12 Targeted knock down of CCL22 and CCL17.pdfVIP

ARTICLE IN PRESS Immunobiology 215 (2010) 153–162 www.elsevier.de/imbio Targeted knock down of CCL22 and CCL17 by siRNA during DC differentiation and maturation affects the recruitment of T subsets a,b, a a a a Shijun Kang , Jianmin Xie , Shudong Ma , Wangjun Liao , Junyi Zhang , Rongcheng Luoa, aDepartment of Oncology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China bDepartment of Immunology, University of Illinois, Chicago 60612, USA Received 1 December 2008; received in revised form 26 February 2009; accepted 3 March 2009 Abstract Chemokines secreted by DC are instrumental for DC to regulate their own migratory capacities and to recruit T lymphocytes during local tumour immune response. Using the recently developed chemokine protein arrays, we analyzed 38 chemokines associated with monocyte-derived DC (MoDC), including the CC family (CCL2, CCL3, CCL4, CCL17, CCL18, CCL22, CCL23, CCL24, CCL27) and the CXC family (CXCL3, CXCL5, CXCL7, CXCL8, CXCL16) chemokines. Our results indicate that MoDC largely inherit the chemokines constitutively expressed by monocytes, with a significant induction of CCL17, CCL22 and CCL23. Spent culture supernatant collected from MoDC exhibited chemotatic abilities to activate CD4 +, CD8 +, and CD25 + Foxp3 + regulatory T cells (Tregs). Selective knock down of CCL22 and CCL17 expression by siRNA decreased the ratios of CD4 + to CD8 +, as well as the frequency of Tregs recruited by MoDC. Intratumoural injection of MoDC transfected with siCCL22 and siCCL17, significantly reduced the number of Tregs while increasing the number of infiltrating CD8 + T cells