title: A sensitive in situ ELISA for quantitative measurements of cytokines and antibodies secreted by culture lymphocytes.
abstract: Cytokines are usually measured using ELISA and ELISPOT. We have developed an in situ ELISA by combining the advantages of the highly sensitive ELISPOT and the quantitative conventional ELISA techniques to measure cytokines and antibodies secreted by culture lymphocytes. In this assay, lymphocytes were cultured on ELISA plates pre-coated with either a special antigen or mAbs against IL-4, IFN-gamma. The caprtured lymphokines or immunoglobulins were subsequently measured by conventional ELISA procedures. As a comparison, a conventional ELISA was carried out in parallel. The in situ ELISA measured a 5-fold increase in antigen-stimulated IL-4 and IFN-gamma, compared to the conventional ELISA. It also rquired 10,000 times fewer lymphocytes to produce a detectable level of antigen-specific antibodies than the conventional ELISA. We conclude that the in situ ELISA is much more sensitive than a conventional ELISA and less costly and easier to perform than ELISPOT, providing a useful tool to quantitatively measure cell-secreted molecules.
authors.affiliation: Department of Paediatrics and Child Health, University of Manitoba, John Buhler Research Centre, Winnipeg, Canada.
authors.forename: Y
authors.initials: Y
authors.lastname: Chen
authors.forename: Z
authors.initials: Z
authors.lastname: Peng
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis; D002478:Cells, Cultured; D016207:Cytokines / Q000032:analysis / Q000096:biosynthesis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D007074:Immunoglobulin G / Q000032:analysis / Q000096:biosynthesis*; D007371:Interferon-gamma / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D007700:Kinetics; D008214:Lymphocytes / Q000276:immunology*; D018942:Macrolides / Q000494:pharmacology; D051379:Mice; D011863:Radioimmunoassay / Q000379:methods
chemical_list: D000906:Antibodies; D016207:Cytokines; D007074:Immunoglobulin G; D018942:Macrolides; D015847:Interleukin-4; C036978:concanamycin A; D007371:Interferon-gamma