SUPPRESSORS OF CYTOKINE SIGNALING AND IL-10 INHIBITORY EFFECT IN LYME DISEASE

这个子项目是众多研究子项目之一

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We recently showed that mouse J774 macrophages incubated with IL-10 and Borrelia burgdorferi spirochetes augmented their SOCS3 expression levels. This correlated with the IL-10 inhibition of expression of several pro-inflammatory cytokines. We hypothesized that SOCS3 expression induced by B. burgdorferi and IL-10 in macrophages is functionally important in the control of inflammation in Lyme disease, with SOCS3 potentially acting as a key mediator of the IL-10 anti-inflammatory activity in macrophages. To begin to address this hypothesis, we silenced socs3 gene expression in J774 cells using SOCS3 small interfering RNA to determine the effect of silencing on IL-10 inhibition of pro-inflammatory mediators induced by B. burgdorferi. Transfected cells were co-stimulated with IL-10 and B. burgdorferi spirochetes; RNA samples were collected and subjected to genome-wide transcriptomics using mouse oligonucleotide microarrays. These studies revealed that the socs3 gene is up-regulated (~40-fold) in cells stimulated with IL-10 and B. burgdorferi. In the presence of SOCS3, IL-10 could down-modulate several pro-inflammatory mediators in B. burgdorferi-stimulated macrophages. However, silencing of the socs3 gene in these macrophages resulted in a marked reversal of the down-modulatory effect exerted by IL-10 on many of these inflammatory mediators including cytokines/receptors, chemokines, IFN-inducible, apoptotic and TNF ligand/receptor genes. This study demonstrates that SOCS3 in part mediates the IL-10 anti-inflammatory effect in macrophages stimulated with B. burgdorferi spirochetes.