Detection of differentially expressed genes in particle disease using array-filter analysis
Project management at the University of Würzburg:
The aim of the study was to search for new gene products induced after contact of macrophages with particles.
Method: Using an established macrophage model THP-1 cells were differentiated by vitamin D and GM-CSF into macrophage-like cells (MLC). MLC were incubated with PE particles and lipopolysaccharides (LPS). RNA was isolated an reverse transcribed into 32 P-labeled cDNA. A cDNA espression array was hybridized and analyzed by autoradiography. Re-evaluation of array data was performed using RT-PCR.
Results: The following gene products were upregulated after particle contact from array analysis: TNF-receptor 2, IL-1 receptor antagonist, bone morphogenic protein 4 (BMP 4) and HM 145. Gene products were reevaluated by RT-PCR and differential expression was found. LPS treatment as well resulted in an upregulation of these gene products, however, HM 145 was downregulated by particle contact.
in RT PCR analysis
Conclusion: Results confirm the value of our model of THP-1 cells for research of particle disease, since the cytokines TNF alpha and IL 1 were markedly upregulated. BMP 4 is a known signaling protein, able to induce bone formation in muscle. It is viewed as a mediator of gene regulation in particle disease. HM 145 belongs to the family of chemotactical peptide receptors. HM 145 is one of the first examples of genes induced in septic process but downregulated in particle disease.
Projekt period: from 01.1999 to 04.2002