CD152 Promotes Migration Figure 1. CD152 enhances chemotaxis of Th1 cells. (A) Migration of unpolarized T cells. Recall response of CD4+ OVA-specific TCRtg T cells from CD1522/2 or CD152+/+ mice was induced by adding 1 mg/ml OVA323–339 and T cell-depleted APCs. On day 6 of recall response cells were analyzed in chemotaxis assays. Bars indicate migrated CD4+ cells as percentage of input cells. (B) Migration of CD1522/2 and CD152+/+ T cells in primary swtiemreualantaiolynz:eCdDi4n+cOheVmA-ostpaexcisifiacsTsaCyRst.gBTarcsesllhsowwertehestcimheumlaotetadcwticitihnd1emxgo/fmClDO4V+Ac3e2l3l–s3.3(C9a)nSpdeTcicfeicllm-diegpralettioedn oAfPaCnst.iOgennd-sapye6ciofifcpsrtiimmaurlyatsetdimTuhl1atcioenllscienllas recall response: Primary stimulation and recall response of CD4+CD62L+ OVA-specific TCRtg T cells were performed under Th1 conditions with 1 mg/ml OVA-peptide in the presence of 200 mg/ml neutralizing anti-CD152 Fab fragments or hamster control Fab fragments. Cells were examined in chemotaxis assay on day 6 of recall response. (D) Chemotactic index of a polyclonally induced recall response of CD1522/2 or CD152+/+ Th1 cells: Primary stimulation and recall response of splenocytes from CD1522/2 and CD152+/+ mice were induced polyclonally (as described in Material and Methods) and CD4+ cells were used for chemotaxis assays on day 4 of recall response. (E) Migration of CD1522/2 and CD152+/+ Th1 cells in a recall response is dose dependent: Primary stimulation and recall response of CD4+ cells from TCRtg CD1522/2 and CD152+/+ mice were induced antigen- specifically using Th1 conditions with indicated amounts of OVA-peptide and cells were analyzed on day 6 of recall response in chemotaxis assays. All data shown represent one out of 3–4 similar experiments. doi:10.1371/journal.pone.0005702.g001 pertussis toxin (PTX), CD152-stimulated migration was reduced to that of background migration (Fig. 2). Thus, CD152-enhanced migration of T cells completely depends on Gai signal transduction. To control for the possibility that increased IFN-c levels of CD1522/2 T cell populations contributed to the reduced migration [11,30], the IFN-c-production was examined during in vitro culture. PLoS ONE | www.plosone.org 3 May 2009 | Volume 4 | Issue 5 | e5702