淋巴管平滑肌瘤病 (LAM) 是一种破坏性疾病，主要见于育龄妇女，会导致肺部囊性破坏。最近的研究表明，LAM 会导致免疫抑制，检查点抑制剂可用作 LAM 治疗。 Toll 样受体 (TLR) 激动剂还可以重新激活免疫力，TLR9 激动剂 CpG-ODN 在动物模型中可有效治疗肺癌。在这里，我们研究了 TLR9 激动剂 CpG-ODN 作为 LAM 免疫疗法与检查点抑制剂、抗 PD1、标准护理雷帕霉素的组合，并确定了治疗效果背后的免疫机制。我们使用生存研究、流式细胞术、ELISA 和组织学来评估转移性 LAM 小鼠模型中 CpG-ODN 鼻内治疗联合雷帕霉素或抗 PD1 治疗后的免疫反应和生存情况。我们发现局部施用 CpG-ODN 可以提高 LAM 小鼠模型的存活率。我们发现，与较高剂量相比，较低剂量可能会导致更长的生存期，因为局部副作用较少，但 LAM 结节数量和大小有所增加。 CpG-ODN 治疗还减少了调节性 T 细胞并增加了 Th17 辅助性 T 细胞以及细胞毒性 T 细胞的数量。这些效应似乎部分是由浆细胞样树突状细胞 (pDC) 介导的，因为 pDC 的耗竭会降低存活率并消除 Th17 T 细胞反应。最后，我们发现 CpG-ODN 治疗对早期和进展性疾病有效，并且与抗 PD1 治疗和雷帕霉素相结合。综上所述，我们发现TLR9激动剂CpG-ODN可以用作LAM免疫疗法，并在LAM中与雷帕霉素和抗PD1疗法有效协同。

Lymphangioleiomyomatosis (LAM) is a devastating disease primarily found in women of reproductive age that leads to cystic destruction of the lungs. Recent work has shown that LAM causes immunosuppression and that checkpoint inhibitors can be used as LAM treatment. Toll-like receptor (TLR) agonists can also re-activate immunity and the TLR9 agonist, CpG-ODN, has been effective in treating lung cancer in animal models. Here we investigate the use of TLR9 agonist CpG-ODN as LAM immunotherapy in combination with checkpoint inhibitor, anti-PD1, standard of care rapamycin and determine the immune mechanisms underlying therapeutic efficacy. We used survival studies, flow cytometry, ELISA, and histology to assess immune response and survival after intranasal treatment with CpG-ODN in combination with rapamycin or anti-PD1 therapy in a mouse model of metastatic LAM. We found that local administration of CpG-ODN enhances survival in a mouse model of LAM. We found that a lower dose led to longer survival likely due to fewer local side effects but increased LAM nodule count and size compared to the higher dose. CpG-ODN treatment also reduced regulatory T cells and increased the number of Th17 helper T cells as well as cytotoxic T cells. These effects appear to be mediated in part by plasmacytoid dendritic cells (pDCs), as depletion of pDCs reduces survival and abrogates Th17 T cell response. Finally, we found that CpG-ODN treatment is effective in early stage and progressive disease and is additive with anti-PD1 therapy and rapamycin. In summary, we have found that TLR9 agonist CpG-ODN can be used as LAM immunotherapy and effectively synergizes with rapamycin and anti-PD1 therapy in LAM.