恙虫病立克次体补体结合抗原的研究

本文介绍应用兔睾丸单层细胞制备恙虫病立克次体补体结合抗原的方法,并初步探讨了影响鸡胚卵黄囊膜抗原效价的一些因紊。鸡胚卵黄囊膜抗原产量大、效价高、特异性较好。但有些恙虫病立克次体株在鸡胚中不易繁殖。而在组织培养中很易繁殖,第一代即能获得大量立克次体,有利于同时制备多株恙虫病立克次体抗原。本文同时介绍了制备豚鼠及家兔免疫血清的简便方法。     

鸡胚卵黄球能否演变为胚胎细胞?

Ο.Б.勒柏辛斯卡娅认为跌入于胚下腔中的卵黄球能够演变为胚胎的内胚层细胞和进入于内外二胚层之间的卵黄球能够演变为血岛的这一假说近年来在细胞学和胚胎学界引起了激烈的争论. 我们从1954年春天开始研究这个问题,工作可分三方面:(1)分别观察鸡胚正常发生过程中的内胚层和血岛的形成过程和卵黄球的变化——包括鸡胚和鸭胚从未孵到孵育20天各期的胚胎和卵黄囊切片,采用各种主要的细胞学上的染色方法和卵黄球的活体染色观察.(2)卵黄球的离体和活体培养——培养材料包括未孵及孵育各期的胚盘下卵黄球.离体培养     

用胰酶消化法浓缩纯化恙虫病立克次体

用不同感染材料和方法进行了浓缩纯化恙虫病立克次体的试验。结果证明以0．25％的胰蛋白酶消化感染鸡胚卵黄囊膜,可获得形态完整的立克次体;浓缩纯化的立克次体保持活性;用于补体结合试验,抗原滴度可达1：128。经冰冻干燥保存后,仍能保持其形态的完整性。     

冻干对恙虫病立克次体抗原性的影响

以含恙虫病立克次体的鸡胚卵黄囊膜材料冻干后,立克次体对小白鼠的ID,。平均下降1．93个对数,但若以鸡胚细胞培养材料进行冻干,则冻干后恙虫病立克次体的抗原性明显遭到破坏,对小白鼠的ID50 平均下降2.73个对数。冻干后的卵黄囊膜材料经4℃冰箱保存六个月至一年,对小白鼠的ID50,一般再下降一个对数左右。     

ELISA法检测鸡胚中抗生素残留量的研究及其应用

用ELISA法测定氨苄青霉素、氯霉素在流感疫苗生产用海兰白鸡鸡胚中的残留。实验通过比较空白组0d龄全鸡胚和给药组0d龄全鸡胚中氨苄青霉素、氯霉素的残留量,说明了饮水给药的方法能够建立抗生素残留的动物模型。通过比较10d龄鸡胚尿囊液、卵黄中氨苄青霉素、氯霉素的残留量,说明尿囊液中的残留量明显高于卵黄。通过绘制残留曲线,可以看出氨苄青霉素、氯霉素在海兰白鸡体内蓄积迅速,却消除缓慢。通过实验初步摸索出了氨苄青霉素和氯霉素在鸡胚中的分布、代谢及残留规律,获得了部分生产用鸡胚尿囊液的抗生素残留量数据,为进一步控制流感疫苗生产用鸡胚质量提供技术保障。     

恙虫热立克次体(宫崎地区分离株)在BSC—1细胞中的增殖

恙虫热立克次氏体的致病性有多种类型。从宫崎地区的恙虫病患者中分离出的立克次氏体对小白鼠致病性弱,小白鼠虽可感染发病,但不死亡,很难在腹腔内细胞或脾中发现立克次氏体。在一般恙虫热立克次氏体的研究中常用的鸡胚卵黄囊和细胞培养法中几乎不繁殖,这就使立克次     

造血干细胞发生学研究——Ⅰ.人胚肝造血干细胞

人卵黄囊的血管内造血已被细致地观察过。卵黄囊的造血停止,肝成为主要的造血器官。肝造血持续到胎生7个月为止。Fukuda,T.在电镜观察中看到了肝造血中出现未分化的单个核细胞,提出预期的干细胞(presumptive stem cell)一词。近来有人对人胚造血提出卵黄囊、肝及骨等组织的造血三阶段。国内进行了胚肝扩散盒培养细胞的电镜观察,开展了各种因素对造血干细胞的调控研究。迄今为止对造血干细胞的发生学尚乏定     

小鼠胚胎黄囊中的造血干细胞

本文比较了卵黄囊造血干细胞与胎儿的肝、脐血管和成人骨髓中的造血干细胞之间的区别,揭示了卵黄囊造血干细胞向各系细胞分化的过程,以及细胞因子、卵黄囊内皮细胞对其增殖,分化的影响。卵黄囊造血干细胞有很强的增殖,分化能力,且不表达组织相容性复合体抗原,故可运用该细胞促进机体造血功能的恢复。     

胚胎发育早期接种人血清白蛋白诱导鸡的免疫耐受

外源基因的引入及表达常会引发宿主动物强烈的免疫应答,导致蛋白产量的下降或引起炎症反应。本研究试图通过在胚胎发育早期接种异源抗原诱导动物产生免疫耐受来解决这些问题。为了诱导鸡产生免疫耐受,将人血清白蛋白（Human serum albumin,HSA）通过胚胎血管微注射的方式接种到发育65—67h的鸡胚血管中,接种剂量为50μg;通过卵黄注射的方式接种到发育3—7d的鸡胚卵黄中,接种剂量为100μg。孵出的小鸡在3周龄时按照常规免疫方法再次接种同种抗原,收集不同时期的血清样本,用酶联免疫吸附试验（Enzyme-linked immunosorbent assay,ELISA）检测血清中抗-HSA抗体水平。结果表明,两种接种方式均能诱导小鸡产生免疫耐受：在65-67h的胚胎中通过血管微注射法接种抗原诱导小鸡产生免疫耐受的比率为64．52％;通过卵黄注射接种抗原诱导小鸡产生免疫耐受的最佳接种时间为发育的第6d,第5、7d接种对后期血清中抗-HSA抗体形成也有一定抑制作用,但是维持耐受的时间较短,第3、4d接种抗原对诱导小鸡免疫耐受的效果不明显[动物学报51（5）：845—851,2005]。     

鸡胚胎期蛋白营养吸收的研究

禽类的胚胎发育,主要在蛋内完成,其受精蛋是一个独立的自给自足的胚胎发育单位。胚胎发育的营养来源,除气体外均由蛋内供给。关于鸡胚胎期,鸡胚营养吸收的途径问题,派登(Patten)曾作过概略性的阐述,认为主要靠循环方式完成的。循环有3:即卵黄囊循环、尿囊     

New serotypes of Morganella morganii.

On the basis of 8 new O and 11 new H antigens determined in 22 strains, the Morganella morganii antigenic schema was supplemented with 8 serogroups (O35-O42) and 13 serotypes. Four strains belonged to O groups described earlier and 2 strains contained new O antigens in combination with known O antigens. Known H antigens were present in one strain as a single factor and in one strain as combination of two factors. New H antigens were demonstrated in 5 serotypes in combination with known H antigens. Six out of the 22 isolates were classified into O group 35. Two isolates contained different B-type surface antigens; these factors were not related to Escherichia coli B antigens and, unlike the latter, their living suspension gave a higher titre agglutination in OK serum as compared to the boild culture.     

Alternative antigens reduce cross-reactions in an ELISA for the detection of Salmonella enteritidis in poultry

Two alternative antigens for the use in detection of antibodies to salmonellas were investigated: firstly, lipopolysaccharide (LPS) from members of the D2 group, having antigens O : 9, 46, and flagella antigens. Whereas LPS from the D2 group did not discriminate sufficiently with control sera, flagella antigens reacted specifically with antibodies directed to serotype specific H antigens. When flagella antigens were used to screen sera from birds of commercial flocks, however, cross-reactivity between flagella antigens was observed. When both LPS and flagella antigens were used to screen sera from chickens infected with Salmonella, enteritidis, the sera gave higher titres with flagella antigens during the early stages of infection, and titres with flagella antigens dropped earlier after infection had ended than titres with lipopolysaccharide.     

Dirofilaria immitis: identification and partial characterization of parasite antigens in the serum of infected dogs

We have recently developed a sensitive and specific immunodiagnostic test for canine Dirofilaria immitis infection based on detection of soluble parasite antigens in dog sera by monoclonal antibody-based enzyme immunoassay. In addition to their importance as markers of infection, these antigens may contribute to the pathogenesis of heartworm disease in dogs. In the present study, a variety of methods were used to identify and characterize circulating D. immitis antigens. Two antigens were identified in infected dog sera that formed lines of identity in rocket-line immunoelectrophoresis with soluble antigens extracted from adult D. immitis. Circulating D. immitis antigens were also demonstrated in infected dog sera by immunoblot analysis with polyclonal and monoclonal antibodies. These antigens had apparent molecular weights that ranged from 50 to 250 kDa. Most of the circulating D. immitis antigens contained the epitope defined by monoclonal antibody 1418BF2.1 which is used in our enzyme immunoassay for circulating D. immitis antigen. Studies of parasite antigens released during in vitro culture indicated that the circulating D. immitis antigens in dog sera that are detected by our enzyme immunoassay are primarily derived from adult female worms.     

Partial purification and characterization of mold antigens commonly found in foods.

Rapid methods are needed for detection of molds in foods; therefore, an enzyme-linked immunosorbent assay was developed. The extracellular and mycelial antigens for Mucor, Aspergillus, Cladosporium, and Geotrichum species were partially purified and characterized. The molecular masses of the mycelial and extracellular antigens, as determined by size exclusion chromatography, ranged from 4.5 x 10(5) to 6.7 x 10(5) Da. There was only one main antigenic peak separated by Sepharose CL-4B and concanavalin A-Sepharose columns for Mucor, Cladosporium, and Geotrichum mycelial and extracellular antigens, but there were two for Aspergillus mycelial antigens and three for Aspergillus extracellular antigens. These antigens contained 10 to 50% protein which was part of the active site since protease digestion significantly decreased antigenic activity. Neutral sugars, ranging from 13 to 75%, made up the rest of the active site, and < 1% phosphate was detected in mycelial antigens. Geotrichum, Cladosporium, and Aspergillus antigens contained mainly glucose, galactose, and mannose. Mucor antigens contained these sugars plus fucose. The percentage of sugars differed between the mycelia and extracellular antigens. Enzymatic digestion and competitive inhibition tests using different sugar derivatives showed that galactosyl residues with beta linkages were immunodominant for Aspergillus, Geotrichum, and Cladosporium antigens and mannosyl residues with alpha linkages were immunodominant for Mucor antigens.     

Erythrocyte diagnostica made from salmonellal phagolysates]

For the first time O antigens obtained from phagolysates were proved to be suitable for use as material for the production of highly specific erythrocyte diagnostic preparations. O antigens obtained from Salmonella by two methods, i.e. phage disintegration and Grasset's method, were subjected to comparative chemical analysis and found to have no essential difference. Nevertheless, the sensitizing potency of O antigens obtained from phagolysates were experimentally shown to be 3 times greater than that of O antigens obtained by Grasset's method. The optimum sensitizing doses established in the passive hemagglutination test for O antigens obtained by both methods indicated that these antigens were highly sensitive and specific.     

ANTIGEN-ANTIBODY CROSSED ELECTROPHORESIS OF RAT BRAIN SYNAPTOSOMES AND SYNAPTIC VESICLES: CORRELATION TO WATER-SOLUBLE ANTIGENS FROM RAT BRAIN

—Antigen-antibody crossed electrophoresis has been applied to the study of rat brain synaptosomes and synaptic vesicles. Several antigens could be visualized. By comparison with previously describéd water-soluble antigens from rat brain, some of the antigens in the synaptosome and the synaptic vesicle preparations were identified; among these were antigens which have been determined as brain-specific. Furthermore, the antisera against the two subcellular fractions were compared with the anti-serum against water-soluble antigens from rat brain.     

Association of mouse adenovirus-induced cell surface antigen(s) with histocompatibility antigens

The topological relationship on the mouse adenovirus (M-Ad)-infected cell surface between virus-induced specific cell surface(s) antigens and serologically defined major histocompatibility antigens (H-2) was analyzed by the cap formation technique. Rhodamine-isothiocyanate (RITC)-labeled anti-S serum failed to stain the surface of virus-infected lymphoid cells which were pretreated with anti-H-2 serum and fluorescein isothiocyanate (FITC)-labeled anti-mouse immunoglobulin serum (anti-M-Ig) to cap the appropriate H-2 antigens. Conversely, the capping of the S antigens by pretreatment with anti-S followed by FITC anti-M-Ig serum induced cocapping of H-2 antigens. The β₂ microglobulins (β₂m) were also shown to be cocapped with S antigens by anti-β₂m or by anti-S serum. The S antigens, however, did not cocap with mouse-immunoglobulins or Thyl. 2 antigens on virus-infected B or T lymphocytes, respectively. To further elucidate the molecular relationship between S and H-2 antigens, radio-iodinated virus-infected cells were solubilized with Nonidet P40 (NP40) and S antigens were precipitated with anti-S serum. When the precipitates were analysed with sodium dodecyl sulfate Polyacrylamide gel electrophoresis, two major peaks were seen at positions of molecules of about 45,000 and 12,000 daltons both of which corresponded with molecules which were observed when NP40 extracts of virus-infected or uninfected cells were precipitated with anti-H-2 serum. Sequential immunoprecipitation analysis of infected cell extracts showed that S antigens were coprecipitated with either H-2K or H-2D antigens. These results suggest that the S antigens are somehow associated with H-2K or H-2D antigens separately.     

Immunodiffusion analysis of water-soluble rat bone marrow antigens

Antisera were obtained against six electrophoretic fractions of the rat bone marrow extract. With their help, 18 tissular antigens and 11 antigens immunologically similar to the blood serum proteins were revealed in the rat bone marrow. All tissular antigens are divided in five groups by the degree of organ specificity: 1) bone marrow organospecific antigens (4 antigens), 2) antigens present in the bone marrow, spleen and lung extracts (2), 3) "granulocytic" antigens (4), 4) antigens common for many rat organs, but not found in the extracts of blood formed elements, skeletal muscle, heart, brain, eyes (3), 5) antigens present in all the organs studied (5). The bone marrow organospecific antigens may be specific antigens of hemopoietic cell precursors. The possibility of utilization of antisera against the bone marrow water soluble proteins for labelling hemopoietic cells of different lines of differentiation is discussed.     

The release of membrane antigens into culture by adult Schistosoma mansoni.

Antigens sharing determinants with surface membranes and soluble proteins of adult Schistosoma mansoni have been detected in culture media after incubation of radioactively labelled worms. The relative quantities of these antigens were measured with specific antisera raised in rabbits and with serum from an immune rhesus monkey. It was found that 12-16% of TCA-precipitable radioactivity in the culture medium consisted of membrane antigens and 6-8% consisted of antigens sharing determinants with proteins found in the soluble fraction of adult worms. Over half the membrane antigens were present in particulate form, while other antigens were present in solution. Surface labelling the adult worms with [125I]confirmed that some of the particles in the culture medium were derived from the surface membrane of the adult worm and electron microscope examination of such particles showed that large membrane fragments were present. These results support the hypothesis that antibodies against schistosome membrane antigens are induced by particulate membrane antigens released by the parasite.     

Comparison of excretory/secretory and circulating antigens of Toxoplasma gondii by enzyme immunoassay and immunoblotting.

, , , , , and 1992. Comparison of excretory/secretory and circulating antigens of Toxoplasma gondii by enzyme immunoassay and immunoblotting. International Journal for Parasitology 22: 1083–1088. Toxoplasma gondii trophozoites (RH strain) were cultured in embryonic fibroblasts in order to study the kinetics of production of excretory/secretory antigens, and the results were compared to the production of circulating antigens in an in vivo mouse model. By capture-ELISA, excretory/secretory antigens were first detected on the fourth day of culture whereas circulating antigens were first detected 1 day after infection. Similar concentrations of antigens were detected in both models as evidenced by comparable absorbance values. By immunoblotting, the excretory/secretory antigens were also detected later compared to circulating antigens (day 4 vs day 1). Seven major polypeptides were detected in both antigen preparations, six of them having the same molecular mass (110, 75,48, 30, 24 and 22 kDa).