Unit

UNIT 14.16 Measuring TLR Function in Transfectants

  1. Prasad Rallabhandi1,2

Published Online: 1 NOV 2010

DOI: 10.1002/0471142735.im1416s91

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Rallabhandi, P. 2010. Measuring TLR Function in Transfectants. Current Protocols in Immunology. 91:14.16:14.16.1–14.16.15.

Author Information

  1. 1

    University of Maryland, Baltimore, Maryland

  2. 2

    U.S. Food and Drug Administration, Laurel, Maryland

Publication History

  1. Published Online: 1 NOV 2010
  2. Published Print: NOV 2010

Abstract

This unit summarizes a combination of methods that can be optimized for measuring toll-like receptor (TLR) function. TLRs serve as primary innate immune sensors and exhibit high specificity towards evolutionarily conserved microbial and viral structures. The unit focuses specifically on TLR4, the principal Gram-negative lipopolysaccharide (LPS) sensor. Methods described include transient transfections, analyses of activation of various promoters in reporter-gene assays, and induction of IL-8 secretion. Other topics that will be briefly discussed include the necessity for the assessment of surface expression of transmembrane receptors (e.g., TLR4) using FACS analysis, and a permutation of the TLR functional analysis approach using site-directed mutagenesis. Curr. Protoc. Immunol. 91:14.16.1-14.16.15. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • toll-like receptors (TLRs);
  • TLR agonists;
  • lipopolysaccharide (LPS);
  • signal transduction;
  • cell activation;
  • transient transfection;
  • reporter or luciferase assays;
  • mutants;
  • FACS analysis