title: ELISA methodology to quantify astrocyte production of cytokines/chemokines in vitro.
abstract: Astrocytes are intimately involved in immunological and inflammatory events occurring in the central nervous system (CNS), due to their ability to secrete and respond to a large number of immunoregulatory cytokines/chemokines such as IL-1β, IL-6, IL-8, IL-10, IL-17, IL-27, TNF-α, TGF-β, IFN-γ, IFN-β, CCL2, CCL3, CCL5, CXCL10, and CXCL12. Although expression of cytokines and chemokines is limited in the normal CNS, elevated expression of these proteins, as seen in disease entities such as multiple sclerosis (MS), HIV-1 associated neurocognitive disorders (HAND), Alzheimer's disease (AD), Parkinson's disease (PD) and amyotrophic lateral sclerosis (ALS), contributes to the development of inflammation and neuronal demise in these diseases. As a potent source of cytokines and chemokines, astrocytes play a pivotal role in the type and extent of neuroinflammatory responses. Astrocytes can be stimulated in vitro to produce numerous cytokines/chemokines, which are secreted and detected in supernatants by a technique known as enzyme-linked immunosorbent assay (ELISA). In this chapter, we describe our experience using ELISAs to detect and quantify cytokines and chemokines secreted by stimulated murine astrocytes, specifically IL-6 and CXCL10.
authors.affiliation: Department of Cell Biology, The University of Alabama at Birmingham, Birmingham, AL, USA.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Qin
authors.forename: Etty N
authors.initials: EN
authors.lastname: Benveniste
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods*; D002490:Central Nervous System / Q000166:cytology* / Q000378:metabolism; D054357:Chemokine CXCL10 / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015850:Interleukin-6 / Q000378:metabolism*; D051379:Mice
chemical_list: D054357:Chemokine CXCL10; D015850:Interleukin-6