Exploration of the potential utility of the luciferase immunoprecipitation system (LIPS) assay for the detection of anti-leptospira antibodies in dogs

IF 1.4 3区农林科学 Q4 IMMUNOLOGY

Veterinary immunology and immunopathology Pub Date : 2023-10-01 DOI:10.1016/j.vetimm.2023.110661

C. Taylor , E. Belin , D. Brodbelt , H.L.B.M. Klaasen , B. Catchpole

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引用次数: 0

Abstract

Canine leptospirosis represents a diagnostic challenge to veterinarians, due to the variability in presenting clinical signs and interpretation of serology test results in dogs that have been vaccinated previously. None of the commercially available serological assays, including the microscopic agglutination test (MAT), have been verified to be capable of differentiating infected from vaccinated animals (DIVA). Recent work identified that half of primary practice attending dogs were up to date with their leptospirosis vaccination and would be expected to have circulating anti-leptospira antibodies (Taylor et al., 2022), indicating that this is a relevant issue for suspected leptospirosis cases in dogs in the UK. This study aimed to explore the utility of three leptospiral outer membrane proteins (OMPs: LipL32, LipL21 and LipL41) as potential DIVA targets in the luciferase immunoprecipitation system (LIPS) assay. N and C terminal nanoluciferase tagged recombinant proteins were generated for each OMP. Differences in reactivity between serum samples from MAT positive dogs (n = 29) and paired samples (n = 6 dogs) taken pre and 21 days post leptospirosis vaccination were assessed against these six constructs. Reactivity was greater towards the N terminal than the C terminal recombinant proteins for all three OMPs. None of the constructs appeared to demonstrate DIVA capability, although two (pNLF1-N-FLAG/LipL32 and pNLF1-N-FLAG/LipL21) were able to detect vaccine seroconversion. The findings of this work suggest that these particular OMP targets do not offer DIVA ability, however LipL32 and LipL21 may be suitable for use in immunoassays for vaccine trials or for detection of infections in humans, where there is no requirement for DIVA capability.

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探索萤光素酶免疫沉淀系统（LIPS）检测狗抗钩端螺旋体抗体的潜在用途。

犬钩端螺旋体病对兽医来说是一个诊断挑战，因为之前接种过疫苗的狗在表现临床症状和血清学测试结果解释方面存在差异。商业上可用的血清学检测，包括显微镜凝集试验（MAT），都没有被证实能够区分感染动物和接种疫苗的动物（DIVA）。最近的研究表明，一半的初级护理犬都接种了最新的钩端螺旋体病疫苗，预计会有循环的抗钩端螺旋抗体（Taylor et al.，2022），这表明这是英国疑似犬钩端螺旋病病例的相关问题。本研究旨在探索三种钩端螺旋体外膜蛋白（OMP:LipL32、LipL21和LipL41）作为潜在DIVA靶标在萤光素酶免疫沉淀系统（LIPS）测定中的作用。为每个OMP产生N和C末端纳米萤光素酶标记的重组蛋白。针对这六种构建体，评估了接种钩端螺旋体病疫苗前和接种后21天采集的MAT阳性犬（n=29）和配对犬（n=6）的血清样本之间的反应性差异。对于所有三种OMP，对N末端的反应性大于对C末端的重组蛋白的反应性。尽管有两种构建体（pNLF1-N-FLAG/LipL32和pNLF1-N-FLAG/LipL21）能够检测疫苗血清转化，但似乎没有一种构建体显示出DIVA能力。这项工作的发现表明，这些特定的OMP靶点不提供DIVA能力，然而LipL32和LipL21可能适合用于疫苗试验的免疫测定或用于检测人类感染，在这些情况下不需要DIVA能力。

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来源期刊

Veterinary immunology and immunopathology 农林科学-免疫学

CiteScore

3.40

自引率

5.60%

发文量

审稿时长

70 days

期刊介绍： The journal reports basic, comparative and clinical immunology as they pertain to the animal species designated here: livestock, poultry, and fish species that are major food animals and companion animals such as cats, dogs, horses and camels, and wildlife species that act as reservoirs for food, companion or human infectious diseases, or as models for human disease. Rodent models of infectious diseases that are of importance in the animal species indicated above,when the disease requires a level of containment that is not readily available for larger animal experimentation (ABSL3), will be considered. Papers on rabbits, lizards, guinea pigs, badgers, armadillos, elephants, antelope, and buffalo will be reviewed if the research advances our fundamental understanding of immunology, or if they act as a reservoir of infectious disease for the primary animal species designated above, or for humans. Manuscripts employing other species will be reviewed if justified as fitting into the categories above. The following topics are appropriate: biology of cells and mechanisms of the immune system, immunochemistry, immunodeficiencies, immunodiagnosis, immunogenetics, immunopathology, immunology of infectious disease and tumors, immunoprophylaxis including vaccine development and delivery, immunological aspects of pregnancy including passive immunity, autoimmuity, neuroimmunology, and transplanatation immunology. Manuscripts that describe new genes and development of tools such as monoclonal antibodies are also of interest when part of a larger biological study. Studies employing extracts or constituents (plant extracts, feed additives or microbiome) must be sufficiently defined to be reproduced in other laboratories and also provide evidence for possible mechanisms and not simply show an effect on the immune system.