A recombinant oncolytic Newcastle virus expressing MIP-3α promotes systemic antitumor immunity
A recombinant oncolytic Newcastle virus expressing MIP-3α promotes systemic antitumor immunity
复制标题
表达 MIP-3α 的重组溶瘤新城疫病毒促进全身抗肿瘤免疫
DOI：
10.1136/jitc-2019-000330
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发表时间：
2020-01-01
影响因子：
10.9
通讯作者：
Tan, Guang-Hong
中科院分区：
文献类型：
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作者：
Huang, Feng-Ying;Wang, Jin-Yan;Tan, Guang-Hong
背景溶瘤性纽卡斯尔病病毒（NDV）具有裂解肿瘤细胞和诱导肿瘤细胞免疫原性细胞死亡（ICD）的特性，是一种优良的基因工程载体。巨噬细胞炎性蛋白-3 α（MIP-3 α）是树突状细胞（DC）的特异性趋化因子。因此，我们构建了表达MIP-3 α的重组NDV（NDV-MIP 3 α）作为用于增强抗肿瘤免疫的体内DC疫苗。方法将MIP-3 α cDNA插入NDV的P和M基因之间，构建重组NDV MIP-3 α。采用Western blotting法和ELISA法检测培养上清和血清中MIP-3 α、HMGB 1、IgG和ATP的含量。Transwell小室检测DC的趋化性。通过流式细胞术分析免疫细胞（例如，DC）的表型。在B16和CT 26荷瘤小鼠中观察NDV-MIP 3 α的抗肿瘤效率。免疫荧光和免疫组化观察DC的外钙网蛋白（CRT）和瘤内粘附。脾细胞和抗体的连续转移和T细胞亚群的耗竭被用来评估抗肿瘤免疫和T细胞亚型的作用之间的关系。结果NDV-MIP 3 α具有与野生型NDV（NDV-WT）相似的溶瘤和诱导ICD的能力。NDV-MIP-3 α分泌的MIP-3 α在体外和体内均能成功地吸引DC。NDV-MIP 3 α感染的B16和CT 26细胞均能明显促进DC的成熟和活化。与NDV-WT相比，瘤内注射NDV-MIP 3 α和过继转移来自注射NDV-MIP 3 α的小鼠的T淋巴细胞导致B16和CT 26肿瘤生长的显著抑制。NDV-MIP 3 α诱导的肿瘤特异性细胞和体液免疫应答的产生依赖于CD 8（+）T细胞，部分依赖于CD 4（+）T细胞。在注射NDV-MIP 3 α的小鼠中发现肿瘤微环境的显著逆转。结论与NDV-WT相比，重组NDV-MIP 3 α作为体内DC疫苗，通过诱导更强的系统免疫和调节肿瘤微环境，具有更强的抗肿瘤活性。这种策略可能是一种潜在的方法，在体内产生的DC疫苗。
Background The oncolytic Newcastle disease virus (NDV) is inherently able to trigger the lysis of tumor cells and induce the immunogenic cell death (ICD) of tumor cells and is also an excellent gene-engineering vector. The macrophage inflammatory protein-3 alpha (MIP-3 alpha) is a specific chemokine for dendritic cells (DCs). Thus, we constructed a recombinant NDV expressing MIP-3 alpha (NDV-MIP3 alpha) as an in vivo DC vaccine for amplifying antitumor immunities. Methods The recombinant NDV-MIP3 alpha was constructed by the insertion of MIP-3 alpha cDNA between the P and M genes. Western blotting assay and ELISA were used to detect MIP-3 alpha, HMGB1, IgG, and ATP in the supernatant and sera. The chemotaxis of DCs was examined by Transwell chambers. The phenotypes of the immune cells (eg, DCs) were analyzed by flow cytometry. The antitumor efficiency of NDV-MIP3 alpha was observed in B16 and CT26 tumor-bearing mice. Immunofluorescence and immunohistochemistry were applied to observe the ecto-calreticulin (CRT) and intratumoral attraction of DCs. Adoptive transfer of splenocytes and antibodies and depletion of T-cell subsets were used to evaluate the relationship between antitumor immunities and the role of the T-cell subtype. Results The findings show that NDV-MIP3 alpha has almost the same capabilities of tumor lysis and induction of ICD as the wild-type NDV (NDV-WT). MIP-3 alpha secreted by NDV-MIP3 alpha could successfully attract DCs in vitro and in vivo. Both B16 and CT26 cells infected with NDV-MIP3 alpha could strongly promote DC maturation and activation. Compared with NDV-WT, intratumoral injection of NDV-MIP3 alpha and the adoptive transfer of T lymphocytes from mice injected with NDV-MIP3 alpha resulted in a significant suppression of B16 and CT26 tumor growth. The NDV-MIP3 alpha-induced production of tumor-specific cellular and humoral immune responses was dependent on CD8(+)T cells and partially on CD4(+)T cells. A significant reversion of tumor microenvironments was found in the mice injected with NDV-MIP3 alpha. Conclusions Compared with NDV-WT, the recombinant NDV-MIP3 alpha as an in vivo DC vaccine demonstrates enhanced antitumor activities through the induction of stronger system immunities and modulation of the tumor microenvironment. This strategy may be a potential approach for the generation of an in vivo DC vaccine.