title: Cytokine detection by multiplex technology useful for assessing antigen specific cytokine profiles and kinetics in whole blood cultured up to seven days.
abstract: Cytokine profile assessment is important to characterize immune responses to pathogens. To identify optimal time points for determination of cytokine profiles, we diluted whole blood 1:10, to enable daily cytokine measurements during one week. Cultures for 10 blood donors were set up in the presence of phytohemagglutinin (PHA), cytomegalovirus (CMV) or Candida. Supernatant levels of interleukin-2 (IL-2), IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, IL-17, interferon-gamma (IFN-gamma), granulocyte/macrophage colony-stimulating factor, and tumor necrosis factor-alpha (TNF-alpha), were determined by multiplex technique, and intracellular cytokine staining (ICS) was employed to detect IFN-gamma, IL-2, IL-4 and IL-13 in CD3+ cells. The multiplex analysis detected representative cytokine profiles for the majority of the cytokines on day 7 by identifying peak levels or good correlation with peak levels, with the exception of IL-2 and TNF-alpha in PHA and CMV cultures and IL-10 in PHA cultures. For these cytokines an extracellular measurement on day 2-3 would be appropriate. The intracellular cytokines showed distinct kinetics for IFN-gamma and IL-2, while IL-4 and IL-13 were not detected at all with ICS. In conclusion, the combination of whole blood cultures with multiplex analysis is a simple and powerful tool that can be used to identify detailed cytokine profiles of specific cell-mediated immune responses.
authors.affiliation: Department of Immunology and Vaccinology, Swedish Institute for Infectious Disease Control, Nobelsv.18, SE-171 82 Solna, Sweden. maria.lagrelius@smi.ki.se
authors.forename: Maria
authors.initials: M
authors.lastname: Lagrelius
authors.forename: Petra
authors.initials: P
authors.lastname: Jones
authors.forename: Kristina
authors.initials: K
authors.lastname: Franck
authors.forename: Hans
authors.initials: H
authors.lastname: Gaines
mesh_terms: D000328:Adult; D000946:Antigens, Fungal / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D002175:Candida / Q000276:immunology; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis / Q000097:blood*; D003587:Cytomegalovirus / Q000276:immunology; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007700:Kinetics; D008875:Middle Aged
chemical_list: D000946:Antigens, Fungal; D000956:Antigens, Viral; D016207:Cytokines