Dey Ayan, Sharma Pawan, Redhu Naresh Singh, Singh Sarman
Department of Laboratory Medicine, All India Institute of Medical Sciences, New Delhi 110029, India.
Clin Vaccine Immunol. 2008 May;15(5):836-42. doi: 10.1128/CVI.00433-07. Epub 2008 Mar 19.
Visceral leishmaniasis (VL) is one of the important parasitic diseases, with approximately 350 million people at risk. Due to the nonavailability of an ideal drug, development of a safe, effective, and affordable vaccine could be a solution for control and prevention of this disease. The present study was carried out to examine the immunological potential of kinesin protein from the microtubule locus of Leishmania donovani as a suitable vaccine candidate. In silico analysis of this region revealed clusters of major histocompatibility complex class I and II binding epitopes in its motor domain region. A recombinant protein was expressed from this region and named rLvacc. The antigenicity and immunogenicity studies of this protein by Western blot analysis revealed that rLvacc is strongly recognized by sera from acute VL patients. To evaluate its immunogenicity, peripheral blood mononuclear cells from cured VL patients were separated, and a lymphocyte proliferation assay was carried out in the presence of rLvacc. After lymphocyte proliferation, the pooled culture supernatant was assayed for anti-rLvacc antibody titers using an enzyme-linked immunosorbent assay. The results showed that immunoglobulin G2 (IgG2) subtype antibodies were predominant, while IgG1 subtype antibodies were produced in very low titers. On the basis of these ex vivo preliminary findings, its immunogenicity was studied in BALB/c mice. Vaccination with the DNA construct generated a good cellular immune response with significant increases in gamma interferon and interleukin-2 (IL-2) cytokine levels (Th1), but no increase in IL-4 levels (Th2). Taken together, our findings suggest the kinesin motor domain region of L. donovani as a potential vaccine candidate against visceral leishmaniasis.
内脏利什曼病(VL)是一种重要的寄生虫病,约有3.5亿人面临感染风险。由于缺乏理想的药物,开发一种安全、有效且价格合理的疫苗可能是控制和预防该病的一种解决方案。本研究旨在检测来自杜氏利什曼原虫微管位点的驱动蛋白作为合适疫苗候选物的免疫潜力。对该区域的计算机分析显示,其运动结构域区域存在主要组织相容性复合体I类和II类结合表位簇。从该区域表达了一种重组蛋白,并将其命名为rLvacc。通过蛋白质印迹分析对该蛋白的抗原性和免疫原性进行研究,结果显示rLvacc能被急性VL患者的血清强烈识别。为评估其免疫原性,分离出治愈的VL患者的外周血单核细胞,并在rLvacc存在的情况下进行淋巴细胞增殖试验。淋巴细胞增殖后,使用酶联免疫吸附测定法检测合并培养上清液中的抗rLvacc抗体滴度。结果表明,免疫球蛋白G2(IgG2)亚型抗体占主导,而IgG1亚型抗体的产生滴度非常低。基于这些体外初步研究结果,在BALB/c小鼠中研究了其免疫原性。用DNA构建体进行疫苗接种产生了良好的细胞免疫反应,γ干扰素和白细胞介素-2(IL-2)细胞因子水平(Th1)显著升高,但IL-4水平(Th2)没有升高。综上所述,我们的研究结果表明杜氏利什曼原虫的驱动蛋白运动结构域区域是抗内脏利什曼病的潜在疫苗候选物。