活的过表达ld_ζ 1结构域的多诺瓦利什曼原虫对实验性人内脏利什曼病的免疫药理学疗效评价。

IF 4.7 2区医学 Q2 IMMUNOLOGY

International immunopharmacology Pub Date : 2025-02-20 DOI:10.1016/j.intimp.2025.114295

Ruby Bansal , Sadat Shafi , Prachi Garg , Aakriti Srivastava , Swati Garg , Neha Jha , Jhalak Singhal , Gajala Deethamvali Ghouse Peer , Ramendra Pati Pandey , Subhajit Basu , Shailja Singh

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引用次数: 0

摘要

目的评价过表达多诺瓦利什曼原虫zeta结构域（ld_ζ 1结构域）作为内脏利什曼病（VL）候选疫苗的有效性和免疫原性。方法利用gfp标记的重组质粒电孔法转化利什曼原虫过表达体ld_ζ 1域（OE）。体外分析产生的过表达细胞，以评估其生长动力学和感染性。我们还研究了这些过表达蛋白在多诺瓦利什曼感染小鼠模型中的免疫保护潜力。免疫反应，包括Th1和Th2途径，通过RT-PCR和ELISA分析进行了全面表征。此外，本研究还对小鼠脾脏和肝脏寄生虫进行了深入的评估，并对组织病理变化进行了定量评估。结果ld_ζ (OE)结构域（sld_ζ 1）免疫小鼠体内的寄生数量明显低于感染LdWT的小鼠，其生存力和复制率明显低于WT。脾脏和肝脏表现出明显的组织学变化，提示保护作用。免疫小鼠脾脏和肝脏的寄生虫载量显著降低。免疫反应显示IFN-γ水平升高，IL-10产生降低，导致IFN-γ/IL-10比值升高，表明寄生虫消除。疫苗接种也引起免疫小鼠显著的IgG体液反应和增加一氧化氮的产生。结论过表达zeta毒素可控制寄生虫的衰减，在保持免疫原性的同时降低致病性。我们的工作确定了zeta过表达蛋白在体外和体内对利什曼原虫攻击的保护功效、免疫原性和增殖反应。这一初步的原型研究表明，ld_ζ 1结构域（OE）寄生虫可能适合用于研制抗利什曼病的减毒疫苗。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Evaluation of Immunopharmacological efficacy of live Leishmania donovani overexpressing Ld_ζ1domain for protection against experimental human visceral Leishmaniasis.

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Evaluation of Immunopharmacological efficacy of live Leishmania donovani overexpressing Ld_ζ1domain for protection against experimental human visceral Leishmaniasis.

Objective

To evaluate the efficacy and immunogenicity of the zeta domain over-expressing Leishmania donovani (Ld_ζ1_domain) as a vaccination candidate against visceral leishmaniasis (VL).

Methods

In this study, Leishmania overexpressor Ld_ζ1_domain (OE) were transformed by electroporation using a GFP-tagged Ld_ζ1_domain recombinant plasmid. The resulting overexpressing cells were analysed in vitro to assess their growth dynamics and infectivity. We also investigated the immune-protective potential of these overexpressor in a mouse model challenged with Leishmania donovani. The immune response, including Th1 and Th2 pathways, was thoroughly characterized using RT-PCR and ELISA assays. In addition, the study conducted a thorough evaluation of the mouse's spleen and liver parasites, as well as quantitative evaluation of tissue pathological changes.

Results

Ld_ζ1 _domain (OE) parasites exhibited significantly lower viability and replication rates than WT parasites, and in vivo studies showed that mice immunized with the Ld_ζ1(OE) _domain had lower parasite numbers than mice infected with LdWT. Spleen and liver showed significant histological changes suggestive of protection. Parasite load in the spleen and liver of vaccinated mice were significantly reduced. The immune response showed increased IFN-γ levels and lower IL-10 production, resulting in a greater IFN-γ/IL-10 ratio, indicating parasite elimination. The vaccination also caused a significant IgG humoral response and increased nitric oxide production in immunized mice.

Conclusion

Our findings demonstrated that overexpressing the zeta toxin resulted in controlled parasite attenuation, lowering pathogenicity while retaining immunogenic features. Our work established the zeta over-expressor's protective efficacy, immunogenicity, and proliferation in response to a Leishmania challenge in vitro and in vivo. This preliminary prototype study suggested that Ld_ζ1_domain (OE) parasites may be suitable for developing an attenuated vaccine against leishmaniasis.

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来源期刊

International immunopharmacology 医学-免疫学

CiteScore

8.40

自引率

3.60%

发文量

935

审稿时长

53 days

期刊介绍： International Immunopharmacology is the primary vehicle for the publication of original research papers pertinent to the overlapping areas of immunology, pharmacology, cytokine biology, immunotherapy, immunopathology and immunotoxicology. Review articles that encompass these subjects are also welcome. The subject material appropriate for submission includes: • Clinical studies employing immunotherapy of any type including the use of: bacterial and chemical agents; thymic hormones, interferon, lymphokines, etc., in transplantation and diseases such as cancer, immunodeficiency, chronic infection and allergic, inflammatory or autoimmune disorders. • Studies on the mechanisms of action of these agents for specific parameters of immune competence as well as the overall clinical state. • Pre-clinical animal studies and in vitro studies on mechanisms of action with immunopotentiators, immunomodulators, immunoadjuvants and other pharmacological agents active on cells participating in immune or allergic responses. • Pharmacological compounds, microbial products and toxicological agents that affect the lymphoid system, and their mechanisms of action. • Agents that activate genes or modify transcription and translation within the immune response. • Substances activated, generated, or released through immunologic or related pathways that are pharmacologically active. • Production, function and regulation of cytokines and their receptors. • Classical pharmacological studies on the effects of chemokines and bioactive factors released during immunological reactions.