Broad-spectrum monoclonal antibodies against chikungunya virus structural proteins: Promising candidates for antibody-based rapid diagnostic test development.

Aekkachai Tuekprakhon,Emi E Nakayama,Tadahiro Sasaki,Natthanej Luplertlop,Koen Bartholomeeusen,Pornsawan Leaungwutiwong,Johan Michiels,Tatsuo Shioda,Juthamas Phadungsombat,Orapim Puiprom,Kevin K Ariën,Ralph Huits,Michael K Meno,Abdallah M Samy

doi:10.1371/journal.pone.0208851

Abstract

In response to the aggressive global spread of the mosquito-borne chikungunya virus (CHIKV), an accurate and accessible diagnostic tool is of high importance. CHIKV, an arthritogenic alphavirus, comprises three genotypes: East/Central/South African (ECSA), West African (WA), and Asian. A previous rapid immunochromatographic (IC) test detecting CHIKV E1 protein showed promising performance for detection of the ECSA genotype. Unfortunately, this kit exhibited lower capacity for detection of the Asian genotype, currently in circulation in the Americas, reflecting the low avidity of one of the monoclonal antibodies (mAbs) in this IC kit for the E1 protein of the Asian-genotype because of a variant amino acid sequence. To address this shortcoming, we set out to generate a new panel of broad-spectrum mouse anti-CHIKV mAbs using hybridoma technology. We report here the successful generation of mouse anti-CHIKV mAbs targeting CHIKV E1 and capsid proteins. These mAbs possessed broad reactivity to all three CHIKV genotypes, while most of the mAbs lacked cross-reactivity towards Sindbis, dengue, and Zika viruses. Two of the mAbs also lacked cross-reactivity towards other alphaviruses, including O'nyong-nyong, Ross River, Mayaro, Western Equine Encephalitis, Eastern Equine Encephalitis, and Venezuelan Equine Encephalitis viruses. In addition, another two mAbs cross-reacted weakly only with most closely related O'nyong-nyong virus. Effective diagnosis is one of the keys to disease control but to date, no antibody-based rapid IC platform for CHIKV is commercially available. Thus, the application of the mAbs characterized here in the rapid diagnostic IC kit for CHIKV detection is expected to be of great value for clinical diagnosis and surveillance purposes.

Highlights

Chikungunya virus (CHIKV), an arthritogenic mosquito-borne virus, was first documented more than six decades ago in Tanzania, East Africa [1]
The observation that Vero cells infected with Sendai virus (SeV) expressing the Asian-genotype chikungunya virus (CHIKV) 6K-E1 reacted positively with CK119 in indirect immunofluorescence test (IIFT) (Fig 1B) confirmed that the SeV system was able to express the CHIKV 6K-E1 envelope protein, which could be used for further immunization
We generated mouse anti-CHIKV monoclonal antibodies (mAbs) that have broad reactivity towards all three genotypes of CHIKV; such mAbs might be of great value for further diagnostic test development

Summary

Introduction

Chikungunya virus (CHIKV), an arthritogenic mosquito-borne virus, was first documented more than six decades ago in Tanzania, East Africa [1]. CHIKV was considered a neglected tropical agent until a massive outbreak was reported on islands of the Indian Ocean in 2005. Following its initial detection in the Indian Ocean islands, ECSA-IOL was identified as the cause of several CHIKV outbreaks in India and South East Asia (including Thailand, Cambodia, and Malaysia), and ECSA-IOL was introduced for the first time into European countries (including Italy and France) [6, 7]. Asian-genotype CHIKV subsequently gave rise to a novel clade, designated as Asian/American [8]. This clade now is considered a public health concern in several countries/territories in the Caribbean and in the Central American mainland [9, 10]. Given these limitations in prevention and control, it is highly likely that CHIKV (along with its insect vector) will continue to spread, increasing the risk of CHIKV infection world-wide

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