Protocol

Allergy Methods and Protocols

Volume 138 of the series Methods in Molecular Medicine pp 121-131

Quantification of Human Chemokine Production in TLR-Stimulated and Antigen-Specific Recall Responses

  • Monique StinsonAffiliated withDepartment of Immunology, University of Manitoba
  • , Renee DouvilleAffiliated withDepartment of Immunology, University of Manitoba
  • , Yuriy LissitsynAffiliated withDepartment of Immunology, University of Manitoba
  • , Melanie BlanchardAffiliated withDepartment of Immunology, University of Manitoba
  • , William StefuraAffiliated withDepartment of Immunology, University of Manitoba
  • , Estelle SimonsAffiliated withDepartment of Immunology, University of Manitoba
  • , Allan BeckerAffiliated withDepartment of Immunology, University of Manitoba
  • , Peter NickersonAffiliated withDepartment of Immunology, University of Manitoba
  • , Kevin CoombsAffiliated withDepartment of Immunology, University of Manitoba
    • , Kent HayGlassAffiliated withDepartment of Immunology, University of Manitoba

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Abstract

Chemokines are primarily low molecular mass proteins that are produced and usually released by a wide variety of cell types. Differential chemokine responses can be excellent early markers of immune dysfunction, allowing clinical intervention prior to expression of full blown undesirable effector responses. Thus, assessment of the nature and intensity of Ag-dependent chemokine production provides a valuable tool for probing human immune regulation.

Here, we provide detailed instructions on approaches we have developed to assess the nature and intensity of recall responses to a wide variety of exogenous and endogenous antigens capable of consistently stimulating chemokine responses by PBMC from adult and pediatric populations. This chapter is divided into two sections. The first is focused on culture techniques for eliciting antigen-driven chemokine responses for a panel of chemokines that are relevant to immune function. The second section details assay systems for their quantitative analysis.

Key Words

Chemokine quantitation chemoattractant human immunoassay ELISA primary culture