Analysis of ovine IL-1β production in vivo and in vitro by enzyme immunoassay and immunohistochemistry

A monoclonal antibody (mAb) specific for ovine IL-1β was produced and, in conjunction with a polyclonal rabbit antiserum, used to develop a sensitive enzyme immunoassay (EIA) for ovine interleukin 1β (IL-1β). The mAb neutralised the activity of recombinant ovine IL-1β (rOvIL-1β) and native OvIL-1 in...

Full description

Saved in:
Bibliographic Details
Published in:Veterinary immunology and immunopathology 1997-07, Vol.57 (3), p.267-278
Main Authors: Rothel, J.S., Hurst, L., Seow, H.-F., Pépin, M., Berthon, P., Corner, L.A., Wood, P.R.
Format: Article
Language:English
Subjects:
Alkaline Phosphatase
Animals
Antibodies, Monoclonal - biosynthesis
Antibodies, Monoclonal - pharmacology
Antibody Specificity
Cell Line
Corynebacterium Infections - immunology
Corynebacterium Infections - pathology
Corynebacterium Infections - veterinary
Corynebacterium pseudotuberculosis
Enzyme immunoassay
Immunoenzyme Techniques - veterinary
Immunohistochemistry
Interleukin 1
Interleukin-1 - antagonists & inhibitors
Interleukin-1 - biosynthesis
Interleukin-1 - immunology
Lymphocyte Activation
Macrophages - metabolism
Mice
Monoclonal antibody
Ovine
Recombinant Proteins - immunology
Sensitivity and Specificity
Sheep - immunology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:A monoclonal antibody (mAb) specific for ovine IL-1β was produced and, in conjunction with a polyclonal rabbit antiserum, used to develop a sensitive enzyme immunoassay (EIA) for ovine interleukin 1β (IL-1β). The mAb neutralised the activity of recombinant ovine IL-1β (rOvIL-1β) and native OvIL-1 in an ovine thymocyte proliferation assay. However, it did not neutralise the biological activity of rOvIL-1β in the murine NOB1 CTLL assay. The mAb did not react with rOvIL-1α, IL-2, IL-4, IL-8, tumor necrosis factor-α, gamma-interferon or recombinant human IL-1β in indirect EIA. Immunohistological staining of activated alveolar macrophages and frozen lymph node sections demonstrated that the mAb detected IL-1β secreted by ovine macrophages (CD11c-positive). The EIA was highly sensitive, detecting less than 50 pg ml −1 of rOvIL-1β and low levels of native IL-1β in supernatants from lipopolysaccharide-stimulated macrophages. The EIA did not detect heat-inactivated IL-1β.
ISSN:0165-2427
1873-2534
DOI:10.1016/S0165-2427(96)05754-6