Detection of interleukin-12 (IL-12)-secreting cells in normal mice with enzyme-linked immunospot (ELISPOT) assay.
To assess mice interleukin-12 (IL-12)-secreting cells at a single cell level, we have developed a murine IL-12 specific enzyme-linked immunospot (ELISPOT) assay. The application of the newly developed method clearly showed the frequency of IL-12-secreting cells in the resident peritoneal exuded cells was higher than other organs of normal DBA/1J mouse. Moreover, we determined the frequency of IL-12-secreting cells in the spleens of five strains of inbred mice, and found the incidence of IL-12 secretors in the strain C57BL/6 to be greatest, and significantly greater than four of the others. These results are compatible with the predicted evidence, supporting this ELISPOT assay for IL-12-secreting cells is accurate. The procedure provides a useful tool for investigating complicated immune responses at a single cell level.
Okamoto
Y
Y
Department of Clinical Pharmacy, Faculty of Pharmacy, Meijo University, Nagoya, Japan. yokamoto@meijo-u.ac.jp
Gotoh
Y
Y
Kobayashi
Y
Y
Mizuno
A
A
Tokui
H
H
Nishida
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000918:Antibody Specificity / Q000187:drug effects / Q000276:immunology; D001202:Ascitic Fluid / Q000166:cytology / Q000378:metabolism*; D001667:Binding, Competitive / Q000276:immunology; D002452:Cell Count; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D016207:Cytokines / Q000032:analysis / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D018664:Interleukin-12 / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D008198:Lymph Nodes / Q000166:cytology / Q000378:metabolism*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012016:Reference Values; D013045:Species Specificity; D013154:Spleen / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D016207:Cytokines; D011500:Protein Synthesis Inhibitors; D018664:Interleukin-12; D003513:Cycloheximide