人醛缩酶A单克隆抗体的制备及应用

纯化并鉴定了人醛缩酶Ａ、Ｂ、Ｃ（ｈＡＬＤ－Ａ、Ｂ、Ｃ）。用ｈＡＬＤ－Ａ免疫Ｂａｌｂ／Ｃ小鼠，将免疫的脾细胞和Ｐ＿３－Ｘ＿（６３）－Ａｇ８．６５３骨髓瘤细胞用ＰＥＧ进行融合，用ＥＬＩＳＡ法筛选，ｈＡＬＤ－Ｂ、Ｃ作阴性对照，将阳性反应的杂交瘤细胞进行克隆，获得３株杂交瘤细胞株。其ＭｃＡｂ的亚类分别为ＩｇＧ２ｂ，ＩｇＧ１、ＩｇＭ，亲合常数分别为７．５×１０￣（１０）、３．５×１０￣９、２．３×１０￣９。３株细胞株分泌的单抗通过Ｉｍｍｕｎｏｂｌｏｔｔｉｎｇ得到证实。用亲合层析法从人肝癌细胞中提纯了ＡＬＤ－Ａ，ＳＤＳ－ＰＡＧＥ显示为单一区带，但其电泳迁移率较ｈＡＬＤ－Ａ滞后。     

重组人红细胞生成素单克隆抗体的制备

本文应用常规淋巴细胞杂交瘤技术制备了４株能稳定分泌抗人重组红细胞生成素（ｒＨｕＥＰＯ）单克隆抗体（ＭｃＡｂ）的小鼠杂交瘤细胞系ＢⅡ１Ｂ５、ＤⅡ６Ｂ９、ＭⅡ１Ｈ４和ＧＩ３Ｅ７。用鼠单克隆抗体分型试剂盒鉴定，其分泌的ＭｃＡｂ的类分别是ＩｇＭ、ＩｇＭ、ＩｇＧ１和ＩｇＧ２ａ。间接ＥＬＩＳＡ法测定细胞上清的效价为１×１０－２～１．２５×１０－４，腹水效价为１×１０－２～１×１０－８。培养上清经ＥＬＩＳＡ鉴定，与ＩＬ－２、ＧＭ－ＣＳＦ、ＩＦＮ－α等细胞因子均无交叉反应，只与ｒＨｕＥＰＯ特异性结合。     

西安地区死胎、流产的孕妇微小病毒B19抗体检测结果分析

用ＥＬＩＳＡ法对孕早期有死胎、流产的５７例孕妇（实验组）与３５例正常孕妇（对照组）血清检测Ｂ１９抗体（ＩｇＭ、ＩｇＧ）其中实验组Ｂ１９－ＩｇＭ阳性率（１２．２８％）与正常组Ｂ１９－ＩｇＭ阳性率比较有显著性差异（ｐ＜０．０５），实验组Ｂ１９－ＩｇＧ阳性率（３３．３３％）与正常组Ｂ１９－ＩｇＧ阳性率（４０％）无统计学差异（Ｐ＞０．０５）。     

抗淋球菌PIA单克隆抗体的制备和应用分析

用淋球菌全菌体免疫ＢＡＬＢ／ｃ小鼠，取脾细胞与Ｓｐ２／０骨髓瘤细胞融合，以纯化ＰＩＡ做ＥＬＩＳＡ间接法筛选，获得５株稳定分泌抗ＰＩＡ的ＭｃＡｂ的杂交瘤细胞株。５株ＭｃＡｂ中３株为ＩｇＭ类（２Ｈ１１，４Ｈ８，４Ｅ１０），另２株分别为ＩｇＧ１（１Ｃ２）和ＩｇＧ２ｂ（５Ａ５）亚类。２Ｈ１１和１Ｃ２为高亲和力抗体，１Ｃ２与５Ａ５识别的抗原表位相同。Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ试验表明，５株ＭｃＡｂ均能从复杂的淋球菌菌体崩解物中特异地识别分子量为３５ＫＤａ的ＰＩＡ抗原，与淋球菌ＰＩＢ无交叉反应。     

两种人化SCID小鼠的建立及其免疫应答性

将人外周血淋巴细胞（ＰＢＬ）和腹腔淋巴结淋巴细胞（ＰＬＮＬ）移植入严重联合免疫缺陷疾病（ＳｅｖｅｒｅＣｏｍｂｉｎｅｄＩｍｍｕｎｏｄｅｆｉｃｉｅｎｔＤｉｓｅａｓｅ，ＳＣＩＤ）小鼠腹腔，建立人化ＳＣＩＤ小鼠（即ＳＣＩＤ－ｈＰＢＬ，ＳＣＩＤ－ｈＰＬＮＬ）。两个月后，两种人化小鼠体内淋巴器官都可以检测到人Ｔ、Ｂ淋巴细胞分布；小鼠血清人源性ＩｇＧ和抗ＥＢ病毒壳抗原ＩｇＧ抗体水平（ＩｇＧ／ＶＣＡ）比较显示，用Ｂ９５８死细胞作为ＶＣＡ来源所诱导的实验组１０只小鼠，ＩｇＧ／ＶＣＡ阳性率为７０％（７／１０），对照组为１７％（２／１２）。１４只ＳＣＩＤ－ｈＰＬＮＬ小鼠血清内人ＩｇＧ／ＶＣＡ的几何平均滴度（ＧＭＴ）和血清ＩｇＧ浓度在１∶１０８和９６．２±５６．４μｇ／Ｌ；在另８只ＳＣＩＤ－ｈＰＢＬ中为１∶７．９和１３．８４±６．０μｇ／Ｌ。实验提示，ＳＣＩＤ－ｈＰＬＮＬ小鼠较ＳＣＩＤ－ｈＰＢＬ小鼠更适合于人源性特异ＩｇＧ的诱导。     

分泌抗金黄色葡萄球菌C1型肠毒素杂交瘤细胞系的建立及初步应用

应用常规方法建立了３株稳定分泌抗金黄色葡萄球菌Ｃ１型肠毒素（ＳＥＣ１）单克隆抗体（ＭｃＡｂ）的小鼠杂交瘤细胞系Ｂ３、Ｃ４和Ｇ８。其中Ｂ３和Ｃ４均为ＩｇＧ１（ｋ），Ｇ８为ＩｇＧ２ａ（ｋ）。Ｂ３和Ｇ８与ＳＥＡ，ＳＥＢ及ＳＥＤ均无交叉反应；Ｃ４虽与ＳＥＡ和ＳＥＤ无交叉反应，但与ＳＥＢ有交叉反应。间接ＥＬＩＳＡ测定小鼠腹水效价为１０＾－５～１０＾－８。应用识别不同表位的ＭｃＡｂ建立了双ＭｃＡｂ夹心ＥＬＩＳ     

新生儿心肌炎柯萨奇B病毒感染的研究

用间接ＥＬＩＳＡ、ＭａｃＥＬＩＳＡ检测２６例新生儿心肌炎血清ＣｏｘＢ病毒特异性ＩｇＧ和ＩｇＭ，其中１４例用聚合酶链反应检测血清肠道病毒（ＥＶ）－ＲＮＡ。结果ＣｏｘＢ病毒ＩｇＧ的阳性检出率为３８％，ＩｇＭ的阳性检出率为５４％，ＥＶ－ＲＮＡ的阳性率为５０％，三者阳性率均明显高于对照组。检测了１０例母婴血清的ＣｏｘＢ病毒的ＩｇＭ，提示ＣｏｘＢ病毒可能通过胎盘传播，１４例做了急性期和恢复期双份血清检测，ＩｇＧ有４倍滴度改变者６例。     

分泌抗金黄色葡萄球菌C1型肠毒素杂交瘤细胞系的建立及初步应用

应用常规方法建立了３株稳定分泌抗金黄色葡萄球菌Ｃ１型肠毒素（ＳＥＣ１）单克隆抗体（ＭｃＡｂ）的小鼠杂交瘤细胞系Ｂ３、Ｃ４和Ｇ８。其中Ｂ３和Ｃ４均为ＩｇＧ１（ｋ），Ｇ８为ＩｇＧ２ａ（ｋ）。Ｂ３和Ｇ８与ＳＥＡ、ＳＥＢ及ＳＥＤ均无交叉反应；Ｃ４虽与ＳＥＡ和ＳＥＤ无交叉反应，但与ＳＥＢ有交叉反应。间接ＥＬＩＳＡ测定小鼠腹水效价为１０－５～１０－８。应用识别不同表位的ＭｃＡｂ建立了双ＭｃＡｂ夹心ＥＬＩＳＡ法检测ＳＥＣ１，敏感性可达１ｎｇ／ｍｌ。     

花粉过敏性哮喘的免疫学诊断研究

对４８例与花粉过敏有关的外源性哮喘病例进行过敏史、病史、体格检查，结合花粉变应原支气管激发试验（ＢＰＴ）、皮肤挑刺试验（ＳＰＴ）、特异性ＩｇＥ（Ｓ－ＩｇＥ）及特异性ＩｇＧ（Ｓ－ＩｇＧ）检查。结果ＢＰＴ阳性率为７０．９％（３５／４８），ＢＰＴ阳性组的ＳＰＴ、Ｓ－ＩｇＥ以及Ｓ－ＩｇＧ阳性率分别为１００％，７１．４％，６２．９％，明显高于ＢＰＴ阴性组。ＳＰＴ，Ｓ－ＩｇＥ，Ｓ－ＩｇＧ与ＢＰＴ的符合率分别为     

rhIL—2调节离体boPBMC增殖和分泌Ig的作用

应用＾３Ｈ－ＴｄＲ掺入法，ＥＬＩＳＡ和ＦＡＣＳ研究了ｒＨＩＬ－２调节离体奶牛外周血单核细胞（ｂｏＰＢＭＣ）免疫应答的特点。ｒｈＩＬ－２能诱导细胞持续性增殖和分泌Ｉｇ，尤其是分泌ＩｇＡ和ＩｇＧ２（与ＰＷＭ相比，Ｐ〈０．０１）；还能增强ＰＷＭ诱导的Ｉｇ的分泌，但不改变ＳＥＢ抑制Ｉｇ分泌的作用。细胞表型也表明，ｒｈＩｌ－２对ＰＷＭ或ＳＥＢ诱导的ｂｏＰＢＭＣｋ中ＣＤ４或ＣＤ８细胞增殖作用无选择性。     

Antibodies to poliovirus detected by immunoradiometric assay with a monoclonal antibody

An immunoradiometric assay (IRMA) for the assay of antibodies to poliovirus antigens is described. Dilutions of the test sera or whole (finger prick) blood samples were incubated with the Poliovirus antigen bound to a solid phase and the specific antibody was detected by the addition of a mouse anti-human IgG monoclonal antibody (McAb), which was itself revealed by iodinated sheep IgG anti-mouse F(ab). We have shown that this technique is suitable for the estimation of IgG anti-poliovirus antibodies induced in children following polio vaccine. The present study shows that SPRIA provides a simple and inexpensive method for serological studies with poliovirus particularly for use in large-scale surveys.     

抗GD2/抗CD16单链双特异性抗体的构建及在大肠杆菌中的表达

利用重叠PCR(overlap-PCR)将抗GD2表面抗原GD2的单链抗体基因m-ScFv和抗CD16的单链抗体基因NM3E2融合在一起,得到新型的单链双特异性抗体基因n-m,双抗的联接肽序列为SerGly4Ser,在肽连的C端引入了6×his以利于纯化。该双特异性抗体基因的序列经测序验证后,连接表达载体pET-22b( ),转化大肠杆菌菌株BL21(DE3),诱导表达,凝胶成像系统扫描显示表达的外源融合蛋白约占菌体总蛋白的27%。表达蛋白分泌于胞质空间,经超声波破胞后收集上清,经Ni 亲和层析柱分离,纯度可达90%以上。经SDS-PAGE及Westernblotting鉴定表达蛋白的分子量为53KD,与预期的相符。     

Immunohistochemical Study of Fiber Types in Human Extraocular Muscles

Fiber types in human extraocular muscle (h-EOM) were examined immunohistochemicaliy with antibodies against slow tonic (anti-ALD) and slow twitch (anti-SOL) myosins. Four types of muscle fiber in h EOM were distinguishable according to their reactivities with these antibodies. Groups 1 and 2 fibers reacted with both antibodies, group 1 fibers showing stronger reactivity than group 2 fibers with anti-ALD. Group 3 fibers reacted only with anti-SOL. Group 4 fibers did not react with either antibody. The latter were the most common, and were the main fibers in both the peripheral (outer orbital) and central zones of h EOM. The next most common were group 1 fibers, which were located mainly in the peripheral layer. Group 2 fibers were less common, but were the second most common type in the central layer. Group 3 fibers were only minor constituents. Multiple innervations were observed in some fibers of groups 1 and 2, and group 1 fibers were suggested to be slow tonic myofibers in h-EOM. These specific immunohistochemical and physiological features of h-EOM seem to be the basis of the low morbidity seen in the usual types of muscular dystrophy. Acta Pathol Jpn 40: 808-814, 1990.     

TLR2和TLR4单克隆抗体对DSS诱导的小鼠结肠炎肠黏膜细胞因子及肠道菌群的影响

Objective To evaluate the effect of TLR2McAb and TLR4McAb on intestinal flora of DSS-induced colitis in mice. Methods Fifty healthy male BALB/c mice (SPF level), were randomly assigned into five groups: the control group( group A), the UC model group( group B), TLR2McAb intervention group( group C), TLR4McAb intervention group( group D) and TLR2McAb + TLR4McAb intervention group(group E). Clinical symptoms were evaluated by the disease activity index(DAI), while tissue sam ples were evaluated by histological scoring(HS). The quantities of mRNA for IFN-γ, IL-4 and IL-17 were determined by real-time PCR. Meanwhile, fecal samples were obtained directly from the cecum for microbiological studies. Results After the treatment with TLR2McAb and TLR4McAb, DAI and HS were decreased significantly. Compared with group A, inflammatory cytokines such as IFN-γ, IL-4 and IL-17 in group B were higher. Compared with group B, expression of these three cytokines in group C to E was all markedly decreased. Group A showed a considerable predominance of Lactobacillus spp and Bifidobacterium spp,while the UC model group showed a conspicuous increase of Escherichia coli and decreases of Lactobacillus spp and Bifidobacterium spp. After treatment with TLR2McAb or/and TLR4McAb, Lactobacillus spp and Bifidobacterium spp increased to the normal level. But counts of E. Coli in the three intervention groups were not changed. Conclusion TLR2McAb and TLR4McAb suppressed the development of DSS-induced colitis and increase cecum counts of Lactobacilli and Bifidobacteria.     

TLR2和TLR4单克隆抗体对DSS诱导的小鼠结肠炎肠黏膜细胞因子及肠道菌群的影响

Objective To evaluate the effect of TLR2McAb and TLR4McAb on intestinal flora of DSS-induced colitis in mice. Methods Fifty healthy male BALB/c mice (SPF level), were randomly assigned into five groups: the control group( group A), the UC model group( group B), TLR2McAb intervention group( group C), TLR4McAb intervention group( group D) and TLR2McAb + TLR4McAb intervention group(group E). Clinical symptoms were evaluated by the disease activity index(DAI), while tissue sam ples were evaluated by histological scoring(HS). The quantities of mRNA for IFN-γ, IL-4 and IL-17 were determined by real-time PCR. Meanwhile, fecal samples were obtained directly from the cecum for microbiological studies. Results After the treatment with TLR2McAb and TLR4McAb, DAI and HS were decreased significantly. Compared with group A, inflammatory cytokines such as IFN-γ, IL-4 and IL-17 in group B were higher. Compared with group B, expression of these three cytokines in group C to E was all markedly decreased. Group A showed a considerable predominance of Lactobacillus spp and Bifidobacterium spp,while the UC model group showed a conspicuous increase of Escherichia coli and decreases of Lactobacillus spp and Bifidobacterium spp. After treatment with TLR2McAb or/and TLR4McAb, Lactobacillus spp and Bifidobacterium spp increased to the normal level. But counts of E. Coli in the three intervention groups were not changed. Conclusion TLR2McAb and TLR4McAb suppressed the development of DSS-induced colitis and increase cecum counts of Lactobacilli and Bifidobacteria.     

McAb检测囊虫病人循环抗原的研究

应用ＭｃＡｂ双抗体夹心－ＥＬＩＳＡ检测脑囊虫病人血清和脑脊液（ＣＳＦ）中循环抗原（ＣＡＧ）。血清经ＰＥＧ沉淀浓缩沸浴和ＣＳＦ经沸浴处理后，ＣＡｇ的阳性率分别为９２．７３％和９０．７４％，总阳性率为９２．０７％，灵敏度６８．７ｎｇ／ｍｌ。对照组中除２例包虫病人血清有交叉反应外，其余均为阴性。平行检测病人ＣＳＦ的ＣＡｇ和ＣＡｂ，以及同步检测血清和ＣＳＦ中的ＣＡｇ，表明它们之间有互补和相互验证的作用，可提高阳性率。对囊虫病人治疗前后血清ＣＡｇ的动态检测，表明血清ＣＡｇ的阴转率随疗程的增加而提高。这些结果说明ＣＡｇ检测不仅可用于囊虫病的诊断，而且可作为疗效考核的重要手段。也说明ＭｃＡｂ双抗体夹心ＥＬＩＳＡ检测ＣＡｇ具有敏感性高、特异性强、重复性好等优点。     

Monoclonal antibody recognizing pre-S(2) epitope of hepatitis B virus: characterization of pre-S(2) epitope and anti-pre-S(2) antibody

A hybrid cell line producing monoclonal antibodies recognizing an epitope encoded by the pre-(S)2 region of hepatitis B virus (HBV) genome was obtained by fusion of mouse myeloma cells with lymphocytes from mice immunized with HBV. The monoclonal antibody Mo-F124 secreted from the hybrid line reacted with the pre-S(2) epitope expressed on the surface of both viral and recombinant HBsAg particles--pre-S(2) and S gene product--localised on 34 kD glycoprotein of the viral envelope. The pre-S(2) epitope was sensitive to digestion with V8 protease from Staphylococcus aureus. The enzyme abolished reactivity with Mo-F124 and polymerized human serum albumin (pHSA) binding activity of recombinant particles. Mo-F124 antibody was used to develop highly sensitive radioimmunoassays for determination of pre-S(2) epitope and anti-pre-S(2) antibody in sera of hepatitis B patients. Detection of a pre-S(2) epitope by the monoclonal antibody-based assay in the early phase of acute HBV infection correlated well with the presence of markers of active viral replication (HBeAg, HBV DNA). The appearance of anti-pre-S(2) antibody, usually in the third month after onset of symptoms, was followed by elimination of circulating HBsAg and seroconversion to anti-HBs in all tested cases of uncomplicated acute hepatitis followed by recovery. Anti-pre-S(2) response was not observed in patients with chronic hepatitis B or acute HBV infection progressing to chronic disease. The observed correlation of anti-pre-S(2) response with recovery suggests that the pre-S(2) epitope may represent one of the epitopes inducing antibodies that neutralize the hepatitis B virus.     

系统性红斑狼疮患者妊娠失败与抗心磷脂抗体间关系的研究

研究６３例ＳＬＥ女患者中有妊娠失败史者与各项自身抗体及其他体液免疫的关系，表明妊娠失败与抗心磷脂抗体（ＡＣＡ）有非常显著的关系（Ｐ＜０．０１）．而与其他自身抗体未见明显关系，认为ＡＣＡ在ＳＬＥ患者中是引起妊娠失败的重要因素。此外ＡＣＡ还与Ｃ＿３、Ｃ＿４免疫指标呈负相关（Ｐ＜０．０５或Ｐ＜０．０１）。     

血清TRAb检测在Graves甲亢治疗中的意义

目的：探讨血清TRAb值在Graves甲亢患者治疗前后的改变及其临床意义。方法：50例正常人和42例Graves甲亢患者在丙基硫氧嘧啶治疗前、后分别用放射免疫分析作了血清TRAb,TSH,FT3,FT4,rT3检测。结果：未经ATD治疗的Graves甲亢患者,血清TRAb阳性率为88％。Graves甲亢患者接受ATD治疗后3个月时,TRAb阳性率为57．14％,其它检测指标除rT3外均与正常组无明显差异（P＞0．05）;6个月时TRAb阳性率为17．50％;12个月时为7．69％;24个月时为5．71％。本组临床缓解停药后有15例复发,该15例中13例（88．66％）血清TRAb原为阳性,显示确有关联。结论：血清TRAb的检测有助于指导Graves甲亢患者的治疗。     

TSH受体抗体测定的临床意义

目的:探讨血清TRAb的测定变化对G raves甲亢的临床意义。方法:应用放射免疫受体分析法(RRA)对302例甲状腺疾病患者及52例正常健康人的血清TRAb的值进行比较。结果:甲亢组TRAb的测定值阳性率为86.3%;甲亢缓解组TRAb的测定值阳性率为74.5%;甲亢治愈组TRAb的测定值阳性率为32.1%;甲亢复发组TRAb的测定值阳性率为90.3%;单纯性甲状腺肿组TRAb的测定值阳性率为0;甲瘤组TRAb的测定值阳性率为0;甲亢组、甲亢缓解组、甲亢治愈组、甲亢复发组与正常对照组相比有显著性差异(P<0.01);单纯性甲状腺肿组、甲瘤组与正常对照组相比无显著性差异(P>0.05)。结论:TRAb的测定对G raves甲亢的治疗具有重要的参考价值。