Detection of interleukin 6-producing cells among various organs in normal mice with an improved enzyme-linked immunospot (ELISPOT) assay.
The enzyme-linked immunospot (ELISPOT) assay is an efficient technique for the enumeration of single cells secreting antibodies or cytokines. To assess mice interleukin 6 (IL-6)-secreting cells at a single cell level, a highly sensitive ELISPOT assay with 5-bromo-4-chloro-3-indolylphosphate p-toluidine salt and nitroblue tetrazolium chloride (BCIP/NBT) as a gel-substrate has been developed. The IL-6 specific ELISPOT assay thus developed with an ordinary plastic ELISA plate is as efficient as the previous IL-6 specific ELISPOT assay system which uses a nitrocellulose coated plate, and is much more economic and convenient. We demonstrated with this newly devised ELISPOT assay that the ratio of IL-6 producing cells in the resident peritoneal exuded cells was highest in normal BALB/c mice, and a few IL-6 secreting cells were detected in all organs we tested, namely the spleen, bone marrow, Peyer's patch, axillary lymph node and thymus.
Okamoto
Y
Y
Department of Clinical Pharmacy, Faculty of Pharmacy, Meijo University, Nagoya, Japan.
Murakami
H
H
Nishida
M
M
D000818:Animals; D001365:Axilla; D001853:Bone Marrow / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D015850:Interleukin-6 / Q000032:analysis / Q000096:biosynthesis* / Q000378:metabolism; D008198:Lymph Nodes / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010581:Peyer's Patches / Q000378:metabolism; D012680:Sensitivity and Specificity; D013154:Spleen / Q000378:metabolism; D013950:Thymus Gland / Q000378:metabolism
D015850:Interleukin-6