Authors:

Brayden Spurlock

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About Paper:

Tertiary lymphoid structures (TLS) are immune aggregates associated with improved prognosis in various human cancers, including invasive urothelial carcinoma (transitional cell carcinoma (InvTCC)) of the urinary bladder. TLS formation is orchestrated by chemokines such as CXCL13 and CCL21, typically induced by pro-inflammatory cytokines, including interleukin-17A (IL-17A), produced by stromal fibroblasts and immune cells. In human and murine models, IL-17A signaling promotes TLS development and enhances anti-tumor immune responses. However, the extent to which this pathway functions in canine tumors remains unclear. Here, we investigated IL-17A-induced expression of TLS-associated chemokines and angiogenic factors in five canine InvTCC cell lines (AxA, AxC, Nk, Original (K9TCC), and Sh) in vitro. Cells were treated with recombinant IL-17A, and quantitative PCR was used to assess expression of IL-17RA, CXCL13, CCL19, CCL21, CXCL12, and VEGF, normalized to 18S RNA. IL-17RA expression was low or undetectable in most lines. Neither CXCL13 nor CCL21 showed significant upregulation following IL-17A treatment, and CCL19 exhibited variable low-level expression. VEGF was consistently expressed, but was unaffected by IL- 17A treatment. These findings suggest IL-17A alone is insufficient to induce TLS-associated chemokine expression in canine tumor cells under monoculture conditions, suggesting that stromal or immune- derived signals are necessary for full TLS pathway activation. In contrast to established IL-17-driven TLS induction models, these results underscore species-specific or context-dependent differences in IL-17 signaling. Our study reveals a knowledge gap in canine cancer immunobiology, supporting the future use of co-culture models incorporating stromal and immune cells to more accurately model TLS- inducing pathways in canine InvTCC anti-tumor responses in vitro. Keywords: IL-17; Canine; Cytokine; TLS; Cancer † Presenting Undergrad Author; ‡ Contributing Undergrad Author; * Undergrad Acknowledgment

Source:

Purdue University / 2025

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Co-authors:

Brayden Spurlock