The inflammatory response is predominantly triggered by immune recognition of microbial products or tissue damage and propagated by inflammatory cytokines inducing effector functions such as activation and recruitment of leukocytes. During inflammation, a plethora of secreted cytokines interact in multiple fashions to induce and control an appropriate response against the encountered threat both the type and strength of the response.
Primary human monocytes were isolated from heparinized blood samples obtained from three healthy donors using a negative selection protocol. Total RNA was isolated, depleted from rRNA, and sequenced after cDNA library generation on a NextSeq sequencer Illumina. For normalization and differential expression analysis of the obtained data set, DESeq2 was used.
These include the four genes ACOD1, INHBA, UBD, and CLEC6A. While primary monocytes seem to differentiate into an activated phenotype mobilizing the immune reaction against intracellular bacteria, other cells may just die to leave the threat to recruited neutrophils. The ubiquitously expressed protein MARCKS myristolated alanine-rich C-kinase substrate was initially identified as a regulator of actin filament dynamics.
However, the involvement of MARCKS in immune cell functions is scarcely characterized. Therefore, we aimed at elucidating the molecular mechanisms by which MARCKS regulates immunologic processes in monocytes.
Both cell types were differentiated towards monocytes using calcitriol nM, 4 d. Differentiation was confirmed by detecting surface marker expression CD14 and CD11b; flow cytometry. Primary human monocytes were isolated from heparinized blood samples by negative selection. Reactive oxygen species ROS were detected in luminometer-based time course experiments and phagocytosis was measured based on the uptake of fluorescent-labelled E.
