PRRSV GP5类特异性肽对CSFV弱毒疫苗免疫猪骨髓中细胞因子分泌的影响

试验从猪骨髓中分离T淋巴细胞,待细胞培养达到相应时间点时,分别用PRRSV GP5类特异性肽进行刺激,继而检测细胞因子IL-10、IL-12以及IFN-γ分泌量的变化情况。结果显示:在PRRSV GP5类特异性肽刺激后,骨髓中IL-10分泌量减少,表明G1、G2肽具有减弱免疫抑制,促进抗炎性细胞因子产生的作用;此外,IL-12的分泌量也减少,提示猪的免疫保护作用减弱,表明PRRSV GP5类特异性肽可能具有双面性;而IFN-γ的分泌量变化不稳定,有待于进一步研究。     

PRRSV N类特异性肽对CSF、PRRS免疫猪肺门淋巴结细胞因子分泌的影响

为研究猪瘟(CSF)、猪繁殖与呼吸综合征(PRRS)免疫猪肺门淋巴结中淋巴细胞对PRRSV N类特异性肽刺激的应答特点,试验从猪肺门淋巴结中分离T淋巴细胞,分别用PRRSV N类特异性肽进行刺激,待细胞培养至24,48h时,离心取细胞上清液,检测上清液中细胞因子TGF-β1、IL-10、IL-12分泌含量的变化。结果显示,PRRSV N类特异性肽能够抑制TGF-β1、IL-10的分泌,表明N1、N2肽具有减弱免疫抑制,促进抗炎性细胞因子分泌的作用;此外IL-12的分泌量也减少,且有逐渐恢复平衡的趋势,提示PRRSV N类特异性肽可能具有双面性,有待于进一步研究。本试验为PRRSV合成肽疫苗及疫苗佐剂的研制提供理论依据。     

PRRSV GP5类特异性肽对CSF、PRRS免疫猪组织淋巴细胞TGF-β1分泌的影响

选择已成功免疫猪瘟(CSF)和猪繁殖与呼吸综合征(PRRS)弱毒疫苗的仔猪作为实验动物,提取其骨髓、胸腺、肺门淋巴结和脾脏,进行T淋巴细胞的分离培养,然后用PRRSV特异性肽GP5-1、GP5-2对培养的细胞进行刺激,再培养至24,48h时间点时,离心取细胞上清液,检测上清液中TGF-β1的分泌含量的变化。结果显示,PRRSV GP5类特异性肽刺激作用后,CSF、PRRS免疫猪组织淋巴细胞中TGF-β1的分泌量减少,表明GP5-1、GP5-2肽具有减弱免疫抑制,恢复宿主抗病毒免疫应答的作用。本试验为PRRSV合成肽疫苗及疫苗佐剂的研制提供了理论依据。     

PRRSV特异性肽对CSFV、PRRSV疫苗免疫猪后T淋巴细胞亚群的影响

用猪瘟疫苗和高致病性猪繁殖与呼吸综合征弱毒疫苗对猪进行免疫,在免疫后的14d和28d采集外周血液,分析特异性抗体表达量和外周血T淋巴细胞表型的变化,并用猪繁殖与呼吸综合征病毒(PRRSV)特异性肽对淋巴细胞进行刺激。结果显示,在PRRSV免疫后14d,机体PRRSV抗体水平较低,其中CSFV低抗组中CD4+细胞百分数明显降低,CD4+/CD8+细胞的比值也明显降低;用PRRSV特异性肽刺激淋巴细胞24h后,CSFV低抗组与高抗组中CD3+、CD8+细胞的百分数都明显升高,CD4+细胞的百分数及CD4+/CD8+细胞的比值都明显降低。在PRRSV免疫后28d,CSFV抗体和PRRSV抗体的产生都比较高,CD3+细胞、CD4+T淋巴细胞百分数及CD4+/CD8+细胞的比值也明显升高,用肽刺激以后,CSFV高抗组中的CD3+细胞百分数降低,CSFV低抗组中的细胞百分数升高,CD4+/CD8+细胞的比值在所有组中均下降。结果表明,PRRSV特异性肽在PRRSV免疫早期可以使CD3+细胞的百分数升高,CD4+/CD8+细胞的比值降低,其详细的机理还有待进一步的探究。     

蚯蚓肽对小鼠非特异性免疫功能的影响

通过用不同剂量的蚯蚓肽(EP)对健康和免疫抑制小鼠灌胃15 d,观察EP对小鼠淋巴细胞增殖反应,巨噬细胞细胞毒效应以及巨噬细胞和脾细胞产生NO的影响,以探讨EP对小鼠非特异性免疫功能的影响。试验发现0.1,0.5 mg/mL组明显提高淋巴细胞增殖率,增强巨噬细胞细胞毒效应,提高巨噬细胞和脾细胞分泌NO的水平(P<0.0l),0.5 mg/mL明显提高免疫抑制小鼠的免疫功能(P<0.01)。结果表明:一定剂量下,EP具有调节免疫功能、拮抗环磷酰胺引起的免疫抑制的作用。     

刺五加多糖对雏鸡脾脏中T、B淋巴细胞定位分布的影响

为了研究雏鸡注射不同浓度刺五加多糖(ASPS)后脾脏中T、B淋巴细胞定位分布的变化情况,将60只1日龄雏鸡随机分为3组:对照组、低剂量组和高剂量组,每组20只,10日龄起对低、高剂量组雏鸡分别皮下注射免疫50和200 mg/m L的ASPS 0.2 m L,对照组雏鸡以相同方式注射等体积的生理盐水,连续注射4 d。于末次免疫后第7天和第14天,从每组中分别随机选取5只雏鸡,取其脾脏制作冰冻切片,采用免疫组织化学方法检测CD3+T淋巴细胞和Bu-1+B淋巴细胞的定位分布。结果显示,与对照组比较,低、高剂量组在注射后的7 d和14 d ASPS均能增加脾脏中CD3+T淋巴细胞和Bu-1+B淋巴细胞的数量,由CD3+T淋巴细胞构成的脾脏特征性结构-动脉周围淋巴鞘(PALS)的面积也显著增加,红髓中CD3+T淋巴细胞也明显增多,分布范围逐渐增加;同时,由Bu-1+B淋巴细胞构成的椭球周围淋巴鞘(EALS)的面积也明显增加,红髓和白髓中的浆细胞数量显著增多,并在注射后14 d时出现生发中心。与对照组比较,高剂量组和注射后14 d时上述变化趋势更为明显。结果表明,ASPS能够增加脾脏中T、B淋巴细胞的数量,并且能够影响其在PALS、EALS等脾脏特征性结构中的定位分布,从而从组织学角度进一步证明ASPS对鸡免疫功能具有显著的增强作用。     

PRRSV特异性肽-N对CSFV弱毒疫苗免疫猪骨髓细胞因子分泌的影响

本试验探讨PRRSV特异性肽-N对CSFV弱毒疫苗免疫猪骨髓细胞因子分泌量的影响。选择已免疫CSFV弱毒疫苗的猪为实验动物,提取四肢长骨骨髓,进行T淋巴细胞的分离培养。分别在培养后的24、72 h,用PRRSV特异性肽N1、N2对所培养细胞进行刺激,再进行培养12 h,经离心方法得到细胞上清液,最终检测上清液中细胞因子IFN-γ、IL-12、IL-10的分泌含量变化。结果显示,PRRSV特异性肽-N对于免疫猪骨髓中IFN-γ的分泌影响较小,能够抑制IL-12的分泌,不利于机体免疫反应,对IL-10的分泌影响复杂,需进一步研究。     

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。     

交感神经对小鼠脾脏中淋巴细胞转化率的影响

神经、内分泌、免疫诸系统间存在着相互作用,这是近30年来新兴的神经免疫学中主要的研究成果。交感神经及其递质去甲肾上腺素(NE)在体内和体外条件下对免疫细胞功能的调节作用是神经免疫学中研究最早、报道最多的内容之一。脾脏中T淋巴细胞和B淋巴细胞转化率分别代表了机体的细胞免疫和体液免疫,但是,对于外周交感神经损毁后,脾脏中淋巴细胞转化率的影响还未见报道。因此,本研究利用6-OHDA损毁小鼠交感神经模型,检测了脾脏中不同浓度ConA诱导的T淋巴细胞转化率,不同浓度LPS诱导的B淋巴细胞转化率,以探讨交感神经对机体免疫状态的影响…     

Nested PCR for diagnosis of canine leishmaniosis in peripheral blood, lymph node and bone marrow aspirates

A nested polymerase chain reaction (PCR) assay was developed using primers selected from the genomic DNA of Leishmania infantum and applied to the diagnosis of leishmaniosis in peripheral blood in dogs. Blood of 39 dogs of different breeds, all sampled in Catalonia (Spain), were tested for leishmaniosis by enzyme linked immunosorbent assay (ELISA), western blotting (WB) and peripheral blood mononuclear cell (PBMC) culture and nested PCR. Twenty negative controls (healthy dogs less than 1-year-old that had not been exposed to a sandfly season) were also studied. Nineteen of the 39 dogs studied were positive by ELISA and/or WB, and 18 of these had a positive PBMC nested PCR. PBMC nested PCR was negative in all the remaining animals that were negative by serological examination, including the 20 negative controls. Parasitological examination and nested PCR of bone marrow and lymph node aspirate from the 19 dogs positive by serological examination, were also positive. These results indicate that PBMC nested PCR is a sensitive and specific tool to diagnose leishmaniosis in dogs. The use of PBMC has the advantage over bone marrow and lymph node aspirates in that it is a less invasive sample.     

PRRSV感染对猪外周血来源的树突状细胞免疫表型及Th1/Th2偏转潜能的影响

为了研究PRRSV感染对猪外周血来源的树突状细胞成熟及Thl及Th2型细胞因子分泌水平的影响,进一步探讨PRRSV调控树状细胞Th1和Th2型免疫应答的潜在机制,揭示PRRSV感染对机体免疫机能的影响。我们从猪外周血分离单核细胞,诱导培养为树突状细胞后接种PRRSV共培养,显微镜观察细胞的形态,流式细胞术分析细胞表面的共刺激分子,半定量RT-PCR法检测Thl型细胞因子IL-12和Th2型细胞因子IL-10的mRNA转录水平。结果显示:经鉴定证实获得形态及表型典型的树突状细胞;流式分析结果表明PRRSV接种组树突状细胞表面的SLA-II-DR类分子、CD40分子、CD80分子均明显低于未接种组,差异极显著(P0.01);RT-PCR半定量结果表明PRRSV接种组的IL-12mRNA转录水平低于未接种组,差异显著(P0.05),而IL-10mRNA转录水平高于未接种组,差异显著(P0.05)。结果表明:PRRSV感染影响树突状细胞的成熟,并能通过高效表达Th2型细胞因子IL-10降低Th2型细胞因子Th1而影响Thl/Th2的平衡。     

Effects of different us isolates of porcine reproductive and respiratory syndrome virus (PRRSV) on blood and bone marrow parameters of experimentally infected pigs

Seventy five-week-old, crossbred, caesarean-derived, colostrum-deprived pigs were randomly divided into five groups of 14 pigs and assigned one of five treatments: the intranasal inoculation of 1 (5.7) TCID50 of one of four plaque-purified isolates of porcine reproductive and respiratory syndrome virus (PRRSV) (VR2385, VR2431, ISU-984 and ISU-22), or uninfected cell culture and media. Haematological variables were measured for 21 days and bone marrow was analysed when the pigs were killed three, seven, 10, 21 or 28 days after the inoculation. The PRRSV-infected pigs had non-regenerative anaemia and markedly increased myeloid:erythroid ratios from three to 21 days after inoculation. There was a significant (P < 0.05) difference in the severity of the anaemia induced by the four PRRSV isolates; the most highly pneumovirulent strains (VR2385, ISU-984 and ISU-22) induced more severe anaemia than the least virulent isolate (VR2431). The anaemia induced by PRRSV was probably due to a direct or indirect effect on erythroid precursor cells in the bone marrow.     

Comparative studies on the distribution and population of immunocompetent cells in bovine hemal node, lymph node and spleen

The distribution and population of immunocompetent cells in bovine hemal node, mesenteric lymph node and spleen were analyzed comparatively by immunohistochemistry and flow cytometry. Many CD8(+) cells, CD172a(+) cells and γδ T cells were found in the lymphatic cord along the sinus of the hemal node and the splenic red pulp. A few CD8(+) cells and γδ T cells were distributed diffusely in the paracortex and medullary cord of the mesenteric lymph node. Many germinal centers were recognized in the lymphatic regions such as the cortex and white pulp of these lymphoid organs. The populations of CD8(+) cells and γδ T cells in the hemal node and the spleen were higher than those of the mesenteric lymph node. In addition, the populations of CD21(+) cells and MHC class II(+) cells in the hemal node and the mesenteric lymph node were higher than those of the spleen. The results suggest that the hemal node has an important role in both cellular and humoral immunity as well as the lymph node and the spleen in cattle.     

Monoclonal antibodies as probes for defining cellular subsets in the bone marrow, thymus, bursa of fabricius, and spleen of the chicken

Using immunohistochemistry, the distribution and characteristics of cells detected by the newly developed monoclonals HIS-CI (B lymphocytes), HIS-C7 (leucocytes), HIS-C12 (IgM), CVI-ChIgM-59.7 (IgM), CVI-ChIgG-47.3 (IgG), and CVI-ChIgA-46.5 (IgA) are described in bone marrow, thymus, bursa of Fabricius, and spleen of chickens of different ages. Furthermore, quantification of cells positive with the described monoclonal antibodies was performed on cytocentrifuge preparations. The specificities of the monoclonal antibodies are discussed.     

Determination of the number of mast cells in lymph node, bone marrow, and buffy coat cytologic specimens from dogs.

Cytologic samples of popliteal lymph node, proximal femoral bone marrow, and the buffy coat fraction of blood were obtained from 56 dogs. The number of mast cells on 1 slide of each sample was determined by microscopic examination. Eleven of 46 slides of lymph node aspirate contained mast cells (range, 1 to 16; mean, 6.4; median, 5 mast cells/slide). Fifty-one bone marrow aspirate slides were evaluated. Two of these contained a single mast cell. None of the 53 buffy coat smear slides examined contained any mast cells. These results indicated that in clinically normal dogs, a few to several mast cells may be encountered in smears of lymph node aspirate, mast cells are rare in smears of bone marrow aspirate, and mast cells are absent from smears of buffy coat.