MHC class I and class II presentation of tumor antigen in retrovirally and adenovirally transduced dendritic cells

Abstract

The unique antigen -presenting capabilities of dendritic cells ( DCs ) make them an attractive means with which to initiate an antitumor immune response. Using DCs transduced with tumor antigens for immunotherapy has several theoretical advantages over peptide -pulsed DCs including the possibility that transduced DCs are capable of presenting epitopes on both class I and class II MHC molecules. To test this theory, we inserted the human tumor antigen gp100 into mouse DCs transgenic for HLA -DRb1*0401 using either adenoviral vector or a VSV -G pseudotyped retroviral vector. DCs transduced with tumor antigen were able to be recognized by both a murine CD8 + T -cell clone and a murine CD4 + T -cell line in a cytokine release assay, thereby demonstrating presentation of both MHC class I and class II gp100 epitopes. This study describes the simultaneous presentation of a tumor -associated antigen to both CD4 + and CD8 + T cells and lends support to the use of gene -modified DCs as a means to initiate both CD4 + and CD8 + antitumor responses.

Dendritic cells transduced with gp100 gene by RGD fiber-mutant adenovirus vectors are highly efficacious in generating anti-B16BL6 melanoma immunity in mice

“…4,28 Owing to these features and establishment of methods for expansion of human and rodent DCs on a large scale from hematopoietic precursors in the presence of appropriate cytokine cocktails, [29][30][31] DCs have attracted great attention as vehicles for the delivery of cancer vaccines. The efficacy of different viral vectors, including retroviral vector, 8,32 Ad, [8][9][10][11][12] and lentiviral vector, 33,34 has been evaluated for genetic modification of DCs. Among them, Ad was confirmed as a good candidate because of its high efficiency and minimum risk associated with insertional mutagenesis.…”

“…Furthermore, TAA gene-modified DCs may also present MHC class IIrestricted epitope(s) to CD4 þ T cells, and can express TAA continuously and MHC-peptide complexes for long periods. [7][8][9] Among various techniques for transferring the TAA gene to DCs, the ex vivo engineering of DCs using adenovirus vector (Ad) provides encouraging results. [10][11][12] However, the low or lack of coxsackie-adenovirus receptor (primary Ad-receptor) expression on the DC surface makes sufficient gene transduction to DCs by conventional Ad difficult.…”

An effective cancer vaccine modality: Lentiviral modification of dendritic cells expressing multiple cancer-specific antigens

“…Transduction of DCs with viral vectors that have been engineered to express TAAs can result in high levels of transgene expression necessary for a strong immune response [16]. Various viral vectors, including poxvirus [40], adenovirus [41], and conventional oncoretrovirus [42], have been used to genetically modify DCs. LVs offer several advantages over other vectors for targeting DCs.…”

Tumor-specific dendritic cells generated by genetic redirection of Toll-like receptor signaling against the tumor-associated antigen, erbB2

“…The amplified DNAs were ligated into the previously described anti-erbB2-CD28-z chimeric construct, 26 as depicted in Figure 1, which was incorporated into the retroviral expression plasmid, CLNCX. 31 The TLR4 transmembrane domain was retained in that construct since CD14 association with TLR4 is necessary for optimal function, 32 and transmembrane domains can be involved in molecular associations.…”

“…31 Briefly, plasmids containing the chimeric receptor and the vesicular stomatitis virus (VSV) envelope were transfected into 293gp cells (30 mg of each per 10 7 cells) using Lipofectamine 2000 according to manufacturer's instructions (Invitrogen, Carlsbad, CA). Following replenishment of media 24 h later, supernatant containing retroviral vector was harvested after a further 24 h and used to transduce JAWS II cells using 8 mg/ml polybrene and centrifugation at 1000 g for 1 hr/RT.…”