Le Noci Valentino, Sommariva Michele, Tortoreto Monica, Zaffaroni Nadia, Campiglio Manuela, Tagliabue Elda, Balsari Andrea, Sfondrini Lucia
Dipartimento di Scienze Biomediche per la Salute, Università degli Studi di Milano, Milan, Italy; Molecular Targeting Unit, Milan, Italy.
Molecular Pharmacology Unit, Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy.
Oncoimmunology. 2016 Sep 20;5(11):e1234571. doi: 10.1080/2162402X.2016.1234571. eCollection 2016.
Due to their constant exposure to inhaled antigens, lungs represent a particularly immunosuppressive environment that limits excessive immune responses; however, cancer cells can exploit this unique environment for their growth. We previously described the ability of aerosolized CpG-ODN combined with Poly(I:C) (TLR9 and TLR3 agonists, respectively) to promote antitumor immunity in a B16 melanoma lung metastasis model. Here, we explored the possibility of improving the therapeutic efficacy of TLR9/TLR3 agonist combinations by including in the inhalant either an antibody directed to both Ly6G and Ly6C markers to locally deplete myeloid-derived suppressive cells (MDSCs) or IFNα to directly activate the natural killer (NK) and macrophage innate immune cells in the lung. Addition of nebulized anti-MDSC antibody RB6-8C5 to aerosolized CpG-ODN/Poly(I:C) resulted in reduced mRNA levels of immunsuppressive molecules (IL10, Arg-1, and Nos2), increased activation of resident NK cells and improved treatment outcome, with a significant reduction in established B16 melanoma lung metastases compared to treatment with CpG-ODN/Poly(I:C) alone. Likewise, addition of aerosolized IFNα led to increased mRNA levels of proinflammatory cytokines (IL15 and IFNγ) in the lung and recruitment of highly activated NK cells, with no evident signs of toxicity and with a significantly improved antitumor effect as compared with aerosolized CpG-ODN/Poly(I:C). Combining both IFNα and RB6-8C5 with CpG-ODN/Poly(I:C) did not produce an additive effect compared to IFNα + CpG-ODN/Poly(I:C) or RB6-8C5 + CpG-ODN/Poly(I:C). Our results indicate that the inhalation therapy is a feasible and non-invasive strategy to deliver immunodulatory molecules, including antibodies and cytokines that reprogram the lung tumor microenvironment to foster immune destruction of tumors.
由于肺部持续暴露于吸入的抗原，因此呈现出一种特别具有免疫抑制作用的环境，可限制过度的免疫反应；然而，癌细胞能够利用这种独特的环境来实现生长。我们之前描述了雾化的CpG-ODN与聚肌胞苷酸（分别为TLR9和TLR3激动剂）联合使用在B16黑色素瘤肺转移模型中促进抗肿瘤免疫的能力。在此，我们探讨了通过在吸入剂中加入一种针对Ly6G和Ly6C标志物的抗体以局部清除髓源性抑制细胞（MDSC）或加入干扰素α以直接激活肺部的自然杀伤（NK）细胞和巨噬细胞固有免疫细胞，从而提高TLR9/TLR3激动剂组合治疗效果的可能性。将雾化的抗MDSC抗体RB6-8C5添加到雾化的CpG-ODN/聚肌胞苷酸中，导致免疫抑制分子（IL10、精氨酸酶-1和一氧化氮合酶2）的mRNA水平降低，驻留NK细胞的活化增加，治疗效果得到改善，与单独使用CpG-ODN/聚肌胞苷酸治疗相比，已形成的B16黑色素瘤肺转移显著减少。同样，添加雾化的干扰素α导致肺部促炎细胞因子（IL15和干扰素γ）的mRNA水平升高以及高活化NK细胞的募集，没有明显的毒性迹象，并且与雾化的CpG-ODN/聚肌胞苷酸相比，抗肿瘤效果显著改善。与干扰素α + CpG-ODN/聚肌胞苷酸或RB6-8C5 + CpG-ODN/聚肌胞苷酸相比，将干扰素α和RB6-8C5两者与CpG-ODN/聚肌胞苷酸联合使用并未产生相加效应。我们的结果表明，吸入疗法是一种可行的非侵入性策略，可用于递送免疫调节分子，包括能够重新编程肺肿瘤微环境以促进肿瘤免疫破坏的抗体和细胞因子。
