藏酋猴类人ABO血型的鉴定与分析

目的:首次对繁育藏酋猴进行类人ABO血型鉴定,了解藏酋猴血型分布。方法:通过商品化的人ABO血型试剂盒,利用ABO血型鉴定方法(反定型法)对藏酋猴血型进行检测,并与恒河猴、食蟹猴血型检测结果进行比较。结果:35只藏酋猴中A型藏酋猴23只(66%)、B型藏酋猴4只(11%)、AB型藏酋猴8只(23%),未发现O型藏酋猴。结论:藏酋猴与恒河猴、食蟹猴血型比较,主要为A型血,具有明显的特异性,为非人灵长类动物在科研领域的使用提供了新的选择。     

犬血型DEA1.1抗原生化特性的研究

为了研究犬血型DEA1.1抗原生化特性,为犬输血前血型检测提供理论依据,试验采用凝聚胺介质试剂调查356只不同地区及品种犬的DEA1.1血型频率分布,用考马斯亮蓝法检测DEA1.1红细胞膜蛋白并通过免疫沉淀鉴定血型蛋白大小,观察多种酶处理对DEA1.1抗原性的影响,通过ABO血型检测试剂和ABO型微柱凝胶检测卡检测人类红细胞ABO血型与犬DEA1.1血型有无交叉反应。结果表明:不同品种犬的血型频率差异较大,DEA1.1血型为50 ku和200 ku膜蛋白;DEA1.1抗原在无花果酶、木瓜酶、链霉蛋白酶处理后抗原性增强,在菠萝酶、糜蛋白酶和胰蛋白酶处理后抗原性未增强;DEA1.1阴、阳性红细胞与ABO血型试剂均未发生凝集。说明为降低输血反应,最好采用DEA1.1血型频率低的犬种作为供血犬,并建议输血前采用木瓜酶介质交叉配血试验进行血型检测。     

试验用山羊血型及临床应用

为了调查研究试验用山羊的ABO、Rh、MN三个血型系统的血型。ABO血型系统采用微柱凝胶卡氏法鉴定血型,Rh、MN血型系统运用经典试管法鉴定血型;山羊的ABO血型结果显示,既无ABO血型系统的A抗原,也无ABO血型系统的B抗原,类似人类O型血型特征;山羊的Rh血型进行了D、E、e、C、c抗原检测,发现6只E抗原山羊;MN血型检测发现了3只M抗原山羊。表明采用不同检测方法,对试验山羊进行了系统研究,既发现了ABO血型系统的O型山羊,又检测到Rh血型系统的E抗原,还印证了MN血型系统的M血型的存在。     

乙肝表面抗体和抗人红细胞单克隆抗体的F(ab′)_2偶联与鉴定

制备人红细胞非凝集型单克隆抗体,形成双功能抗体试剂,用于临床诊断。用人O型红细胞膜蛋白作为抗原免疫Balb/c小鼠,采用淋巴细胞杂交瘤技术进行细胞融合,制备非凝集型单克隆抗体,通过化学偶联法将非凝集性单克隆抗体和乙肝表面抗体的F(ab′)2片段偶联,制备出双功能抗体,用直接凝集法检测乙肝表面抗原。获得5株分泌非凝集型杂交瘤细胞,利用戊二醛一步法制备出双功能抗体,形成全血凝集检测试剂,并进行了初步鉴定。     

亚洲黑熊(Ursusthibetanus)血型检测

本试验旨在分析亚洲黑熊（Ursusthibetanus）是否具有不同血型。采集中国福建省的40只亚洲黑熊全血样本,使用试管法检测黑熊的血型抗原及血型抗体,抗体的同种型使用2-巯基乙醇鉴定,并对黑熊血浆进行质谱分析。4只黑熊的血浆中检出抗体,该抗体可能是IgM型,可凝集30只亚洲黑熊的红细胞。10只亚洲黑熊血液使用人类血型试剂检测,4只黑熊具有类似人类O型血型,6只具有类似人类B型血型。质谱结果显示,黑熊血清蛋白与大熊猫（Ailuropodamelanoleuca）血清类白蛋白同型异构体1（serum albumin-like isoform 1）有很高同源性。上述结果提示,亚洲黑熊具有至少1个血型系统,该系统至少具有2种血型。如果患病黑熊需要接受输血治疗,交叉配血试验是必需的。此外,在紧急情况下,大熊猫可接受亚洲黑熊血液。     

野生猕猴STLV-1和B病毒抗体的检测

为了调查保护区野生猕猴是否感染猴的3种主要病毒,从广西某自然保护区一只濒死的野生猕猴抽取血液分离血清,分别应用猴3种病毒斑点酶免疫检测试剂盒检测,结果猴T淋巴细胞白血病病毒1型(STLV-1)和猕猴疱疹病毒I型(B病毒)抗体呈阳性,而猴免疫缺陷病毒抗体呈阴性.     

犬红细胞抗原1.1血型多克隆抗体的制备与鉴定

为了制备犬红细胞抗原1.1(DEA1.1)血型多克隆抗血清,用犬血型快速检测卡分别鉴定DEA1.1阳性犬和DEA1.1阴性犬,用DEA1.1阳性犬完整红细胞免疫DEA1.1阴性犬。多次免疫后采集阴性犬血清,用抗球蛋白血凝试验验证多抗血清的符合率。结果显示,制备的多抗血清与商品化的多抗血清符合率为98%。抗球蛋白试验检测抗体血凝效价为211。试验表明,制备的犬血型DEA1.1多克隆抗体能达到血型鉴定的要求,能满足国内犬临床输血中DEA1.1血型检测的要求。     

实验动物山羊血型鉴定初探

目的:旨在分析研究山羊是否具有与人类相似的不同血型。方法:用一次性EDTA-K2真空管,采集河北省邢台县当地山羊全血样本。采用微柱凝胶卡鉴定血型,正定型检测山羊A B O血型的A抗原、B抗原及Rh系统的D抗原;反定型检测抗-A、抗-B及A B O血型系统以外抗体。结果:正定型抗-A、抗-B及抗-D试剂血清与山羊红细胞在微柱凝胶血型卡中,凝集反应均为阴性;反定型山羊血清与试剂A细胞、B细胞均为阴性反应。山羊血型正定型为O型,Rh(D)血型为阴性;反定型也似O型;但对照管与O型红细胞发生凝集反应,推测山羊血清中存在着抗人红细胞的种族抗体。结论:山羊红细胞上无抗原,正反定型类似人类红细胞A B O系统的O型、Rh(D)阴性;但山羊血清中有抗人类红细胞的种族抗体。     

猕猴B病毒抗体的检测报告

用人单纯疱症病毒1型(HSV-1)为抗原片,玻片免疫酶(IEA)检测抗体的方法,对全场猕猴B病毒抗体阴性动物进行分区隔离。B病毒抗体阴性动物连续3年检测,B病毒感染1.05%,B病毒抗体阴性猴再次感染率不高,为日后群养动物提供可行依据。将分区隔离后B病毒抗体阴性动物每隔2周进行检测,表明潜在病毒携带者并非随时排毒。用IEA和ELISA两种方法检测72只B病毒抗体阳性母猴所生仔猴抗体,71只B病毒抗体阴性,阴性率98.6%。结果说明,母猴抗体阳性垂直传染给仔猴几率极低。     

血型分析技术及其在养牛业中的应用

近几年来 ,随着我国奶牛业的飞速发展 ,细管冷冻精液的普及 ,胚胎移植技术的应用 ,使以遗传标记进行的牛亲子鉴定技术显得尤为重要。目前国际上奶牛亲子判定技术主要有 4种 :红血球抗原型检查、蛋白多态型检查、多价抗血清法、DNA指纹技术。本文主要对血型分析技术的原理及其在奶牛业中的应用做一综述。我国迄今为止还没有正式开展中国荷斯坦奶牛的登记工作 ,也未曾普及血型分析技术 ,延边农学院曾用日本家畜改良事业团检查课赠送的抗血清 ,对黑龙江省家畜繁育指导站的6 2头种公牛的血型进行过红细胞抗原型的检查 ,现在也未再延续工作。目…     

Cloning, expression and characterization of monkey (Macaca fascicularis) CD137

CD137 plays an important role as a co-stimulatory molecule in activated T cells. Agonistic CD137 specific antibodies have been investigated as therapeutic agents to promote tumor-specific immune responses by direct activation of T cells. As part of the pre-clinical pharmacological evaluation of cynomolgus monkeys, monkey CD137 was cloned and characterized. The deduced amino acid sequence encoded a full-length gene of 254 amino acids 95% identical to human CD137. Sequence variants identified in monkey CD137 include four splice variants lacking the transmembrane domain. These variants were detectable in human including two previously unreported variants. Two missense single nucleotide polymorphisms were detected present in 42 and 50% of 36 monkeys tested. In both monkey and human, mRNA expression of full-length CD137 and splice variants were significantly increased in peripheral blood mononuclear cells (PBMCs) upon stimulation by anti-CD3 antibodies. Recombinant monkey CD137 protein was bound with high affinity by an agonistic anti-human CD137 antibody but not by an anti-mouse CD137 antibody. In summary, compared to human, monkey CD137 showed distinct extracellular domain amino acid sequence and sequence polymorphisms. Thus, antibodies directed against epitopes in this extracellular domain could have differences in pharmacologic activity between cynomolgus monkeys and human or across individual cynomolgus monkeys.     

Characterization of a new fimbrial antigen present in Escherichia coli strains isolated from calves.

Thirteen Escherichia coli strains isolated from calves with diarrhoea, supposed to carry a common antigen were examined for their hemagglutinating activity and compared by bacterial agglutination, double diffusion in two dimensions and by crossed immunoelectrophoresis (CIE). Two of the strains were examined also in the electron microscope. Most of the strains agglutinated red blood cells of horse, ox, guinea pig and chicken, of which the agglutination of ox erythrocytes was mannose-resistant (MRHA). None of the strains agglutinated human erythrocytes. All strains with MRHA of ox red blood cells, regardless to their O:K:H antigens could be agglutinated in unabsorbed or absorbed antisera produced against cultures C1209 (020:K-:H9) and C1213 (09:K36:H-) when live cells as antigens were used. None of these sera agglutinated reference strains carrying K88, K99, 987P, F41 or FY (Att25) antigen respectively. By the double gel diffusion test and by CIE in extracts (60 degrees C) of the strains a common heat labile antigen, responsible for the MRHA of ox red blood cells was identified. Electron microscopy revealed that this common antigen was represented by thin, long, hair-like fimbriae on cells of E. coli C1213, and that specific homologous antibodies attached to these fimbriae.     

神经退行性疾病转基因猴模型的构建及相关技术体系建立

非人灵长类动物因与人有最近亲缘关系而成为研究人类生命领域最高级模式动物。本研究采用慢病毒载体技术结合胚胎工程技术,拟建立一套高效简便转基因猴制作技术体系,建立小脑共济失调转基因猴模型,为该病研究提供最为理想动物模型,同时为建立其他疾病转基因猴模型提供技术路线。利用卵泡浆内单精子显微注射技术(Introeytoplasmiesperm injection,ICSI)技术总共构建88个体外受精胚胎,经过体外培养共计32枚发育到原核期并进行病毒注射后移入7只受体猴输卵管内。B超妊娠检测,怀孕3只,其中:1只未能发育到期中间流产,1只发育到期生1死胎,1只顺产生1只活试管猴,经过人工喂养奶粉后健康存活,但经检测转基因为阴性。本研究虽然未能通过病毒注射获得阳性转基因猴,但对食蟹猴(Macaca fascicularis)超数排卵及B超监测卵泡发育技术、卵母细胞回收及体外成熟培养技术、电刺激采精及精子获能技术、食蟹猴体外受精技术和胚胎体外培养技术以及胚胎移植等技术体系进行多次探索和优化,并成功建立相关技术平台,将为后续利用试管猴技术结合高效的TALEN和CRISPR/Cas9等基因编辑工具获得各种疾病模型奠定基础。     

Natural infection with canine distemper virus in a Japanese monkey (Macaca fuscata)

A case of encephalitis in a Japanese monkey (Macaca fuscata) was examined histopathologically and serologically. The animal had brain lesions consisting of perivascular cuffs, malacia, inclusion bodies and giant cells. Monoclonal antibody to the nucleoprotein of canine distemper virus (CDV) stained the inclusions, and the distribution of the virus antigen was closely associated with that of the histological lesions. Serologically, all the 22 monkeys in the same group as the diseased monkey had relatively high titers of neutralizing antibody to CDV, but not to measles virus (MV). The pattern of the antibody titers to CDV and MV closely resembled that of cynomolgus monkeys experimentally inoculated with CDV, but differed from that of monkeys inoculated with MV. These findings suggest that an epidemic of CDV occurred in these Japanese monkeys, associated with one case of fatal viral encephalitis. This is believed to be the first report of a natural infection by CDV in non-human primates.     

Canine Dal blood type: A red cell antigen lacking in some Dalmatians

BACKGROUND: Based upon alloantibodies produced after sensitizing dogs with transfused blood, more than a dozen blood group systems have been recognized thus far, and some have been classified as dog erythrocyte antigens (DEA). HYPOTHESIS: A new canine red cell antigen was suspected, based on the development of specific alloantibodies in a Dalmatian previously sensitized by blood transfusions. ANIMALS: Twenty-six Dalmatians (including 1 Dalmatian in need of blood compatibility studies); 55 canine blood donors. METHODS: Serologic tests, including blood typing, crossmatching, and direct Coombs' test were performed by standard tube techniques and a novel gel column technology adapted from human blood banking. RESULTS: By day 40 after transfusion of an anemic Dalmatian, all major crossmatch tests to 55 non-Dalmatian dogs were incompatible. The 2 initial donors, who were compatible before transfusion, were also now incompatible, suggesting the development of an alloantibody to a common red cell antigen. No siblings were available, but 4 of 25 unrelated Dalmatians were crossmatch compatible, suggesting that they were missing the same red cell antigen. The patient was blood typed DEA 1.1, 3, 4, and 5 positive, but DEA 7 negative. Further blood typing and crossmatching results did not support an association to any of these known blood types. The alloantibodies produced were determined to be of the immunoglobulin G class. CONCLUSIONS AND CLINICAL IMPORTANCE: Based upon the identification of an acquired alloantibody in a Dalmatian, a presumably new common blood type named Dal was identified. Dalmatians lacking the Dal antigen are likely at risk of delayed and acute hemolytic transfusion reactions.     

Complement-fixation and Hemagglutination Antigens from a Chlamydial (Ornithosis) Agent Grown in Cell Cultures

Methods are described for the preparation of complement-fixation (CF) and hemagglutination (HA) antigens from the Texas turkey ornithosis agent grown in McCoy cell culture monolayers. The particulate antigens prepared for this study were satisfactory for testing mammalian sera by direct CF tests and avian sera by indirect CF or modified direct CF tests. Comparison of titers were made on human, bovine and ovine sera using direct CF tests employing antigen prepared for this study, 6 BC yolk sac antigen, and a commercially available antigen.

The HA antigen agglutinated mouse erythrocytes, but it was not of value in hemagglutination inhibition tests because of “nonspecific” inhibitors in both mammalian and avian sera.

     

Spontaneously occurring hepatocellular neoplasia in adolescent cynomolgus monkeys (Macaca fascicularis)

Spontaneous hepatic neoplasms were identified in two adolescent (<5 years of age) male cynomolgus monkeys (Macaca fascicularis). Monkey No. 1 had a solitary hepatocellular carcinoma (HCC). Monkey No. 2 had multiple discrete tumors consisting of several poorly circumscribed HCCs and a mixed hepatocholangiocellular carcinoma (MHC). Metastases were not evident in either monkey. Histochemical and immunohistochemical stains were used to assess phenotypic alterations in the tumors. Many or most neoplastic hepatocytes (NHs) of both monkeys stained positive for low-molecular-weight cytokeratin (LMWCK), cytokeratin (CK) 8, and CK 18. In monkey No. 1, small aggregates of NHs were positive for carcinoembryonic antigen (CEA), glutathione S-transferase-pi (GST), and alpha-fetoprotein (AFP), but NHs were uniformly negative for CK 7. NHs in monkey No. 2 were negative for CEA and AFP but were multifocally positive for GST and CK 7. Broad-spectrum cytokeratin (BSCK), high-molecular-weight cytokeratin (HMWCK), and CK 19 did not react with NHs of either animal. Neoplastic cells forming ductlike structures in the MHC of monkey No. 2 stained with LMWCK, CK 7, CK8, CK 18, BSCK, and GST but not with HMWCK or CK 19. Tumors in both monkeys had enhanced pericellular fibronectin staining. Nonneoplastic parenchyma of both monkeys contained multiple discrete foci of cellular alteration and scattered aggregates of hepatocytes with strong cytoplasmic staining for fibronectin. Staining patterns of these tumors demonstrate immunophenotypic heterogeneity of the neoplastic cells within individual tumors and variability among tumors. This information may serve as a useful reference for others encountering similar lesions in primates.     

3种食蟹猴早孕诊断方法的比较

本试验旨在对3种食蟹猴早孕诊断方法进行比较,找出比较准确、可靠的诊断方法。在食蟹猴怀孕第20天时用直肠触诊、B超检查和孕酮值检测3种方法进行早孕诊断,在怀孕第30天时B超确诊是否怀孕。结果表明,B超检查的准确率为96.4%;直肠触诊的准确率为92.9%;孕酮值检测的准确率为64.3%。因此B超检查准确率高,是食蟹猴怀孕诊断的金标法;直肠触诊准确率较高且方便、经济,适合基层生产中进行大批量诊断;孕酮值检测确诊率较低,但可作为早孕诊断的辅助方法。     

Detection of Echinococcus multilocularis infection in a colony of cynomolgus monkeys (Macaca fascicularis) using serology and ultrasonography.

Five animals in a colony of cynomolgus monkeys (Macaca fascicularis) died or were euthanatized because of alveolar echinococcosis, during a period of 5 years. The remainder of the colony was screened for possible infection with Echinococcus multilocularis, using serology and ultrasonography. A total of 46 animals out of a group of 55 were examined. The presence of anti-Em2 antibodies analyzed with enzyme-linked immunosorbent assay was demonstrated in 3 monkeys. In 2 of these 3 monkeys, multilocular structures compatible with metacestodal cysts in the liver were identified, using ultrasonography. The presence of alveolar echinococcosis was subsequently confirmed at postmortem examination in 1 animal. The other animals are still alive. Two other monkeys were negative in the serological examination but had cystic structures in the liver, which were identified as bile duct cysts at postmortem examination in 1 animal. The other monkey is still alive. These findings suggest that serology for antibodies against the Em2 antigen may represent a useful method in identifying animals that might be infected with E. multilocularis and are therefore at risk of developing fatal alveolar echinococcosis.     

A case of spontaneous autoimmune skin disease in a cynomolgus monkey (Macaca fascicularis)

Pemphigus is an autoimmune blistering disease characterized by lesions on the skin and mucous membranes. To date, no spontaneous cases of this disease have been reported in cynomolgus monkeys. This report describes the histopathological characteristics of spontaneous pemphigus in a cynomolgus monkey. Macroscopically, redness and scaling with pruritus were observed on the skin of the entire body. Histopathologically, the epidermis showed intercellular edema, and eosinophils and mononuclear cells infiltrated the epidermis. There was no obvious acantholysis in the epidermis. The perivascular area showed edema, and eosinophils and mononuclear cells infiltrated the vessels in the dermis. Immunohistochemically, the intercellular area in the epidermis was positive for Immunoglobulin G and Complement component 3. Serologically, anti-desmoglein 1 and desmoglein 3 antibodies in the serum were negative. From these findings, this case was diagnosed as an autoimmune skin disease, suspected to be pemphigus, and concluded as lesions being similar to those in human “pemphigus herpetiformis”.