Das Nabarun Chandra, Gupta Parth Sarthi Sen, Panda Saroj Kumar, Rana Malay Kumar, Mukherjee Suprabhat
Integrative Biochemistry & Immunology Laboratory, Department of Animal Science, Kazi Nazrul University, Asansol 713 340, West Bengal, India.
School of Biosciences & Bioengineering, D. Y. Patil International University, Akurdi, Pune 411044, India.
Int Immunopharmacol. 2023 Feb;115:109639. doi: 10.1016/j.intimp.2022.109639. Epub 2022 Dec 29.
Proteases are the critical mediators of immunomodulation exerted by the filarial parasites to bypass and divert host immunity. Cystatin is a small (∼15 kDa) immunomodulatory filarial protein and known to contribute in the immunomodulation strategy by inducing anti-inflammatory response through alternative activation of macrophages. Recently, Wuchereria bancrofti cystatin has been discovered as a ligand of human toll-like receptor 4 which is key behind the cystatin-induced anti-inflammatory response in major human antigen-presenting cells. Considering the pivotal role of cystatin in the immunobiology of filariasis, cystatin could be an efficacious target for developing vaccine. Herein, we present the design and in-silico analyses of a multi-epitope-based peptide vaccine to target W. bancrofti cystatin through immune-informatics approaches. The 262 amino acid long antigen construct comprises 9 MHC-I epitopes and MHC-II epitopes linked together by GPGPG peptide alongside an adjuvant (50S ribosomal protein L7/L12) at N terminus and 6 His tags at C terminus. Molecular docking study reveals that the peptide could trigger TLR4-MD2 to induce protective innate immune responses while the induced adaptive responses were found to be mediated by IgG, IgM and Th1 mediated responses. Notably, the designed vaccine exhibits high stability and no allergenicity in-silico. Furthermore, the muti epitope-vaccine was also predicted for its RNA structure and cloned in pET30ax for further experimental validation. Taken together, this study presents a novel multi-epitope peptide vaccine for triggering efficient innate and adaptive immune responses against W. bancrofti to intervene LF through immunotherapy.
蛋白酶是丝虫寄生虫发挥免疫调节作用以绕过和转移宿主免疫的关键介质。胱抑素是一种小分子量（约15 kDa）的免疫调节性丝虫蛋白，已知其通过巨噬细胞的替代激活诱导抗炎反应，从而在免疫调节策略中发挥作用。最近，班氏吴策线虫胱抑素被发现是人类 Toll 样受体 4 的配体，这是胱抑素在主要人类抗原呈递细胞中诱导抗炎反应的关键所在。鉴于胱抑素在丝虫病免疫生物学中的关键作用，它可能是开发疫苗的有效靶点。在此，我们通过免疫信息学方法展示了一种基于多表位的肽疫苗的设计和计算机模拟分析，该疫苗旨在靶向班氏吴策线虫胱抑素。这个由262个氨基酸组成的抗原构建体包含9个MHC-I表位和MHC-II表位，通过GPGPG肽连接在一起，在N端有一个佐剂（50S核糖体蛋白L7/L12），在C端有6个组氨酸标签。分子对接研究表明，该肽可以触发TLR4-MD2诱导保护性先天免疫反应，而诱导的适应性反应则由IgG、IgM和Th1介导的反应介导。值得注意的是，所设计的疫苗在计算机模拟中表现出高稳定性且无致敏性。此外，还预测了多表位疫苗的RNA结构，并将其克隆到pET30ax中进行进一步的实验验证。综上所述，本研究提出了一种新型的多表位肽疫苗，用于触发针对班氏吴策线虫的高效先天和适应性免疫反应，通过免疫疗法干预淋巴丝虫病。
