Gene analysis of tumor cells from mice with the Treg-specific deletion of Blimp1 compared to WT mice (NanoString Cancer)

We observed that mice with the Treg-specific deletion of Blimp1 had delayed and smaller tumor growth associated with the activation of tumor-infiltrating immune effector cells. To determine what extent disruptions of Treg suppressive activity by a specific deletion of Blimp1 could impact on the tumor, we assessed the gene expression of sorted tumor cells by NanoString analysis. Prdm1fl/flFoxP3YFP-Cre (KO) and FoxP3YFP-Cre (WT) mice were subcutaneously inoculated with B16.OVA at day 0, and intraperitoneally immunized with NP-OVA in CFA at day 0 and NP-OVA in IFA at day 7. When the tumor reached ≤ 1.5 cm3, CD45-live tumor cells were FACS-sorted from the tumors of these mice. RNA was prepared and analyzed using NanoString PanCancer Pathways Panel.

我们观察到，调节性T细胞（Treg）特异性缺失Blimp1的小鼠，其肿瘤生长延迟且瘤体更小，该表型与肿瘤浸润免疫效应细胞的活化密切相关。为明确Blimp1特异性敲除所破坏的Treg抑制活性可在多大程度上影响肿瘤进程，我们通过纳米String（NanoString）分析对分选获得的肿瘤细胞进行了基因表达检测。我们于第0天对Prdm1fl/flFoxP3YFP-Cre（敲除组，KO）及FoxP3YFP-Cre（野生型组，WT）小鼠皮下接种B16.OVA细胞，并于第0天采用完全弗氏佐剂（CFA）乳化的NP-OVA进行腹腔免疫，第7天时以不完全弗氏佐剂（IFA）乳化的NP-OVA再次实施腹腔免疫。当肿瘤体积≤1.5 cm³时，我们从上述小鼠的肿瘤组织中通过荧光激活细胞分选术（FACS）分选出CD45阳性活肿瘤细胞。提取总RNA后，采用纳米String泛癌通路基因分析面板（NanoString PanCancer Pathways Panel）完成了检测与分析。