Serum Antibodies Against the Oncogenic Merkel Cell Polyomavirus Detected by an Innovative Immunological Assay With Mimotopes in Healthy Subjects

Front Immunol. 2021 Jun 8:12:676627. doi: 10.3389/fimmu.2021.676627. eCollection 2021.

Abstract

Merkel cell polyomavirus (MCPyV), a small DNA tumor virus, has been detected in Merkel cell carcinoma (MCC) and in normal tissues. Since MCPyV infection occurs in both MCC-affected patients and healthy subjects (HS), innovative immunoassays for detecting antibodies (abs) against MCPyV are required. Herein, sera from HS were analyzed with a novel indirect ELISA using two synthetic peptides mimicking MCPyV capsid protein epitopes of VP1 and VP2. Synthetic peptides were designed to recognize IgGs against MCPyV VP mimotopes using a computer-assisted approach. The assay was set up evaluating its performance in detecting IgGs anti-MCPyV on MCPyV-positive (n=65) and -negative (n=67) control sera. Then, the ELISA was extended to sera (n=548) from HS aged 18-65 yrs old. Age-specific MCPyV-seroprevalence was investigated. Performance evaluation indicated that the assay showed 80% sensitivity, 91% specificity and 83.9% accuracy, with positive and negative predictive values of 94.3% and 71%, respectively. The ratio expected/obtained data agreement was 86%, with a Cohen's kappa of 0.72. Receiver-operating characteristic (ROC) curves analysis indicated that the areas under the curves (AUCs) for the two peptides were 0.82 and 0.74, respectively. Intra-/inter-run variations were below 9%. The overall prevalence of serum IgGs anti-MCPyV in HS was 62.9% (345/548). Age-specific MCPyV-seroprevalence was 63.1% (82/130), 56.7% (68/120), 64.5% (91/141), and 66.2% (104/157) in HS aged 18-30, 31-40, 41-50 and 51-65 yrs old, respectively (p>0.05). Performance evaluation suggests that our indirect ELISA is reliable in detecting IgGs anti-MCPyV. Our immunological data indicate that MCPyV infection occurs asymptomatically, at a relatively high prevalence, in humans.

Keywords: IgGs; MCPyV; Merkel cell carcinoma; Merkel cell polyomavirus; antibodies; indirect ELISA; polyomavirus; serology.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Antibodies, Viral / blood*
  • Antibodies, Viral / immunology
  • Asymptomatic Infections
  • Capsid Proteins / immunology
  • Computer Simulation
  • Data Accuracy
  • Diagnosis, Computer-Assisted
  • Enzyme Assays / methods*
  • Enzyme-Linked Immunosorbent Assay / methods
  • Epitopes / immunology
  • Female
  • Healthy Volunteers
  • Humans
  • Immunoglobulin G / blood*
  • Immunoglobulin G / immunology
  • Male
  • Merkel cell polyomavirus / immunology*
  • Middle Aged
  • Oncogenic Viruses / immunology*
  • Polyomavirus Infections / blood*
  • Polyomavirus Infections / diagnosis*
  • Polyomavirus Infections / virology
  • Sensitivity and Specificity
  • Seroepidemiologic Studies
  • Tumor Virus Infections / blood*
  • Tumor Virus Infections / diagnosis*
  • Tumor Virus Infections / virology

Substances

  • Antibodies, Viral
  • Capsid Proteins
  • Epitopes
  • Immunoglobulin G