Detection of Cardiotrophin-like Cytokine Factor 1 (CLCF1) by flow cytometry

Abstract

Abstract Cardiotrophin-like Cytokine Factor 1 (CLCF1) belongs to the IL6 family of cytokines and possesses pro-neurotrophic and immuno-modulating functions. Coding mRNA for CLCF1 has been detected in primary and secondary lymphoid organs (i.e. lymph nodes, spleen and bone marrow), as well as in the lungs and feminine reproductive organs. Modulation of CLCF1’s mRNA levels has been associated with the Th17 polarization in CD4+ T cells. However, little information is available regarding CLCF1 protein levels in these tissues or the nature of the immune cells responsible for its production. This can be explained by a lack of in situ detection options for CLCF1. We have therefore developed a methodology for the detection of human and murine CLCF1 by flow cytometry in permeabilized cells. This technique has been validated using derivatives of the Ba/F3 cell line in which cDNAs coding for human and murine CLCF1 were introduced by transduction with recombinant retroviruses. We are currently using this approach to study CLCF1 production by human and murine immune cells. Preliminary results indicate a production of CLCF1 by Th17 T cells. The development of a method to detect CLCF1 by flow cytometry will be beneficial for the study of CLCF1’s functions in the regulation of the immune response.