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Determination of Herpes Simplex Virus‐Induced Alpha Interferon‐Secreting Human Blood Leucocytes by a Filter Immuno‐Plaque Assay
Author(s) -
RÖNNBLOM L.,
CEDERBLAD B.,
SANDBERG K.,
ALM G. V.
Publication year - 1988
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1988.tb02335.x
Subject(s) - polyclonal antibodies , monoclonal antibody , herpes simplex virus , microbiology and biotechnology , antibody , virology , biology , virus quantification , amnion , alpha interferon , interferon , virus , immunology , pregnancy , fetus , genetics
A filter immuno‐plaque assay was developed which detects alpha interferon (IFN‐α) secreting human peripheral blood leucocytes (PBL) Polyclonal anti‐IFN‐α antibodies were fixed to the nitrocellulose membrane bottoms of 96‐well Millititer plates, which also contained monolayers of glutaraldehyde fixed human WISH amnion cells infected by Herpes simplex virus type 1 (HSV). Such cell are potent IFN inducers and during a 16 h cocultivation with PBL, IFN‐α was absorbed by the membrane‐bound polyclonal antibodies around IFN‐α‐secreting cells. This IFN‐α was detected with murine monoclonal antibodies against IFN‐α and peroxidase labelled antibodies against murine immunoglobulin, using diaminobenzidine as substrate Distinctly stained plaques were seen, the frequency of which gave a minimal estimate of approximately 10 IFN‐α‐producing cells in 10 4 PBL (range in 12 blood donors 2.95–25.1). Fewer plaques than expected were seen at low PBL numbers per culture, one explanation being that cell interactions then limit the IFN‐α response. The immuno‐plaque assay should be useful in further studies of the cellular basis of the IFN‐α response.