异种胸腺修饰和全身照射在猪-猴心脏移植模型中对抗体产生的影响

为观察胸腺修饰和全身照射预处理受体对猕猴抗猪抗体产生的影响,将受体中国猕猴随机分成空白组(A组)、全身照射组(B组)、照射+胸腺注射组(C组),用流式细胞仪法监测猕猴外周血细胞表面IgM、IgG阳性细胞的百分比在预处理前后及移植后的变化。结果表明:全身照射后,IgM类天然抗体阳性细胞的百分比水平较空白组踢显下降(P<0.01),而IgG类天然抗体阳性细胞的百分比水平无明显变化;在移植后,照射+胸腺注射组的IgM类诱生抗体阳性细胞的百分比水平及IgG类诱生抗体阳性细胞的百分比水平较空白组及全身照射组上升延缓。提示异种胸腺修饰能抑制IgM及IgG类诱生异种抗体的产生。     

小鼠胸腺雌激素受体α与β亚型的免疫组织化学研究

目的:探讨雌激素受体α与β亚型在不同年龄段小鼠胸腺内的表达。方法:免疫组化分析雌激素受体α与β亚型在3、8及15月龄段小鼠胸腺内的表达水平;双色免疫荧光分析雌激素受体α与Treg细胞标志Foxp3表达的关系。结果:3月龄小鼠的胸腺皮质区及髓质区内有大量的雌激素受体β及α亚型的阳性细胞。然而,8月龄及15月龄的小鼠胸腺则缺乏雌激素受体β亚型表达而雌激素受体α亚型阳性细胞仍然可发现于这些胸腺的皮质区内。使用雌激素受体α/β及Foxp3(调节性T细胞的标志)抗体对标本进行双色标记结果证实雌激素受体α/β阳性的细胞为Foxp3阴性。结论:成年小鼠胸腺缺乏雌激素受体β,但雌激素受体α为阳性,提示雌激素受体α是介导雌激素引发的胸腺萎缩的关键激素受体。同时也证实雌激素受体α/β不参与Foxp3调节性T细胞的发育。     

体外循环与抗血栓形成血浆C蛋白的激活

目的　我们假设血浆激活的抗血栓形成 C蛋白在冠状动脉缺血及缺血后再灌注过程中起防御性的抗血栓形成功能。方法及结果　测定 2 0例冠状动脉旁路术患者体外循环及冠状动脉再灌注期间 C蛋白的激活程度 ,在心肺转流期间及主动脉开放后 10 min,血浆 C蛋白水平从正常均数 ( M SE)的 12 3 %± 7%降至 74%± 5 % ;相反 ,血浆中激活 C蛋白水平从 12 2 %± 8%上升至 15 9%± 2 1% ,激活 C蛋白 /C蛋白比例从 1.0 4± 0 .0 8增加到 2 .2 9± 0 .3 1( P=0 .0 0 6)。以再灌注 10 min后冠状窦血浆激活 C蛋白 /C蛋白比例 1.5作人为标准 ,对患者激活 …     

恒河猴对小鼠单克隆抗体的抗抗体应答

本研究对感染日本脑炎病毒的恒河猴注射单克隆抗体后血液及脑脊液中产生的抗抗体进行了测定.第1次注射单克隆抗体后,仅从少数猴的血液和脑脊液中检出IgM类抗抗体或其免疫复合物;最早的第3天出现,最晚为第2周,大多数在1个月内消失.有几只猴的血液和脑脊液中在1周内可检出IgG类抗抗体或其免疫复合物,最晚的于第3周才出现。IgG的维持时间较长.大多数标本可同时检出抗抗体和免疫复合物;随着时间的推移,免疫复合物的检出率减少,这可能与其被清除有关.抗独特型抗体与抗抗体有同步消长的关系。多次注射单克隆抗体,使IgM、IgG类抗抗体及抗独特型抗体的产生增多,但IgE水平却无明显变化,免疫复合物的形成也无明显增多。     

异种胸腺修饰对大鼠抗豚鼠天然抗体产生的影响

为观察异种胸腺修饰对大鼠抗豚鼠天然抗体产生的影响 ,将供体豚鼠和受体SD大鼠随机分成对照组 (a组 )、全身照射组 (b组 )、照射 +胸腺注射组 (c组 ) ,a组大鼠腹腔内注射经处理的豚鼠脾淋巴细胞 (5× 10 6个 ) ,7d后流式细胞仪法检测抗豚鼠IgM及IgG类天然抗体水平的变化 ,b、c组在腹腔注射异种抗原前 17d接受全身照射 ,c组大鼠另在全身照射后 3d接受胸腺内豚鼠脾淋巴细胞注射。结果表明 :a组和b组中 ,腹腔注射后两类天然抗体的水平均明显上升 (P <0 0 1) ,c组中 ,腹腔注射后IgM类的水平亦明显上升 (P <0 0 1) ,而IgG类上升受抑。提示异种胸腺修饰能抑制IgG类天然抗体的生成 ,但对IgM类天然抗体的产生无明显影响。     

自身免疫性疾病患者血清抗bFGF自身抗体的检测及对疾病诊断价值的临床分析

为探讨自身免疫性疾病(autoimmune disease,AID)患者血清中抗碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)自身抗体水平及对患者的诊断价值,回顾性收集2015年3月至2017年5月医院收治的AID患者80例为观察组,取同期入院健康体检者80例为对照组。采用ELISA检测2组患者血清中抗bFGF抗体IgM和IgG水平。结果显示80例AID患者经过检测血清中抗bFGF自身抗体均得到确诊,排在前3位的分别为SLE、RA及慢性肾炎,分别占28.75%、22.50%和18.75%;观察组bFGF水平高于对照组(t=7.572,P0.05);观察组中SLE、不明原因发热患者抗bFGF IgM抗体与对照组比较差异无统计学意义;观察组RA、皮肌炎及特发性血小板减少性紫癜患者抗bFGF IgG抗体阳性率高于对照组(P0.05);观察组RA、慢性肾炎、皮肌炎、特发性血小板减少性紫癜患者抗bFGF IgM抗体水平高于对照组(P0.05)。由此AID患者血清中存在bFGF自身抗体水平升高现象,加强抗bFGF自身抗体水平测定有助于AID的诊断,值得推广应用。     

BXSB小鼠血清抗DNA抗体水平及其对巴细胞转化的影响

用酶联免疫吸附剂测定法(ELISA)和3H-胸腺嘧啶核苷掺入法,观察一种自发性系统性红斑狼疮(SLE)好发小鼠(BXSB小鼠)在不同月龄时,血中抗DNA抗体水平的变化,并观察该血清对正常LACA小鼠淋巴细胞转化的影响.结果显示,自3月龄起,BXSB雄性鼠血中的抗DNA抗体水平开始升高,4月龄上升至高峰,5月龄仍保持较高水平,各月龄BXSB血清与对照血清相比,P<0.05(ANOVA).同时,该血清对正常LACA小鼠淋巴细胞转化也有抑制作用,以4月龄血清的抑制作用最大.如果将同一只小鼠血清抗DNA抗体水平的变化与该血清对淋巴细胞转化的影响做回归和相关分析,发现两者有明显的正相关关系(r=0.232,n=53, P<0.05).提示雄性BXSB小鼠可能从3月龄开始表现SLE.发病后,血清中产生某种能够抑制淋巴细胞转化的物质,它的产生与抗DNA抗体有平行关系.     

胎盘部位滋养细胞肿瘤免疫组织化学分析

目的 :探讨免疫组织化学在胎盘部位滋养细胞肿瘤 (PSTT)病理诊断中的意义。方法 :应用免疫组织化学检测 8例PSTT中HPL、HCG及PCNA表达 ,并与胎盘部位过度反应及绒毛膜癌比较。结果 :HPL染色 8例PSTT均为阳性 ,阳性细胞指数为 6 2 2 5 %± 10 95 % ,高于绒毛膜癌的 11 13%± 9 2 6 %和EPS的 40 30 %± 2 3 90 % (P <0 0 1,P <0 0 5 ) ;6例PSTT细胞HCG染色阳性 (75 % ) ,阳性细胞指数为 10 87%± 8 31% ,EPS为 2 5 0 %± 4 40 % ,绒毛膜癌为 5 3 0 0 %± 2 2 10 % ;PSTT增殖细胞指数为 31 2 5 %± 8 86 % ,EPS为 10 37%± 5 2 8% ,绒毛膜癌为 71 10 %± 5 12 %。PSTT的HCG和增殖细胞指数与EPS和绒毛膜癌相比差异均有显著性 (P <0 0 1)。结论 :免疫组织化学检测HPL、HCG及PCNA对PSTT的诊断和鉴别诊断具有重要意义 ,但需结合临床与常规病理形态     

不同月龄大鼠垂体前叶滤泡星形细胞S100蛋白的表达

目的:观察不同月龄大鼠垂体重量、垂体前叶3类细胞比例及滤泡星形细胞(FS细胞)S100蛋白表达的形态学变化。方法:选取2~3月(青年组)、10~12月(中年组)、18~20月(老年组)3组大鼠垂体经分析天平称重后,行HE染色和免疫组化ABC法观察垂体前叶S100蛋白的表达。结果:中年组垂体重量最大;垂体前叶嫌色细胞(其中多为滤泡星形细胞)所占比例以中年组最高(39.26%±0.10%);3年龄组S100蛋白阳性细胞的分布方式不同;细胞阳性率以中年组最高(27.13%±0.16%),与其它两组比较有显著差异。结论:中年组大鼠垂体重量、垂体前叶FS细胞的数量、分布与其它两组比较变化最大。在大鼠垂体前叶增龄过程中,FS细胞发挥着重要作用。     

SHR大鼠血压升高前后脑内VEGF的变化及其意义

为探讨自发性高血压大鼠(SHR)血压升高前后脑内血管内皮生长因子(VEGF)表达水平的变化及其意义,本研究应用免疫组化SP法及计算机图像分析技术,对幼年(6周龄)和成年(12月龄)SHR大鼠及幼年(6周龄)和成年(12个月龄)同品系正常血压对照组京都Wistar大鼠(WKY)脑切片VEGF蛋白的表达水平进行了定量分析。结果发现在4组大鼠大脑皮质的广泛区域、纹状体、室管膜上皮及脉络丛等处均观察到散在分布的VEGF样免疫阳性细胞,阳性反应产物呈棕色颗粒状,位于胞浆中,其中幼年和成年WKY及幼年SHR三组间阳性细胞数及其灰度差异无显著性(P>0.05),而12月龄SHR组阳性细胞数及其灰度均高于其它三组(P<0.05)。以上结果提示:随着成年SHR大鼠血压的升高,脑内VEGF蛋白的表达水平上调,高表达的VEGF可能对高血压时脑缺血缺氧诱导的血管内皮细胞凋亡及微血管的存活有保护意义,同时也可能参与了高血压所致的脑损伤。     

老年人T细胞膜型CD28及血清中可溶性CD28的表达及其意义

分析老年人(年龄在60岁以上)外周血T细胞膜型CD28(mCD28)及血清中可溶性CD28(sCD28)的表达水平,探讨该分子在老年人免疫功能改变中的意义。分别收集肝素抗凝及不抗凝的老年人外周血,抗凝血经分离获取单个核细胞,间接免疫荧光及流式细胞仪分析,老年人T细胞mCD28的阳性表达率为男性(41.56%±11.23%)及女性(42.97%±12.02%)。与年轻人[男性(61.51%±17.45%)及女性(62.38%±16.22%)]相比,差别具有显著性(P<0.01)。酶联免疫吸附法(ELISA)检测,老年人血清中sCD28的含量为男性(1.08±0.45)及女性(0.98±0.47)ng/ml,与青年人[男性(0.55±0.16)及女性(0.52±0.13)ng/ml]相比,差别具有显著性(P<0.01)。老年人T细胞mCD28的表达水平下降,而血清中sCD28的含量升高。     

Detection of circulating toxocaral antigens in dogs by sandwich enzyme-immunoassay.

This study describes the presence of circulating toxocaral antigens (CTA) in the sera of dogs infected with Toxocara canis (T. canis) by using a sandwich enzyme-immunoassay (SEIA). A specificity of this assay with different antigens was observed, i.e. the EIA values, which express the antigen concentration, of excretory-secretory antigen from T. canis larvae were higher than those of other antigens (Ascaris lumbricoides, Dirofilaria immitis and Fasciola hepatica). The variability in intra-assay was below 10%. In age distribution of CTA levels, the highest level was observed at 1 month of age. Thereafter, the levels decreased gradually until 6 months of age and then the same levels were maintained until adult age. Also, slightly elevated levels were found in the sera of foetuses. A significant correlation was obtained between age and CTA levels. The positive correlation between the number of worms and CTA levels was significant. As for the IgG, IgM and IgA antibodies, a significant correlation was observed between the IgM antibody activities and CTA levels, but this was not observed with IgG and IgA antibodies. From these results, it was indicated that the immunological response to T. canis infection in dogs may not be reached until 1 or 2 months after birth, although detectable CTA levels were observed in foetal and early life. It was also suggested that the immunological stimulation for canine toxocariasis may be maintained by the excretory-secretory materials from the larvae through life and as a result, IgM antibody production may be observed even in chronically infected adult dogs. The SEIA technique reported in this study may be useful as a diagnostic tool of human toxocariasis, since the CTA can be directly demonstrated by the technique.     

Natural antibodies to treponemal antigens in four strains of guinea-pigs.

A total of 185 serum samples obtained from healthy male and female guinea-pigs of inbred strains 2 and 13 and outbred strains C4D and Hartley A were examined for natural antibodies to treponemal antigens by ELISA using Treponema pallidum (TP), T. phagedenis biotype Reiter (TR) and T. vincentii (TV) antigens and by the FTA test. The prevalence and titres of natural antibodies depended on the age and strain of guinea-pig and the treponemal antigen used. One- and 7-day-old guinea-pigs contained significantly (P less than 0.001) higher levels of natural antibodies than did animals 1 or 3-6 months old. The similar high levels of natural antibodies in newborn guinea-pigs and their mothers (12-30 months old) and the sharp drop observed at the age of 1 month suggested maternal transfer as the mechanism of acquisition. In young adults 3-6 months old, the age group most susceptible to TP infection, antibodies to TP and TR were at their lowest levels, but antibodies reacting to TV had already begun to rise. Natural antibodies were of the IgG1 and IgG2 but not of the IgM class. The highest levels of natural antibodies were in the C4D guinea-pigs; the lowest were in the Hartley A strain. Natural antibody activity was inhibited or adsorbed by TR antigens.     

Naturally Occurring Immune-Complex Glomerulonephritis in Monkeys (Macaca irus): I. Light, Immunofluorescence and Electron Microscopic Studies

Light, immunofluorescence and electron microscopic studies were carried out on renal biopsies from 32 randomly selected adult monkeys (Macaca irus). Histopathology was limited to glomeruli and consisted of mild to moderate segmental increases in mesangial cells, mesangial matrix, and/or glomerular basement membrane (GBM) thickness in 41% of the animals. Granular deposits of IgM were present in the mesangial region and along the GBM in 72% of the monkeys, whereas IgG, C1q, C4 and C3 were detected in approximately 30%. Electrondense deposits were seen predominantly in epithelial foot processes adjacent to the GBM and, to a lesser extent, in the mesangium. Those monkeys with the heaviest IgM deposition were found to have decreased serum levels of C3, IgM and IgA. Follow-up biopsies over a period of 3 to 11 months revealed that the disease process was persistent yet nonprogressive. No correlation with age or sex was noted. All animals examined were clinically healthy and had normal renal function. This is the first documented occurrence of spontaneous immune-complex glomerulonephritis in a large monkey population. It appears to be a persistent disease which does not progress to renal insufficiency and which may serve as an investigative model for mild nonprogressive forms of human glomerulonephritis.     

Kinetics of specific IgA, IgD, IgE, IgG, and IgM antibody responses in rubella

Rubella-specific IgD and IgE antibodies were determined with a solid-phase enzyme immunoassay using enzyme-labeled heavy-chain specific anti-immuno-globulins, and the antibody responses in rubella infection were compared to IgM, IgA, and IgG antibodies. IgD and IgE antibodies increased rapidly after the onset of infection, remained at a high level for at least 2 months, and declined slightly by 6 months. In comparison, the IgM antibodies decreased more rapidly, whereas the IgG antibodies persisted longer at a steady level. By 6 months the mean levels of the different antibodies had declined from their maximal mean levels as follows: IgM, 52%; IgA, 42%; IgE, 35%; IgD, 29%; and IgG, 8%. Thus IgD and IgE antibodies, in spite of their known short half lives, persisted longer than IgM and IgA antibodies, which limits their diagnostic value. The IgA antibody responses were found too variable to substitute for IgM antibody determination in diagnosis of a recent rubella virus infection from a single serum specimen. Comparison of maternal and cord blood sera indicated that, in addition to IgG antibodies, rubella-specific IgD antibodies were found to cross the placenta.     

REACTION OF COMPLEMENT WITH ENDOTHEUAL CELLS IN A MODEL OF XEN0TRANSPLANTATION

We review our studies on the role of complement (C) as mediator of xenograft hyperacute rejection using an in vitro model consisting of porcine endothelial cells as target and human serum as source of natural antibodies and C. Cytotoxicity of endothelial cells required IgM antibodies to porcine endothelial cells, and the classical pathway and membrane attack complex of C. These findings correlated with in vivo results of porcine organs transplanted into rhesus monkeys, which showed a) co-deposition of IgM, C3, C4 and C9, along blood vessels of rejecting organs, with trace deposits of factors B or P, and b) minimal deposition of IgM and C components in transplants with prolonged survival that were performed in rtiesus monkeys depleted of natural antibodies but with normal C levels. Human serum causes activation of porcine endothelial cells manifested by release of heparan sulfate proteogiycan. Heparan sulfate release was induced by C5a alone. A new approach to avert xenograft hyperacute rejection was tested. To inhibit cytotoxicity of porcine endothelial cells by human C, the membrane-associated C inhibitor decay-accelerating factor (DAF) of human origin was incorporated into endothelial cells. Human DAF was able to efficiently inhibit C-mediated killing of porcine endothelial cells, suggesting that the use of DAF and other C inhibitors could be used to interfere with C-mediated xenograft hyperacute rejection.     

微循环荧光造影的安全性及其控制

用围堤式颅窗软脑膜微循环观察方法探讨了微循环荧光造影的安全性。发现：５０Ｗ超高压汞灯的激发光（波长峰值４９０ｎｍ）照射９０ｍｉｎ对血管内皮和平滑肌均有损伤作用。可使微动脉对乙酰胆碱和硝普钠的扩张反应减弱（血管内径变化分别由３７．５％±７．９％降至２３．８％±１１．９％，由４１．１％±２０．８％降至３６．７％±１１．１％，（Ｐ＜０．０５）。单独注射荧光素钠对血管反应性无影响。在激发光持续照射下注射５％荧光素钠０．２ｍｌ进行微循环荧光造影６次（１次／１５ｍｉｎ），血管明显损伤，微动脉对乙酰胆碱和硝普钠的扩张反应分别由４３．５％±１６．９％降至２２．０％±１３．３％，５５．４％±１７．９％降至２５．１％±１４．０％，Ｐ＜０．０５，且伴有严重的血栓形成。认为：控制激发光照射时间以及荧光素钠的浓度和剂量，可以防止上述损伤的发生。     

Loss of NMDA, but not GABA-A, binding in the brains of aged rats and monkeys.

In this quantitative neurochemical study we investigated age-related changes in the GABAergic, glutamatergic, and cholinergic neurotransmitter systems in rats and rhesus monkeys. Sixteen young (5 months) and 20 aged (24 months) rats and seven "young" (4-9 years), six "adult" (20-25 years), and five "aged" (29-34 years) monkeys were studied. NMDA-displaceable 1-[3H]glutamate binding was significantly decreased in many neocortical and subcortical regions examined in aged rats and monkeys. The level of choline acetyltransferase (ChAT) activity and [3H]muscimol binding were unchanged in aged animals.     

重症肌无力患者外周血CD5~+B细胞和CD4~-CD8~-T细胞的变化

研究重症肌无力 (MG )患者外周血CD5 + B细胞和CD4 CD8 T细胞的变化 ,以探讨这两种细胞与MG的关系。采用流式细胞仪分析MG患者和对照组外周血中CD5 + B细胞和CD4 CD8 T细胞的频率 ,同时以ELISA方法检测这些患者的血清AchR、PsmR抗体水平。结果 :2 8例MG患者的CD5 + B细胞为 19 75 %± 10 8% ,高于对照组的 15 4 %± 9 6 7% (P <0 0 1) ;胸腺未切除MG患者的CD5 + B细胞为 2 2 31%± 7 4 7% ,显著高于对照组 (P <0 0 0 1) ;两种抗体阳性MG患者的CD5 + B细胞为 2 4 96 %± 13 1% ,显著高于对照组 (P <0 0 0 1) ;以上各组MG患者的CD4 CD8 T细胞与对照组均无显著区别 ;两种抗体阴性组的CD5 + B细胞和CD4 CD8 T细胞亦与对照组均无显著区别 ;两种抗体阴性组的CD5 + B细胞和CD4 CD8 T细胞亦与对照组无明显差异。本研究提示MG患者外周血的CD5 + B细胞频率增高 ,与胸腺切除与否以及突触前后膜抗体的阳性程度密切相关 ,而CD4 CD8 T细胞是否与MG有关还需进一步研究证实。     

糖皮质激素吸入对哮喘患儿外周血CD4+CD25+ 调节性T细胞水平的影响

目的： 探讨糖皮质激素吸入对哮喘患儿外周血CD4+CD25+ 调节性T细胞（Tr）水平的影响。方法：采用糖皮质激素吸入治疗70例发作期哮喘患儿,应用流式细胞术检测患儿外周血的Tr细胞数。结果：糖皮质激素规则治疗后患儿外周血CD4+CD25+Tr水平（7.05%±1.61%）明显高于治疗前（5.62%±1.29%）,P<0.01。完全控制组患儿外周血CD4+CD25+Tr水平最高（7.56%±1.88%）,部分控制组患儿次之（7.09%±1.23%）,控制不佳组患儿最低（6.11%±1.96%）,差异均显著,P<0.05。经激素规则治疗的患儿外周血Tr水平（7.05%±1.61%）明显高于不规则治疗组患儿（5.91%±1.76%）,P<0.01。结论：规则使用糖皮质激素吸入疗法可明显提高哮喘患儿外周血Tr水平,Tr水平与哮喘的激素治疗效果有关。