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。     

鲫、草鱼外周血中淋巴细胞和单核细胞对鲤血管内皮细胞的粘附

借鉴哺乳动物内皮细胞体外培养技术，结合鱼类细胞自身特点，纯化培养鲤血管内皮细胞并传至3代，以此作为粘附材料。应用淋巴细胞分离液离心技术并结合玻璃粘附法分离纯化鲫、草鱼外周血中淋巴细胞、单核细胞，并与上述3代内皮细胞进行免疫粘附试验，同时进行免疫粘附动力学观察。结果显示，鲫、草鱼两类细胞对内皮细胞的粘附率分别为0.09±0.013，0.20±0.018；0.11±0.015，0.21±0.023。表明鲤科鱼类不同种的同类细胞粘附率差异不大，而同种不同类细胞则差异显著。动力学观察分析表明，淋巴细胞粘附较快，60min进入平台期；单核细胞粘附慢，120min进入平台期。初步证明鱼类内皮细胞具有免疫介导作用。     

Response to soy: T-cell-like reactivity in the intestine of Atlantic salmon, Salmo salar L

T-cell-mediated hypersensitivity could be central in soybean meal (SBM)-induced intestinal changes in salmon. However, tools for immunohistochemical detection of T cells have been lacking in teleosts, including Atlantic salmon. Application of a specific histochemical protocol allowed demonstration of T-cell-like reactivities in formalin-fixed, paraffin-embedded tissues using an antibody reacting to a conserved region of human CD3epsilon (Dako A0452). Characteristic staining was observed in cells of the thymus as well as distal intestine, skin, gills and spleen. These cells were negative for immunoglobulin M (IgM). Intestinal intraepithelial leucocytes were CD3epsilon positive. During the SBM-induced enteropathy, the mixed inflammatory infiltrate in the lamina propria of the distal intestine included many lymphocytes with a T-cell-like reactivity. Real-time polymerase chain reaction revealed significantly increased expression of a complex polypeptide (CD3pp), CD4 and CD8beta (P < 0.05) in the distal intestine of SBM-fed fish compared to fish meal-fed reference fish. Increased reactivity for extracellular IgM in the lamina propria and a positive material between the epithelial cells at the tips of the folds was observed, possibly due to leakage of IgM through an abrogated epithelial barrier. In conclusion, a T-cell-like response appears to be involved in this example of a food-sensitive enteropathy.     

The effect of feeding the leucine metabolite β-hydroxy-β-methylbutyrate (HMB) on cell-mediated immunity and protection against Yersinia ruckeri in pikeperch (Sander lucioperca)

The present study examined the influence of leucine metabolite β‐hydroxy‐β‐methylbutyrate (HMB), a natural product HMB, on nonspecific cell‐mediated defence mechanisms and protection against enteric redmouth disease (ERM) in pikeperch (Sander lucioperca). β‐hydroxy‐β‐methylbutyrate was fed in a pelletted ration at 50 mg kg^?1 of the feed for 8 weeks. The phagocytic ability and potential killing activity of blood and pronephric phagocytes were examined in HMB‐ and control‐fed fish before and after 8 weeks of feeding HMB. Simultaneously, the proliferative response of blood and pronephric lymphocytes stimulated by mitogens concanavaline A and lipopolisaccharide were examined in experimental and control groups. Following 8 weeks of HMB feeding, a challenge test was performed by injecting the fish with live pathogenic bacteria Yersinia ruckeri. β‐hydroxy‐β‐methylbutyrate applied in the diet for 8 weeks prompted a statistically significant (P<0.05) increase in phagocytic ability and potential killing activity of the blood and pronephric phagocytes and the proliferative response of blood and pronephric lymphocytes. The changes in these mechanisms correlated with protection against Y. ruckeri, the ERM pathogen. The results showed that feeding HMB increased the nonspecific cell‐mediated defence mechanisms and protection against ERM by reducing cumulative mortality (30%) following the challenge with pathogenic bacteria. Future studies will include determination of optimal doses and protocols of oral application of HMB to maximize the immunomodulatory effects and protection against viral diseases in intensive pikeperch culture.     

Effect of Different Mulberry Leaf Extracts on Spleen Lymphocyte Proliferation in Mice

This experiment was aimed to compare the effects of different mulberry leaf extract on spleen lymphocyte proliferation in mice. Mulberry leaf polysaccharides (MLP-1, MLP-2, MLP-3, MLP-4 and MLP-5) were prepared with mulberry leaf aqueous extract (MLAE) by different concentration of alcohol deposit, polysaccharides concentration of all extracts were determined by phenol sulfuric acid method. Five different MLPs and MLAE were as the experimental drug, when polysaccharide concentration was 15.625, 31.25, 62.5, 125 and 250 μg/mL, the changes of spleen B and T lymphocytes proliferation in mice which were stimulated by MLPs and MLAE in single or synergistical stimulation of drugs with LPS and PHA were determined by MTT method. The results showed that when polysaccharide concentration was in the range of 15.625 to 250 μg/mL, MLPs and MLAE in single or synergistical stimulation with LPS and PHA could stimulate the spleen B and T lymphocytes proliferation, MLP-1 at low concentration could stimulate significantly spleen B lymphocyte proliferation remarkably (P<0.05), and MLP-3 and MLP-5 at low concentrations showed strong stimulation ability of T lymphocyte proliferation. When MLAE was in synergistical stimulation with LPS at most concentrations points, the spleen B lymphocytes proliferation in mice were significantly higher than those of the cell control and LPS control groups, which revealed that MLAE could significantly improve the humoral immune function (P<0.05).     

酵母壁多糖对断奶仔猪外周血免疫和肠道免疫的影响

本试验旨在探讨饲粮中添加酵母壁多糖对断奶仔猪外周血免疫和肠道免疫的影响。采用单因素试验设计方法,选取21日龄遗传胎次、体重接近的断奶仔猪180头,随机分为4个组,每组5个重复,每个重复9头猪。4组分别饲喂饲粮中添加0(对照组)、0.15%、0.30%和0.45%酵母壁多糖的试验饲粮。试验期21 d。结果表明:1)与对照组相比,饲粮中添加0.15%、0.30%和0.45%酵母壁多糖显著提高了断奶仔猪血清免疫球蛋白A(IgA)含量(P0.05),饲粮中添加0.30%和0.45%酵母壁多糖显著提高了断奶仔猪血清免疫球蛋白G(IgG)含量(P0.05)。2)与对照组相比,饲粮中添加0.15%、0.30%和0.45%酵母壁多糖显著降低了断奶仔猪血清干扰素-γ(IFN-γ)含量(P0.05),饲粮中添加0.30%酵母壁多糖显著提高了断奶仔猪血清白细胞介素-10(IL-10)的含量(P0.05)。饲粮中添加酵母壁多糖对断奶仔猪血清白细胞介素-2(IL-2)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)含量均无显著影响(P0.05)。3)与对照组相比,饲粮中添加0.30%和0.45%酵母壁多糖显著提高了断奶仔猪回肠CD4~+淋巴细胞含量(P0.05),饲粮中添加酵母壁多糖能一定程度提高断奶仔猪回肠CD8~+和CD20~+淋巴细胞含量,但差异不显著(P0.05)。由此可知,酵母壁多糖能一定程度提高断奶仔猪外周血免疫和肠道免疫,缓解断奶应激。     

树舌荧光多糖的制备及在小鼠脾淋巴细胞中的定位

为确定树舌多糖在小鼠脾淋巴细胞上的定位,应用胺化还原法在树舌多糖(GAP)的还原性末端连接异硫氰酸荧光素(FITC),用荧光分光光度计测定其荧光取代度,xCELLigence RTCA DP全自动实时细胞分析仪检测GAP荧光标记前后生物学稳定性.流式细胞仪检测小鼠脾T淋巴细胞、B淋巴细胞和NK细胞中的GAP荧光强度,荧光显微镜观察GAP在小鼠脾淋巴细胞中的定位.结果显示:在荧光标记GAP (FITC-GAP)前后的细胞毒活性不变,并且其荧光取代率为0.90％;流式细胞仪检测发现小鼠脾T淋巴细胞、B淋巴细胞和NK细胞上的荧光信号强度与FITC-GAP加入量成正相关,荧光显微照片显示GAP定位于小鼠脾淋巴细胞表面并可转运至细胞核.     

银耳菌丝体多糖对小鼠免疫功能的影响

本文研究了银耳菌丝体多糖对小鼠免疫功能的影响。实验结果表明:每天灌胃0.5或1.0g/kg TMP,连续20天,不仅可明显地促进小鼠特异性抗体的形成,亦可明显地促进小鼠腹腔巨噬细胞的吞噬功能,增加外周血T淋巴细胞数量,延缓胸腺萎缩,并可对抗由CY所致的细胞免疫和体液免疫低下的作用。     

滑鼠蛇出生至50日龄身体大小和免疫功能的变化

为探究滑鼠蛇（Ptyas mucosus）生后发育不同阶段身体大小和免疫功能的变化,在室温为（30±1）℃、相对湿度为70%~75%条件下,监测了3、10、20、30、40和50日龄滑鼠蛇身体大小及各型白细胞的百分比、嗜中性粒细胞与淋巴细胞的比值（N/L）和血清杀菌能力的变化。体重、体全长、重长比和尺度化的质量指数3至20日龄增长缓慢,之后陡增,50日龄最高（P<0.05）;淋巴细胞和嗜中性粒细胞的百分比居前两位,但两者的百分比、N/L的比值和血清杀菌能力均无组间差异（P>0.05）;嗜碱性粒细胞的百分比10日龄组与3日龄组接近,但高于其他日龄组（P<0.05）;50日龄内,滑鼠蛇的身体大小开口前增长缓慢,开口后增加迅速,蜕皮后嗜碱性粒细胞的百分比显著提升,建议可根据体重和嗜碱性粒细胞百分比的变化来预判蛇场内幼蛇的健康状态。