Screening the differentially expressed genes of immature and LPS-induced mature dendritic cells using cDNA microarray / 解放军医学杂志

ABSTRACT

Objective To study the changes in gene expression profiles of dendritic cells(DCs) during their maturation processes using cDNA microarray.Method DCs were generated from human peripheral blood monocytes induced by GM-CSF and IL-4,and lipopolysaccharide(LPS) was used to induce the maturation of DCs.After inducing for 48h,cells were collected to extract the general RNA.Genes expressions of mature DCs(maDC) and immature DCs(imDC) were analyzed using cDNA microarray,and immature DCs without induction served as control.Results DCs expressed highly surface markers of mature DCs after LPS induction.The studied data showed that among those 165 target genes,a total of 50 genes(30.3%) exhibited differential levels of expression in LPS-induced mature DCs.Thirty-five genes were up-regulated and fifteen genes were down-regulated.Maturation-dependent up-regulation,defined by a differential expression ratio of ≥2,included twelve cytokine and chemokine genes,ten antigen uptake and presentation genes and thirteen signal transduction molecule genes.Reciprocally,maturation-dependent down-regulation,defined by a differential expression ratio of ≤0.5,occurred with two cytokine,five antigen uptake and presentation genes and eight signal transduction molecule genes.Semi-quantitative reverse transcription polymerase chain reaction(RT-PCR) was employed to further confirm the results of cDNA microarray.Conclusion It is suggested that LPS has significant influence on many genes of DCs in their maturing processes.Those changed genes include cytokines,chemokines and their receptors,antigen uptake and presentation molecules and signal transduction molecules.Analysis on these differentially expressed genes is helpful in understanding the maturing processes of DCs.