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.     

Peripheral Blood Lymphocyte Subpopulations and Immunoglobulin Concentrations in Healthy Foals and Foals with Rhodococcus equi Pneumonia

Infectious diseases are common in foals aged 1-5 months. The objectives of this investigation were to evaluate immunologic parameters in foals from birth to weaning to establish reference values for the proportion of circulating lymphocytes that were helper (CD4+) or cytotoxic (CD8+) T cells, or B cells; to measure serum immunoglobulin (IgM and IgG) concentrations; and to compare these immunologic parameters to values in foals with naturally occurring Rhodococcus equi pneumonia and in adult horses. Peripheral blood lymphocyte subpopulations were determined by flow cytometric analysis, and serum IgG and IgM concentrations were determined by radial immunodiffusion. Flow cytometric analysis of lymphocyte subpopulations suggested age-related changes in the cell-mediated immune system in horses. Absolute circulating CD4+ and CD8+ T lymphocytes and B cells increased linearly up to 3 months of age. Circulating B cell concentrations from birth to 6 months of age were greater than values in adult horses and the lymphocyte differences among the age groups are mainly due to variation in B lymphocytes. Both absolute and proportional B cell concentrations were greater in foals with R equi pneumonia than in healthy foals at the same age. The increase in absolute cell counts of each subpopulation was dependent on the increase of absolute peripheral blood lymphocyte count. Serum IgG concentration increased linearly from 1 to 3 months of age, and serum IgM concentrations increased from 1 to 6 months of age. These data suggest age-dependent cell-mediated and humoral development in young foals.     

高致病性猪繁殖与呼吸综合征 GD强弱毒株感染对猪外周血淋巴细胞亚群的影响

为了从细胞水平探讨PRRSV GD强毒株和PRRSV弱毒疫苗接种仔猪后对其外周血淋巴细胞亚群的影响,运用血常规技术和流式细胞技术分析了猪体外周血淋巴细胞亚群的动态变化。将30日龄20头SPF猪分成3组,分别感染HP-PRRSV GD第5代强毒株、GDr180弱毒疫苗株及留作对照。于感染后第0、3、7、10、14、21、28及35天采血进行外周血淋巴细胞亚群测定,分析淋巴细胞亚群的变化。结果表明:HP-PRRSV GD强毒株感染可导致白细胞、淋巴细胞数量、单核细胞、粒细胞、B细胞、Tc细胞、Th细胞、Tm细胞、和γδT细胞数量的下降;而弱毒疫苗GDr180株免疫则表现为白细胞、淋巴细胞、单核细胞、引起粒细胞、B细胞、Tc细胞、Th细胞和Tm细胞的上升,对γδT细胞的影响不大;阴性对照组在整个检测期间各种细胞基本上保持稳定状态。从实验结果可以看出:与HP-PRRSV GD强毒株破坏免疫细胞不同,PRRSV弱毒疫苗GDr180株免疫接种能刺激猪免疫细胞增殖,给试验猪提供良好的免疫反应。     

Effect of Chronic Administration of Oxytocin on Corpus Luteum Function in Cycling Mares

The objective of this study was to determine if intramuscular administration of 60 units of oxytocin once daily for 29 days, regardless of when treatment was initiated during the estrous cycle (i.e., without monitoring estrous behavior and/or detecting ovulation), would induce prolonged corpus luteum (CL) function in cycling mares. Mares were randomly assigned to two groups: (1) saline-treated control (n = 7) and (2) oxytocin-treated (n = 9) subjects. Control mares received 3 cc of saline, and oxytocin-treated mares received 60 units (3 cc) of oxytocin intramuscularly for 29 consecutive days. Treatment was initiated in all mares on the same day (day 1), independent of the day of the cycle. Jugular blood samples for determination of progesterone concentration were collected three times weekly (M, W, and F) for 21 days before treatment was initiated to confirm that all mares had a luteal phase of normal duration immediately before treatment. Beginning on the first day of treatment, blood samples were collected daily for eight days and then three times weekly through day 80. Mares were considered to have prolonged CL function if serum progesterone remained >1.0 ng/mL continuously for at least 25 days after the end of the treatment period. The proportion of mares with prolonged CL function was higher in the oxytocin-treated group than in the saline-treated group (7/9 vs. 1/7, respectively; P < .05). Three of the seven oxytocin-treated mares that developed prolonged CL function initially underwent luteolysis within 4–7 days of the start of oxytocin treatment and then developed prolonged CL function after the subsequent ovulation during the treatment period. In the other four oxytocin-treated mares that developed prolonged CL function, progesterone remained >1.0 ng/mL throughout the treatment period and into the post-treatment period. All mares with prolonged CL function maintained elevated progesterone concentrations through at least day 55 of the study. In conclusion, intramuscular administration of 60 units of oxytocin for 29 consecutive days effectively prolonged CL function in mares, regardless of when treatment was initiated during the estrous cycle. Importantly, this represents a protocol for using oxytocin treatment to prolong CL function that does not require detection of estrous behavior or day of ovulation.     

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.     

鸭病毒性肝炎对雏鸭外周血淋巴细胞转化的影响

鸭病毒性肝炎(Duck Virus Hepatitis,DVH)是雏鸭的一种传播迅速且致死率极高的传染病,世界主要养鸭国家均有本病的流行[1]。国内已开展了大量的有关DVH防治方面的研究工作,并在DVH特异性预防方面发挥了重要的作用。在治疗上,多采用高免血清和高免鸡或鸭蛋黄匀浆进行被动免疫和防治,亦有用中草药治疗的报道,如邹海棠等用田基黄治疗雏鸭DVH取得了一定疗效。近年来常有鸭肝炎并发病毒或细菌病的报道[2]。     

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。     

小花棘豆生物碱对小白鼠外周血T淋巴细胞亚群CD4+、CD8+免疫功能的影响

试验采用小花棘豆生物碱苦马豆素(SW)、黄花碱(Ts)以及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天、第14天、第21天、第28天、第35天,从各组中随机抽取4只小白鼠,自其眼球取抗凝血,利用流式细胞术检测外周血T淋巴细胞亚群CD4+、CD8+的百分检率,并计算出CD4+与CD8+的比值。结果表明,低浓度的SW具有提高机体免疫机能的作用,而中等浓度和高浓度的SW在短时间内仍然可以提高机体的免疫机能,长时间使用却会严重降低机体的免疫机能;Ts在低浓度下不会影响机体的免疫机能,中等浓度和高浓度能够降低机体的免疫机能;混合生物碱组在低浓度时对机体的免疫机能影响不大,但在高浓度时仍然能够显著降低机体的免疫机能。     

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。     

新鱼腥草素钠对鸡外周血T淋巴细胞增殖的影响

采集健康鸡肝素抗凝血,按常规方法分离外周血淋巴细胞并于体外培养24 h后,加入不同剂量的新鱼腥草素钠/刀豆素A(ConA),继续培养44 h后,以MTT法测定新鱼腥草素钠对淋巴细胞增殖的影响。结果表明,新鱼腥草素钠在体外具有刺激淋巴细胞增殖的能力,且与ConA具有协同作用;进一步分析表明,不同给药剂量的新鱼腥草素钠对淋巴细胞的影响不同,其中以终浓度为250 mg/L效果最明显。     

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。     

甲硝唑对鸡离体外周血淋巴细胞的作用

对咪唑类化合物甲硝唑对离体鸡外周血淋巴细胞(PBL)增殖及伴刀豆球蛋白A(ConA)诱导细胞(CIC)活性的作用并与咪唑类化合物左旋咪唑进行了比较；甲硝唑和左旋咪唑均以剂量依赖性方式促进鸡PBL的增殖；甲硝唑和左旋咪唑与ConA对促进鸡PBL增殖均呈现协同作用；甲硝唑和左旋咪唑对脂多糖(LPS)激活鸡PBL均具有明显促增殖作用；甲硝唑和左旋咪唑均可逆转磷酸组胺和肾上肾对ConA和nLPS激活鸡PBL增殖的抑制作用；高浓度左旋咪唑(100～400μg／mL)可明显逆转氢化可的松对ConA激活鸡PBL增殖的抑制作用，各种浓度甲硝唑和左旋咪唑均可明显逆转氢化可的松对LPS激活鸡PBL增殖的抑制作用；甲硝唑和左旋咪唑均可促进CD^ 4细胞增殖，抑制CE^ 8细胞增殖，使CD^ 4／CD^ 8细胞比值升高，CIC活性增强。     

Omega-3 Fatty Acid Supplementation Affects Selected Phospholipids in Peripheral White Blood Cells and in Plasma of Full-Sized and Miniature Mares

Dietary omega-3 fatty acids (n-3 PUFA), notably eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), impart health benefits in humans and animals. In horses, dietary n-3 PUFAs elevate EPA and DHA and may promote anti-inflammatory effects. No reports document effects of dietary n-3 PUFA on fatty acyl components of circulating and cellular phospholipids in horses nor whether responses to dietary n-3 PUFA are similar among horse breeds. Ten Quarter Horse and 10 American Miniature Horse mares were assigned to n-3 PUFA (64.4 mg· kg body weight [BW]⁻¹·d⁻¹) or control diet for 56 days. Blood was sampled at 0, 28, and 56 days. Apparent phospholipid molecular species from several classes (phosphatidylcholine [PC]; “ether-linked” phosphatidylcholine [i.e., alk(en)yl, acyl glycerophosphocholine] [ePC]; phosphatidylethanolamine [PE]; phosphatidylinositol [PI]; and phosphatidylserine [PS]) were determined in plasma and peripheral blood mononuclear cells (PBMC) by mass spectrometry. Statistical analysis showed that six phospholipid species had diet × day interactions (P < .05) for both plasma and PBMC. Further evaluation of these species demonstrated that the mole percentage of PC(38:6), PC(40:7), PC(42:10), PE(38:5), PE(40:6), and PE(40:7) (where x:y represents total acyl carbon:total carbon-carbon double bonds) in both plasma and PBMC phospholipids was elevated in horses fed n-3 PUFA (P < .001 for all). Analysis of the acyl product ions revealed that these contained an acyl chain of mass consistent with an n-3 PUFA. Thus, supplementation increased n-3 PUFA in selected plasma and PBMC phospholipids. The absence of breed effects suggests that miniature and full-size horses responded similarly to dietary treatment.     

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。