Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Dec;70(12):3629-3642.
doi: 10.1007/s00262-021-02932-5. Epub 2021 May 5.

The human anti-CD40 agonist antibody mitazalimab (ADC-1013; JNJ-64457107) activates antigen-presenting cells, improves expansion of antigen-specific T cells, and enhances anti-tumor efficacy of a model cancer vaccine in vivo

Adnan Deronic  1 , Anneli Nilsson  1 , Mia Thagesson  1 , Doreen Werchau  1 , Karin Enell Smith  1 , Peter Ellmark  2   3
Affiliations

The human anti-CD40 agonist antibody mitazalimab (ADC-1013; JNJ-64457107) activates antigen-presenting cells, improves expansion of antigen-specific T cells, and enhances anti-tumor efficacy of a model cancer vaccine in vivo

Adnan Deronic et al. Cancer Immunol Immunother. 2021 Dec.

Abstract

Non-responders to checkpoint inhibitors generally have low tumor T cell infiltration and could benefit from immunotherapy that activates dendritic cells, with priming of tumor-reactive T cells as a result. Such therapies may be augmented by providing tumor antigen in the form of cancer vaccines. Our aim was to study the effects of mitazalimab (ADC-1013; JNJ-64457107), a human anti-CD40 agonist IgG1 antibody, on activation of antigen-presenting cells, and how this influences the priming and anti-tumor potential of antigen-specific T cells, in mice transgenic for human CD40. Mitazalimab activated splenic CD11c+ MHCII+ dendritic cells and CD19+ MHCII+ B cells within 6 h, with a return to baseline within 1 week. This was associated with a dose-dependent release of proinflammatory cytokines in the blood, including IP-10, MIP-1α and TNF-α. Mitazalimab administered at different dose regimens with ovalbumin protein showed that repeated dosing expanded ovalbumin peptide (SIINFEKL)-specific CD8+ T cells and increased the frequency of activated ICOS+ T cells and CD44hi CD62L- effector memory T cells in the spleen. Mitazalimab prolonged survival of mice bearing MB49 bladder carcinoma tumors and increased the frequency of activated granzyme B+ CD8+ T cells in the tumor. In the ovalbumin-transfected tumor E.G7-OVA lymphoma, mitazalimab administered with either ovalbumin protein or SIINFEKL peptide prolonged the survival of E.G7-OVA tumor-bearing mice, as prophylactic and therapeutic treatment. Thus, mitazalimab activates antigen-presenting cells, which improves expansion and activation of antigen-specific T cells and enhances the anti-tumor efficacy of a model cancer vaccine.

Keywords: CD40 agonist antibody; Cancer immunotherapy; Cancer vaccine; Dendritic cell activation.

PubMed Disclaimer

Conflict of interest statement

All authors are current employees of, and hold stocks or stock options in, Alligator Bioscience AB.

Figures

Fig. 1
Fig. 1
Mitazalimab activates splenic DC and B cells—Naïve hCD40tg mice were administered a single dose of 100 µg mitazalimab or Ctr IgG i.p. and spleens collected for flow cytometry from 6 h up to 7 days after treatment. a Frequency of activated CD80+ CD86+ splenic DC (CD11c+ MHCII+), b representative FACS plots from the 24 h time point in a, c frequency of activated CD80+ CD86+ splenic B cells (CD19+ MHCII+). MB49 tumor-bearing mice were administered repeated dosing of 100 or 300 µg mitazalimab i.p. on day 7, 10 and 13 post-inoculation, and 24 h after the final dose, the mice were anaesthetized and blood collected via vena cava for flow cytometry, d number of circulating B cells per ml of blood, e frequency of activated CD86+ circulating B cells. n = 3 per group in a–c; n = 8 per group in d–e. Statistical significance was analyzed by ordinary two-way ANOVA and Šídák’s multiple comparisons test in a and c, and by Mann–Whitney U test in d and e. Error bars for all data points are included, although not always visible, and indicate ± SEM
Fig. 2
Fig. 2
Mitazalimab induces the release of proinflammatory cytokines and chemokines in the blood—MB49 tumor-bearing hCD40tg mice were administered a single dose of 10, 30, 100 or 300 µg mitazalimab i.p. on day 7 post-inoculation, and 24 h after dosing, blood was collected via vena saphena for MSD analysis. The V-PLEX Mouse Cytokine 19-Plex Kit was used to measure CXCL1 (KC/GRO), IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12p70, IL-15, IL-17A/F, IL-27p28/IL-30, IL-33, IP-10, MCP-1, MIP-1α, MIP-2 and TNF-α. Only the cytokines/chemokines that showed a consistent change across two experiments are shown. n = 2–5 per group. Statistical significance was analyzed by Mann–Whitney U test. The lower limit of detection (LLOD) is defined by a horizontal dotted line. All error bars indicate ± SEM
Fig. 3
Fig. 3
Repeated dosing of mitazalimab in OVA-rechallenged mice results in expansion of OVA-specific CD8+ T cells—Naïve hCD40tg mice were immunized with 200 µg OVA protein i.v. on 3 occasions, 7 days between. The mice were divided into different cohorts, wherein each cohort was given 100 µg mitazalimab i.p. at different dose regimens. One cohort received no mitazalimab treatment (OVA only). A subset of mice from each cohort was sacrificed once weekly for 6 weeks after the first immunization and spleens and blood collected for flow cytometry. a Overview of the experimental set-up, b frequency of OVA-specific (SIINFEKL-MHCI tetramer+) splenic CD8+ T cells, c representative FACS plots from two groups from the day 14 time point in b, d frequency of activated CD80+ CD86+ splenic DC (CD11c+ MHCII+), e frequency of activated CD80+ CD86+ splenic B cells (CD19+ MHCII+), f frequency of activated CD86+ circulating B cells. n = 3–4 per group in a–f. Statistical significance was analyzed by ordinary two-way ANOVA and Šídák’s multiple comparisons test in b, d–f. No relevant comparisons were significant in b. All error bars indicate ± SEM
Fig. 4
Fig. 4
Prophylactic treatment with mitazalimab and OVA protein prolongs the survival of E.G7-OVA tumor-bearing mice and induces immunological memory against OVA—Naïve hCD40tg mice were immunized twice with 200 µg OVA protein or PBS control i.v., 7 days between. Mitazalimab or Ctr IgG (100 µg) were given i.p. either every 2–3 days or every 7 days during a 12-day period, starting from the first OVA injection. On day 14, 1 × 106 E.G7-OVA cells were inoculated s.c. a Overview of the experimental setup, b E.G7-OVA tumor growth throughout the study. Tumor growth is displayed until the point where the first mouse in each group approaches the ethical tumor volume limit of 2 cm3 and is sacrificed, c frequency of survival throughout the study. Pooled data from two identical experiments are shown in b and c. The complete responders in c were again inoculated with 1 × 106 E.G7-OVA cells s.c. on one flank and 1 × 106 EL-4 cells on the other flank, d E.G7-OVA and EL-4 tumor growth of the complete responders. Tumor growth is displayed until the point where the first complete responder is sacrificed. n = 8–10 per group, per experiment, in a–c; n = 5 in total in d. Statistical significance was analyzed by the log-rank test in c. All error bars indicate ± SEM
Fig. 5
Fig. 5
Mitazalimab treatment results in improved T cell activation—A cohort of mice from each treatment group in Fig. 4a was sacrificed before tumor inoculation on day 14 and spleens collected for flow cytometry. a Frequency of OVA-specific (SIINFEKL-MHCI tetramer+) splenic CD8+ T cells, b frequency of ICOS+ splenic CD8+ T cells, c frequency of CD44hi CD62L− splenic CD8+ T cells, d representative FACS plots from two of the treatment groups in c. n = 3–4 per group in a–d. Statistical significance was analyzed by Mann–Whitney U test. All error bars indicate ± SEM
Fig. 6
Fig. 6
Therapeutic treatment with mitazalimab prolongs the survival of MB49 tumor-bearing mice and, combined with OVA peptide, also prolongs the survival of E.G7-OVA tumor-bearing mice—Mice (hCD40tg) were inoculated with 0.25 × 106 MB49 cells s.c. and administered 100 µg mitazalimab or Ctr IgG p.t. or i.p. on day 7, 10 and 13 post-inoculation. a Frequency of survival throughout the study. Mice were sacrificed once their tumor volumes approached the ethical tumor volume limit of 2 cm3. Alternatively, the MB49 tumor-bearing mice were administered 100 or 300 µg mitazalimab i.p. on day 7, 10 and 13 post-inoculation and 24 h after the final dose, tumors were collected for flow cytometry, b frequency of granzyme B+ CD8+ T cells in the tumor, c frequency of CD44hi CD62L− CD8+ T cells in the tumor. Mice were inoculated with 1 × 106 E.G7-OVA cells s.c. on one flank and administered 100 µg mitazalimab and/or 10 µg OVA peptide (SIINFEKL) s.c. on the other flank. The treatments were administered on the day of tumor inoculation and once more, 7 days later, d E.G7-OVA tumor growth throughout the study, e frequency of survival throughout the study. n = 9–10 per group in a; n = 8 per group in b–c; n = 10 per group in d–e. Statistical significance was analyzed by the log-rank test in a and e, and by Mann–Whitney U test in b and c. All error bars indicate ± SEM
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Cogdill AP, Andrews MC, Wargo JA. Hallmarks of response to immune checkpoint blockade. Br J Cancer. 2017;117:1–7. doi: 10.1038/bjc.2017.136. - DOI - PMC - PubMed
    1. Kim TK, Herbst RS, Chen L. Defining and understanding adaptive resistance in cancer immunotherapy. Trends Immunol. 2018;39:624–631. doi: 10.1016/j.it.2018.05.001. - DOI - PMC - PubMed
    1. Darvin P, Toor SM, Sasidharan Nair V, Elkord E. Immune checkpoint inhibitors: recent progress and potential biomarkers. Exp Mol Med. 2018;50:1–11. doi: 10.1038/s12276-018-0191-1. - DOI - PMC - PubMed
    1. Galon J, Bruni D. Approaches to treat immune hot, altered and cold tumours with combination immunotherapies. Nat Rev Drug Discov. 2019;18:197–218. doi: 10.1038/s41573-018-0007-y. - DOI - PubMed
    1. Bonaventura P, Shekarian T, Alcazer V, Valladeau-Guilemond J, Valsesia-Wittmann S, Amigorena S, Caux C, Depil S. Cold tumsors: a therapeutic challenge for immunotherapy. Front Immunol. 2019;10:168. doi: 10.3389/fimmu.2019.00168. - DOI - PMC - PubMed

MeSH terms