山羊和水牛感染肝片吸虫前后部分细胞免疫指标动态变化

1.LDH 短程释放法测定 NK 细胞活性动态变化本实验以 SP2/0细胞为靶细胞(即效应细胞),采用乳酸脱氢酶(LDH)短程释放法测定人工感染肝片吸虫的水牛和山羊的外周血液中淋巴细胞的 NK 活性动态变化。发现山羊在感染后 NK 活性有两次高峰,推测与虫体的发育或淋巴细胞的减少或嗜酸性粒细胞的增多有关;两个感染组(Ⅰ,200个囊蚴/只;Ⅱ,500个囊蚴/只)变化趋势相近,表明感染量与 NK 活性无相     

CD158受体表位封闭对急性髓系白血病患者NK细胞杀伤活性的影响

目的 研究急性髓系白血病患者NK细胞CD158受体表位封闭对自身白血病细胞杀伤活性的影响.方法 分离急性髓系白血病患者及健康志愿者外周血NK细胞,以自身白血病细胞及K562细胞为靶细胞,用CCK-8试剂盒检测CD158a、CD158b单克隆抗体封闭前后NK细胞在1：1、5：1、10：1效靶比下对靶细胞的杀伤活性.结果 患者与正常人NK细胞对K562细胞均有高度杀伤活性,且随效靶比增大而增高（P＜0.01）.效靶比为1：1、5：1、10：1时,患者NK胞在封闭前对白血病细胞杀伤活性分别为（1.5±0.3）%、（5.6±0.8）％、（11.8±0.6）%,封闭后分别为（21.8±0.7）％、（38.6±0.9）％、（53.9±1.4）％,各效靶比组封闭前后差异有统计学意义（P＜0.01）.结论 急性髓系白血病患者NK细胞CD158受体表位封闭可提高NK细胞对自身白血病细胞的体外杀伤能力.     

重组牛白细胞介素-2增强小鼠对伊氏锥虫可溶性抗原免疫反应的研究

实验使用伊氏锥虫可溶性抗原免疫小鼠,研究了重组牛白细胞介素-2(rBIL-2)对小鼠 NK 细胞杀伤活性及特异性抗体水平的影响.试验鼠随机分为4个组:Ag 组、Ag+rBIL-2组、rBIL-2组和 PBS 对照组.以LDH 释放法和间接 ELISA 法分别检测各组小鼠 NK 细胞杀伤活性和抗体水平.结果表明,首次免疫后24d·Ag 组 NK 细胞杀伤活性为31.6%,rBIL-2组为99.66%,Ag+rBIL-2组为98.98%,对照组为19.73%,抗体效价 Ag 组为2~4,Ag+rBIL-2组为2~6.说明 rEIL-2可显著提高 NK 细胞杀伤活性及促进抗体生成,提高宿主免疫力.     

杀伤细胞免疫球蛋白样受体在输血中的作用

目的 初步探讨杀伤细胞免疫球蛋白样受体在输血中的作用.方法 用免疫磁珠法分离正常人外周血NK细胞,再用PCR-SSP方法进行KIR分型.将分型的NK细胞与已知HLA分型的全血混合,37℃培养0、6、12、24h,ELISA法检测IL-2、IL-4、前列腺素E2(PGE2)、TNF-α、TNF-β、IFN-γ、粒细胞-巨...     

胶体金对小鼠细胞免疫的影响

实验设计了四种直径和三种剂量12个组合的胶体金(CG)对小白鼠进行注射处理;采用MTT法、放射性免疫法、LDH短程释放法,测定了T、B淋巴细胞增殖活性及其腹腔巨嗜细胞活性,外周血血清和脾脏淋巴细胞诱导白介素2含量和脾脏NK细胞活性,结果表明:不同直径和剂量的CG对机体有不同程度的影响,其中直径15nm、10nmCG的0.3mL和0.2mL剂量组不仅对T、B淋巴细胞增殖活性和血清中IL-2含量有显著促进,也可显著提高腹腔巨噬细胞活性及脾脏NK细胞杀伤活性,0.1mL剂量组虽有提高但不显著;直径5nm、20nm各个剂量组效果的波动较大,中、高剂量组对四项指标起促进作用但不显著,低剂量组则没有促进作用。证明了15nm和10nm中高剂量的胶体金可显著提高小鼠机体特异性和非特异性细胞免疫功能。     

神经介素U在体外对猪淋巴细胞活性及自然杀伤细胞活性的影响

[目的]本文旨在研究神经介素U(NMU)对猪外周血淋巴细胞(PBL)的免疫调节作用。[方法]以小梅山猪为研究对象,用半定量RT-PCR方法研究了猪PBL中NMU及其受体(NMUR1和NMUR~2)基因的表达,用免疫组织化学染色方法检测了猪PBL中NMU及其受体蛋白的分布,用Cell Counting Kit-8(CCK-8)细胞计数法研究了NMU对猪PBL细胞和NK细胞活性的影响。[结果]NMUR1 mRNA在猪PBL中表达,NMU和NMUR1蛋白在猪PBL的胞膜及胞浆均有分布。用不同浓度(0.01、0.1、1、10、100和1 000 nmol·L~(-1))NMU处理体外培养的PBL,与对照组相比,0.1、1和10 nmol·L~(-1)NMU极显著提高猪PBL的细胞活性(P0.01),且当NMU浓度为10 nmol·L~(-1)时,PBL活性的水平达到峰值(179.4±3.0)%。用不同浓度(0.01、0.1、1、10、100和1 000 nmol·L~(-1))NMU处理体外培养的PBL细胞作为效应细胞,以K562细胞作为靶细胞。与对照组相比,在效靶比为15∶1时,0.1、1和10 nmol·L~(-1)的NMU均能显著提高NK细胞的细胞活性(P0.01);效靶比为30∶1时,1和10 nmol·L~(-1)的NMU均能显著提高NK细胞的细胞活性(P0.01);在效靶比为60∶1时,10 nmol·L~(-1)NMU能显著提高NK细胞的细胞活性(P0.01)。[结论]NMU及NMUR1在猪PBL中均有表达,且NMU在一定浓度范围内能提高猪NK细胞对K562细胞的杀伤能力。     

肝癌患者外周血HBV DNA 与T细胞亚群、NK细胞活性关系的研究

目的:探讨肝癌患者外周血中HBV DNA与T细胞亚群、NK细胞活性的关系.方法: 选择肝癌90例,应用PCR技术和流式细胞术分别检测外周血中HBV DNA和T细胞亚群、NK细胞活性.结果: 外周血HBV 5项标志物均阴性的30例肝癌患者中16例 HBV DNA 阳性,阳性率53%.肝癌患者与正常对照组比较,CD4、CD8和NK细胞活性显著降低,但HBV DNA 阳性者比阴性者降低更明显,两者差异有显著性(P＜0.05).结论:T细胞亚群及NK细胞活性影响HBV的致癌作用.     

HLA-G1的基因表达、纯化及其活性分析

在获得HLA-G1cDNA克隆序列的基础上,构建了pGEX-4T2-HLA-G1原核表达载体,对HLA-G1的诱导表达发现,融合蛋白以包涵体形式存在于表达菌中。进一步溶解包涵体,改善复性条件,最终获得高纯度的GST/HLA-G1融和蛋白。51Gr释放试验表明,纯化的HLA-G1具有抑制NK细胞对靶细胞的杀伤活性。     

低温诱导淡水白鲳细胞凋亡的研究

为探讨10℃低温导致淡水白鲳尾鳍细胞系CBT死亡的原因及其可能机制.采用CCK-8试剂盒法检测细胞存活率,荧光染料H2DCFDA染色测定细胞内活性氧(ROS)含量,亚二倍体峰和Tunel分析检测细胞凋亡,琼脂糖凝胶电泳进行DNA片段分析,Hoechst33258染色观察凋亡细胞核形态变化,比色法测定乳酸脱氢酶(LDH)释放率.结果表明随CBT细胞受10℃低温作用时间的延长,细胞存活率显著下降,细胞内ROS含量和LDH释放率显著增加(P＜0.01).低温处理3 d的CBT细胞经流式细胞仪检测出现亚二倍体峰,凋亡率为13%.再经32℃复温12 h后,琼脂糖凝胶电泳显示DNA凋亡梯带;Tunel检测,低温处理3、4和5 d的CBT细胞分别有23.49%,27.72%和35.10%发生凋亡.低温处理5 d的CBT细胞核分裂成多个小核,呈现出典型的凋亡小体.因此10℃低温能诱导CBT细胞凋亡.ROS参与了10℃低温致CBT细胞损伤及凋亡过程.     

CD3抗体对EB病毒特异性T细胞杀伤作用的影响

用CD3抗体(UCH—T1)与EB病毒特异性效应细胞共同孵育,检测靶细胞的~(51)Cr释放率,以观察其阻断EB病毒特异性T细胞及其克隆细胞对EB病毒转化类淋巴母细胞株(LCLs)和NK细胞杀伤敏感细胞株     

气管滴注大肠杆菌对肉仔鸡肝脏抗氧化能力的影响

为研究呼吸道大肠杆菌对肉仔鸡肝脏抗氧化能力的影响,选取24只初始体质量相近、健康状况良好的固始肉仔鸡,随机分为2组,对照组肉仔鸡气管滴注1 m L无菌培养液,大肠杆菌组肉仔鸡气管滴注1 m L大肠杆菌培养液(1×10~9cfu/m L)。滴注后2 h采集血液和肝脏组织,测定血清中细菌脂多糖(LPS)、谷丙转氨酶(ALT)和谷草转氨酶(AST)含量,以及肝脏组织中丙二醛(MDA)含量、谷胱甘肽过氧化物酶(GSH-PX)和超氧化物歧化酶(SOD)活性。结果表明,气管滴注大肠杆菌后2 h,大肠杆菌组肉仔鸡血清中LPS含量为33.86 U/mg,较对照组升高了26.15%;AST和ALT含量较对照组分别增加了50.89%(P0.05)和298.77%(P0.01);大肠杆菌组肉仔鸡肝脏组织中SOD活性和GSH-PX活性分别为186.60 U/mg和13.20 U/mg,较对照组分别降低了43.83%(P0.01)和31.18%(P0.05);MDA含量增加了51.72%(P0.05)。综上,气管滴注大肠杆菌后2 h,大肠杆菌大量繁殖,释放出的LPS通过血液循环进入肝脏,肝脏中抗氧化酶活性降低,引起脂质过氧化物堆积,破坏肝脏细胞,导致血清中转氨酶水平升高。     

肉桂醛替代肉用仔鸡日粮中抗生素的使用效果

为研究肉桂醛替代肉用仔鸡日粮中抗生素的使用效果,选用1日龄健康青脚麻肉用雏鸡360只,随机分为4组:试验1(对照)、2、3和4组。每组设3个重复,每个重复30只雏鸡。在基础日粮中添加抗生素(1～3周龄:吉他霉素预混剂50mg/kg和盐霉素预混剂75mg/kg;4～8周龄:吉他霉素预混剂10mg/kg)后,饲喂对照组鸡;在基础日粮中分别添加肉桂醛预混剂40、80、120mg/kg后,分别饲喂试验2、3和4组鸡。饲养试验周期56d,观测鸡的健康状况、增重、血清生化指标、肠道菌群和肉质等。结果表明:1)在整个饲养周期,试验3组鸡的平均日增重较对照组提高5.68%;试验3组的料重质量分数(2.14±0.17)最低,但与其他组的差异不显著;2)与对照组比较,试验3、4组鸡血清谷丙转氨酶活性分别降低43.66%、61.97%,试验4组鸡血清谷草转氨酶活性降低9.27%,试验2组鸡血清总蛋白提高14.34%,试验2、4组鸡血清尿素氮都降低24.32%,试验3组鸡血清IgA提高31.03%,试验3、4组鸡血清IgG分别提高61.47%和66.97%,试验2、3、4组鸡血清IgM分别提高169.09%、283.64%和300.00%;3)试验3、4组鸡胸肌失水率分别较对照组降低17.21%和14.10%。本研究证实:肉桂醛能促进肉用仔鸡的生长,改进营养生化代谢,增强鸡体免疫功能,提高鸡肉品质;肉桂醛在肉用仔鸡中的应用效果好于抗生素;肉桂醛预混剂在肉用仔鸡日粮中的适宜添加量为80～120mg/kg。     

大豆黄酮对肉鸡血液胰岛素、胰高血糖素和肝脏苹果酸脱氢酶的影响

试验选用 2 1日龄艾维茵商品代肉鸡 4 8只 ,随机分成 2组 ,分别为基础日粮组和添加大豆黄酮 (daidzein Da)组 ,饲喂 5周。结果表明 ,外周血液中的胰岛素对照组与 Da组分别为 3.94± 0 .2 0 ,4 .30± 1.2 8︼ IU· m L- 1 ,比对照组增加 8.30 % ;胰高血糖素分别为 319.90± 32 .72 ,4 73.13± 77.0 8pg· m L- 1 ,比对照组增加了 37.37% (P <0 .0 1) ;肝中的苹果酸脱氢酶 (MDH)分别为 331.14± 17.99,30 8.12± 79.90 U· mg- 1 。试验结果提示 Da具有调控肉鸡脂肪代谢作用     

MHC class I deficiency: susceptibility to natural killer (NK) cells and impaired NK activity.

The role of major histocompatibility complex (MHC) class I expression in natural killer (NK) cell target recognition is controversial. Normal T cell blasts from MHC class I-deficient mutant mice were found to serve as target cells for NK cells in vitro, which suggests that MHC class I molecules are directly involved in NK cell recognition. Spleen cells from the mutant mice were deficient in their ability to lyse MHC class I-deficient target cells or NK-susceptible tumor targets, and mutant mice could not reject allogeneic bone marrow. Thus, class I molecules may participate in the positive selection or tolerance induction of NK cells.     

尖锐湿疣患者自然杀伤细胞活性及其与血清白介素—12关系？…

目的：研究尖锐湿疣（ＣＡ）患者自然杀伤（ＮＫ）细胞活性及其与血清白介素１２（ＩＬ－１２）水平的关系,探讨其发病机理。方法：应用ＮＫ细胞毒性试验乳酸脱氢酶释放法（ＬＤＨ－释放法）测定３２例ＣＡ患者ＮＫ细胞的活性;和抗体夹心ＥＬＩＳＡ法检测其血清中ＩＬ－１２水平;并对其ＮＫ细胞活性与血清ＩＬ－１２水平作相关分析。结果：ＣＡ患者ＮＫ细胞活性明显低于正常对照组（Ｐ〈０．０５）;血清ＩＬ－１２水平明显低于正     

五味子CO_2超临界提取物对肉仔鸡免疫功能的影响

文章通过以五味子CO2超临界提取物作为添加剂,研究其对肉鸡免疫功能的影响。试验选用1日龄的健康艾维茵肉仔鸡200只,随机分为5个处理组,空白组饲喂基础日粮,试验组分别在基础日粮中添加黄霉素5mg·kg-1、五味子CO2超临界提取物0.1%、0.15%、0.3%。于21、42、56日龄进行五味子提取物的免疫功能调控试验。结果表明,日粮中添加五味子提取物可显著提高肉仔鸡后期的胸腺指数,在日粮中添加0.3%五味子提取物可显著提高肉仔鸡血清NDV-HI抗体效价和血清溶菌酶含量,并可显著提高肉仔鸡血清IgM含量和肉仔鸡外周淋巴细胞转化率。结果显示,五味子能够增强肉鸡的免疫功能。     

Homeostasis of the antibody response: immunoregulation by NK cells

When injected into mice, the synthetic double-stranded polynucleotide poly(inosinic) X poly(cytidylic) acid induces high natural killer (NK) cell activity within 4 to 12 hours. Induction of NK activity in mice immunized 2 or 3 days previously, or the addition of NK cells to cultures immunized in vitro 2 or 3 days previously, promotes early termination of the ongoing primary immunoglobulin M antibody response. A target for NK cells is a population of accessory cells that has interacted with antigen and is necessary for sustaining the antibody response. The inference is strong that NK cells induced normally by immunization also terminate the usual antibody response in vivo by elimination of antigen-exposed accessory cells.     

硒对雏鸡T淋巴细胞转化和自然杀伤细胞活力的影响

将１日龄罗曼系商品代蛋用健康公雏３６０羽随机均分成３组。对照组饲喂基础日粮,补硒Ⅰ组和补硒Ⅱ组在饲喂基础日粮的同时分别添加０．３和０．６ｍｇ／ｋｇ硒。分别于７,２１,３５和４９日龄采用微量全血培养３Ｈ－ＴｄＲ掺入法测定雏鸡外周血Ｔ淋巴细胞转化率,ＬＤＨ释放法测定ＮＫ细胞活力,荧光分析法测定血硒含量,化学法测定ＧＳＨ－Ｐｘ活性。定期称重并进行临床观察。结果表明,补硒能显著并持续地增强雏鸡外周血淋巴细胞对ＰＨＡ的应答能力和ＮＫ细胞活力。另外,还对全血淋转试验３Ｈ－ＴｄＲ掺入法进行了讨论。     

Effects of selenium source and level on growth performance,antioxidative ability and meat quality of broilers

This experiment was conducted to investigate the effects of selenium(Se) source and level on growth performance, carcass traits, antioxidative ability and meat quality of broilers. A total of 320 one-d-old Arbor Acres commercial broilers were randomly assigned to 1 of 5 treatments with 8 replicates in a completely randomized design involving a 2×2 factorial arrangement of treatments plus one Se-unsupplemented basal diet control for 42 d. The two Se sources were sodium selenite and Se yeast, and the two supplemental Se levels were 0.20 and 0.40 mg Se kg–1. The results showed that broilers fed the Se-supplemented diets had higher(P0.05) average daily gain and average daily feed intake from 22 to 42 d of age, eviscerated yield and abdominal fat percentages, Se concentrations and glutathione peroxidase(GSH-Px) activities in breast and thigh muscles on d 42, and lower(P0.05) feed/gain from 1 to 21 and 22 to 42 d of age, mortality from 22 to 42 d of age and malondialdehyde(MDA) concentration in thigh muscle on d 42 than those fed the control diet. Broilers fed the diets supplemented with Se yeast had higher(P0.05) p H value and lower(P0.05) shear force in thigh muscle than those fed the diets supplemented with sodium selenite. Additionally, broilers fed the diets supplemented with 0.40 mg Se kg–1 had lower(P0.05) shear force in thigh muscle and higher(P0.05) GSH-Px activities in breast and thigh muscles than those fed the diets supplemented with 0.20 mg Se kg–1. Furthermore, broilers fed the diet supplemented with Se yeast at 0.40 mg Se kg–1 had higher(P0.05) Se concentrations in breast and thigh muscles than those fed the diet supplemented with Se yeast at 0.20 mg Se kg–1, but no differences(P0.05) were observed in these indices of broilers fed the diets supplemented with sodium selenite between 0.20 and 0.40 mg Se kg–1. The results from the present study indicated that supplemental Se could increase the growth performance, muscle Se concentration and antioxidative ability of broilers; and the Se from Se yeast was more effective than the Se from sodium selenite in improving meat quality of broilers.     

Natural killer cell signaling pathways

Natural killer (NK) cells are lymphocytes of the innate immune system that are involved in the early defenses against foreign cells, as well as autologous cells undergoing various forms of stress, such as microbial infection or tumor transformation. NK cell activation is controlled by a dynamic balance between complementary and antagonistic pathways that are initiated upon interaction with potential target cells. NK cells express an array of activating cell surface receptors that can trigger cytolytic programs, as well as cytokine or chemokine secretion. Some of these activating cell surface receptors initiate protein tyrosine kinase (PTK)-dependent pathways through noncovalent associations with transmembrane signaling adaptors that harbor intracytoplasmic ITAMs (immunoreceptor tyrosine-based activation motifs). Additional cell surface receptors that are not directly coupled to ITAMs also participate in NK cell activation. These include NKG2D, which is noncovalently associated to the DAP10 transmembrane signaling adaptor, as well as integrins and cytokine receptors. NK cells also express cell surface inhibitory receptors that antagonize activating pathways through protein tyrosine phosphatases (PTPs). These inhibitory cell surface receptors are characterized by intracytoplasmic ITIMs (immunoreceptor tyrosine-based inhibition motifs). The tyrosine-phosphorylation status of several signaling components that are substrates for both PTKs and PTPs is thus key to the propagation of the NK cell effector pathways. Understanding the integration of these multiple signals is central to the understanding and manipulation of NK cell effector signaling pathways.