Immunopathology of vesicular cutaneous lupus erythematosus in the rough collie and Shetland sheepdog: a canine homologue of subacute cutaneous lupus erythematosus in humans

Clinical and histological features of an erosive disease in the rough collie and Shetland sheepdog are most consistent with a vesicular variant of cutaneous lupus erythematosus (VCLE). This paper reports the immunopathological findings of canine VCLE using samples from 17 affected dogs. Lesional skin sections were stained with monoclonal antibodies specific for CD3 (11 dogs) or a panel of monoclonal antibodies specific for leukocyte antigens (two dogs). Apoptotic cells were detected using the TUNEL method in 12 cases. Direct (14 dogs) and indirect immunofluorescence tests (five dogs) were also performed. Circulating antibodies to extractable nuclear antigens (ENA) were surveyed in 11 dogs by immunoblotting and ELISA. The predominant cells at the dermal-epidermal interface were identified as CD3(+) T lymphocytes expressing CD4 or CD8 and CD1(+) dendritic antigen presenting cells. In 7/12 dogs (58%), apoptosis of basal keratinocyte nuclei was present. Up-regulation of MHCII and ICAM-1 was observed on basal keratinocytes from the two dogs examined. Direct immunofluorescence revealed deposition of immunoglobulins bound to the cytoplasm of keratinocytes (6/14 dogs; 43%), to the dermal-epidermal junction (7/14 dogs; 50%), or to superficial dermal venules (13/14 dogs; 93%). Circulating IgG auto-antibodies targeting one or more ENA were detected in nine (82%) and eight (73%) of 11 dogs by immunoblotting and ELISA, respectively. These auto-antibodies recognized Ro/SSA and/or La/SSB in four (36%) and six (55%) of 11 dogs respectively by these two methods. Altogether, results of these studies provide evidence supporting the hypothesis that canine VCLE is an immunological homologue of subacute cutaneous lupus erythematosus in humans.     

Seroprevalence of neutralizing antibodies to human adenovirus type 5, human adenovirus type 26 and chimpanzee adenovirus type 68 in patients with chronic hepatitis B and patients with primary liver cancer

AIM: To investigate the seroprevalence of neutralizing antibodies to human adenovirus type 5 (AdHu5), human adenovirus type 26 (AdHu26) and chimpanzee adenovirus type 68 (AdC68) in the patients with chronic hepatitis B (CHB) and the patients with primary liver cancer (PLC), and to provide guidance for developing safe and effective biotherapy vectors against CHB and PLC. METHODS: The blood samples from 196 patients with CHB and 193 patients with PLC were examined to assess the presence of neutralizing antibodies against AdHu5, AdHu26 and AdC68 by adenovirus neutralization assays. RESULTS: The seroprevalence rates of neutralizing antibodies to AdHu5, AdHu26 and AdC68 in the CHB patients were 84.7%, 58.2% and 39.8%, respectively. Among the patients with PLC, the prevalence rates of neutralizing antibodies were as follows: AdHu5, 75.1%; AdHu26, 66.8%; AdC68, 32.1%. CONCLUSION: The prevalence rates and titers of neutralizing antibodies against AdC68 were the lowest among the 3 adenoviruses. Therefore, AdC68 serves as more suitable biological therapy vectors for CHB and PLC than AdHu5 and AdHu26.     

抗猪大肠杆菌功能性卵黄粉的制备及对小鼠的攻毒保护作用

通过制备抗致病性大肠杆菌特异性IgY及富含特异性IgY的功能性卵黄粉,进行小鼠的攻毒保护试验,探讨功能性卵黄粉对由致病性大肠杆菌引起的疾病的防治作用.结果显示:(1)致病性大肠杆菌与特异性IgY混合后,菌体出现凝集,且病原菌的运动力明显受抑制;(2)浓度为1.3×109CFU·mL-1的致病性大肠杆菌K88、K99和9...     

传染性法氏囊卵黄抗体不同提取工艺的比较

本文对硫酸酯葡聚糖法、PEG6000与水稀释法三种不同方法进行比较,通过记录分层的时间、上清液的体积、清澈程度及测定抗体效价、蛋白质含量等,选择去脂效果较好,效价和蛋白质含量较高的方法作为提取传染性法氏囊卵黄免疫球蛋白的最佳途径。     

猪瘟母源抗体衰减及保护水平分析

为分析仔猪体内猪瘟母源抗体的衰减规律及母源抗体保护水平,测定了断奶时母猪和仔猪抗体水平,并监测仔猪母源抗体水平至81日龄;分别对12头81日龄仔猪（来自5头母猪）和25头35日龄仔猪（来自5头母猪）进行了攻毒试验,并用RT—nPCR进行了跟踪检测,对7头攻毒耐过猪用兔体中和法测定了中和效价。结果表明,母猪抗体水平不均衡会导致仔猪群体母源抗体水平不均衡;仔猪群体抗体到60日龄时仍然有67％（12／18）的个体用ELISA检测阻断率在50％以上：81日龄攻毒组有3头（3／12）耐过,35日龄攻毒组能全部耐过（25／25）,但有2头（2／25）在攻毒27d后仍然呈PCR阳性,此时中和抗体水平从低于1：4到1：16不等,推测中和抗体水平1：8～1：16是母源抗体保护的临界线。     

猪SIgA分泌片的分离纯化及其多克隆抗体的制备

为制备抗分泌片多克隆抗体,采用分子筛凝胶层析、离子交换层析和硫酸铵盐析法从猪初乳中分离纯化分泌片和SIgA。SDS—PAGE鉴定表明,纯化的SIgA呈现分泌片、重链和轻链三条带,大小约为70ku;凝胶薄层扫描分析SIgA和分泌片纯度分别为90％和89％;琼脂扩散鉴定表明,SIgA和分泌片与抗分泌片单克隆抗体发生了特异反应。以纯化分泌片为抗原,制备抗分泌片多克隆抗体,琼脂扩散检测多抗的效价为1：32。高纯度分泌片的提取及高效价抗分泌片多抗的制备为粘膜负．疫研究奠定了物质基础。     

一种具有对映选择性催化萘普生乙酯水解的多克隆抗体

研究了一种具有对映选择性催化萘普生乙酯水解的多克隆抗体。先设计并合成消旋膦酸酯Ｐ１，与载体蛋白偶联得到适于动物免疫的抗原体系Ｐ２和Ｐ３，通过适当的分离和离子交换色谱处理，得到一种新的免疫球蛋白（ＩｇＧ）－多克隆抗体。它能加速Ｒ－构型萘普生乙酯水解成为Ｓ－构型，且具有高度专一性，此多克隆抗体为超分子识别。     

Maternal transfer of antibodies specific for avian influenza viruses in captive whooper swans (Cygnus cygnus)

The transfer of maternal antibodies to offspring can effectively protect against avian influenza virus (AIV) infection during early life in chickens and can prevent AIV spread by decreasing the overall percentage of the avian population susceptible to this pathogen. Herein, we evaluated maternal antibody transfer dynamics in whooper swans (Cygnus cygnus), which represent an important AIV host species. In total, 57 eggs from 19 nests were collected to study the relationship between egg yolk AIV-specific antibody concentrations and factors including egg size, laying order, maternal serum AIV antibody titer, and maternal body condition. Overall, we found that AIV-specific antibodies were present in the serum of 63.2 % of surveyed female swans and were transferred to 50.8 % of analyzed eggs. We found maternal AIV-specific antibody concentration and body weight to be positively correlated with egg yolk AIV antibody concentration, whereas egg laying order was negatively correlated with yolk antibody titer. Overall, these findings maternal transfer of AIV-specific antibodies may function as a key mechanism governing the dynamics of AIV infection in swan populations.     

H9N2亚型禽流感病毒中和单克隆抗体的制备与应用

为建立H9N2亚型禽流感病毒（Avian influenza virus,AIV）免疫层析快速检测技术,本研究以差速离心法纯化H9N2亚型AIV免疫BALB/c小鼠,将免疫小鼠脾细胞与骨髓瘤细胞SP2/0进行细胞融合和HAT选择性培养;以H9N2亚型AIV感染MDCK细胞建立异源免疫过氧化物酶单层细胞试验（IPMA）的单克隆抗体检测方法,通过对杂交瘤细胞的IPMA筛选和连续克隆化筛选鉴定抗H9N2亚型AIV中和性单克隆抗体;以胶体金标记HA单克隆抗体,配对HA单克隆抗体和羊抗小鼠IgG为检测线和质控线,制备H9N2亚型AIV快速检测试纸条,测定其特异性和敏感性。结果显示,获得了11株稳定分泌抗H9N2亚型AIV单克隆抗体的杂交瘤细胞,其单克隆抗体腹水IPMA效价在1.28×10^-4至2.56×10^-5之间。单克隆抗体3A2、5H6、6B8、7E10和9G12血凝抑制试验（HI）显示血凝抑制活性,其（HI）效价在6log2~9log2之间。单克隆抗体3A2、6B8和9G12在病毒中和试验中对H9N2亚型AIV有显著病毒中和活性,中和效价分别1∶6 400、1∶25 600和1∶25 600。Western blotting结果提示,该中和单克隆抗体识别HA蛋白线性抗原表位。利用配对单克隆抗体3A2和9G12研制的H9N2亚型AIV检测试纸条检测H9N2亚型AIV尿囊液的效价为9log2,灵敏度与经典血凝试验（HA）相当,与其他亚型AIV （H1、H3、H5、H7）,以及新城疫病毒和鸡传染性法氏囊病病毒等相关病毒均无交叉反应。本研究制备了具有病毒中和活性的抗H9N2亚型AIV单克隆抗体,并初步研制了H9N2亚型AIV检测试纸条,为H9N2亚型AIV新型疫苗研制和快速检测奠定良好的研究基础。