Immunological blockade of endogenous thyrotropin-releasing hormone impairs recovery from hyperglycemia in mice

Administration of antiserum to thyrotropin-releasing hormone (TRH) into the lateral cerebral ventricle of mice significantly attenuated recovery from hyperglycemia induced by treatment with 2-deoxyglucose but had no effect on the plasma glucose of saline-treated mice. TRH, injected centrally together with the anti-TRH antibody, reversed the effect of the antiserum and blocked the development of hyperglycemia. These findings suggest that activation of TRH neurons in the central nervous system may be a physiological event influencing recovery from hyperglycemia.     

Increased reactivity to chemical but not to heat noxious stimuli in mice producing anti-idiotypic antibodies for substance P

Mice immunized against anti-substance P (anti-SP) monoclonal antibodies produced anti-SP anti-idiotypic antibodies (SPAb2). In a previous report. SPAb2 antibodies were found to have in vitro biological activity i.e. to behave either as agonists or as antagonists for substance P (SP) depending on the biological test. In this study, the involvement of SPAb2 in vivo biological activity has been tested. Because of the possible implication of SP in the generation and transmission of nociceptive information, we have tested the responsiveness of SPAb2 responding mice in behavioral nociceptive tests. SPAb2 mice showed very small behavioral variations in the hot plate test as compared with a control group of mice immunized against an unrelated monoclonal antibody. In the formalin test, however, SPAb2 mice displayed a significant increase in paw licking time, which was significantly correleted with SPAb2 serum concentration. These results are discussed in terms of the use of SPAb2 as pharmacological tools for studying the biological properties of SP receptors and more generally of auto anti-idiotypic antibodies in modulating behavior responses.     

肾小球系膜细胞的分离和培养

目的分离、培养和鉴定正常人肾小球系膜细胞。方法筛网滤过分离正常人肾小球系膜细胞,同时采用免疫荧光技术,利用异硫氰酸荧光素(FITC)间接标记单克隆抗体:抗Ⅷ因子相关抗原抗体、抗结蛋白抗体、抗角蛋白抗体、抗细胞角蛋白抗体、抗Ⅳ型胶原抗体、抗纤维连接蛋白抗体、抗层粘连蛋白抗体对系膜细胞特有的中间微丝进行鉴定。并采用同期培养的同一来源的肾小球内皮细胞作对照。结果抗Ⅷ因子相关抗原、抗角蛋白和抗细胞角蛋白抗体表达均为阴性,而抗结蛋白抗体、抗Ⅳ型胶原抗体、抗纤维连接蛋白抗体、抗层粘连蛋白抗体为阳性,说明培养的细胞为系膜细胞。结论该种方法分离培养的细胞经过免疫荧光染色鉴定为肾小球系膜细胞。     

Extremely High Association Between Appearance of HLA Antibodies and Failure of Kidney Grafts in a Five-Year Longitudinal Study

Longitudinal studies were conducted over a five-year period for HLA antibodies on 493 sera tested from 54 kidney transplant patients. HLA single antigen beads were employed to establish donor specificity of the antibodies. Only 3 of 22 patients without antibodies rejected a graft in contrast to 17 out of 32 patients with posttransplant antibodies (p = 0.003). Using a serum creatinine value of 4.0 mg/dL as the cut-off for a failed graft, 4 of 22 patients without antibodies failed compared to 21 of 32 with antibodies (p = 0.0006). Among patients with donor-specific antibodies (DSA) 13 of 15 failed (p = 0.000004). Even among patients with non-donor specific antibodies (NDSA), 8 of 17 failed (p = 0.05). Among patients who could be identified as making de novo antibodies (since they developed antibodies while not having antibodies for more than six months after transplantation), 6 of 11 failed (p = 0.03). Sequential testing for HLA antibodies shows that antibodies appear prior to a rise in serum creatinine and subsequent graft failure. The very strong association between the production of HLA antibodies after transplantation and graft failure indicates the importance of monitoring for posttransplant HLA antibodies.     

大鼠抗GBM肾炎模型的建立及不同病期的生化指标和肾组织病理学观察

目的：为研究人类新月体抗肾炎的发病机制，建立大鼠抗肾小球基底膜（BGM）肾炎的动物模型，方法：提取S－D大鼠GBM抗原，将此抗原免疫新西兰白兔获得抗血清，再将此抗血清从尾静脉一次性注射给S－D大鼠。实验分2组：肾炎模型组及正常对照组。定期于第4天、第14天第第21天检测大鼠24h尿蛋白，血肌酐及血尿素的含量和肾组织病理学改变。结果：大鼠肾炎模型组：大鼠注射抗血清后于第4天出现大量蛋白尿，第14天血肌酐，血尿素显著升高，并持续上升，肾组织病理表现为肾小球内细胞数明显增加，大量新月体形成及蛋白管型，GBM呈不规则增厚，足突融合，内皮细胞脱落，坏死，免疫荧光检查见IgG、鼠IgG沿GBM线形分布，大鼠正常对照组以上指标均无明显改变。结论：通过动态观察大鼠抗GBM肾炎的病变变化，证实该模型病变与人类新月体性肾炎的病变较为一致，该模型可用于探讨人类新月体体肾炎的实验研究。     

单克隆抗体对溶组织内阿米巴吞噬红细胞的影响

用具有抗细胞膜活性和抗细胞核、胞浆活性的单克隆抗体３Ｆ７和４Ｇ６与致病性溶组织内阿米巴ＨＭ－１∶ＩＭＳＳ株滋养体３７℃孵育，在孵育即刻、５ｍｉｎ、１０ｍｉｎ加入红细胞，检测滋养体吞噬红细胞的能力，并以正常小鼠血清和抗克氏锥虫抗体ＴＣＥ０４作为对照。结果，经单克隆抗体３Ｆ７孵育即刻的ＨＭ－１∶ＩＭＳＳ虫株滋养体吞噬红细胞能力明显减弱（Ｐ＜０．００１），而４Ｇ６抑制滋养体吞噬红细胞的作用不如前者明显。提示，抗细胞膜单克隆抗体对抑制滋养体吞噬红细胞有显著作用，但随着时间的推移，抑制作用明显减弱。     

Antibodies against dengue virus E protein peptide bind to human plasminogen and inhibit plasmin activity

Both mice and rabbits immunized with dengue virus E protein peptide spanning amino acids 100–119 (D4E) produced antibodies that reacted not only with the D4E peptide itself but also with human plasminogen, as shown by ELISA and Western blot. Sera from dengue virus-hyperimmunized mice and dengue patients also contained antibodies against D4E and plasminogen. Furthermore, such sera all contained plasmin inhibitory activity. Using affinity-purified anti-D4E antibodies and free D4E peptide for competitive inhibition, we demonstrated that the inhibition of plasmin activity was due to anti-D4E antibodies rather than other substances in the sera. Taken together, these results suggest dengue virus E protein amino acids 100–119 are a cross-reactive immunogenic region, and antibodies against this region may interfere with human fibrinolysis.     

对硫磷人工抗原合成及其鉴定

目的 制备对硫磷人工抗原和抗血清，为建立酶联免疫吸附法提供技术储备。方法 还原对硫磷硝基成为氨基，合成半抗原氨基对硫磷；重氮化偶联大分子载体牛血清白蛋白制备人工抗原；免疫新西兰白兔。进行多抗制备。结果 合成的半抗原核磁共振图谱鉴定结果为6：6．9～7(m，2H，CH2)6．4～6．5(m，2H，CH2)4．1～4．2(q，4H-CH2)3．4(s，2H，NH2)1．3～1．4(t，6H，CH3)；氨基对硫磷与牛血清白蛋白的结合比为35～36：1；抗血清经间接酶联免疫吸附法检测，效价达1：128000，双向免疫扩散实验测得抗血清与人工抗原形成沉淀；间接竞争酶联免疫吸附法测得抗血清对对硫磷最低抑制浓度为0．16ng／mL；经间接酶联免疫吸附法检测人工抗原与羊抗人血清、兔抗人血清、正常兔血清无交叉反应。结论 合成对硫磷半抗原及人工抗原纯度高、结合比高。具有较好的免疫原性。     

单克隆抗体反向IHAT检测囊虫病循环抗原

应用抗猪囊虫抗原的单克隆抗体反向间接血凝试验（ＩＨＡＴ），对临床确诊的囊虫病患者进行了血清循环抗原（ＣＡｇ）的检测，阳性率为５１．１４％（６７／１３１）。对１９例脑型囊虫病患者同时测定了血清和脑脊液中的ＣＡｇ，阳性率分别为４７．３６％和７８．９４％，总阳性率为８４．２１％。对治疗０．５３后的１９例患者检测时，血清和脑脊液中ＣＡｇ均呈阴性反应，而检测抗体仍呈阳性反应。５０例正常人、４９例其它寄生虫病及５例其它脑部疾病患者脑脊液的ＣＡｇ均呈阴性反应。结果提示：测定ＣＡｇ对囊虫病活动性感染的诊断及疗效判定均具有一定的实用价值。     

Passively transferable protection against Schistosoma japonicum induced in the mouse by multiple vaccination with attenuated larvae: the development of immunity, antibody isotype responses and antigen recognition

Vaccination of mice with attenuated S. japonicum cercariae induces protection against secondary infection which can be transferred to naive mice with serum (VMS). The presence of antibody does not per se impart protection as serum from mice carrying non-attenuated infections (CIS), contains high levels of specific antibody, but confers no protection. Here we describe the increased protection transferred (20 to 68%) with increased number of vaccinations (one to five) given to the donors, and its decline with time after the final vaccination. We also describe the development of IgM, IgA, IgE, total IgG and IgG subclass responses in VMS, giving different levels of protection and CIS, directed against sodium periodate-sensitive and -resistant epitopes in ‘skin-stage’, ‘lung-stage’and ‘liver-stage’schistosomula, adult worms and eggs. In addition, antibody affinity maturation, development of S. japonicum species-specific responses, and vaccination-specific responses were examined. No response developed in parallel with serum-mediated immunity, suggesting immunity may be due to responses against individual antigens. Preliminary examination of antigens recognized in Western blot showed that two schistosomal membrane antigens, of 13 and 40 kDa, were recognized by VMS from mice vaccinated five times (68% protection), but not by twice vaccinated VMS (27% protection). Neither antigen was recognized by non-protective CIS.