Department of Radiology, The Sixth Affiliated Hospital of Wenzhou Medical University, Lishui, Zhejiang, China.
Department of Respiratory, The Sixth Affiliated Hospital of Wenzhou Medical University, Lishui, Zhejiang, China.
Immun Inflamm Dis. 2023 Sep;11(9):e1011. doi: 10.1002/iid3.1011.
Fibroblast activation protein-α (FAP) and livin α are considered as cancer-associated fibroblasts (CAFs) and tumor-specific targets, respectively, for immunogenic tumor vaccines. This study is designed to decipher the antitumor effect of double-gene modified dendritic cells (DCs) on Lewis lung carcinoma (LLC).
By encoding mouse FAP cDNA and human livin α (i.e., hlivin α) cDNA into recombinant adenoviral vector (rAd), rAd-FAP, rAd-hlivin α, and rAd-FAP/hlivin α were constructed, which were then transduced into mouse DCs. LLC-bearinig mice were immunized with the infected DCs (5 × 10 cells/mouse), followed by calculation of tumor volume and survival rate. The identification of CAFs from mouse LLC as well as the determination on expressions of FAP and livin α, was accomplished by western blot. Cytotoxic T lymphocyte assay was harnessed to assess the effect of the infected DCs on inducing splenic lymphocytes to lyse CAFs.
DCs were successfully transduced with rAd-FAP/hlivin α in vitro. FAP was highly expressed in CAFs. CAFs were positive for α-SMA and negative for CD45 and CD31. Livin α level was upregulated in mouse LLC. Immunization with rAd-FAP/hlivin α-transduced DCs suppressed LLC volume and improved the survival of tumor-bearing mice. Immunization with rAd-FAP/hlivin α-transduced DCs enhanced the cytotoxic effect of splenic lymphocytes on LLC tumor-derived CAFs.
Injection with rAd-FAP/hlivin α-transduced DCs promotes immune-enhanced tumor microenvironment by decreasing CAFs and suppresses tumor growth in LLC mouse models.
成纤维细胞激活蛋白-α（FAP）和 livin α 分别被认为是癌症相关成纤维细胞（CAFs）和肿瘤特异性的免疫原性肿瘤疫苗靶点。本研究旨在阐明双基因修饰树突状细胞（DC）对 Lewis 肺癌（LLC）的抗肿瘤作用。
通过将小鼠 FAP cDNA 和人 livin α（即 hlivin α）cDNA 编码到重组腺病毒载体（rAd）中，构建了 rAd-FAP、rAd-hlivin α 和 rAd-FAP/hlivin α，然后将其转染到小鼠 DC 中。用感染的 DC（5×10 个细胞/只）免疫 LLC 荷瘤小鼠，计算肿瘤体积和存活率。通过 Western blot 鉴定小鼠 LLC 中的 CAFs 以及 FAP 和 livin α 的表达情况。细胞毒性 T 淋巴细胞试验用于评估感染的 DC 对诱导脾淋巴细胞溶解 CAFs 的影响。
体外成功转染 rAd-FAP/hlivin α 到 DC 中。FAP 在 CAFs 中高表达。CAFs 呈 α-SMA 阳性，CD45 和 CD31 阴性。Livin α 水平在小鼠 LLC 中上调。用 rAd-FAP/hlivin α 转染的 DC 免疫抑制 LLC 体积，提高荷瘤小鼠的存活率。rAd-FAP/hlivin α 转染的 DC 免疫增强了脾淋巴细胞对 LLC 肿瘤衍生 CAFs 的细胞毒性作用。
注射 rAd-FAP/hlivin α 转染的 DC 通过减少 CAFs 促进免疫增强的肿瘤微环境，并抑制 LLC 小鼠模型中的肿瘤生长。
