Simultaneous detection of human IL-17, FoxP3 and other Th cytokines and surface markers by multi-color flow cytometry (47.4)

Abstract Naïve CD4+ T helper cells can be induced to differentiate towards at least four types of committed helper T cells, namely T helper 1 (Th1), Th2, Th17 and regulatory T (Treg) cells according to the local cytokine milieu. The committed cells are characterized by the expression of specific transcription factors and by the cytokines released. Th17 cells are acknowledged to be instrumental in the response against microbial infection, but are also very efficient inducers of tissue inflammation and crucial initiators of organ specific autoimmunity. FoxP3+ Treg cells are necessary and sufficient to prevent autoimmunity throughout the life span of an individual. Immune and autoimmune responses are regulated by a fine balance between effector and regulatory T cells. There is a reciprocal relationship between pathogenic Th17 cells and Treg cells. Therefore the need to simultaneously detect multiple transcription factors and cytokines to define these populations is becoming increasingly important. In this report, we describe the simultaneous detection of IL-17 and FoxP3 as well as other Th cytokines (IFN-gamma and IL-4) and cell surface markers (CD3, CD4, CD25, CCR4, CCR6, CD45RA) using optimized buffer systems and protocols with multi-color flow cytometry.