An Antibody Fusion Protein for Cancer Immunotherapy Mimicking IL-15 <i>trans</i>-Presentation at the Tumor Site

Kermer, Vanessa; Baum, Volker; Hornig, Nora; Kontermann, Roland E.; Müller, Dafne

doi:10.1158/1535-7163.mct-12-0019

Cited by 40 publications

(42 citation statements)

References 36 publications

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“…The therapeutic potential of the murine scDuokines was investigated in vivo using a syngeneic lung tumor model with human FAP-expressing B16 cells in C57BL/6N. The treatment protocol comprised a suboptimal dose of a bispecific single-chain diabody targeting human FAP and mouse CD3 (scDb332C11) 17,22,23 as primary MHC-independent T cell activation signal and the murine scDuokines as potential co-stimulatory signal. Groups of six C57BL/6N mice challenged with B16-FAP (i.v.)…”

Section: Resultsmentioning

confidence: 99%

Duokines: a novel class of dual-acting co-stimulatory molecules acting in cis or trans

et al. 2018

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Co-stimulatory signals induced by ligands of the tumor necrosis factor superfamily (TNFSF) play a central role in T cell activation and have emerged as a promising strategy in cancer immunotherapy. Here, we established a novel class of bifunctional co-stimulatory fusion proteins with the aim to boost T cell activation at the level of T cell – antigen-presenting cell (APC) interaction. These novel dual-acting cytokine fusion proteins were created by connecting two different homotrimeric TNFSF ligands to form homotrimeric bifunctional molecules (Duokines) or by connecting single-chain derivatives of two different homotrimeric TNFSF with a single, flexible linker (single-chain Duokines, scDuokines). By linking the TNFSF ligands 4-1BBL, OX40L and CD27L in all possible combinations, cis-acting Duokines were generated that act on the same or adjacent T cells, while combining CD40L with 4-1BBL, OX40L and CD27L resulted in trans-acting Duokines acting simultaneously on APCs and T cells. In vitro, co-stimulation of T cells was seen for cis- and trans-acting Duokines and scDuokines in an antigen-independent as well as antigen-specific setting. Trans-acting molecules furthermore activated B cells, which represent a subclass of APCs. In a pilot experiment using the syngeneic B16-FAP mouse tumor model scDuokines displayed antitumoral activity in vivo in combination with a primary T cell-activating bispecific antibody, evident from reduced number of lung metastasis compared to the antibody-only treated group. Our data show that the bifunctional, co-stimulatory duokines are capable to enhance T cell-mediated anti-tumor immune responses, suggesting that they can serve as a new class of immuno-stimulatory molecules for use in cancer immunotherapy strategies.

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Section: Resultsmentioning

confidence: 99%

Duokines: a novel class of dual-acting co-stimulatory molecules acting in cis or trans

et al. 2018

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“…RD_IL-15_scFv_m4-1BBL and scFv_m4-1BBL were obtained by replacing the extracellular domain of human 4-1BBL (aa 71-254) by the corresponding mouse 4-1BBL (aa 104-309). All recombinant proteins were produced in stably transfected HEK293 cells and were purified by immobilized metal ion affinity chromatography (IMAC) as described elsewhere (13). In brief, producer cells were expanded and grown to 90% confluence in RPMI 5% FBS before switching to serum-free Opti-MEM I medium (Life Technologies).…”

Section: Generation Of Recombinant Antibody Fusion Proteinsmentioning

confidence: 99%

“…Therefore, 2 Â 10 4 CTLL-2 cells per well were seeded and starved for 4 hours, before addition of the respective fusion proteins. After 3 days, cell proliferation was measured by MTT assay (13).…”

Section: Proliferation Assay With Ctll-2mentioning

confidence: 99%

“…Also, a fusion protein composed of IL-15, IL-15Ra's sushi domain, and apolipoprotein A-I (Apo A-I) has been reported, inducing therapeutic effects in lung and liver metastases mouse models (11). Furthermore, tumor targeting by antibody fusion proteins with IL-15 (12) or RD_IL-15 (13,14) resulted in enhanced antitumor effects in different mouse models. On the other hand, improved therapeutic effects could be achieved by the combination of IL-15 with other immunomodulatory approaches, for example, cytokines (IL-21, IL-7, IL-12; refs.…”

Section: Introductionmentioning

confidence: 99%

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Combining Antibody-Directed Presentation of IL-15 and 4-1BBL in a Trifunctional Fusion Protein for Cancer Immunotherapy

Kermer

Hornig

Harder

et al. 2014

Molecular Cancer Therapeutics

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Influencing the cytokine receptor network that modulates the immune response holds great potential for cancer immunotherapy. Although encouraging results have been obtained by focusing on individual members of the common g-chain (gc) receptor family and TNF receptor superfamily so far, combination strategies might be required to further improve the effectiveness of the antitumor response. Here, we propose the combination of interleukin (IL)-15 and 4-1BBL in a single, tumor-directed molecule. Therefore, a trifunctional antibody fusion protein was generated, composed of a tumor-specific recombinant antibody, IL-15 linked to a fragment of the IL-15Ra chain (RD) and the extracellular domain of 4-1BBL. In soluble and targeted forms, the trifunctional antibody fusion protein RD_IL-15_scFv_4-1BBL was shown to stimulate activated T-cell proliferation and induce T-cell cytotoxicity to a similar degree as the bifunctional scFv_RD_IL-15 fusion protein. On the other hand, in targeted form, the trifunctional fusion protein was much more effective in inducing T-cell proliferation and IFN-g release of unstimulated peripheral blood mononuclear cells (PBMC). Here, the additional signal enhancement could be attributed to the costimulatory activity of 4-1BBL, indicating a clear benefit for the simultaneous presentation of IL-15 and 4-1BBL in one molecule. Furthermore, the trifunctional antibody fusion protein was more effective than the corresponding bifunctional fusion proteins in reducing metastases in a tumor mouse model in vivo. Hence, the targeted combination of IL-15 and 4-BBL in the form of a trifunctional antibody-fusion protein is a promising new approach for cancer immunotherapy.

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“…18 IL-15 agonists (e.g., IL-15/IL15Ra-Fc complex and IL-15/IL-15Ra fusion protein) reduce the dose of delivered IL-15 required to reach biologicallymeaningful levels in vivo. [19][20][21][22][23] However, cell (particularly DC) contact-dependent trans-presentation of membrane-bound IL-15/IL-15Ra appears required for optimal IL-15-mediated signaling in vivo. 24,25 In vivo, IL-15 derived from DC (DCIL-15) can "auto"-activate DC and substitute for the functional licensing events normally associated with DC interaction with CD4…”

Section: Introductionmentioning

confidence: 99%

Dendritic cell-derived interleukin-15 is crucial for therapeutic cancer vaccine potency

et al. 2014

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IL-15 supports improved antitumor immunity. How to best incorporate IL-15 into vaccine formulations for superior cancer immunotherapy remains a challenge. DC-derived IL-15 (DCIL-15) notably has the capacity to activate DC, to substitute for CD4 C Th and to potentiate vaccine efficacy making IL-15-based therapies attractive treatment options. We observed in transplantable melanoma, glioma and metastatic breast carcinoma models that DCIL-15-based DNA vaccines in which DC specifically express IL-15 and simultaneously produce tumor Aghsp70 were able to mediate potent therapeutic efficacy that required both host Batf3C DC and CD8 C T cells. In an inducible Braf V600E /Pten-driven murine melanoma model, DCIL-15 (not rIL-15)-based DNA vaccines elicited durable therapeutic CD8C T cell-dependent antitumor immunity. DCIL-15 was found to be superior to rIL-15 in "licensing" both mouse and human DC, and for activating CD8 C T cells. Such activation occurred even in the presence of Treg, without a need for CD4 C Th, but was IL-15/IL-15Ra-dependent. A single low-dose of DCIL-15 (not rIL-15)-based DC vaccines induced therapeutic antitumor immunity. CD14C DC emigrating from human skin explants genetically-immunized by IL-15 and Aghsp70 were more effective than similar DC emigrating from the explants genetically-immunized by Aghsp70 in the presence of rIL-15 in expressing membrane-bound IL-15/IL-15Ra and activating CD8 C T cells. These results support future clinical use of DCIL-15 as a therapeutic agent in battling cancer.

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An Antibody Fusion Protein for Cancer Immunotherapy Mimicking IL-15 trans-Presentation at the Tumor Site

Cited by 40 publications

References 36 publications

Duokines: a novel class of dual-acting co-stimulatory molecules acting in cis or trans

Duokines: a novel class of dual-acting co-stimulatory molecules acting in cis or trans

Combining Antibody-Directed Presentation of IL-15 and 4-1BBL in a Trifunctional Fusion Protein for Cancer Immunotherapy

Dendritic cell-derived interleukin-15 is crucial for therapeutic cancer vaccine potency

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