8 +/- 11.9%, 4.7 +/- 3.1%, 40.7 +/- 13.2%, and 0.33 +/- 0.12, respectively, while those in peripheral blood lymphocytes were 85.4 +/- 6.5%, 9.3 +/- 6.1%, 5.3 +/- 1.8%, and 1.64 +/- 0.36, respectively. These results indicated that the constitution of hepatic lymphocytes quite differed from that of peripheral blood lymphocytes in dogs, and suggested that the regional immunity in canine liver might be specific.     

Lymphatic organ development in dogs: major lymphocyte subsets and activity

In the present study, we have characterized lymphocyte subsets and activity in peripheral blood, spleen, mesenteric and popliteal lymph nodes in pups from birth till the age of one month and compared the results with the situation in the group of three adult dogs. In neonatal pups, lower numbers of CD3(+) T-cells were detected in both the spleen and peripheral blood than in lymph nodes. In contrast to the other compartments, CD21(+) B-cells prevailed in the spleen, which resulted in low values (<1) of the CD3(+)/CD21(+) ratio. Low numbers of CD8(+) lymphocytes were characteristic in all compartments immediately after birth; consequently a high CD4(+)/CD8(+) ratio has been calculated. Postnatal development was characterized by an increasing frequency of CD8(+) lymphocytes in all organs studied. Another typical feature of the early period of life was a relative decrease of B-cell numbers, which was compensated by an increasing proportion of T-lymphocytes, particularly in the peripheral blood and spleen. DNA synthesis in newborn pups' cells as measured by in vitro thymidine incorporation was surprisingly high in non-stimulated control samples, notably in the spleen. Further development of lymphocyte activity was characterized by the decline in spontaneous activity in all organs. Stimulation indices upon mitogen-induced proliferation increased proportionally to the decrease in spontaneous activity. Based on our experimental data, we have concluded that pups are born with a relatively competent immune system the structure of which, however, markedly develops during a few postnatal weeks.