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. 2018 Mar 21;3(2):e00003-18.
doi: 10.1128/mSphere.00003-18. eCollection 2018 Mar-Apr.

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

Jessica L Smith  1 , Christine L Pugh  1 , Emily D Cisney  1 , Sarah L Keasey  1   2 , Carolina Guevara  3 , Julia S Ampuero  3 , Guillermo Comach  4 , Doris Gomez  5 , Margarita Ochoa-Diaz  5 , Robert D Hontz  3 , Robert G Ulrich  1
Affiliations

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

Jessica L Smith et al. mSphere. .

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.

Keywords: alphavirus; humoral immunity; protein microarray; viral antigen.

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Figures

FIG 1
FIG 1
Schematic representations of structural proteins included in the alphavirus protein microarray. (A) Schematics of ECSA CHIKV structural polyprotein (top) and domains that were cloned and expressed (bottom) with residue numbers indicated. The expressed domains are full-length C (purple) and truncated E1 (light blue) and E2 (royal blue). (B) Structural models of chikungunya virus trimeric spike and C molecules from cryoelectron microscopy (PDB code 3J2W). The E1-E2 heterodimer (colored as in panel A) is represented by a combination of space fill and ribbon structures to show domains that were (space fill) or were not (ribbon) included in the expression construct. The other two heterodimers (gray) are represented as ribbon structures and show the orientation in the trimeric spike. The full-length C protein (purple) was expressed for the protein microarrays, but only a partial C structure is available and is shown in the model.
FIG 2
FIG 2
Mouse polyclonal antibody recognition of alphavirus microarray proteins. Mouse polyclonal antibodies raised against whole alphaviruses of the Semliki Forest complex (A: CHIKV, MAYV, ONNV, and RRV, top to bottom) or equine encephalitis viruses (B: VEEV, EEEV, and WEEV, top to bottom). The mouse polyclonal antibody used is indicated in the upper left corner of each graph. The ratio of the antigen signal to the cutoff value was determined for each replicate spot. Cutoff values were determined as the mean signal of the control proteins plus 3 standard deviations. The average ratio for replicate antigen spots was determined, and ratios >1 are shown, with error bars representing the standard deviation. Vertical lines separate antigens into C, E1, and E2 groups.
FIG 3
FIG 3
Recognition of MAYV antigens by antibodies from human MAYV infections. Serum samples from patients (n = 10) with PCR-confirmed MAYV infections were used to probe protein microarrays consisting of MAYV and VEEV antigens to detect IgG binding. Acute-phase, convalescent-phase, and follow-up serum samples were analyzed for subjects 1 to 9, while acute-phase and follow-up serum samples were analyzed for subject 10. For each individual, antigen signal levels were plotted relative to the acute-phase signal level.
FIG 4
FIG 4
Antibody specificity of human alphavirus infections to alphavirus structural antigens. Serum samples from patients with alphavirus or influenza A virus (Flu A) infections were used to probe alphavirus protein microarrays to detect specific antigen-IgG binding. All serum samples, except one MAYV follow-up sample, were collected within 10 to 30 days of infection confirmation by RT-PCR. The signal levels of the alphavirus-infected subjects were compared to the mean signal level of each antigen in the influenza A virus group. Signal ratios were calculated as described in Materials and Methods. (A) Graphs showing individual and mean signal ratios for 10 patients per alphavirus infection group for MAYV (top), VEEV (middle), and CHIKV (bottom) with error bars representing the standard error of the mean. Vertical lines separate antigens into C, E1, and E2 groups. Horizontal dotted lines indicate an alphavirus infection signal to influenza A virus signal ratio of 1. (B) M statistics were used to identify antigens with significant recognition by antibodies in the alphavirus versus influenza A virus infection groups. Antigens with significant signal levels in the alphavirus infection groups, prevalence, and statistical significance are shown.
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References

    1. Pan American Health Organization 2017. Chikungunya: data, maps and statistics. Pan American Health Organization, Washington, DC: http://www.paho.org/hq/index.php?option=com_topics&view=readall&cid=5927.... Accessed: 5 September 2017.
    1. Lumsden WH. 1955. An epidemic of virus disease in Southern Province, Tanganyika Territory, in 1952–53. II. General description and epidemiology. Trans R Soc Trop Med Hyg 49:33–57. - PubMed
    1. Robinson MC. 1955. An epidemic of virus disease in Southern Province, Tanganyika Territory, in 1952–53. I. Clinical features. Trans R Soc Trop Med Hyg 49:28–32. - PubMed
    1. Ross RW. 1956. The Newala epidemic. III. The virus: isolation, pathogenic properties and relationship to the epidemic. J Hyg (Lond) 54:177–191. doi:10.1017/S0022172400044442. - DOI - PMC - PubMed
    1. Centers for Disease Control and Prevention 2016. Chikungunya virus: geographic distribution. Centers for Disease Control and Prevention, Atlanta, GA. https://www.cdc.gov/chikungunya/geo/index.html. Accessed: 5 September 2017.

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