Detection of Cytomegalovirus Interleukin 10 (cmvIL-10) by Enzyme-Linked Immunosorbent Assay (ELISA)

Methods Mol Biol. 2021:2244:291-299. doi: 10.1007/978-1-0716-1111-1_15.

Abstract

Since its introduction in 1971, the enzyme-linked immunosorbent assay (ELISA) has revolutionized medicine by enabling detection of both antigens and antibodies in a variety of samples. We describe here a customized sandwich ELISA developed for the detection of Human Cytomegalovirus interleukin-10 (cmvIL-10). CmvIL-10 is a virally encoded cytokine and ortholog of human interleukin 10 (hIL-10). While cmvIL-10 and hIL-10 are similar in structure and function, overall amino acid sequence identity is only 27%, resulting in antigenically distinct proteins. The cmvIL-10 ELISA is specific and does not detect hIL-10. The assay is sensitive enough to detect cmvIL-10 in both culture supernatants and patient serum. The ability to quantify cmvIL-10 levels during HCMV infection could provide valuable information about immune evasion strategies and viral control of host signaling pathways.

Keywords: CMV; Cytokine; ELISA; Human cytomegalovirus; Immunoassay; cmvIL-10.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Viral / metabolism
  • Cytokines / metabolism
  • Cytomegalovirus / immunology*
  • Cytomegalovirus / metabolism
  • Cytomegalovirus Infections / immunology
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Humans
  • Interleukin-10 / analysis*
  • Interleukin-10 / immunology
  • Protein Binding
  • STAT3 Transcription Factor / metabolism
  • Signal Transduction
  • Viral Proteins / metabolism

Substances

  • Antibodies, Viral
  • Cytokines
  • IL10 protein, human
  • STAT3 Transcription Factor
  • Viral Proteins
  • Interleukin-10