Blood Lymphocyte Subpopulations, Neutrophil Phagocytosis and Proteinogram During Late Pregnancy and Postpartum in Mares

The aim of this study was to evaluate peripheral blood lymphocyte subpopulations, neutrophil phagocytic capacity and proteinogram characteristics in mares, during the last trimester of pregnancy and in postpartum. Measurement of phagocytosis and quantification of T‐lymphocyte subsets were done by flow cytometry. Quantification of T‐lymphocyte subsets was performed with monoclonal antibodies specific for CD2, CD3, CD4 and CD8 cell markers. Natural killer and B‐cell counts were estimated mathematically. Serum proteinogram was obtained by electrophoresis. No significant differences were observed between gestation and postpartum on CD4⁺, CD8⁺ and NK⁺ lymphocyte subsets, CD4 : CD8 ratio and phagocytosis. The percentage of cells expressing CD3 (64.2 ± 1.8) and CD2 (68.4 ± 1.7) (Mean ± SEM) was reduced during gestation vs postpartum (69.7 ± 1.5 and 73.8 ± 1.4 respectively) (p < 0.05). During pregnancy, CD19⁺ (31.6 ± 1.7) was higher than in postpartum (26.2 ± 1.4) (p < 0.05). Total T cells (2911 ± 227 cells/μl), T helper cells (2144 ± 169 cells/μl) and T‐cytotoxic cells (767 ± 68 cells/μl) were depressed in pregnancy, when compared with postpartum (4093 ± 337 cells/μl; 3004 ± 276 cells/μl; 1089 ± 94 cells/μl respectively) (p < 0.01). Total white blood cell count was reduced during pregnancy (8815 ± 427 cells/μl) with respect to postpartum (10742 ± 446 cells/μl) (p < 0.01), while neutrophil count did not change. Total proteins, albumin, α₁,α₂,β₁, β₂, γ globulins and albumin : globulin did not differ. Our results suggest that the physiological immune depression occurring in mares, during gestation might be due to T‐helper and T‐cytotoxic lymphocytes reduction.     

绵羊妊娠期外周血T细胞及其亚群的观察

应用酸性α-萘酯酶(ANAE)组化反应标记绵羊妊娠期外周血T细胞,与间情期相比,T细胞总数和辅助性T细胞(Th)无明显变化(P>0.05),而抑制性T细胞(Ts)明显增加(P<0.05).间情期和妊娠期Th/Ts分别为4.6和2.6.结果提示,妊娠期羊可能通过增加Ts细胞来压低母体免疫水平.     

日粮高硒对雏鸡外周血T淋巴细胞增殖功能和亚群的影响

300只1日龄艾维茵肉鸡健雏随机分为5组,分别喂以基础日粮(日粮Se含量0.2 mg·kg-1,对照组)和高硒日粮(Se 1 mg·kg-2,高硒Ⅰ组;Se 5 mg·kg-1,高硒Ⅱ组;Se 10 mg·kg-1,高硒Ⅲ组;Se 15 mg·kg-1,高硒Ⅳ组)6周,用流式细胞术和MTT法研究高硒对外周血T淋巴细胞亚群、淋巴细胞增殖功能的影响.结果显示,当雏鸡日粮硒水平达5 mg·kg-1以上时,外周血CD3+ T细胞、CD3+CD4+T细胞和CD3+CD8+T细胞百分比不同程度地降低;高硒Ⅲ、Ⅳ组的T淋巴细胞增殖能力下降,血清IL-2含量降低,与对照组间差异显著或极显著(P<0.05或P<0.01).结果表明,5 mg·kg-1以上的日粮硒降低雏鸡细胞免疫功能.     

黄芪多糖对雏鸡外周血T淋巴细胞转化功能的影响

将１０８只１日龄伊莎系蛋用公雏均分为３组：一组为对照,其余２组在３日龄时,于背侧颈部皮下分别注射０．２、０．４ｍＬ黄芪多糖注射液（０．０１ｇ／ｍＬ）１次,再分别于７、２１、３５、４９日龄时采用ＭＴＴ比色法及微量全血培养３Ｈ－ＴｄＲ掺入法检测外周血Ｔ淋巴细胞转化率的动态变化。结果表明,黄芪多糖对２１、３５日龄雏鸡Ｔ淋巴细胞转化功能有增强作用,且与剂量有相关性,而对７、４９日龄雏鸡的作用不明显。ＭＴＴ比色法与３Ｈ－ＴｄＲ掺入法的检测结果无显著差异（Ｐ＜０．０５）。     

连城白鸭外周血淋巴细胞染色体核型研究

本试验通过对连城白鸭外周血淋巴细胞培养、秋水仙素处理后制备染色体标本,并进行染色体核型分析。结果表明,连城白鸭体细胞数目为2n=78±,1号常染色体为亚中央着丝粒染色体;2号常染色体为中央着丝粒染色体;3～9号常染色体及W性染色体为端着丝粒染色体;Z性染色体为亚端着丝粒染色体。本文通过对外周血淋巴细胞染色体核型分析,为进一步研究连城白鸭品种选育奠定基础。     

Efficacy of Medroxyprogesterone Acetate in Suppression of Estrus in Cycling Mares

The effects of compounded medroxyprogesterone acetate (MPA) on follicular activity and estrous behavior were evaluated. Eighteen cycling mares were assigned to one of three treatment groups. Mares in the MPA group (n = 6) were injected intramuscularly with 1,600 mg MPA (week 1), then 400 mg weekly for the next 5 weeks. Saline mares (n = 6) were injected intramuscularly weekly for 6 weeks. Altrenogest mares (n = 6) received 10 mL orally daily for 7 weeks. Mares were teased daily for 60 days and categorized as displaying estrous, diestrous, or neutral behavior. Transrectal ultrasound examinations were performed three times weekly, or daily when a 30-mm follicle was identified, until ovulation. Blood samples were harvested weekly for analysis of progesterone concentration and daily from days 14 to 23 for analysis of luteinizing hormone (LH) concentration. Mares treated with saline or MPA showed normal intervals of diestrus and estrus during the study. All altrenogest mares showed behavioral diestrus during treatment. All mares in the saline and MPA groups showed normal follicular development and ovulations. No altrenogest mares ovulated during treatment; four mares returned to estrus and resumed normal follicular development after treatment ceased. Progesterone analyses agreed with transrectal ultrasonographic ovarian activity for all mares. LH levels were lower for altrenogest-treated mares compared with MPA-treated and saline-treated mares during the treatment period. In conclusion, compounded MPA at dose rates and intervals used in this study was not effective in suppression of estrus, follicular development, or LH secretion in mares.     

临床健康牛外周血淋巴细胞亚类分布的研究

以流式细胞术研究了临床健康牛外周血中CD3、CD4、CD8和γδT淋巴细胞的分布情况.结果表明,牛外周血淋巴细胞中各T淋巴细胞亚类分布差异较大.其中CD3平均值为53.88±8.71%,CD4为17.13±3.22%,CD8为15.10±5.81%,γδT为18.77±8.07%.同时检测了广泛存在于各种淋巴细胞表面的白细胞介素-2α受体(IL-2Rα),发现IL-2Rα百分率与γδT细胞百分率具有正相关关系,即γδT细胞百分率高时,IL-2Rα也高,反之亦然.跟踪3头牛外周血各淋巴细胞亚类分布百分率的实验结果,证明牛外周血中各淋巴细胞亚类分布的差异,与牛的生物节律有着密切关系.     

牛流行热病毒人工感染牛外周血淋巴细胞T淋巴细胞亚类的分布研究

通过人工接种，使牛患牛流行热，并以流式细胞术分析了患牛高热期外周血中CD3^ 、CD4^ 、CD8^ 、γδT淋巴细胞和B淋巴细胞的分布情况。结果表明，高热期患牛外周血中CD4^ 细胞数明显升高，说明细胞免疫在牛流行热中的重要性；其它淋巴细胞亚类百分率无显著变化，但CD3^ 、CD8^ 和B淋巴细胞也有一定程度的提高；在反刍动物中起重要作用的γδT淋巴细胞则表现为下降，其作用尚需进一步探讨。本研究首次证明，BEFV可显著刺激牛外周血淋巴细胞CD4^ 的急剧增殖，证明牛流行热病程中细胞免疫可能起着重要作用。     

激动素对衰老模型小鼠外周血T淋巴细胞亚群及免疫力的影响

为了研究激动素对衰老模型小鼠外周血T淋巴细胞亚群及免疫力的影响,将昆明系小白鼠分为青年对照组、衰老模型组和激动素低、中、高处理组,衰老模型组和各激动素处理组小鼠背部皮下注射125 mg/（kg·BW）D-半乳糖制造衰老模型,低、中、高激动素处理组分别腹腔注射浓度为5、10、20mg/（kg·BW）的激动素溶液,青年对照组不作处理。检测各组小鼠外周血T淋巴细胞亚群的变化、红细胞结合C3b受体（C3b R）和免疫复合物受体（ICR）的能力以及胸腺、脾淋巴细胞的凋亡情况。结果显示,不同浓度的激动素可提高衰老模型小鼠的外周血CD3＋、CD4＋、CD8＋T淋巴细胞亚群的百分比,提高RBC-C3b R花环率,降低RBC-ICR花环率,降低胸腺、脾淋巴细胞凋亡率,表明激动素能够提高小鼠机体免疫力,延缓免疫衰老。     

Effect of Recombinant Canine Granulocyte Colony-stimulating Factor on Peripheral Blood Neutrophil Counts in Normal Cats

Recombinant canine granulocyte colony-stimulating factor (rcC-CSF) was administered subcutaneously at a dosage of 5 μg/kg/day to five healthy, young adult cats for 42 days. Mean neutrophil counts ± standard deviation increased significantly ( P > 0.001) from 10,966/μL ± 2324 to 30,688/μL ± 5296 within 24 hours after administration of the first dosage of rcG-CSF. Mean neutrophil counts reached 52,978/μL ±11,207 on day 6, representing a second significant increase ( P > 0.01) over the previous 5 days. Mean neutrophil counts continued to increase, reaching 66,994/μL ± 12,419 on day 14, then remaining within a range of 66,994 to 87,839/μL throughout the remainder of the study. The maximum mean neutrophil count was 87,839/μL ± 8,695 on day 42. Neutrophil counts remained high until the administration of recombinant canine granulocyte colony-stimulating factor was discontinued 42 days after initiation of therapy. Once the rcG-CSF administration was discontinued, neutrophil counts returned to pretreatment values within 5 days. There were no significant changes in numbers of any of the other cell lines. There was no clinically significant toxicosis associated with the administration of rcGCSF.     

牛流行热病毒灭活疫苗免疫后牛外周血淋巴细胞亚类分布的研究

对牛流行热病毒灭活疫苗免疫并攻每后，牛外周血淋巴细胞亚类的变化进行了研究。结果表明，灭活苗免疫后，牛的CD4^ 显著升高，可能与参与辅助B淋巴细胞合成抗体有关，攻毒后，γδT细胞均显著上升，免疫组在攻毒后3周仍保持在相当的高水平，IL－2Rα阳性细胞在攻毒后高热期也有显著升高，而CD8^ 细胞没有明显变化。     

感染李氏杆菌鸡外周血淋巴细胞rDNA转录活性分析

李氏杆菌是引起多种动物体内组织器官单核细胞增多的一种人畜共患传染病,其致死率较高.自1926年首次分离到病原以后,现已在世界广泛分布.80年代以来食源性李氏杆菌的感染不断增多,越来越引起人们的关注.顾玉芳等[1]对山鸡发生李氏杆菌病做了详细的病理学观察,为了探讨发病机体的免疫状态,本试验对感染李氏杆菌的鸡外周血淋巴细胞rDNA转录活性做了分析,以此来评估机体免疫水平.     

叶尔羌高原鳅外周血淋巴细胞的分离

以体质健康的叶尔羌高原鳅为研究对象,分离其外周血淋巴细胞。结果表明,采用浓度为1.085g/mL的淋巴细胞分离液在2000r/min离心35min的情况下,可将叶尔羌高原鳅外周血淋巴细胞有效地分离出来。该研究不仅为叶尔羌高原鳅淋巴细胞功能研究提供了依据,还可为叶尔羌高原鳅的疾病防治、人工养殖提供重要参考。     

一氧化氮介导内毒素所致的外周血淋巴细胞凋亡及阳离子A的保护作用

采用内毒素（ET）所致家兔ET休克模型，分别在相应处理后3、4、8、12h检测血浆中NO含量的动态变化；用荧光显微镜观察外周血淋巴细胞（PBL）的凋亡情况并计算凋亡指数；用琼脂糖凝胶电泳检测PBL凋亡的特征性DNA降解，并观察阳离子A（CA）注射液对上述指标的影响。结果显示，模型组中各时间段的血浆中NO含量均显著高于对照组（P〈0．01），随着NO含量的增加，PBL凋亡指数明显上升（P〈0．01），其基因组DNA在处理后3、4、8h均发生180-200bp及其整数倍的“阶梯状”断裂，12h出现“弥散状”条带；CA保护组中NO水平和PBL凋亡指数均出现不同程度的降低（P〈0．01，P〈0．05），并且在电泳结果中无明显的“梯状”条带。提示ET可使血液中NO含量升高。NO参与了ET休克的发病机制，介导PBL凋亡，在晚期介导PBL坏死；而CA可有效地中和血液循环中的ET，明显降低机体内NO的含量，抑制PBL凋亡和坏死，对ET休克有一定的保护作用。     

沙葱、油料籽实对绵羊外周血单核细胞和T淋巴细胞的影响

研究选用28只内蒙古羯羊,依照体重相近的原则随机分为4组,分别为对照组、4%沙葱组(占精料干物质的比例,以下相同)、4%沙葱+油料籽实(8%胡麻籽+2%麻籽+2%葵花籽)组、油料籽实(8%胡麻籽+2%麻籽+2%葵花籽)组,在试验期45d内采4次血,研究以上4个处理对绵羊血液细胞免疫、非特异性免疫功能的影响。结果表明:试验组单核细胞、CD2+和CD4+T淋巴细胞数均显著高于对照组(P<0.05);对照组CD8+T淋巴细胞数高于试验组,差异不显著(P>0.05)。从结果来看,沙葱和油料籽实显著提高了绵羊机体的非特异性免疫和细胞免疫机能,为以后开发舍饲肉羊新型的免疫添加剂以及进一步研究沙葱影响免疫机能的机理提供了理论依据。     

Changes in Peripheral Blood Leukocyte Percentages and Function of Neutrophils in Stressed Channel Catfish

Abstract

Channel catfish Ictalurus punctatus were subjected to a simulated handling (including bleeding) and transport stress. Using flow cytometry in conjunction with monoclonal antibodies to channel catfish B and T lymphocytes (minimal cross-reaction with thrombocytes) and neutrophils coupled with routine hematological methods, we demonstrated significant decreases in the percentage of B lymphocytes and increases in T lymphocytes and neutrophils of transported fish, yet there was no change in neutrophil phagocytic function. These data suggest that the length of the stressing event or the duration of exposure of the neutrophils to cortisol could determine if immune responses become suppressed. These data prompted us to expose channel catfish in vivo, and isolated neutrophils in vitro, to cortisol at various concentrations and for various time periods. Exposure of neutrophils to cortisol concentrations of 100 μg/dL for 2 h did not suppress phagocytosis; however, significant decreases in the percent phagocytosis and bacterial killing and increases in total number of neutrophils isolated were obtained after repeated in vivo administration of cortisol. The results of these experiments indicate (1) that bleeding, handling, and transport of channel catfish induce a characteristic relative lymphopenia and neutrophilia, (2) that cortisol does not act alone to induce suppression of phagocytic function (based on the in vitro administration of cortisol), and (3) that high physiological concentration of cortisol in vivo can initiate phagocytic suppression. In general, the intraperitoneal cortisol injection experiments suggested that stress on channel catfish that produces a concomitant high serum concentration of cortisol can suppress neutrophil phagocytic function, perhaps providing an avenue for the onset of infection.     

Study on Change of Lymphocyte Subpopulations in Swine after Immunization with Inactivated Vaccine

In order to investigate the change of lymphocyte subpopulations in the peripheral blood after injection of foot-and-mouth disease inactivated vaccine, blood samples were collected after vaccination and assayed with flow cytometry.The results showed that the proportion of dendritic cells (DCs) increased significantly in a short time after vaccination, after which the number of B cells, helper T (Th) cells and cytotoxic lymphocytes (CTLs) also raised with significance.The changing trend of B cells proportion was highly relevant with Th cells and a same correlation also appeared in the antibody titers.This indicated that the innate immune system could be activated rapidly after vaccination with the inactivated vaccine, and the adaptive immune response including cellular immune response and humoral immune response could be invoked subsequently.The high level of neutralizing antibodies was of great importance in evading FMDV and the cellular immune response also exerted a crucial role.     

Study on the Effect of Ricin on Human Peripheral Blood B Lymphocyte and the Expression of Related Genes

This study was amied to learn the toxic effects of ricin on human peripheral blood B lymphocytes（IM-9）and the expression of related genes.The extracted ricin of different concentrations were added to culture cells for 6 and 12 h to make sure the median inhibitory concentration,then culture cells at the median inhibitory concentration for 2,6,10 and 12 h,at each time point detect the expression of immune related genes CD40,IL-1β,TNF-α and apoptosis related genes Bcl-2,Bax,Caspase-3.The results showed that the inhibitory effect of ricin on IM-9 cells was increased with the increase of concentration and time.The expression of TNF-α and CD40 genes increased significantly in experimental group than control group at 6 h（P<0.05).10 and 12 h reached extremely significant level（P<0.01);There was no significance of IL-1β gene expression between experimental and control groups（P>0.05).About apoptosis gene,the Bax gene expression decreased extremely significantly in experimental group at 10 h（P<0.05）and decreased significantly at 12 h（P<0.05);Bcl-2 gene had reached significant levels of four time periods（P<0.05), and 2,12 h were extremely significant（P<0.01);The expression of Caspase-3 reduced at 6 h,the other time points increased significantly（P<0.05),at 2 h reached extremely significant level（P<0.01).It showed that the gene expression of IM-9 cells could be significantly affected by ricin,TNF-α and CD40 genes were two potential genes to evaluate the toxicity by IM-9 cells.     

3种植物多糖对鸡外周血和脾脏淋巴细胞体外增殖作用的影响

本试验旨在比较3种植物多糖对鸡外周血T淋巴细胞和脾脏B淋巴细胞体外增强免疫活性的研究.通过酶学检测法测定3种植物多糖对鸡胚成纤维细胞的安全浓度,MTT法比较3种植物多糖单独刺激和PHA协同刺激对鸡外周血T淋巴细胞增殖指数的影响,同时比较3种植物多糖单独刺激及与LPS协同刺激对鸡脾脏B淋巴细胞增殖率的影响.结果显示,黄芪多糖和桑叶多糖具有较高的细胞安全浓度,在64~512 μg/mL时,3种植物多糖对鸡外周血T淋巴细胞和脾脏B淋巴细胞的增殖作用较强.结果表明,3种植物多糖对鸡外周血和脾脏淋巴细胞都有不同程度的体外增殖作用,其中黄芪多糖的增殖作用最强,桑叶多糖其次,山药多糖最差.