Detecting Immune Responses to Type III Secretion Systems.

Measuring immune responses against type III secretion and related molecules has been made easier with the advent of reporter cell lines. For example, the THP-1-XBlue and HEK-Blue cells from InvivoGen provide easy detection of gene activation under NF-kB and AP-1 control. In addition, many of these cells have been engineered to express specific pattern recognition receptors (PRRs) helping to elucidate activation pathways. In addition, the THP-1 parent cell line from ATCC provides a useful tool for evaluating cytokine release upon stimulation. This chapter will outline using X-blue reporter cell lines, setting up experiments to measure cytokine secretion, and general guidelines for cytokine quantification.