Extended Data Fig. 6. (related to Sub-heading, CAN-3110 increases T cells in tumours.) Representative immunohistochemistry (IHC) Images.

(a) upper panel: Nestin IHC was carried out in a rGBM (IDHwt) resected 279 days after CAN-3110 injection (Subject 044, 10¹⁰ pfus in 1 ml); lower panel: Nectin-1/CD111 was carried out in a rGBM (IDHwt) resected 253 days after CAN-3110 (Subject 028, 10⁹ pfu in 1 ml). (b) Time course of Nestin and Nectin-1 IHC. Subject 046 (IDHmut astrocytoma) was injected on day 0. Because of an SAE consisting of multiple seizures 2 days after injection (Table 1b), he was treated with antivirals with resolution of the event. He was then brought back for re-resection twice showing both times persistence of both nestin and nectin-1 expression with tumour progression. Non tumour brain shows nestin expression peri-vascularly with no nectin-1 expression. (c) Representative example of CD8+ T cell immunohistochemistry (IHC) from Pre- and Post-CAN-3110 injected tumour. Subject 016 was injected with 3 × 10⁷ PFUs of CAN-3110 and post-injection tumour was resected 24 days after injection due to MRI evidence of continued progression. Areas shown were relatively far from the area of injection. (d) Representative examples of perivascular CD3+ T cell accumulation from 3 subjects (016, 019, 021) after CAN-3110 injection. Dose and time of post-injection tumour harvest are indicated for each. (e) Representative examples of perinecrotic accumulation of CD20+ B and CD8+ and CD4+ T cells from subject 034. Zoomed images of regions outlined in red are shown in bottom right corner of respective plots. Scale bars are 25 µm (panel c), 100 µm (panels a, b, and d), and 500 µm (panel e).