title: Monitoring of human immunological responses to vaccinia virus.
abstract: For the last 30 yr, interest in vaccinia virus immune monitoring has focused on the use of the vaccinia virus as a recombinant vaccine vector and the potential detrimental effect of antivector immunity on subsequent vaccination with a recombinant vaccinia virus. However, interest in this area has intensified after the publication of reports suggesting that smallpox may be a major pathogen selected for bioterrorist activities. Owing to the unacceptably high incidence of complications induced by previous effective smallpox vaccine strains, alternative safer strains (e.g., modified vaccinia Ankara [MVA]) are being assessed for their antigenicity in clinical trials. The exact immune effector mechanism responsible for vaccine-induced protection to smallpox infection has not been fully elucidated, although it is believed that neutralizing antibody plays a major role. This chapter describes a simple enzyme-linked immunosorbent assay (ELISA) to quantify vaccinia virus antibody titer. Additionally, to define serum-neutralizing activity, both a classical plaque reduction assay and a high-throughput 96-well plate method based on reduction of recombinant vaccinia virus expressed beta-galactosidase is described. Furthermore, details are given for a T-cell proliferation assay, primarily for monitoring T-helper CD4 activity and an enzyme-linked immunospot (ELISPOT) assay for CD8 analysis. The use of reliable immunological assays is vital in assessing the potential efficacy of new vaccines to protect against smallpox infection.
authors.affiliation: Oxford BioMedica (UK) Ltd, Oxford, United Kingdom.
authors.forename: Richard
authors.initials: R
authors.lastname: Harrop
authors.forename: Matthew G
authors.initials: MG
authors.lastname: Ryan
authors.forename: Hana
authors.initials: H
authors.lastname: Golding
authors.forename: Irina
authors.initials: I
authors.lastname: Redchenko
authors.forename: Miles W
authors.initials: MW
authors.lastname: Carroll
mesh_terms: D000914:Antibodies, Viral / Q000097:blood*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007107:Immune System; D009500:Neutralization Tests; D012900:Smallpox Vaccine / Q000008:administration & dosage / Q000276:immunology*; D014615:Vaccinia / Q000276:immunology*; D014616:Vaccinia virus / Q000276:immunology* / Q000502:physiology
chemical_list: D000914:Antibodies, Viral; D012900:Smallpox Vaccine