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Chikungunya virus (CHIKV) is an Alphavirus that causes chronic and incapacitating arthralgia in humans. Although patient cohort studies have shown the production of CHIKV specific antibodies, the fine specificity of the antibody response against CHIKV is not completely defined.

The macaque model of CHIKV infection was established due to limitations of clinical specimens. More importantly, its close relation to humans will allow the study of chronic infection and further identify important CHIKV targets.

In this study, serum samples from CHIKV-infected macaques collected at different time-points post infection were used to characterize the antibody production pattern and kinetics. Results revealed that anti-CHIKV antibodies were neutralizing and the E2 glycoprotein, Capsid, nsP1, nsP3 and nsP4 proteins were targets of the anti-CHIKV antibody response in macaques. Furthermore, linear B-cell epitopes recognized by these anti-CHIKV antibodies were identified, and mapped to their structural localization. This characterizes the specificity of anti-CHIKV antibody response in macaques and further demonstrates the importance of the different regions in CHIKV-encoded proteins in the adaptive immune response.

Information from this study provides critical knowledge that will aid in the understanding of CHIKV infection and immunity, vaccine design, and pre-clinical efficacy studies. Introduction Chikungunya virus (CHIKV) was first described during an epidemic in 1952 in Tanzania, East Africa as the causative agent of Chikungunya fever (CHIKF),. CHIKV belongs to the genus Alphavirus of the family Togaviridae and is an enveloped virus with a single-stranded positive-sense RNA genome. The 12kb RNA genome is capped at the 5′ end and polyadenylated at the 3′ end and consists of two open reading frames coding for four non-structural proteins (nsP1–4), three major structural proteins (Capsid, E1, and E2) and two small cleavage products (E3 and 6K),.

The E1 and E2 glycoproteins form heterodimers that associate as trimeric spikes on the virion surface while E3 and 6K were demonstrated to act as helper proteins in the budding and maturation process of the virion envelope –. In the last decade, multiple CHIKF epidemics have occurred in East Africa, the Indian Ocean Islands, and many parts of South East Asia –. More recently, new episodes of CHIKF have been reported in the Americas, further broadening the geographical spread of the disease. The Aedes species of mosquito has been the major arthropod vector associated with CHIKV transmission to humans. CHIKV infection usually leads to the development of CHIKF and is characterized by an abrupt onset of fever, headache, fatigue, nausea, vomiting, rash, myalgia, and severe arthralgia. Similar to other arthralgia-causing arbovirus infections, a fraction of patients developed chronic symptoms lasting from several weeks to months,,.

Currently, there are no licensed vaccines or antiviral drugs against CHIKV infection for human use. Therapy for CHIKV infection is often limited to supportive care. Despite the development of several animal models, few have met the requirement to be used in pre-clinical study to assess potential therapeutics.

Recent epidemiological data showed the increasing importance of antibody-mediated protection against CHIKV –, highlighting the feasibility of using anti-CHIKV antibodies as therapeutics or as a prophylactic treatment. However, information about the exact target of the adaptive immune response either in human or in animal models remains limited, although B-cell epitopes have been identified within the E1/E2 glycoproteins,. Due to the close lineage relationship between humans and macaques, macaque models of CHIKV infection have been developed –. These models allow comparison of the adaptive immunity between humans and macaques. Furthermore, information obtained from macaque studies will be valuable for the design of future therapeutics. In this study, we aimed to investigate the kinetics and specificity of anti-CHIKV antibodies induced after experimental infection in cynomolgus macaques ( Macaca fascicularis). In addition to the anti-E2 glycoprotein responses, we also identified new linear B-cell epitopes that were recognized by anti-CHIKV antibodies from CHIKV-infected macaques.

These epitopes can potentially be used for future development of vaccine candidates. Cells and Virus Isolates HEK 293T cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) (Gibco, Invitrogen).