ARUP Institute for Clinical and Experimental Pathology
Jaskowski, Troy D.
Martins, Thomas B.; Pasi, Brian M.; Pickering, Jerry W.; Litwin, Christine M.; Hill, Harry R.
Determination of cytokine responses using a multiplexed fluorescent microsphere immunoassay
We used a multiplexed fluorescent microsphere immunoassay to develop a sandwich capture assay to assess simultaneously the production of thymus helper (TH) 1- and TH2-type cytokines in tissue culture supernatant obtained from stimulated peripheral blood mononuclear cells. The assay then was used to assess the cytokine production of patients with hyperimmunoglobulinemia E syndrome and in cord blood from neonates. The multiplexed assay has a reportable range of less than 10 to 50,000 pg/mL. For linearity and recovery studies, R2 values for the 6 cytokines ranged from 0.988 to 0.999 for samples spiked with known concentrations of recombinant cytokine standards and for patient samples. The assay showed good specificity, with little cross-reactivity between cytokines. Results from supernatants of Staphylococcus aureus–stimulated peripheral blood mononuclear cells obtained from 6 patients with hyperimmunoglobulinemia E syndrome showed significantly less interferon (IFN)-gamma production than cells from healthy control subjects. Cord blood cells from neonates produced significantly less interleukin 12 and IFN-gamma than cells from adults in group B streptococci–stimulated mononuclear cells. The fluorescent multiplexed microsphere immunoassay can be used to quantitate multiple cytokines from 1 sample and should be useful for further understanding of the cytokine role in disease.
American Society for Clinical Pathology
Multiplexed fluorescent microsphere immunoassay;
Cytokines; Immunoassay; Immunodeficiency;
Martins, T. B., Pasi, B. M., Pickering, J. W., Jaskowski, T. D., Litwin, C. M., & Hill, H. R. (2002). Determination of cytokine responses using a multiplexed fluorescent microsphere immunoassay. American Journal of Clinical Pathology, 118(3), 346-53.