Journal: Cancer Research Communications
Article Title: Intratumoral Virus-Like Particles Containing a TLR9 Agonist Combined with Systemic αPD-1 Activate Tumor-Specific CD8 + T Cells
doi: 10.1158/2767-9764.CRC-26-0175
Figure Lengend Snippet: Vidu decreases proliferation of tumor-specific CD8 + T cells while enhancing activation of cells that divided. OT-1 splenocytes were labeled with a proliferation tracking dye (CellTrace Violet) and cultured with either media (unstimulated) or E.G7-OVA cells (stimulated) for 1 hour, followed by no additional treatment (untreated) or the addition of Vidu and αQβ (treated). After 3 days of coculture, proliferation and activation marker expression by OT-1 CD8 + T cells was analyzed by multicolor spectral flow cytometry. A, Percent divided and ( B ) proliferation index of OT-1 CD8 + T cells ( n = 10 mice/group). C, Representative histograms showing OT-1 CD8 + T-cell proliferation. D, MdFI of IFNγ expression by undivided and divided OT-1 CD8 + T cells ( n = 10 mice/group) and ( E ) representative dot plot and histogram in unstimulated (blue), stimulated/untreated (black), and stimulated/treated (red) samples. F, MdFI of TNFα expression by undivided and divided OT-1 CD8 + T cells ( n = 4 mice/group) and ( G ) representative dot plot and histogram. H, MdFI of PD-1 expression by undivided and divided OT-1 CD8 + T cells ( n = 10 mice/group) and ( I ) representative dot plot and histogram. J, MdFI of CD25 expression by undivided and divided OT-1 CD8 + T cells ( n = 7 mice/group) and ( K ) representative dot plot and histogram. E , G , I , and K, Red arrows indicate undivided population, and vertical dashed lines indicate separation of negative (left) from positive (right) marker expression. Each symbol connected by a line represents an individual mouse in untreated and Vidu/αQβ conditions and the mean of the group, with error bars indicating the SEM. Statistical significance was determined using a paired t test ( A and B ) or two-way ANOVA with Sidak multiple comparisons test ( D–J ): *, P < 0.05; **, P < 0.01; ****, P < 0.0001; ns, not significant.
Article Snippet: EL4 (cat. #TIB-39, RRID: CVCL_0255) and E.G7-OVA (cat. #CRL-2113, RRID: CVCL_3505) cell lines were obtained from the American Type Culture Collection and stored in liquid nitrogen tanks in the Weiner Laboratory.
Techniques: Activation Assay, Labeling, Cell Culture, Marker, Expressing, Flow Cytometry