Accurate and simple measurement of the pro-inflammatory cytokine XXXX using a whole blood stimulation assay.

@entity1 @entity1356 @entity74 @entity10400 @entity855
Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues as well as altered cytokine homeostasis. The innate immune system plays a crucial role in recognizing pathogens and other endogenous danger stimuli. One of the major cytokines released by innate immune cells is Interleukin (IL)-1. Therefore, we utilize a whole blood stimulation assay in order to measure the secretion of inflammatory cytokines and specifically of the pro-inflammatory cytokine @entity1356 (1, 2, 3). @entity1 with @entity74 of the innate immune system causing autoinflammatory syndromes show an exaggerated release of mature @entity1356 upon stimulation with LPS alone. In order to evaluate the innate immune component of @entity1 who present with inflammatory-associated pathologies, we use a specific immunoassay to detect cellular immune responses to pathogen-associated molecular patterns (PAMPs), such as the gram-negative bacterial endotoxin, lipopolysaccharide (LPS). These PAMPs are recognized by @entity10400 ( @entity10400 ), which are found on the cells of the innate immune system (4, 5, 6, 7). A primary signal, LPS, in conjunction with a secondary signal, @entity855 , is necessary for the activation of the inflammasome, a multiprotein complex that processes @entity1356 to its mature, bioactive form (4, 5, 6, 8, 9, 10). The whole blood assay requires minimal sample manipulation to assess cytokine production when compared to other methods that require labor intensive isolation and culturing of specific cell populations. This method differs from other whole blood stimulation assays; rather than diluting samples with a ratio of RPMI media, we perform a white blood cell count directly from diluted whole blood and therefore, stimulate a known number of white blood cells in culture (2). The results of this particular whole blood assay demonstrate a novel technique useful in elucidating @entity1 cohorts presenting with autoinflammatory pathophysiologies.
