Fig. 5

Asparaginase treatment increased NKTCL cell sensitivity to anti-PD-1 antibody.

(a) SLC1A1 expression on NK-92 cells transfected with SLC1A1 vector or control vector (upper panel) and SNK-6 cells transfected with SLC1A1 shRNA or scramble (lower panel).

(b) Ki-67 and TIM-3 positivity of CD3+/CD8+ T cells in PBMC co-cultured with NK-92 cells (upper panel) or SNK-6 cells (lower panel) transfected with indicated vectors or shRNAs in medium with or without extra glutamine (2mM).

(c) PD-L1 mRNA expression in NK-92 cells transfected with SLC1A1 vector or control vector (upper panel) and SNK-6 cells transfected with SLC1A1 shRNA or scramble (lower panel) upon asparaginase (10 IU/mL) treatment. The control vector or scramble values were normalized to 1, respectively.

(d and e) Median fluorescence intensity of PD-L1 (d) on NK-92 cells (upper panel) or SNK-6 cells (lower panel), as well as Ki-67 and TIM-3 positivity of CD3+/CD8+ T cells in PBMC co-cultured with NK-92 cells (upper panel) or SNK-6 cells (lower panel) upon indicated treatment.

(f) Gene expression correlation of tumor SLC1A1 with PD-L1 in NKTCL patients (n=128).

(g) Tumor EAAT3 expression according to the TSIM, HEA, and MB subtypes in NKTCL patients (n=100).

(h) PD-L1 mRNA expression of NK-92 cells transfected with TP53 R248Q, TP53 R273H, EP300 vector, or MGA shRNA upon indicated treatment.

(i) Ki-67 positivity of CD3+/CD8+ T cells in PBMC co-cultured with NK-92 cells transfected with TP53 R248Q, TP53 R273H, EP300 vector, or MGA shRNA upon indicated treatment.

Assays in (a), (b), (c), (d), (e), (h), and (i) were set up in triplicate. Data in (a), (b), (c), (d), (e), (h), and (i) were represented as mean ± SD. P values in in (a), (b), (c), (d), (e), (h), and (i) were calculated with unpaired t-test. P value in (f) was calculated with Pearson correlation test. P values in (g) were calculated by Pearson's chi-square test.