Data represent mean SEM of measurements made from total 10 mice for each group

Data represent mean SEM of measurements made from total 10 mice for each group. was go through at 400 nm. The percentage of hexosaminidase released is definitely indicated with mean SEM. *denotes the p value 0.05. ns is not significant. Suppl. Fig. 3. Antigen focusing on to FcRI does not enhance antigen demonstration to CD8+ T cells in hFcRI-Tg mice. (A) Schematic of SIINFEKL (OVA (257C264))-Fc. (BCC) hFcRI-Tg mice (Tg+, top panel) and Tg-negative control Rabbit Polyclonal to MOK mice (Tg?, lesser panel) were adoptively transferred with CTV-labeled CD45.1+CD8+ OTI T cells one day before iv injection with 0.2 g or 0.02 g SIINFEKL-Fc. Three days later, spleens were harvested and cells were stained and analyzed by circulation cytometry. The percentage of proliferating CD45.1+TCRV2+CD8+ OTI T cells was determined by gating CTV-diluted cells. 1-Methylinosine Demonstrated in (B) are data from one representative mouse for each group. Demonstrated in (C) are data from 5 mice for each group injected with 0.2 g SIINFEKL-Fc with mean SEM. ns denotes not significant. NIHMS698938-supplement-supplement_1.pdf (701K) GUID:?E7C5011E-DBA4-404D-AB6E-F58D550CB1A3 Abstract Dendritic cells (DCs) play an important role in immune homeostasis through their ability to present Ags at constant state and mediate T cell tolerance. This characteristic renders DCs a stylish restorative target for the induction of tolerance against auto-antigens or allergens. Accordingly, Ag-conjugated DCCspecific Abs have been proposed to be an excellent vehicle to deliver Ags to DCs for demonstration and tolerance induction. However, this approach requires laborious reagent generation methods and entails unpredictable side effects resulting from Ab-induced crosslinking of DC surface molecules. In this study, we examined whether IgE, a high-affinity, nonCcross-linking natural ligand of FcRI, could be used to target Ags to DCs and to induce Ag-specific T cell tolerance. We found that Ag-conjugated human being IgE Fc website (Fc) effectively delivered Ags to DCs and enhanced Ag demonstration by 1000- to 2500-collapse in human being FcRI-transgenic mice. Importantly, this demonstration resulted in a systemic deletion of Ag-specific T cells and prevented these mice from developing delayed-type hypersensitivity, which is definitely critically dependent on Ag-specific T cell immunity. Thus, focusing on FcRI on DCs via Ag-Fc fusion protein may serve an alternative method to induce Ag-specific T cell tolerance in humans. Dendritic cells (DCs) perform an important role in immune tolerance (1). Mice lacking DCs spontaneously develop fatal autoimmunity (2), assisting the significant contribution of DCs to the development or maintenance of tolerance. The tolerogenic part of DCs is dependent on constant state self-antigen demonstration. At rest, DCs continually endocytose and present self-antigens (3C5). This demonstration results in the unresponsiveness or deletion of self-reactive T cells (3, 6). It also mediates the 1-Methylinosine development of regulatory T cells, a unique T cell subset equipped with potent immune-suppressive functions 1-Methylinosine (7, 8). Focusing on Ags to resting DCs using a DC-specific Ab has been suggested like a potential restorative strategy for the induction of tolerance against auto-antigens (9, 10). Injection of nonobese diabetic (NOD) mice having a -cell Ag-fused DEC-205 mAb offers been shown to prevent diabetes (11, 12). Injection with myelin oligodendrocyte glycoprotein Ag fused with DEC205 or Langerin mAbs offers been shown to suppress experimental autoimmune encephalomyelitis in mice (13, 14). However, it is not known whether these Abs would target DCs in humans as efficiently as with mice, because the protein manifestation pattern differs significantly between varieties. Indeed, human being DEC-205 is indicated on more 1-Methylinosine leukocyte populations than mouse DEC-205, including B cells, T cells, monocytes, macrophages, and NK cells (15). In addition, it is hard to forecast the adverse effects elicited by Ab binding. Because Abs are bivalent, their binding.