CD40-targeted adenoviral gene transfer to dendritic cells through the use of a novel bispecific single-chain Fv antibody enhances cytotoxic T cell activation

Brandão, Joana G.; Scheper, Rik J.; Lougheed, Sinéad M.; Curiel, David T.; Tillman, Bryan; Gerritsen, Winald R.; Eertwegh, Alfons J.M. van den; Pinedo, H.M.; Haisma, Hidde J.; Gruijl, Tanja D. de

doi:10.1016/s0264-410x(03)00050-1

Cited by 62 publications

(46 citation statements)

References 25 publications

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“…The reason for this poor transduction efficiency is thought to be due to the lack of expression of the primary receptor for the virus, CAR, on dendritic cells (9). This observation has prompted a number of investigators to investigate whether one can increase the efficiency of rAd5-mediated gene transfer in DC by targeting the virus to alternate cell surface receptors on this cell type (10)(11)(12)(13)(14).…”

Section: Introductionmentioning

confidence: 99%

“…These approaches have been used to target a small subset of DC-expressed surface receptors, including RGD-binding integrins and CD40 (13,(18)(19)(20)(21)(22), which were chosen in part because they are endocytosing molecules that are known to play a role in DC activation and maturation. RAd5 vectors that have been targeted in this manner not only have an improved capacity to transduce cultured DC, but the virally-transduced DC have also been shown to elicit enhanced immune responses in vivo (18,21,23).…”

Section: Introductionmentioning

confidence: 99%

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Recombinant adenovirus type 5 vectors that target DC-SIGN, ChemR23 and αvβ3 integrin efficiently transduce human dendritic cells and enhance presentation of vectored antigens

Maguire

Sapinoro

Girgis

et al. 2006

Vaccine

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Recombinant adenoviruses (rAds) represent a promising system for vaccine delivery but transduce dendritic cells (DC) relatively poorly. To address this concern, we used a biotin-avidin linkage to conjugate rAd vectors to ligands which bind with high affinity to selected receptors on DC (ChemR23, α v β 3 integrin, and DC-SIGN). The targeted vectors had an enhanced ability to transduce human monocyte-derived DC compared to untargeted virus. In addition, DC transduced with targeted rAd vectors were more efficient at stimulating cytokine production by autologous memory CD8+ T cells, against a vector-encoded antigen. These results expand the range of cell surface receptors that can be used to target rAd5 vectors to DC, and may facilitate future development of rAd-based vaccines.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Recombinant adenovirus type 5 vectors that target DC-SIGN, ChemR23 and αvβ3 integrin efficiently transduce human dendritic cells and enhance presentation of vectored antigens

Maguire

Sapinoro

Girgis

et al. 2006

Vaccine

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“…Hence, refolding techniques or mammalian expression systems are routinely used to produce soluble taFv (25,26). Researchers have engineered linker length (27,28) and optimized linkers using phage display technology (15) to promote the expression of soluble taFv; however, whether these linker sequences are applicable to other taFv is unclear. Additionally, the peptide tag usually required for purification may affect the function of small BsAbs in samples prepared using bacterial or mammalian expression systems.…”

Section: Discussionmentioning

confidence: 99%

Cytotoxic Enhancement of a Bispecific Diabody by Format Conversion to Tandem Single-chain Variable Fragment (taFv)

Asano

Ikoma

Shimomura

et al. 2011

Journal of Biological Chemistry

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Diabodies (Dbs) and tandem single-chain variable fragments (taFv) are the most widely used recombinant formats for constructing small bispecific antibodies. However, only a few studies have compared these formats, and none have discussed their binding kinetics and cross-linking ability. We previously reported the usefulness for cancer immunotherapy of a humanized bispecific Db (hEx3-Db) and its single-chain format (hEx3-scDb) that target epidermal growth factor receptor and CD3. Here, we converted hEx3-Db into a taFv format to investigate how format affects the function of a small bispecific antibody; our investigation included a cytotoxicity assay, surface plasmon resonance spectroscopy, thermodynamic analysis, and flow cytometry. The prepared taFv (hEx3-taFv) showed an enhanced cytotoxicity, which may be attributable to a structural superiority to the diabody format in cross-linking target cells but not to differences in the binding affinities of the formats. Comparable cross-linking ability for soluble antigens was observed among hEx3-Db, hEx3-scDb, and hEx3-taFv with surface plasmon resonance spectroscopy. Furthermore, drastic increases in cytotoxicity were found in the dimeric form of hEx3-taFv, especially when the two hEx3-taFv were covalently linked. Our results show that converting the format of small bispecific antibodies can improve their function. In particular, for small bispecific antibodies that target tumor and immune cells, a functional orientation that avoids steric hindrance in cross-linking two target cells may be important in enhancing the growth inhibition effect. Bispecific antibodies (BsAbs)2 are recombinant antibodies that can bind to two different antigenic epitopes. Bispecificity can be used in cancer immunotherapy to cross-link tumor cells to immune cells such as cytotoxic T cells, natural killer cells, and macrophages. This cross-linking accelerates the destruction of the tumor cells by the immune cells, so that the high potency of BsAb may translate into improved antitumor therapy and lower treatment costs by decreasing the necessary doses (1, 2). However, the use of BsAbs in clinical studies has been hampered by difficulties in producing them on a large scale. Conventional chemical conjugation has been used, but the quality of the antibodies produced is inconsistent (3). The production of BsAbs by somatic fusion of two hybridomas to form a quadroma yields BsAbs of more consistent quality but results in the formation of various chain-shuffled antibodies; for instance, 10 different antibodies can be generated after random association of two heavy and two light chains (4, 5).Advances in recombinant technology have made it feasible to generate small recombinant BsAbs constructed from two different variable antibody fragments, such as variable fragments (Fv) and single-chain Fv (scFv). These recombinant BsAbs include bispecific diabodies (Dbs) (6), single-chain diabodies (scDbs) (7), tandem scFv (taFv) (8), and minibodies (dimeric scDb-CH3 fusion proteins) (9). Compared with classic BsAb...

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“…Alternatively, DC-targeting motifs that simultaneously effect maturation induction may be used, such as the TLR4-binding extra domain A of fibronectin (14) or CD40L. Indeed, our in vitro and in situ studies have shown that the genetic targeting of antigens to the CD40 receptor efficiently and selectively delivers antigens to both human and murine DCs and simultaneously induces their stable maturation, obviating the need for additional maturation stimuli for proper T-cell activation (15)(16)(17). Genetic targeting of TAAs ensures durable presentation through MHC-I to specific CTLs (18).…”

Section: Introductionmentioning

confidence: 99%

“…22). We previously showed that this limitation could be overcome by retargeting Ad5 to CD40 by using bispecific adaptor molecules that simultaneously neutralized the Ad5 fiber knob and agonistically bound CD40 on the DC surface (16,23). This approach facilitated the induction of T-cell-mediated immune responses against nonself transgene products (17, 24) and protected mice from tumor growth through vaccination with ex vivo modified DCs (17).…”

Section: Introductionmentioning

confidence: 99%

Potent Antitumor Immunity Generated by a CD40-Targeted Adenoviral Vaccine

et al. 2011

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In situ delivery of tumor-associated antigen (TAA) genes into dendritic cells (DC) has great potential as a generally applicable tumor vaccination approach. Although adenoviruses (Ad) are an attractive vaccine vehicle in this regard, Ad-mediated transduction of DCs is hampered by the lack of expression of the Ad receptor CAR on the DC surface. DC activation also requires interaction of CD40 with its ligand CD40L to generate protective T-cell-mediated tumor immunity. Therefore, to create a strategy to target Ads to DCs in vivo, we constructed a bispecific adaptor molecule with the CAR ectodomain linked to the CD40L extracellular domain via a trimerization motif (CFm40L). By targeting Ad to CD40 with the use of CFm40L, we enhanced both transduction and maturation of cultured bone marrow-derived DCs. Moreover, we improved transduction efficiency of DCs in lymph node and splenic cell suspensions in vitro and in skin and vaccination site-draining lymph nodes in vivo.

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CD40-targeted adenoviral gene transfer to dendritic cells through the use of a novel bispecific single-chain Fv antibody enhances cytotoxic T cell activation

Cited by 62 publications

References 25 publications

Recombinant adenovirus type 5 vectors that target DC-SIGN, ChemR23 and αvβ3 integrin efficiently transduce human dendritic cells and enhance presentation of vectored antigens

Recombinant adenovirus type 5 vectors that target DC-SIGN, ChemR23 and αvβ3 integrin efficiently transduce human dendritic cells and enhance presentation of vectored antigens

Cytotoxic Enhancement of a Bispecific Diabody by Format Conversion to Tandem Single-chain Variable Fragment (taFv)

Potent Antitumor Immunity Generated by a CD40-Targeted Adenoviral Vaccine

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