Use of the whole leucocyte population in the study of the NFκB pathway

Abstract

The nuclear factor NF-{kappa}B (NFκB) is involved in the regulation of innate immunity and in particular, inflammatory genes. It is associated with the pathogenesis of many chronic diseases such as coronary heart disease (CHD). It is believed that individual susceptibility to CHD might be affected by differences in gene transcription and therefore gene expression in circulating cell populations such as leucocytes is of interest. The aim of this study was to investigate whether the total white blood cell population (leucocytes) could be used to study the effect of lipopolysaccharide (LPS) treatment on the expression of genes of the NFκB pathway. Gene expression of the NFκB pathway was examined in total leucocyte, monocyte and neutrophil populations. The majority of the 84 genes examined were up-regulated after treatment with LPS for 12 h in all cell populations examined. The total leucocyte population behaved in a similar manner to both neutrophils and monocytic cells, indicating that it alone could be used in studies, therefore avoiding cell separation, which is time-consuming and can result in cell activation. Furthermore, in clinical studies, it enables a larger number of patient samples to be studied simultaneously, while also reducing the amount of blood required from each. This will provide enough starting material for use with molecular techniques, such as chromatin immunoprecipitation (ChIP) and ChIP-sequencing, and allow large-scale gene expression studies of the NFκB pathway in patients with chronic and acute inflammation with established CHD.