Analysis of ovine IL-1 beta production in vivo and in vitro by enzyme immunoassay and immunohistochemistry.
A monoclonal antibody (mAb) specific for ovine IL-1 beta was produced and, in conjunction with a polyclonal rabbit antiserum, used to develop a sensitive enzyme immunoassay (EIA) for ovine interleukin 1 beta (IL-1 beta). The mAb neutralised the activity of recombinant ovine IL-1 beta (rOvIL-1 beta) and native OvIL-1 in an ovine thymocyte proliferation assay. However, it did not neutralise the biological activity of rOvIL-1 beta in the murine NOB1/CTLL assay. The mAb did not react with rOvIL-1 alpha, IL-2, IL-4, IL-8, tumor necrosis factor-alpha, gamma-interferon or recombinant human IL-1 beta in indirect EIA. Immunohistological staining of activated alveolar macrophages and frozen lymph node sections sections demonstrated that the mAb detected IL-1 beta secreted by ovine macrophages (CD11c-positive). The EIA was highly sensitive, detecting less than 50 pg ml-1 of rOvIL-1 beta and low levels of native IL-1 beta in supernatants from lipopolysaccharide-stimulated macrophages. The EIA did not detect heat-inactivated IL-1 beta.
Rothel
J S
JS
CSIRO Division of Animal Health, Animal Health Research Laboratory, Parkville, Vic., Australia.
Hurst
L
L
Seow
H F
HF
Pépin
M
M
Berthon
P
P
Corner
L A
LA
Wood
P R
PR
D000469:Alkaline Phosphatase; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000494:pharmacology; D000918:Antibody Specificity; D002460:Cell Line; D003354:Corynebacterium Infections / Q000276:immunology / Q000473:pathology / Q000662:veterinary; D016925:Corynebacterium pseudotuberculosis; D007124:Immunoenzyme Techniques / Q000662:veterinary; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000276:immunology*; D008213:Lymphocyte Activation; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D012756:Sheep / Q000276:immunology*
D000911:Antibodies, Monoclonal; D007375:Interleukin-1; D011994:Recombinant Proteins; D000469:Alkaline Phosphatase