Background Pancreatic ductal adenocarcinoma (PDAC) does not respond to immune checkpoint inhibitors (ICI) therapy as single agent treatments including anti-PD-1 antibody. One of the mechanisms for the resistance of PDAC to ICI is now attributed to the immunosuppressive microenvironment (TME) in PDAC. Myeloid cells are thought to be the predominant immunosuppressive cells in the TME. Human interleukin-8 (IL-8) is a pro-inflammatory chemokine in the CXC family and has the capability of recruiting myeloid cells into the TME to promote tumor progression and immune escape. Therefore, several anti-IL-8 blockade antibodies were developed including HuMax-IL8 and B108-IL8, which both are fully human IgG1 kappa monoclonal antibodies. We therefore tested whether anti-IL-8 antibodies can potentiate anti-tumor activity of anti-PD-1 antibody in a humanized model of PDAC.
Materials and Methods We reconstituted the immune system of the NGS mice with ex vivo activated human T cells and a combination of CD14+ and CD16+ myeloid cells after the mice were orthotopically implanted with human PDAC cells. 10x single nuclei RNA-Seq data processing was further performed to analyze differentially expressed genes among certain cell clusters.
Results Our results showed that anti-PD-1 antibody alone had a minimal anti-tumor activity when mice was reconstituted with ex vivo activated T cells. Interestingly, the infusion of the combination of CD14+ and CD16+ myeloid cells together with anti-PD-1 antibody resulted in a modest anti-tumor activity. Adding either HuMAX-IL8 or B108-IL8 led to a significantly enhanced anti-tumor activity. Both CD14+ and CD16+ myeloid cells appeared to be needed for the full anti-tumor activity of IL-8 blockade because mice infused with only CD14+ myeloid cells did not respond to IL-8 blockade and mice infused with only CD16+ myeloid cells responded partially to IL-8 blockade. This result suggested that the target of IL-8 is mainly present in CD16+ myeloid cells and is likely to be granulocytes. Tumor infiltrating immune cells were isolated and demonstrated that IL-8 blockade increases CD45+CD11b+CD15+CD14- myeloid cells, which is known to comprise neutrophils and granulocytic myeloid derived suppressive cells (G-MDSC), in the tumors. Reconstitution of the mice with myeloid cells led to a decrease of CD8+ T cells in the tumors; however, IL-8 blockade brought the CD8+ T cell number back to the baseline. Consistent with an effect of IL-8 blockade on the increase of CD15+CD14- myeloid cells, single nuclear RNA sequencing analysis of the tumor tissues showed that the innate immune response and cytokine response pathways in the myeloid cell cluster were activated by IL-8 blockade.
Conclusions This result suggested that IL-8 blockade did not simply inhibit myeloid cells as previously anticipated, but potentiated myeloid cells for the innate immune response and concomitant production of type I cytokines. Such immune responses may subsequently activate the effector T cells as the single nuclear RNA sequencing analysis demonstrated enhanced activation signals in the T cell cluster from the tumors treated by anti-IL-8 antibodies. Taken together, this study supports further testing of anti-IL-8 antibodies including B108-IL8 and HuMax-IL8 in combination with anti-PD-1 antibodies for PDAC treatment.
Disclosure Information P. Li: None. N. Rozich: None. J. Wang: None. J. Gai: None. J. Wang: None. Y. Xu: None. B. Herbst: None. R. Yu: A. Employment (full or part-time); Significant; NovaRock. S. Muth: None. N. Niu: None. K. Li: None. V. Fune: None. A. Osipov: None. C. Wolfgang: None. M. Lei: A. Employment (full or part-time); Significant; NovaRock. T. Liang: None. L. Zheng: B. Research Grant (principal investigator, collaborator or consultant and pending grants as well as grants already received); Significant; Bristol-Meyer Squibb, Merck, iTeos, Amgen, NovaRock, Inxmed, Halozyme. E. Ownership Interest (stock, stock options, patent or other intellectual property); Significant; Alphamab, Mingruzhiyao. F. Consultant/Advisory Board; Significant; Biosion, Alphamab, NovaRock, Akrevia/Xilio, Ambrx, Novagenesis, Datarevive, Snow Lake Capitals, Mingruzhiyao. Other; Significant; Aduro Biotech.
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