Development of a neutralizing monoclonal antibody against rabbit IL-1 receptor antagonist and utilization for ELISA and measurement of masked IL-1 activity in biological materials.
We developed a monoclonal antibody with neutralizing activity against the rabbit IL-1 receptor antagonist (IL-1ra) and a sandwich ELISA for rabbit IL-1ra. Using this ELISA, we measured IL-1ra contents in inflammatory exudate cells and culture supernatants of LPS-stimulated macrophages. The production of IL-1ra occurred rapidly after the onset of inflammation in vivo and after stimulation of macrophages in vitro and was sustained for a long period. The time interval for detection of IL-1 activity overlapped with the above mentioned period of IL-1ra production. In the presence of this neutralizing monoclonal IgG, the effect of IL-1ra in these biological specimens was nil and heretofore unrecognized IL-1 activity in the samples was measurable; the rate of this increment in activity ranging between 1.5- and 4.8-fold. We conclude that underestimations of IL-1 activity have been made and previously reported results on IL-1 activities in many biological materials have to be re-examined.
Matsukawa
A
A
Department of Pathology, Kumamoto University School of Medicine, Japan.
Furukawa
S
S
Ohkawara
S
S
Takagi
K
K
Yoshinaga
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D002478:Cells, Cultured; D005260:Female; D007074:Immunoglobulin G / Q000276:immunology; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1 / Q000032:analysis / Q000276:immunology*; D017737:Macrophages, Peritoneal / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D017472:Receptors, Interleukin-1 / Q000037:antagonists & inhibitors*; D011994:Recombinant Proteins / Q000276:immunology; D012795:Sialoglycoproteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; C507764:Il1rn protein, mouse; D007074:Immunoglobulin G; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins; D012795:Sialoglycoproteins