Keywords: Cancer r Cross-presentation r Long peptides r Plasmacytoid dendritic cells r Viral infectionAdditional supporting information may be found in the online version of this article at the publisher's web-site IntroductionDeveloping efficient therapeutic strategies for cancer and viral infection remains challenging. Successful immunotherapies should elicit robust cell-mediated immunity that is able to eliminate tumour-or virus-infected cells, since strong and multi-specific cytotoxic T lymphocytes (CTLs) responses are often associated with better clinical outcomes in cancer [1] and chronic infectious diseases [2,3]. In the past, the main focus of immunotherapies was on eliciting cytotoxic CD8 + T-cell responses; however, accumulatCorrespondence: Dr. Caroline Aspord e-mail: carolineaspord@yahoo.com ing evidence suggests that CD4 + T cells also play an important role in sustaining immune responses against cancer and infection [4,5]. CD4 + T cells provide critical help to CD8 + T cells, promoting their survival, expansion, and acquisition of effector functions [6,7]. CD4 + T cells also display direct antitumour cytolytic activity [8]. In melanoma, more favourable clinical outcomes are associated with immune responses towards many MHC class I and class II epitopes derived from several tumour antigens [9][10][11]. T-cell immunity, particularly when both CD4 + and CD8 + arms of immunity are involved, has also been shown to protect against cytomegalovirus (CMV) reactivation and disease [12]. Therefore, both CD4 + and CD8 + T cells play a pivotal role in inducing and maintaining therapeutic anti-tumour and anti-viral immunity.C 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.eji-journal.eu Eur. J. Immunol. 2014. 44: 2880-2892 Antigen processing As their size prevents a direct binding to MHC class I molecules, SLPs must be taken up and processed before they can be presented to T cells. Such cross-presentation involves dendritic cells (DCs) [21,22], which go on to cross-prime CTL and Th-cell immune responses [23]. Myeloid DCs (mDCs) have already been shown to cross-present SLPs and subsequently induce immune responses [22,24]. Plasmacytoid DCs (pDCs) are another interesting DC subset, also playing a critical role in shaping and regulating immune responses [25,26]. pDCs appear as novel vectors for immunotherapy for both cancer and infections [27][28][29][30], but their ability to cross-present SLP has never been investigated. Nonetheless, there is increasing evidence that pDCs effectively capture, process, and present antigens to T cells [31] Fig. 1C and D). Interestingly, despite some variations depending on the donors used, cross-presentation of long peptides significantly improved with a longer processing time (24 h) with both the pDC line ( Fig. 1E) and primary pDCs (Fig. 1F). To exclude any bystander effects of the peptivator pools, we used three individual long peptides encompassing the cross-presented epitope. All of them were well processed and cross-presented by pDCs (Fig. 2). The pDC line induced a sligh...
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