Human Antibody Responses to Emerging Mayaro Virus and Cocirculating Alphavirus Infections Examined by Using Structural Proteins from Nine New and Old World Lineages

Abstract

Mayaro virus (MAYV), Venezuelan equine encephalitis virus (VEEV), and chikungunya virus (CHIKV) are vector-borne alphaviruses that cocirculate in South America. Human infections by these viruses are frequently underdiagnosed or misdiagnosed, especially in areas with high dengue virus endemicity. Disease may progress to debilitating arthralgia (MAYV, CHIKV), encephalitis (VEEV), and death. Few standardized serological assays exist for specific human alphavirus infection detection, and antigen cross-reactivity can be problematic. Therefore, serological platforms that aid in the specific detection of multiple alphavirus infections will greatly expand disease surveillance for these emerging infections. In this study, serum samples from South American patients with PCR-and/or isolation-confirmed infections caused by MAYV, VEEV, and CHIKV were examined by using a protein microarray assembled with recombinant capsid, envelope protein 1 (E1), and E2 from nine New and Old World alphaviruses. Notably, specific antibody recognition of E1 was observed only with MAYV infections, whereas E2 was specifically targeted by antibodies from all of the alphavirus infections investigated, with evidence of cross-reactivity to E2 of o'nyong-nyong virus only in CHIKV-infected patient serum samples. Our findings suggest that alphavirus structural protein microarrays can distinguish infections caused by MAYV, VEEV, and CHIKV and that this multiplexed serological platform could be useful for high-throughput disease surveillance.IMPORTANCE Mayaro, chikungunya, and Venezuelan equine encephalitis viruses are closely related alphaviruses that are spread by mosquitos, causing diseases that produce similar influenza-like symptoms or more severe illnesses. Moreover, alphavirus infection symptoms can be similar to those of dengue or Zika disease, leading to underreporting of cases and potential misdiagnoses. New methods that can be used to detect antibody responses to multiple alphaviruses within the same assay would greatly aid disease surveillance efforts. However, possible antibody cross-reactivity between viruses can reduce the quality of laboratory results. Our results demonstrate that antibody responses to multiple alphaviruses can be specifically quantified within the same assay by using selected recombinant protein antigens and further show that Mayaro virus infections result in unique responses to viral envelope proteins.

Detection of chikungunya virus‐specific IgM on laser‐cut paper‐based device using pseudo‐particles as capture antigen

“…These viruses were chosen because of their recurrent cocirculation with CHIKV in tropical countries and the capacity of the flavivirus genus to induce antibody cross‐reactivity . We also evaluated the sample from a patient infected with O'Nyong'Nyong viruses (ONNV), an alphavirus known to cross‐react with CHIKV . Thus, a total of 26 specimens, consisting of recently infected DENV ( n = 11), ZIKV ( n = 14), and ONNV ( n = 1) patient samples, were evaluated in parallel using the CHIKV MAC‐ELISA (ie the reference method), the CTK Biotech anti‐CHIKV IgM LFA (CTK Biotech, San Diego, CA), and the PAD.…”

“…53 We also evaluated the sample from a patient infected with O'Nyong'Nyong viruses (ONNV), an alphavirus known to cross-react with CHIKV. 54,55 Thus, a total of 26 specimens, consisting of recently infected DENV (n = 11), ZIKV (n = 14), and ONNV (n = 1) patient samples, were evaluated in parallel using the CHIKV MAC-ELISA (ie the reference method), the CTK Biotech anti-CHIKV IgM LFA (CTK Biotech, San Diego, CA), and the PAD. In addition, 18 healthy samples were tested as negative controls.…”

Role of T Cells in Chikungunya Virus Infection and Utilizing Their Potential in Anti-Viral Immunity

“…The geographical range of mosquitoes that transmit the alphaviruses is projected to increase in the future due to climate change (84). Moreover, CHIKV shares some homology with other alphaviruses with the potential to cause unprecedented outbreaks (85)(86)(87)(88)(89). This shift may ultimately lead to the spread of CHIKV and other alphaviruses and cause future epidemics.…”

Virus-Like Particles and Magnetic Microspheres Provide a Flexible and Sustainable Multiplexed Alphavirus Immunodiagnostic Platform

“…Herein, we outline the design and implementation of a novel diagnostic reagent, which pairs the sustainability of a VLP with the sensitivity of the MAGPIX® platform to serve as a versatile tool for detection of anti-viral glycoprotein humoral responses in a serum sample (Figure 1). More specifically, we focused on production, characterization, and optimization of alphavirus diagnostic reagents, due to previously documented problems associated with recombinant expression of their E1/E2 heterodimeric glycoproteins and because these glycoproteins play a dominant role in host immune response during alphavirus infection (19-21). As a proof of concept, Venezuelan equine encephalitis virus (VEEV) E1/E2 glycoproteins were expressed on a retroviral core VLP and conjugated to fluorescent, magnetic microspheres to create VLP-conjugated microspheres (VCMs).…”