Alpha-Fetoprotein- and CD40Ligand-Expressing Dendritic Cells for Immunotherapy of Hepatocellular Carcinoma

Annabelle Vogt; Farsaneh Sadeghlar; Tiyasha H Ayub; Carlo Schneider; Christian Möhring; Taotao Zhou; Robert Mahn; Alexandra Bartels; Michael Praktiknjo; Miroslaw T Kornek; Marieta Toma; Ingo G H Schmidt-Wolf; Vittorio Branchi; Hanno Matthaei; Jörg C Kalff; Christian P Strassburg; Maria A Gonzalez-Carmona

doi:10.3390/cancers13133375

Alpha-Fetoprotein- and CD40Ligand-Expressing Dendritic Cells for Immunotherapy of Hepatocellular Carcinoma

Cancers (Basel). 2021 Jul 5;13(13):3375. doi: 10.3390/cancers13133375.

Authors

Annabelle Vogt¹, Farsaneh Sadeghlar¹, Tiyasha H Ayub¹, Carlo Schneider¹, Christian Möhring¹, Taotao Zhou¹, Robert Mahn¹, Alexandra Bartels¹, Michael Praktiknjo¹, Miroslaw T Kornek¹, Marieta Toma², Ingo G H Schmidt-Wolf³, Vittorio Branchi⁴, Hanno Matthaei⁴, Jörg C Kalff⁴, Christian P Strassburg¹, Maria A Gonzalez-Carmona¹

Affiliations

¹ Department of Internal Medicine I, University Hospital of Bonn, 53127 Bonn, Germany.
² Department of Pathology, University Hospital of Bonn, 53127 Bonn, Germany.
³ Department of Integrated Oncology (CIO), University of Bonn, 53127 Bonn, Germany.
⁴ Department of Visceral Surgery, University Hospital of Bonn, 53127 Bonn, Germany.

Abstract

Dendritic cells (DC) as professional antigen presenting cells are able to prime T-cells against the tumor-associated antigen α-fetoprotein (AFP) for immunotherapy of hepatocellular carcinoma (HCC). However, a strong immunosuppressive tumor environment limits their efficacy in patients. The co-stimulation with CD40Ligand (CD40L) is critical in the maturation of DC and T-cell priming. In this study, the impact of intratumoral (i.t.) CD40L-expressing DC to improve vaccination with murine (m)AFP-transduced DC (Ad-mAFP-DC) was analyzed in subcutaneous (s.c.) and orthotopic murine HCC. Murine DC were adenovirally transduced with Ad-mAFP or Ad-CD40L. Hepa129-mAFP-cells were injected into the right flank or the liver of C3H-mice to induce subcutaneous (s.c.) and orthotopic HCC. For treatments, 10⁶ Ad-mAFP-transduced DC were inoculated s.c. followed by 10⁶ CD40L-expressing DC injected intratumorally (i.t.). S.c. inoculation with Ad-mAFP-transduced DC, as vaccine, induced a delay of tumor-growth of AFP-positive HCC compared to controls. When s.c.-inoculation of Ad-mAFP-DC was combined with i.t.-application of Ad-CD40L-DC synergistic antitumoral effects were observed and complete remissions and long-term survival in 62% of tumor-bearing animals were achieved. Analysis of the tumor environment at different time points revealed that s.c.-vaccination with Ad-mAFP-DC seems to stimulate tumor-specific effector cells, allowing an earlier recruitment of effector T-cells and a Th1 shift within the tumors. After i.t. co-stimulation with Ad-CD40L-DC, production of Th1-cytokines was strongly increased and accompanied by a robust tumor infiltration of mature DC, activated CD4⁺-, CD8⁺-T-cells as well as reduction of regulatory T-cells. Moreover, Ad-CD40L-DC induced tumor cell apoptosis. Intratumoral co-stimulation with CD40L-expressing DC significantly improves vaccination with Ad-mAFP-DC in pre-established HCC in vivo. Combined therapy caused an early and strong Th1-shift in the tumor environment as well as higher tumor apoptosis, leading to synergistic tumor regression of HCC. Thus, CD40L co-stimulation represents a promising tool for improving DC-based immunotherapy of HCC.

Keywords: CD40Ligand; alpha-fetoprotein; dendritic cells; hepatocellular carcinoma; subcutaneous and orthotopic murine HCC.

Abstract

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