小鼠膜性肾小球肾炎复制方法及免疫荧光定量研究

目的：介绍实验性小鼠膜性肾小球肾炎(MGN)的复制方法，并探讨其免疫荧光定量分析在肾小球肾炎研究中的应用价值。方法：制备阳离子化牛血清白蛋白(GBSA)并复制小鼠MGN，对各组小鼠进行电镜及免疫荧光观察，并进行免疫荧光定量研究。结果：电镜、免疫荧光观察均显示病理Ⅰ组(PⅡ组)具有典型的MGN病变，病理Ⅱ组(PⅡ组)病变轻微且不典型。免疫荧光定量研究证实PⅠ、PⅡ组与对照组差异有显著性；PⅠ、PⅡ组间差异无显著性。结论：C—BSA可作为复制小鼠MGN的良好抗原，隔日2mg／只尾静脉注射4w即可复制出稳定的小鼠MGN模型。免疫荧光定量分析不仅能直接而准确地反映MGN病理变化，而且在MGN早期即具有诊断价值。     

Identification of Babesia bigemina infected erythrocyte surface antigens containing epitopes conserved among strains

The presence of previously uncharacterized antigens (new antigens) on the surface of intact erythrocytes infected with three strains of Babesia bigemina from Kenya and one each from Puerto Rico, Mexico, St. Croix, and Texcoco-Mexico was demonstrated by indirect immunofluorescent antibody (IFA) reactions. These antigens were not strain specific because antibodies in bovine immune serum to either the Mexico or Kenya isolates reacted with all seven strains tested. Homologous and heterologous immune serum antibodies bound a maximum of 83% and 55%, respectively, of intact erythrocytes infected with the Kenya-Ngong strain but not uninfected erythrocytes. Both sera caused agglutination of only infected erythrocytes. Antibodies eluted from the surface of glutaraldehyde (0.25%) fixed infected erythrocytes had IFA reaction patterns among strains similar to those of immune sera before elation. Eluted antibodies were used to determine if these antigens were protein and encoded by B. bigemina. Eluted antibodies bound seven parasite-encoded proteins of 240, 220, 66, 62, 58, 52 and 38 kDa in an erythrocyte surfacespecific immunoprecipitation reaction of ³⁵-methionine labelled proteins. It was concluded that the surface of B. bigemina infected erythrocytes had parasite-encoded proteins and that these proteins had surface exposed epitopes that were conserved among the seven strains examined which were from two continents.     

Protective immunity to malaria: studies with cloned lines of Plasmodium chabaudi chabaudi in CBA/Ca mice. III. Protective and suppressive responses induced by immunization with purified antigens

The protective effect of affinity purified antigen has been investigated in an experimental model for malaria which shows a well marked recrudescence of parasitaemia, a feature of the disease in man. A monoclonal antibody (MoAb) recognizing an epitope common to two genetically distinct cloned lines of Plasmodium chabaudi (AS and CB), was used to purify a Mr250,000 polymorphic schizont antigen (PSA) from these parasites. The purified preparations were then examined for the presence of specific and cross-reactive epitopes by immunoprecipitation with a panel of MoAb raised against P. chabaudi AS. When tested previously on smears of parasitized blood by immunofluorescence, or against lysates of parasitized erythrocytes by immunoprecipitation, most of these MoAb had been found to be AS specific. When either AS or CB affinity purified Mr250,000 PSA was used as the target, these same MoAb immunoprecipitated both antigens, and in some cases, a number of associated polypeptides (AP) which copurify with the Mr250,000 PSA. Subsequently, mice were immunized with either the purified AS or CB antigens in Freund's complete adjuvant (FCA). Prechallenge sera were compared by indirect immunofluorescence and immunoprecipitation. Sera from mice immunized with AS antigen reacted strongly with AS and cross-reacted with CB parasite preparations. Pre-challenge serum from CB antigen immunized mice reacted well with CB, but only faintly with AS preparations. In mice immunized with the AS antigen and then challenged with either AS or CB parasites, the initial parasitaemias were delayed in appearance and the height of the peak parasitaemia reduced, an effect which was most pronounced after challenge with homologous parasites. Only homologous challenge of the mice immunized with CB antigen produced statistically significant modification of the initial parasitaemia. In the immunized mice challenged with homologous parasites, the delayed appearance and slightly reduced peak of the primary parasitaemia was associated with delayed resolution of the patent parasitaemia and significant enhancement of the recrudescence.     

The correlation of ureaplasma urealyticum infection with infertility

Summary. A prevalence study of Ureaplasma urealyticum (UU) infection of the male genital tract was carried out in Shanghai between March 1992 and June 1995. Significantly higher frequency of UU infection was found among infertile males (549/1416) as compared to fertile controls (34/375). Examination of 8 specimens each from infertile men and fertile subjects by electron microscopy, immunogold and immunofluorescence techniques, demonstrated adhesion of Ureaplasma urealyticum to the membrane of spermatozoa and exfoliated germ cells. In addition, gold particles on Ureaplasma urealyticum were found to be adhered to the sperm surface in 4 of the 8 samples. Strong specific anti-UU fluorescence was detected in 6 of 8 samples, mainly on the midpieces and post-acrosomal regions of the spermatozoa.
To further study the effects of Ureaplasma urealyticum on fertility, 47 male Sprague-Dawley (SD) rats were infected artificially with Ureaplasma urealyticum serotype 8 (T960). Morphological changes in the seminiferous tubules were observed 3–5 weeks after inoculation in the sacrificed animals. Dramatic impairment of spermatogenesis of both testes was found in 11 rats. Mating experiment confirmed infertility in 12 of 40 rats. Offsprings of the infected rats were significantly smaller than those of controls in terms of prenatal weights and birthweights.     

抗中性粒细胞胞浆抗体在肾小球疾病中的检测及临床意义

目的:检测各种肾小球疾病患者血清中抗中性粒细胞胞浆抗体(ANCA)并探讨其临床意义。方法:应用间接免疫荧光法(IIF)检测20例正常人及131例各种肾小球疾病患者血清中ANCA。结果:131例患者中有11例ANCA阳性性(阳性率为8.4％),且均为P-ANCA阳性,而20例正常对照组无1例阳性(P<0.05)。ANCA阳性中以新月体性肾炎、狼疮性肾炎、紫癜性肾炎的阳性率较高,分别为67％、25％、16.7％。新月体性肾炎与IgA肾病等原发性肾小球疾病组比较有显著性差异(P<0.05)。结论:ANCA在各种肾小球疾病中并不少见,尤其是新月体性肾炎、狼疮性肾炎、紫癜性肾炎的阳性率较高,推测ANCA可能在这些疾病的发病机理中起了一定的作用。     

Localization of nerve-specific protein antigens on the surface membrane of neurons and glial cells of Helix pomatia

The existence of cross protein antigens common to several species of invertebrates and vertebrates on the membrane of neurons and glial cells of Helix pomatia was demonstrated in vitro by Coons' immunofluorescence method. The presence of nerve-specific protein S-100 on the membrane of these cells was established. The antigenic heterogeneity of membranes of a population of neurons also was observed. Differences were found in the concentrations of antigens on the somatic and axon membranes. The character of distribution of specific fluorescence indicates possible qualitative and (or) quantitative differences in the content of nervespecific proteins in different areas of the neuron membrane.Laboratory of Central Mechanisms of Regulation and Control, Institute of Clinical and Experimental Medicine, Siberian Branch, Academy of Medical Sciences of the USSR, Novosibirsk. (Presented by Academician of the Academy of Medical Sciences of the USSR V. P. Kaznacheev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 88, No. 9, pp. 299–301, September, 1979.     

Monoclonal antibodies against human granulocytes and myeloid differentiation antigens

Monoclonal antibodies (MCA) were obtained by immunizing BALB/c mice with 99% pure granulocytes from normal donors or with a whole leukocyte suspension obtained from a chronic myelogenous leukemia (CML) patient, and then fusing the mouse spleen cells with a 315–43 myeloma cell clone. Four MCA were selected and studied using ELISA, immunofluorescence, cytotoxicity assays, and FACS analysis. Antibodies 80H.1. 80H.3. and 80H.5 (from normals) and 81H.1 (from CML) detected antigens expressed on neutrophils. Antibodies 80H.1 and 80H.3 (lgG) also reacted with monocytes but not with other blood cell subsets. Antibodies 80H.5 and 81H.1 (lgM) were cytotoxic and reacted strongly with most of the cells of the neutrophil maturation sequence. i.e., myeloblasts, promyelocytes, myelocytes, and mature granulocytes. Antibodies 80H.5 and 81H.1 also inhibited BFU-GM and CFU-E. Antigens recognized by 80H.3. 80H.5, and 81H.1 were expressed both on a proportion of cells from HL.60, KG.1, ML.1, and K562 myeloid cell lines, and on a proportion of blast cells isolated from patients with acute myelogenous leukemia. They were not found on lymphoid cell lines or lymphoid leukemia cells. These MCA recognize either late differentiation antigens expressed on mature neutrophils and monocytes (80H.1 and 80H.3) or early differentiation antigens (80H.5 and 81H.1) specific to the granulocytic lineage. They may be useful for a better definition of those antigens specific to hematopoietic stem cells and their relationship with normal or neoplastic hematopoiesis.     

PENA方法的建立及与ELISA IgM检测CMV的比较

目的　介绍一种敏感、稳定、快速、简便的实验室检测CMV的方法 ,同时探讨该种新方法与ELISA检测CMV方法的优、缺点。方法　对 5 5 2例病人应用间接荧光免疫法测定细胞核中的特异病毒早期抗体 (PENA)和ELISA法测定IgM抗体。结果　PENA方法 :强阳性 88例 ,阳性率 15 4 9% ,弱阳性 2 73例 ,阳性率 5 9 4 6 % ;ELISA -IgM方法 :阳性 34例 ,阳性率6 16 %。结论　PENA方法操作简便 ,与ELISA方法相比较 ,可对CMV感染进行早期测定及诊断 ,并可区分既往感染和即时感染 ,具有敏感性和稳定性 ,是测定小儿CMV感染的一种较好的方法     

Analysis of thymic epithelial cell proliferation in vitro by combining bromodeoxyuridine and keratin labeling in an immunofluorescence assay

A simple method of analyzing thymic epithelial cell (TEC) proliferation has been developed by combining bromodeoxyuridine (BrDU) and keratin labeling in an immunofluorescence assay. The first reagent specifically visualizes the cells entering the S phase of the cell cycle, whereas the second immunostaining reveals which of the proliferating BrDU-positive cells actually belong to the epithelial lineage. This method, besides being rapid and free of radioactivity, appears to be reliable in view of the minor variations in the percentages of BrDU+ TEC observed in several distinct experiments. Thus, BrDU/keratin immunolabeling appears to represent a useful tool for the analysis of in vitro TEC proliferation.     

Demonstration of T4, T8, M1 and B7 determinants on human T cells with a rosette test: implications for the assay specificity of monoclonal antibodies

Under optimal test conditions significantly more freshly isolated human T cells reacted with OKT4, OKT8, OKM1 and OKB7 monoclonal antibodies (Mabs) in the indirect antiglobulin rosetting reaction (IARR) than by indirect immunofluorescence. Rabbit erythrocytes (E) coated with anti-mouse immunoglobulin were more sensitive indicator cells in the IARR than similarly coated sheep E. Treatment of T cells with neuraminidase further enhanced T cell reactivity in the IARR with each Mab so that an average of 60% or more of T cells were T4+, T8+ and M1+ and at least 40% had the T4+ T8+ phenotype. The various findings suggest that the rosette assay detects determinants on T cells that are expressed below the detection threshold of immunofluorescence. Moreover, these findings indicate that the cellular specificities of a particular Mab may change when one assay system is substituted for another or when the protocol of a particular assay is altered.