Fig 2: Comparison of the effectiveness of OV-A4-IgG2b versus OV- A4-IgG3 in a fully immunocompetent mouse GBM model.a Survival of CT2A tumor-bearing C57BL/6 immunocompetent mice treated with vehicle control, OV-Q1, OV-A4-IgG2b, or OV-4-IgG3. b Survival of CT2A tumor-bearing C57BL/6 immunocompetent mice treated with vehicle control, OV-Q1, or OV-A4-IgG2b in the presence or absence of NK cell depletion. c Survival of CT2A tumor-bearing C57BL/6 immunocompetent mice treated with vehicle control, OV-Q1, or OV-A4-IgG2b in the presence or absence of macrophage depletion. Survival was estimated by the Kaplan–Meier method and compared by two-side log-rank test. d-f Intracranial infiltration of NK cells (CD3-NKp46+) (d), macrophages (F4/80+CD45highCD11b+) (e), and T cells (CD3+NKp46-) (f) in CT2A-bearing mice measured by flow cytometry 3 days after virus injection (n = 5 animals). g Viral production of OV-Q1-, OV-A4-IgG2b-, and OV-A4-IgG3-treated mice. h CD69 expression in NK cells isolated from mouse brains 3 days after virus injection. Representative flow cytometric plots of each group (n = 5 animals). i Quantification of CD69 expression in NK cells in (h). j In vivo macrophage phagocytosis of GBM-expressing CT2A cells after oHSV treatment. Representative flow cytometric plots of each group (n = 6 animals). k Quantification of macrophage phagocytosis of GBM-expressing CT2A cells in (j). l Red blood cell counts in peripheral blood after i.c. injection of OV-A4-IgG2b or i.p. injection of purified A4-IgG2b (n = 6 mice). For (d-g), (i) and (k, l), one-way ANOVA with P values corrected for multiple comparisons by the Bonferroni test. Data are presented as mean values ± SD.

Fig 3: DHA feeding affected NK cell frequency, maturation and Ly49H+ NK cell expansion during the response to MCMV infection. A Flow cytometric analysis and enumeration of NK cells (CD45+CD3-CD19- NK1.1+NKp46+) in the BM, spleens, pLNs and livers of control versus DHA-fed mice. B Flow cytometric analysis and cumulative frequencies of subpopulations of NK cell (CD3-CD19-NK1.1+NKp46+) subsets based on CD11b and CD27 expression in the BM, spleens, pLNs and livers of control versus DHA-fed mice. C Flow cytometric analysis of the KLRG1+ subsets of NK cells (CD3-CD19-NK1.1+NKp46+) in the BM, spleens, pLNs and livers of control versus DHA-fed mice at day 7 postinfection. D Flow cytometric analysis and enumeration of Ly49H+ subsets of NK cells (CD3-CD19-NK1.1+NKp46+) in the BM, spleens, pLNs and livers of control versus DHA-fed mice at day 7 postinfection. E Cumulative frequencies of NK cell (CD3-CD19-NK1.1+NKp46+) subsets found in the spleens, and livers of control versus DHA-fed mice at day 21 postinfection. For each experiment, n = 4 to 9 pooled from 2–3 independent experiments (A–E). Each symbol represents an individual mouse, and the blue dots and red square represent control and DHA-enriched diet-fed mice, respectively. Error bars represent interquartile ranges; *p < 0.05; **p < 0.01; ***p < 0.001

Fig 4: Systemic, long-term, DLN-independent memory is formed with tumor only SBRT.A Tumor growth curves of mice rechallenged with LY2 tumor cells in both the buccal and in the flank (Cured, n = 3; Naïve, n = 8). B t-SNE clustering and FlowSOM clusters superimposed of CD45+ immune cells in the DLN and Blood. CD8 T cells (CD3+CD8+), CD4 T cells (CD3+CD4+), NK cells (CD3-NKp46+), replicating cells (Ki-67+), and MHCII+ cells are highlighted. C–H Quantification of activation markers expressed by T cells in the DLN and in the blood of naive mice or mice rechallenged with LY2 cells in the buccal (<6 months after initial eradication) (Cured, n = 3; Naïve, n = 5). I–L Quantification of immune cells in the blood of mice before and after rechallenging with LY2 cells in the buccal or implanting LY2 cells into naïve mice (<6 months after initial eradication) (Cured, n = 4; Naïve, n = 4). M Tumor growth curves of naïve mice or cured mice treated with either a CD8 or CD4 T cell depleting antibody (anti-CD4, n = 5; anti-CD8, n = 3; naïve, n = 5). N Tumor growth curves of mice treated with neoadjuvant SBRT and anti-CD25 with a neck dissection occurring on day 19 post-tumor implantation. Individual tumor growth curves are provided to the right (Tumor and LN removal, n = 6; Tumor removal, n = 6; sham surgery, n = 6; no surgery, n = 4). A two-tailed student’s t test was used to examine group differences. Significance was determined if the p-value was <0.05*, <0.01**, <0.001***, and <0.0001****. The error bars represent the standard error of the mean (±SEM). Source data are provided as a Source Data file. p-values are indicated for figures C. *0.011, D. ***0.0002, *0.0114, E. CD4+CD44+***0.0001, *0.0137, CD4+IL-2+ ***0.0009, F. ***0.0008, **0.0014, G. CD4***0.0007, CD8***0.0003, H. CD103**** < 0.0001, CD80**** < 0.0001, CCR7***0.0006, Ki-67***0.0004, I. **0.0011, J. *0.0423, K. control ****<0.0001, cured ****<0.0001, and L. *0.0341.

Fig 5: Differences in the cellular metabolic status and mitochondrial activity of splenic NK cells between DHA-fed and control mice at day 7 post-MCMV infection. A–E Flow cytometric analysis and cumulative results depicting the gMFI of CD71 (A) and CD98 (B) and the uptake of 2-NBDG (C), MitoTracker (D) and tetramethylrhodamine, methyl ester (TMRM) (E) by splenic NK cells (CD3-CD19-NK1.1+NKp46+) from control versus DHA-fed mice at day 7 postinfection. For each experiment, n = 7 pooled from 2 independent experiments. Each symbol represents an individual mouse, and the blue dots and red squares represent control and DHA-enriched diet-fed mice, respectively. Error bars represent interquartile ranges; *p < 0.05; **p < 0.01; ***p < 0.001