Table_1_Inhibition of MC38 colon cancer growth by multicomponent chemoimmunotherapy with anti-IL-10R antibodies, HES-MTX nanoconjugate, depends on application of IL-12, IL-15 or IL-18 secreting dendritic cell vaccines.docx

BackgroundThe tumor microenvironment (TME) provides a conducive environment for the growth and survival of tumors. Negative factors present in TME, such as IL-10, may limit the effectiveness of cellular vaccines based on dendritic cells, therefore, it is important to control its effect. The influence of IL-10 on immune cells can be abolished e.g., by using antibodies against the receptor for this cytokine - anti-IL-10R. Furthermore, the anticancer activity of cellular vaccines can be enhanced by modifying them to produce proinflammatory cytokines, such as IL-12, IL-15 or IL-18. Additionally, an immunomodulatory dose of methotrexate and hydroxyethyl starch (HES-MTX) nanoconjugate may stimulate effector immune cells and eliminate regulatory T cells, which should enhance the antitumor action of immunotherapy based on DC vaccines. The main aim of our study was to determine whether the HES-MTX administered before immunotherapy with anti-IL-10R antibodies would change the effect of vaccines based on dendritic cells overproducing IL-12, IL-15, or IL-18.MethodsThe activity of modified DCs was checked in two therapeutic protocols - immunotherapy with the addition of anti-IL10R antibodies and chemoimmunotherapy with HES-MTX and anti-IL10R antibodies. The inhibition of tumor growth and the effectiveness of the therapy in inducing a specific antitumor response were determined by analyzing lymphoid and myeloid cell populations in tumor nodules, and the activity of restimulated splenocytes.Results and conclusionsUsing the HES-MTX nanoconjugate before immunotherapy based on multiple administrations of anti-IL-10R antibodies and cellular vaccines capable of overproducing proinflammatory cytokines IL-12, IL-15 or IL-18 created optimal conditions for the effective action of these vaccines in murine colon carcinoma MC38 model. The applied chemoimmunotherapy caused the highest inhibition of tumor growth in the group receiving DC/IL-15/IL-15Rα/TAg + DC/IL-18/TAg at the level of 72.4%. The use of cellular vaccines resulted in cytotoxic activity increase in both immuno- or chemoimmunotherapy. However, the greatest potential was observed both in tumor tissue and splenocytes obtained from mice receiving two- or three-component vaccines in the course of combined application. Thus, the designed treatment schedule may be promising in anticancer therapy.

背景：肿瘤微环境（TME）为肿瘤的生长与存活提供了有利条件。存在于TME中的负面因素，如IL-10，可能限制基于树突状细胞的细胞疫苗的效果，因此，控制其影响至关重要。通过使用针对该细胞因子受体的抗体——抗-IL-10R，可以消除IL-10对免疫细胞的影响。此外，通过修改细胞疫苗以产生促炎细胞因子，如IL-12、IL-15或IL-18，可以增强其抗癌活性。此外，免疫调节剂甲氨蝶呤与羟乙基淀粉（HES-MTX）纳米共轭物可能刺激效应免疫细胞并消除调节性T细胞，从而增强基于DC疫苗的免疫疗法的抗肿瘤作用。本研究的主要目的是确定在免疫疗法中使用抗-IL-10R抗体之前给予HES-MTX是否会改变过表达IL-12、IL-15或IL-18的树突状细胞疫苗的效果。方法：通过检查两种治疗方案的修改后的DCs活性——抗-IL10R抗体联合免疫疗法和HES-MTX与抗-IL10R抗体联合的化免疫疗法，来评估其活性。通过分析肿瘤结节中的淋巴和髓细胞群，以及重刺激的脾细胞活性，确定肿瘤生长的抑制和疗法诱导的特异性抗肿瘤反应的有效性。结果与结论：在基于多次给予抗-IL-10R抗体和能够过表达促炎细胞因子IL-12、IL-15或IL-18的细胞疫苗的免疫疗法之前使用HES-MTX纳米共轭物，为这些疫苗在鼠结肠癌细胞株MC38模型中的有效作用创造了最佳条件。应用化免疫疗法导致接受DC/IL-15/IL-15Rα/TAg + DC/IL-18/TAg治疗的组别肿瘤生长抑制水平最高，达到72.4%。使用细胞疫苗在免疫疗法或化免疫疗法中均导致细胞毒性活性增加。然而，在肿瘤组织和从接受两或三组分疫苗联合应用的小鼠中获得的脾细胞中观察到最大的潜力。因此，设计的治疗方案在抗癌疗法中具有潜在的可行性。