Manjunathachar Haranahally Vasanthachar, Kumar Binod, Parthasarathi Balasamudram Chandrasekhar, Chigure Gajanan M, Saravanan Buddhi Chandrasekaran, Sankar Muthu, Harish Darasaguppe Ramachandra, de la Fuente José, Ghosh Srikanta
Entomology Laboratory, Division of Parasitology, Indian Council of Agricultural Research (ICAR)- Indian Veterinary Research Institute (IVRI)-Izatnagar, Bareilly, Uttar Pradesh, India.
Division of Animal Biotechnology, Indian Council of Agricultural Research (ICAR)-Indian Veterinary Research Institute (IVRI)-Izatnagar, Bareilly, Uttar Pradesh, India.
Front Immunol. 2024 Nov 19;15:1471317. doi: 10.3389/fimmu.2024.1471317. eCollection 2024.
Globally, ticks rank second only to mosquitoes as vectors of deadly pathogens affecting humans and first in transmitting animal pathogens, presenting a significant challenge to human wellness and sustainability of livestock-based industries. Traditional tick control via chemical acaricides impacts on the environment and has led to the emergence of multi-acaricide-resistant tick populations. Use of immunoprophylactic, along with other components of integrated tick management, holds the potential to mitigate tick infestations in a sustainable manner. To control multi-species tick infestations, the concept of a cocktail vaccine comprising of more than one antigens has emerged as a viable solution due to the inconsistent efficacy of single antigen-based immunization protocol.
In this study, a dual antigen cocktail immunization protocol was developed targeting ferritin2 (FER2) and tropomyosin (TPM) proteins, which are associated with ticks' essential cellular and physiological functions, like blood iron homeostasis and muscle contractions.
Dual gene silencing of FER2 and TPM genes in resulted in a 75.3% reduction in infested ticks, a 95.4% decrease in egg masses, and a complete loss of egg hatching when compared to control ticks. Microscopically, an altered ovarian cellular architecture, marked by vacuolation and reduced nucleus-to-cytoplasmic ratio were noted in the gene knocked down ticks. An immunization with cocktails of 300 µg dose of each protein, rHaFER2 and rHaTPM was standardized in a rat model and was used to immunize cross-bred ( x ) male cattle with Montanide ISA 50V2 adjuvant on days 0, 28, and 49. A significant ( < 0.001) IgG and IgG2 antibody response was observed in the immunized animals with high IgG levels sustained until day 119 post-primary immunization, showing a 4.1-fold increase over the pre-immunization period. The animals were challenged with larvae and adults of and larvae of . Immunization with the cocktail antigen resulted an efficacy of 70% and 76% against larvae and adults, respectively, and 54% against infestations. Compared to single-antigen immunization, the immunization with cocktail antigens demonstrated higher protection against and ticks. The results advance the development of cocktail vaccines to control multiple tick species.
在全球范围内,蜱作为影响人类的致命病原体的传播媒介,其危害性仅次于蚊子,在传播动物病原体方面则位居首位,这对人类健康和以牲畜为基础的产业的可持续发展构成了重大挑战。传统的通过化学杀螨剂控制蜱虫的方法对环境有影响,并导致了对多种杀螨剂产生抗性的蜱虫种群的出现。使用免疫预防措施以及综合蜱虫管理的其他组成部分,有望以可持续的方式减轻蜱虫侵扰。为了控制多种蜱虫的侵扰,由于基于单一抗原的免疫方案效果不一致,由多种抗原组成的联合疫苗的概念已成为一种可行的解决方案。
在本研究中,开发了一种针对铁蛋白2(FER2)和原肌球蛋白(TPM)蛋白的双抗原联合免疫方案,这些蛋白与蜱虫的基本细胞和生理功能相关,如血中铁离子稳态和肌肉收缩。
与对照蜱虫相比,FER2和TPM基因的双基因沉默导致受侵染的蜱虫减少了75.3%,卵块减少了95.4%,并且卵孵化完全丧失。在显微镜下,在基因敲除的蜱虫中观察到卵巢细胞结构发生改变,其特征为空泡化和核质比降低。在大鼠模型中对每种蛋白质(重组哈氏铁蛋白2(rHaFER2)和重组哈氏原肌球蛋白(rHaTPM))300μg剂量的联合疫苗进行了标准化,并在第0、28和49天使用Montanide ISA 50V2佐剂对杂交(×)雄性牛进行免疫。在免疫动物中观察到显著(<0.001)的IgG和IgG2抗体反应,高IgG水平一直持续到初次免疫后第119天,比免疫前时期增加了4.1倍。用微小牛蜱的幼虫和成虫以及长角血蜱的幼虫对动物进行攻击。用联合抗原免疫分别对微小牛蜱的幼虫和成虫产生了70%和76%的效力,对长角血蜱侵染产生了54%的效力。与单一抗原免疫相比,用联合抗原免疫对微小牛蜱和长角血蜱表现出更高的保护作用。这些结果推动了用于控制多种蜱虫种类的联合疫苗的开发。