Anti-DEF6 (aa 499-528) polyclonal antibody

Although GM-CSF has been widely used in dendritic cell (DC) research, the mechanisms, factors, and signals regulating steady-state differentiation and maturation of GM-CSF-dependent DCs are insufficiently known. We found that the absence, individually or combined, of the related proteins DEF6 and SWAP-70 strongly enhances differentiation of murine GM-CSF-derived DCs. Contrasting SWAP-70, control through DEF6 does not depend on RHOA activation. DEF6 deficiency leads to expression of the DC-specific transcription factor ZBTB46 and prolonged STATS activation in GM-CSF cultures. SWAP-70 and DEF6-mediated restriction of DC differentiation converges mechanistically at the NF-kappa B pathway. DEF6 acts at early stages of DC differentiation in CD115(-) cKIT(+) myeloid DC progenitors, whereas SWAP-70 acts subsequently. SWAP-70 and DEF6 regulate steady-state DC cytokine expression as well as in vivo accumulation in lymphatic tissue of migratory DCs. Our studies thus elucidate previously unknown roles of two closely related factors with distinct and complementary activities in DC differentiation and steady-state DC function.

Many proteins that are aberrantly expressed in malignant tumors play important roles in promoting tumorigenesis, metastasis and immune escape. IFN regulatory factor 4-binding protein (IBP), which is a novel PH-DH-like protein related to SWAP-70, and functions as an upstream activator of Rho GTPases. It is widely expressed in cells of the immune system and is involved in coupling activated cell receptors to downstream signaling events that mediate cell proliferation, differentiation and polarization. Although IBP was detected in human chondrosarcoma, its function in tumor cells remains unknown. In this study, newly generated monoclonal anti-IBP antibodies were employed and they detected higher level expression of IBP in some human invasive breast carcinoma tissues and in two breast cancer cell lines that form highly invasive tumors in nude mice. In contrast, the levels of IBP mRNA and protein were low or undetectable in normal human breast tissues, benign breast lesions or low-tumorigenic breast cancer cell lines. over-expression of wild-type IBP in an IBP-negative breast cancer cell line markedly increased its proliferation and invasiveness in vitro. Conversely, RNA interference-mediated knockdown of IBP expression in an IBP-positive breast cancer cell line significantly reduced cell growth and invasiveness. Our results indicate that IBP is expressed in more highly invasive human breast cancer cells, such as MCF-7 and MDA-MB-231, with lower expression in normal breast tissue, benign tumors and less aggressive breast cancer cells, such as SKBR3 and MDA-MB-453. Thus, expression of IBP is correlated with the degree of malignant breast tumors. Nevertheless, it should be pointed our that further study with more tumor types is required to fully elucidate the role of IBP in tumorigenesis and the potential of IBP as a marker for more highly malignant tumors. (C) 2009 Elsevier B.V. All rights reserved.