Genes cytokine expression analysis

Gene expression analysis provides a window into the molecular events that initiate cytokine synthesis in response to various stimuli encountered by... 

Alvaro-Gracia, J. M., Zvaifler, N. J., Brown, C. B., Kaushansky, K., Firestein, G. S. (1991). Cytokines in chronic inflammatory arthritis VI. Analysis of the synovial cells involved in granulocyte-macrophage colony-stimulating factor production and gene expression in rheumatoid arthritis and its regulation by IL-1 and tumor necrosis factor-a J. Immunol 146,3365-71. 

As stated earlier, activation of endothelial cells by pro-inflammatory stimuli leads to the expression of cell adhesion molecules and cytokines such as IL-6 and IL-8. The expression and hence modulation of surface expressed adhesion molecules by e.g. targeted delivery of inhibitors of NFkB, can be measured using flow cytometric analysis or whole cell ELISA techniques. Cytokine production can be measured in the supernatant of cultured cells or in biological fluids. Furthermore, competitive or quantitative RT-PCR analysis of mRNA levels of cell adhesion molecules or cytokines, allows the transcriptional activity of the genes of interest to be estimated. 

Reverse transcription-polymerase chain reaction (RT-PCR) has been widely used for the detection of cytokine gene expression in clinical samples (W18, K5). However, conventional RT-PCR only offers a semiquantitative analysis. Recently, the Perkin-Elmer Corporation (Wellesley, MA) developed the TaqMan cytokine gene expression plate for real-time, in vitro quantitative evaluation of a panel of human cytokine gene expression using fluorescence detection. 

Fourteen-Membered-Ring Macrolide Antibiotics Suppress Gene Expression of Inflammatory Cytokines in Human Bronchial Epithelial Cells We evaluated the effects of macrolides on steady-state levels of IL-6 and IL-8 mRNA by Northern blot analysis [17, 60]. As shown in Fig. 13, bronchial epithelial cells expressed constitutive IL-8 mRNA, which was significantly upregulated by the cytokines such as IL-la, IL-1[3, and TNF-a. We reported that EM, CAM, and RXM inhibited steady-state levels of IL-6 and IL-8 expression in normal and immortalized bronchial epithelial cells [60, 62, 63]. This action appeared to be unique, because other antibiotics including a 16-membered ring macrolide JM... 

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