抗抗体

To characterize the isotype of human xenoreactive nat ural antibodies against porcine pancrease islet cells, cyostat sections of the p ancrease were incubated with the sera from 18 cases of IDDM or 20 human controls respectively, further with FITC-conjugated goat antihuman IgG or IgM secondary antibodies. IgG was found to bind to pancrease islet cells in sera from 8 cases of IDDM and 8 human controls, while IgM in 4 IDDM and 6 human controls.

为检测人抗猪胰岛细胞天然抗体的同种型，采取18份胰岛素依赖型糖尿病患者血清及20份正常人血清分别与杂种猪胰腺组织的冰冻切片孵化后，再用免疫荧光标记的羊抗人IgG、IgM单克隆抗体染色。8份IDDM血清和8份正常人血清中含有直接针对猪胰岛细胞的IgG型抗体；4份IDDM血清和6份正常人血清中含有IgM 型抗猪胰岛细胞抗体。

We optimized reaction system for labelling-antibody in which the optimal amount of the purified anti-Stx2B IgG conjugated with colloidal gold beads was 60μg/mL, the purified antibody for E.coli O157 was 57μg/mL, the optimal pH was 8.2, the size of colloidal gold partical was 20nm, the optimization of stabilizing agent was BSA, the optimization buffer was boracic acid buffer with pH 8.2, the optimization of preserving fluid and eluant was boracic acid buffer with pH 8.2 including 5mM NaCl-1%BSA, confining liquid for NC membrane was 0.01% PBST with 3%BSA, the amount of polyclonal antibody against E.coli O157 and Stx2 conjugated with colloidal gold beads for conjugate pad was 3μg respectively, the amount of anti-Stx monoclonal antibody for test line was 0.1μg and 1μg for E.coli O157, the amount of goat anti rabbit IgG for control line of both GICA were 1μg.

测定了胶体金颗粒的最优化反应体系：胶体金颗粒的大小为20nm；抗体与胶体金溶液结合的最佳pH约为8.2；最佳蛋白结合量分别为抗大肠杆菌O157 IgG为57μg/mL，抗重组Stx2B IgG为60μg/mL；最佳稳定剂为BSA；最佳缓冲液为pH8.2硼酸溶液；最佳金标保存液和洗涤液为5mM NaCl-1%BSA的pH 8.2的硼酸缓冲液；NC膜的封闭液为3%BSA的0.01mol/L PBST；Stx2试纸条和大肠杆菌O157试纸条的质控线上的羊抗兔IgG的多克隆抗体最佳点样量均为1μg，其检测线的抗Stx2的单克隆抗体的点样量为0.1μg、抗大肠杆菌O157的单克隆抗体的点样量为1μg，结合垫上的金标抗大肠杆菌O157和重组Stx2B多克隆抗体点样量均为3μg。

Methods VEGF in the serum of 60 patients with RA and 28 healthy control subjects was quantified by our highly sensitive enzyme-linked immunoad sorbent assay, and its relationship to clinical and laboratory variables was analyzed . Synovial specimens were obtained from 1 RA and 1 OA patients undergoing synovectomy.

体内实验：用酶联免疫吸附方法检测了60例RA和28例健康对照血清中的VEGF值，并检测其他活动性指标及血清中特异性自身抗体：抗核周因子抗体、抗角蛋白抗体、抗环瓜氨酸肽抗体，分析它们之间的关系。

And the different concentrations of natural monoclonal anti-keratin antibody IgM 3B4 were incubated with the mixed suspenions of Candida albicans yeast phase with malpighian cells, human buccal epithelial cells, endothelial cells of fetal umbilical vein, respectively, to observe the action of natural monoclonal anti-keratin antibody IgM 3B4 to the adhesion of Candida albicans to malpighian cells, human buccal epithelial cells and endothelial cells of fetal umbilical vein.

结果：与对照组比较，不同浓度的单克隆天然抗角蛋白抗体IgM3B4均能抑制白念珠菌芽管生成，并且抗体浓度越高，抑制白念珠菌芽管形成的作用越明显；同时单克隆天然抗角蛋白抗体IgM3B4能够显著减少白念珠菌与角质形成细胞、上皮细胞、内皮细胞的粘附，其抑制作用与单克隆天然抗角蛋白抗体IgM3B4的浓度呈正相关。

The blots were probed with anti-Cdx-2, anti-JNK (Santa Cruz Biotechnology, Santa Cruz, California), anti-ERK, anti-phospho -ERK, antip38MAPK,anti-p-p38MAPK, anti-p-JNK (Cell Signaling, Beverly, Massachusetts) and anti-actin (Amersham Life Sciences,Piscataway, New Jersey) antibodies, and then incubated with appropriate species specific secondary antibodies labeled with horseradish peroxidase.

Blots分别与抗CDX-2抗体，抗JNK抗体（圣克鲁斯生物技术，圣克鲁斯，加州），抗ERK抗体，抗phospho -ERK抗体，抗p38MAPK抗体，抗p-p38MAPK抗体，抗p-JNK抗体（Cell Signaling， Beverly，美国马萨诸塞州）和抗actin抗体（ Amersham生命科学，皮斯卡塔韦，新泽西州）作用，然后在与辣根过氧化物酶标记的第二抗体孵育。

Results The level of antibodies against monocytes (34.94%) were higher than that of the antibodies against lymphocytes (26.67%) in pretransplant patients. The level of antibodies in first time transplant patients (32.5%) was lower than that of antibodies in retransplant patients (59.7%). The patients with intensity of antibodies against monocytes PRA more than ≥50% only accounted for 1.87%, 10%＜ MPRA ＜50% for 17.62% and MPRA ＜10% 15.44%,respectively.

结果 肾移植术前患者的抗单核细胞抗体水平较抗淋巴细胞抗体水平高，其百分比分别为34.94%和26.67%；而从患者初次和再次肾移植前的比较中可见，初次比再次产生的抗单核细胞抗体水平要低，其百分比分别为32.05%和59.7%；对于抗单核细胞抗体群体反应性抗体的强度，MPRA≥50%的只占1.87%，而50%＜MPRA＞10%以及MPRA＜10%的分别占17.62%和15.44%。

Results The level of antibodies against monocytes (34.94%) were higher than that of the antibodies against lymphocytes (26.67%) in pretransplant patients. The level of antibodies in first time transplant patients (32.5%) was lower than that of antibodies in retransplant patients (59.7%). The patients with intensity of antibodies against monocytes PRA more than ≥50%only accounted for 1.87%, 10%MPRA50%for 17.62%and MPRA10%15.44%,respectively.Conclusion Before renal transplantation, the detection of MPRA level could predict the occurrence of rejection in renal transplantation.

结果 肾移植术前患者的抗单核细胞抗体水平较抗淋巴细胞抗体水平高，其百分比分别为34.94%和26.67%；而从患者初次和再次肾移植前的比较中可见，初次比再次产生的抗单核细胞抗体水平要低，其百分比分别为32.05%和59.7%；对于抗单核细胞抗体群体反应性抗体的强度，MPRA≥50%的只占1.87%，而50%10%以及MPRA结论在肾移植术前检测患者MPRA水平，对肾移植后的排斥反应能起到预测目的。

To inestigate sensitisation against food antigen, mice were intragastrically administered with oalbumin eery other day for nine weeks, and antioalbumin antibody titres were measured weekly.

为了研究对食物抗原的致敏反应，小鼠予胃内灌注卵白蛋白，隔天1次，共9周，每周测试卵白蛋白抗抗体滴度。

PART Ⅱ IMMUNOPATHOLOGICAL EVIDANCE OF SIALIC ACID STRUCTURE IN CAMPYLOBACTER JEJUNI AS THE CRITICAL ANTIGEN TO INDUCE PERIPHERAL NEUROPATHY To demonstrate the critical role of NANA structure in the pathogenesis of allergic peripheral neuropathy induced by Campylobacter jejuni and provide immunopathological evidence to confirm the supposition of molecular mimicry and cross-immunity between CJ LPS and gangliosides in nerve.

LPS免疫后第2周实验豚鼠的免疫血清中，抗LPS IgG抗体滴度均明显增高，第3周达高峰，第5周仍维持在峰水平，野生株和突变株LPS免疫血清中的抗-野生株LPS IgG抗体滴度较各自免疫前分别增高8倍和4倍，抗-突变株LPS IgG抗体滴度则较各自免疫前分别增高6.5倍和15倍；(2)全身免疫后第3周和第5周，野生株LPS免疫血清中检测到较免疫前增高6倍的抗-GM1 IgG抗体，而NANA缺失的突变株LPS免疫血清中一直未检测到抗-GM1 IgG抗体；(3)野生株LPS免疫组中有17.3％的坐骨神经原纤维发生以轴索变性为主（占65％）的免疫性损伤，与突变株LPS免疫组和对照组比较均有显著性差异。

Pneumophila serogroup 5 antibody to develop two immuo-colloidal gold tests respectively. Main works listed below as:1、Preparation and identification of polysaccharide antigen of L. pneumophila Bacteria of LP1 ~ 7,9 and 10 were cultured on yeast agar buffer activated carbon for three to five days at 37℃, 5%CO2. Protein-free polysaccharide antigens were obtained after harvest in cells, extraction, deproteinization, dialysis and other steps. Their immunogenicities were verified by ultraviolet spectrophotometer full-wavelength scanning and Western blotting.2、Preparation and identification of rabbit anti-LP1 antibodies and rabbit anti-LP5 antibody Rabbit anti-LP1 and anti-LP5 antibodies were purified after rabbits were immuned with antigens isolated as described above. The purities of both antibodies were above 80% and the titer of blood serum 1:32 tested by double antibody sandwich assay.3、Development of colloidal gold immunochromatographic assay kit The size of colloidal gold particles in the kit was 25nm. The optimal concentrations for antibodies were 30μg/ml and the sensitized concentrations of NC membrane were 5 mg/ml.

主要研究工作从以下几个方面进行：1、LP1~7、9和10型多糖抗原的制备与鉴定将LP1~7、9和10型菌株分别接种在缓冲活性炭酵母琼脂培养基上，37℃、5%CO2的条件下培养，3~5天后洗下菌苔，经抽提、除蛋白、透析等步骤后得到基本无蛋白的LP多糖抗原，经紫外分光光度仪全波长扫描及Western blotting验证其抗原良好。2、兔抗LP1抗体和兔抗LP5抗体的制备与鉴定分别用LP1、LP5型多糖抗原免疫家兔，采用琼脂糖双向扩散试验检测，两种抗体血清效价均为1：32；饱和硫酸铵法提取抗体，SDS-PAGE检定其抗体纯度均达到80%以上。3、胶体金免疫层析检测试剂的初步研制采用柠檬酸三钠还原法制备约25nm大小的胶体金颗粒；分别制备兔抗LP1抗体、兔抗LP5抗体的金标探针，两种抗体的最适标记量均为30μg/ml；选择适当孔径的微孔滤膜为载体包被两种抗体，NC膜包被浓度均为5mg/ml。

This one mode pays close attention to network credence foundation of the businessman very much.

这一模式非常关注商人的网络信用基础。

Cell morphology of bacterial ghost of Pasteurella multocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysi.

扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率。