肿瘤微环境（TME）为肿瘤的生长和存活提供了有利的环境。TME中存在的负性因素，如IL-10，可能限制基于树突状细胞的细胞疫苗的效果，因此，控制其影响是很重要的。可以通过使用针对该细胞因子受体的抗体—抗IL-10R来消除IL-10对免疫细胞的影响。此外，通过改造细胞疫苗以产生促炎性细胞因子，如IL-12、IL-15或IL-18，可以增强细胞疫苗的抗癌活性。此外，免疫调节剂剂量的甲氨蝶呤和羟乙基淀粉（HES-MTX）纳米复合物可以刺激效应免疫细胞并消除调节性T细胞，从而应该增强基于树突状细胞疫苗的免疫治疗的抗肿瘤作用。我们研究的主要目标是确定在具有过表达IL-12、IL-15或IL-18的树突状细胞的疫苗上，HES-MTX在免疫治疗之前是否会改变使用抗IL-10R抗体的疫苗的效果。通过分析肿瘤结节中的淋巴细胞和髓系细胞群体以及再刺激脾细胞的活性，确定了抑制肿瘤生长和诱导特异性抗肿瘤反应的疗法的有效性。在小鼠结肠癌MC38模型中，使用HES-MTX纳米复合物在多次给予抗IL-10R抗体和过表达促炎细胞因子IL-12、IL-15或IL-18的细胞疫苗的免疫治疗之前，为这些疫苗的有效作用创造了最佳条件。应用的化疗免疫治疗在接受DC/IL-15/IL-15Rα/TAg + DC/IL-18/TAg两者治疗的组中以72.4%的水平抑制了肿瘤生长。使用细胞疫苗导致了免疫治疗或化疗免疫治疗中的细胞毒作用增加。然而，无论是在肿瘤组织还是从小鼠中获得的脾细胞中，都观察到了两种或三种成分疫苗联合应用时的最大潜力。因此，设计的治疗方案在抗癌治疗中可能是有希望的。版权所有 © 2023 Węgierek-Ciura, Mierzejewska, Szczygieł, Rossowska, Wróblewska, Świtalska, Goszczyński, Szermer-Olearnik and Pajtasz-Piasecka。

The 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.The 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.Using 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.Copyright © 2023 Węgierek-Ciura, Mierzejewska, Szczygieł, Rossowska, Wróblewska, Świtalska, Goszczyński, Szermer-Olearnik and Pajtasz-Piasecka.