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首页> 外文期刊>Vaccine >Leishmania donovani: Identification of stimulatory soluble antigenic proteins using cured human and hamster lymphocytes for their prophylactic potential against visceral leishmaniasis
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Leishmania donovani: Identification of stimulatory soluble antigenic proteins using cured human and hamster lymphocytes for their prophylactic potential against visceral leishmaniasis

机译：Leishmania donovani：使用治愈的人和仓鼠淋巴细胞对内脏利什曼病的预防潜力，鉴定刺激性可溶性抗原蛋白

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Most of the studies for the identification of prophylactic antigens that elicit T cell responses were concentrated on membrane proteins of Leishmania donovani. This study was taken up to assess L. donovani soluble promastigote antigens for their ability to stimulate proliferation of peripheral blood mononuclear cells (PBMCs) from cured visceral leishmaniasis (VL) patients, endemic and non-endemic controls and lymphocytes/peritoneal macrophages of cured hamsters. The soluble protein was subjected to sequential precipitation with saturated ammonium sulphate (20%, 40%, 60% and 80%), of which largely 80% fractioned protein showed significant cellular responses in cured patients and hamsters. This fraction was further fractionated into five sub fractions by preparative SDS-PAGE and subjected to re-evaluation for their ability to induce cellular responses. Out of these, only F2 sub fraction belonging to the MW of 97.4-68kDa stimulated remarkable lymphoproliferative and IFN-gamma responses in cured VL patients and in endemic controls. Similarly, significant lymphoproliferative responses and nitric oxide production were also noticed in cured Leishmania infected animals indicating an element of uniformity in responses between hamster and human. F2 sub fraction, when evaluated for its prophylactic efficacy with BCG against L. donovani challenge in hamster exhibited significant parasite inhibition in spleen (71.1%; p<0.001) and liver (68.2%; p<0.001) as compared to their unvaccinated counterpart. The vaccinated animals showed significant lymphoproliferative response and nitric oxide production but leishmania specific IgG level were suppressed. The results indicate the presence of immunostimulatory and protective molecules in F2 sub fraction which may further be exploited for the development of a vaccine against VL, hitherto an unrealized goal.
机译：鉴定引起T细胞反应的预防性抗原的大多数研究都集中在多形利什曼原虫的膜蛋白上。开展这项研究是为了评估多诺瓦氏乳杆菌可溶性前鞭毛体抗原刺激已治愈内脏利什曼病（VL）患者，地方病和非地方病对照以及已治愈仓鼠的淋巴细胞/腹膜巨噬细胞刺激外周血单个核细胞（PBMC）增殖的能力。可溶性蛋白先后用饱和硫酸铵（20％，40％，60％和80％）沉淀，其中80％的分离蛋白在治愈的患者和仓鼠中显示出明显的细胞反应。通过制备的SDS-PAGE将该级分进一步分为五个亚级，并对其细胞诱导细胞应答的能力进行重新评估。在这些当中，只有9 MW-9kkDa的F2子部分在治愈的VL患者和地方性对照中刺激了显着的淋巴增生和IFN-γ反应。类似地，在治愈的利什曼原虫感染的动物中也观察到显着的淋巴增生反应和一氧化氮的产生，表明仓鼠和人之间的反应是均匀的。当评估B2组分与BCG对抗仓鼠L. donovani攻击的预防效果时，与未接种疫苗的F2组分相比，脾脏（71.1％; p <0.001）和肝脏（68.2％; p <0.001）表现出明显的寄生虫抑制作用。接种的动物表现出显着的淋巴增生反应和一氧化氮生成，但利什曼原虫特异性IgG水平被抑制。结果表明在F2亚部分中存在免疫刺激和保护性分子，可以进一步利用其开发针对VL的疫苗，这迄今为止尚未实现。

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