克拉玛依地区的利什曼病Ⅷ.白蛉人工感染大沙鼠体内利什曼原虫的实验

蒙古白蛉和安氏白蛉在叮咬了感染当地大沙鼠耳组织内利什曼原虫的BALB/c小鼠后,当白蛉的胃皿仔在时,利什曼前鞭毛体的感染率分别为37.6％和52.9％,在胃血完全消化后,则感染率下降为83％和6.9％,其原因是白蛉吸血后形成的围食膜限制了大多数蛉胃内前鞭毛体的发展;但这二种白蛉在胃血消化过程中约有1/4的围食膜可以破裂,从膜内逸出的前鞭毛体即可继续繁殖和向蛉的消化道前端移行,并且不再受到下一次吸血后形成的围食膜的限制。硕大白蛉吴氏亚种在吸了感染利什曼原虫的小鼠的血后也形成围食膜,但于吸血后第三天膜即破裂,故白蛉感染的前鞭毛体的繁殖和发展不受限制,感染率在蛉胃内有血时为62.8％,无血时为55.6％(P>0.05),无甚差异。     

亚历山大白蛉传播黑热病的研究

在新疆吐鲁番煤窑沟黑热病疫区内,亚历山大的白蛉数量大,亲人,叮咬杜氏利什曼病鼠后,前鞭毛体在蛉胃内大量繁殖并进入喙部。野外和人房内该种白蛉具有杜氏利什曼原虫的自然感染。传播试验使正常仓鼠发生内脏利什曼病,从而首次证实亚历山大白蛉可作为黑热病的传播媒介。并对煤窑沟疫区的性质,作了讨论。     

克拉玛依地区的利什曼病Ⅻ.利什曼原虫单克隆抗体对大沙鼠及白蛉体内利什曼原虫的检测

本文报道建立的抗杜氏利什曼L.donovani,抗硕大利什曼L.major,抗热带利什曼L.tropica及抗新疆克拉玛依大沙鼠体内的利什曼原虫的4种单克隆抗体(McAb),以免疫荧光法的交叉反应显示出有较强的特异性,应用McAb的dot-ELISA试验,对新疆大沙鼠体内分离的利什曼原虫及白蛉自然感染的利什曼原虫前鞭毛体进行检测,证实新疆克拉玛依鼠体、蛉体的利什曼原虫仅与当地大沙鼠体内利什曼原虫制备的McAb呈阳性反应。     

应用单克隆抗体鉴定利什曼原虫的斑点ELISA试验

本文报道用单克隆抗体鉴定自蛉体内自然感染利什曼原虫的虫种,获得较好效果,其中L9E4单克隆抗体对新疆杜氏利什曼前鞭毛体抗原的最高阳性滴度达1:655 360;对吐鲁番亚历山大白蛉自_(10)及自_(11)的阳性滴度为1:81 920,比IFA法提高3～20倍。此法敏感,可用作鉴别白蛉体内利什曼原虫虫株的一种手段.     

克拉玛依地区的利什曼病 Ⅸ.白蛉自然感染前鞭毛体的鉴定

本文报道新疆克拉玛依地区的大沙鼠洞、旷野及人房等不同场所内白蛉自然感染利什曼原虫的情况、蛉体内原虫对实验动物的致病特征以及用利什曼原虫McAb的dot-ELISA法对白蛉自然感染的利什曼前鞭毛体的检测结果;并结合4年来对当地白蛉的生态和大沙鼠体内的利什曼原虫对白蛉的感染性等一系列的调查研究,证实蒙古白蛉和安氏白蛉为克拉玛依大沙鼠体内利什曼原虫的传播媒介。     

克拉玛依地区的利什曼病XIV．硕大白蛉吴氏亚种自然感染前鞭毛体的鉴定

硕大白蛉吴氏亚种是新疆克拉玛依地区的主要蛉种之一，具有强的亲人性，在野外和居民点内常能查见该蛉有前鞭毛体的自然感染。本文结果表明，白蛉自然感染的前鞭毛体能使仓鼠及ＢＡＬＢ／ｃ小鼠发生内脏利什曼病；在感染仓鼠内脏涂片上的无鞭毛体，由蛉体而来的明显较由大沙鼠而来的都兰利什曼原虫为小；白蛉自然感染的前鞭毛体在ＮＮＮ培养基内生长不良；用￣（３２）Ｐ标记的ｇｐ￣（６３）基因为探针，与婴儿利什曼原虫、都兰利什曼原虫及白蛉自然感染的前鞭毛体的ＤＮＡ进行杂交，证实蚌体自然感染的原虫与婴儿利什曼原虫同源。克拉玛依无内脏利什曼病人，但人群中有皮肤利什曼病流行。关于硕大白蛉吴氏亚种自然感染的来源以及当地的皮肤利什曼病究竟是由都兰利什曼原虫抑或婴儿利什曼原虫所致，尚待阐明。     

溴氰菊酯药浴阳性鼠阻断内脏利什曼病传播的实验研究

用中华白蛉分别饲吸两组染有杜氏利什曼原虫的背纹仓鼠（Ｃｒｉｃｅｔｕｌｕｓｂａｒａｂｅｎｓｉｓ），一组系经澳氰菊酯药浴后９－４５ｄ的仓鼠，另一组系未经药浴的仓鼠。实验结果表明饲吸药浴鼠的白岭在２４ｈ内全部中毒致死，饲吸未经药浴鼠的白蛉存活率很高。在剖检的吸血白蛉中，有６９．１％的白蛉，在其消化道内发现有前鞭毛体的感染，这些前鞭毛体不仅在白蛉消化道内发育良好，且随饲吸时间的推移，前鞭毛体侵入食道、咽和喙部。背纹仓鼠是杜氏利什曼原虫理想的保虫宿主，经药浴的仓鼠进行人工感染，已失去感染能力。     

我国不同地区的杜氏利什曼原虫对亚历山大白蛉感染性的观察

用白蛉人工感染利什曼原虫的方法,观察从新疆荒漠、甘肃山区及河南平原三地自人体成蛉体分离出来的杜氏利什曼原虫对新疆亚历山大白蛉的感染性。以白蛉的感染率、感染程度以及原虫在白蛉消化道内的进展等项指标,来衡量原虫对白蛉的适应性。结果发现,新疆荒漠的原虫对亚历山大白蛉有高度的感染性,甘肃的原虫居次,河南的原虫对白蛉的感染性很差。结合以往用单克隆抗体检测法和K-DNA杂交法对我国一些地区杜氏利什曼原虫的研究,以及我国荒漠、山区和平原地区的黑热病具有不同的流行病学特征等的结果分析,认为我国的杜氏利什曼原虫很可能存在不同的地域株。     

溴氰菊酯药浴阳性鼠阻断内脏利什曼病传播的实…

用中华白蛉分别饲吸两组染有杜氏利什曼原虫的背纹仓鼠（Ｃｒｉｃｅｔｕｌｕｓ ｂａｒａｂｅｎｓｉｓ），－组系经溴氰菊酯药浴后９－４５ｄ的仓鼠，另一组系未药浴的仓鼠。实验结果表明饲吸药浴鼠的白蛉在２４ｈ内全部中毒致死，饲吸未经药浴鼠的白蛉存活率很高。在剖检的吸血蛉中，有６９．１％的白蛉，在其消化道内发现有前鞭毛体的感染，这些前鞭毛体不仅在白蛉消化道内发育良好，且随饲吸时间的推移，前鞭毛体侵入食道，咽和喙     

克拉玛依地区的利什曼病ⅩⅣ.硕大白蛉吴氏亚…

硕大白蛉吴氏亚种是新疆克拉玛依地区的主要蛉种之一，具有强的亲人性，在野外和居民点内常能查见该蛉有前鞭毛体的自然感染。本文结果表明，白蛉自然感染的前鞭毛体能使仓鼠及ＢＡＬＢ／ｃ小鼠发生内脏利什曼病；在感染仓属内脏涂片上的无鞭毛体，由蛉体而来的明显较由大沙鼠而来的都兰利什曼原虫为小；白蛉自然感染的前鞭毛体在ＮＮＮ培养基内生长不良，用３２Ｐ标记的ｇｐ＾６＾３基因为探针，与婴儿利什曼原虫同源。克拉玛依无内     

A method for the purification of Leishmania promastigotes from infected phlebotomine sandflies

We describe a method for the purification of Leishmania promastigotes, isolated from infected sandflies (Lutzomyia longipalpis) using a discontinuous density centrifugation gradient (Percoll/Homem). The sandflies, infected seven days previously with Leishmania donovani chagasi or Leishmania mexicana mexicana from culture, were homogenized and centrifuged on a Percoll discontinuous gradient. Five interface bands were formed, and most of the promastigotes settled out at the interface between the (30% and 40%) Percoll concentrations. An extraction of 3.5 x 10(4) promastigotes from 90 female flies was achieved using this technique.     

Vectorial efficacy of Phlebotomus argentipes in Kala-azar endemic foci of Bihar (India) under natural and artificial conditions

Ability of Phlebotomus argentipes to acquire Leishmania donovani the causative agent of Indian Kala-azar was evaluated in the laboratory. Flies were fed artificially on infected blood suspensions, using a chick-skin-membrane feeding apparatus, and naturally on Leishmania donovani infected mice. In addition flies collected from different endemic areas were dissected and examined for natural infection. Flies fed on infected mice showed significantly higher feeding rate (14.4%, p < 0.01) compared to that of other experiments (9%, 8.75%) but the percentage of infection was very low (2.43%). No Chi-square comparison was made between infection rate and feeding rate because of low value in infection rate (less than 5). Flies dissected for natural infection showed only 0.1% infection. Not much difference was observed in the intensity of Leishmania donovani infection in the mid gut of sandflies examined from any of these experiments. These observations have confirmed that Phlebotomus argentipes has ability to acquire infection and it provides the final piece of evidence that Phlebotomus argentipes is the vector of Leishmania donovani in Bihar State.     

单克隆抗体检测砂鼠利什曼原虫的研究

应用淋巴细胞杂交瘤技术,制备出高特异性、高效价的抗砂鼠利什曼原虫单克隆抗体,选择其中2株对我国西北荒漠地区大砂鼠体内的分离物给予检测。从IFA,斑点-ELISA的实验结果,均证明为砂鼠利什曼原虫,同时以抗杜氏利什曼单克隆抗体测定则为阴性。从而进一步阐明砂鼠利什曼原虫为大砂鼠体内特有的一种亲皮肤但对人不致病的原虫。     

Surface antigenic change during differentiation of a parasitic protozoan, Leishmania mexicana: Identification by monoclonal antibodies.

The fusion of SP2/0 myeloma cells with spleen cells from mice immunized with Leishmania mexicana amazonensis promastigotes produced hybridoma clones. Indirect immunofluorescent antibody assay with live leishmanias showed that the monoclonal antibody 6H12 recognized only the antigens bound to the surface of L. mexicana amazonensis promastigotes. It also showed that the antibody bound to neither amastigotes of this species nor to other Leishmania species--i.e., L. braziliensis braziliensis, L. tropica, and L. donovani. Monoclonal antibodies from three other clones (4D11, 4H9, and 6A11) were found to compete with 6H12 for binding to L. mexicana promastigotes. With lysates of [35S]methionine-labeled promastigotes, all four monoclonal antibodies precipitated the same triplet set of protein bands at the approximately equal to 68,000-dalton region, whereas another monoclonal antibody (6G5) precipitated a different band at approximately equal to 90,000 daltons. During differentiation of L. mexicana amazonensis from amastigotes to promastigotes, there was a 4- to 8-fold increase above the initial level in the binding of 6H12 monoclonal antibody to leishmanias, as detected by enzyme-linked immunosorbent assay and quantitative fluorometric assay, respectively. Thus, we have demonstrated the use of monoclonal antibodies as probes for antigens that change during leishmanial differentiation.