Had a great time at AAD 2025. Particularly proud of our medical students and fellows who presented.

Night #2 at Kīlauea Volcano. Spraying Lava from the central bright spot!

Hiked down crater rim drive…awe inspiring

So cool to be here at the Kilaeua volcano in Hawaii!

Using Seurat for clustering and phenotyping, we identified better mixing between CD8 T cells and macrophages in the 4-1BBL/IL-12 group and mixing of CD8 T cells with PD-L1 positive tumor cells as potentially drivers of the lack of improvement seen with addition of IFNg.

Using Seurat for clustering and phenotyping, we identified better mixing between CD8 T cells and macrophages in the 4-1BBL/IL-12 group and mixing of CD8 T cells with PD-L1 positive tumor cells as potentially drivers of the lack of improvement seen with addition of IFNg.

4-1BBL/IL-12 delivery resulted in improved antigen presentation by tumor, macrophages, and dendritic cells, with IFNg delivery further boosting antigen presentation and beta-2-microglobulin expression, while also further upregulating tumor PD-L1 expression.

…including CD4 and CD8 T cells, dendritic cells, and macrophages. However, 4-1BBL/IL-12 alone induced significant pro-inflammatory M1 skewing which was reduced by the addition of IFNg.

We applied this panel to coverslips for 1stgeneration CODEX imaging and to tumor arrays using the 2ndgeneration PhenoCycler Fusion from AkoyaBio. Most markers showed functional equivalence, but some low intensity markers could only be reliably visualized by the Fusion.

So, we set out to validate by single color IF a number of antibodies to profile the tumor immune microenvironment, ending with 28 validated antibodies to identify immune cell populations, macrophage polarization, antigen presentation, and immune checkpoints; see Table 1.