Figure 5: Intratumor fungus accelerates PDAC tumor growth.

(A) Schematic showing fungal depletion strategy using amphotericin B (200 ug/dose) followed by orthotopic transplantation of PDAC cells and tumor progression studies.

(B) Representative MRI images with their relative volumes (n=5) on day 26, showing orthotopically transplanted shCtrl and shIL33 PDAC tumors with or without amphotericin B treatment. Red dotted circles define the tumor boundaries.

(C) Bar graph showing tumor volume of orthotopically transplanted shCtrl and shIL33 PDAC tumors (n=14) and shIL33 (#1 [n=9] and #2 [n=10]) with or without amphotericin B treatment.

(D) Kaplan–Meier survival curves of mice orthotopically transplanted with shCtrl and shIL33 PDAC tumors with or without amphotericin B treatment (n=10). Statistical analysis was done using the Gehan-Breslow Wilcoxon test.

(E) Frequency of ILC2 in orthotopically transplanted shCtrl and shIL33 PDAC tumors with or without amphotericin B treatment relative to total Lin⁻ cells.

(F) Frequency of T_H2 cells in orthotopically transplanted shCtrl and shIL33 PDAC tumors with or without amphotericin B treatment relative to total CD4⁺ cells.

(G) Schematic showing strategy for fungal transplantation followed by orthotopic transplantation of PDAC cells and tumor progression studies.

(H) Representative IHC (top) and immunofluorescence (middle) images of PDAC tumors showing IL33 expression in control and fungal transplanted mice by oral gavage (n=10), Magnification 40x. Scale bar 100 μm. FISH (bottom) showing fungal colonization of PDAC tumors in fungal transplanted mice by oral gavage. Scale bar 10 μm.

(I) 18S rRNA sequence showing fungal species in PDAC tumors of the Alternaria alternata and Malassezia globosa transplanted mice by oral gavage. Also, shown are the 18S rRNA sequencing in stool samples of the Alternaria alternata and Malassezia globosa transplanted mice by oral gavage. Positive control-Alternaria is a sample of pure Alternaria alternata culture extract.

(J) Bar Graph showing the wet weight of PDAC tumors of the control, amphotericin B and fungal transplanted mice (n=10).

(K) Frequency of ILC2/total Lin⁻ cells in PDAC tumors of control, amphotericin B and fungal transplanted mice.

Data are presented as mean ±SD. p-values were calculated using the Student t-test. ns, no significance. Individual p-values are indicated in the figures.

See also Figure S5.