Abstract Melanoma is a tumor that forms as a result of malignant transformation of melanin-producing pigment cells (melanocytes). It is the most aggressive form of skin cancer and is characterized by high resistance to chemotherapeutic drugs, which results in a need to explore alternative methods for this disease therapy. Currently, new approaches to cancer treatment are being intensely developed, oncolytic immunotherapy being one of them. This approach consists in using viruses as targeted tumor-specific cytolytic agents capable of stimulating both tumor-specific and nonspecific immune responses. A considerable body of research is currently aimed on improving the immunostimulatory properties of viruses by inserting the genes encoding immunomodulatory proteins or tumor-specific antigenic determinants into viral genomes. For melanoma, the highest number of tumor-associated antigens (TAAs) has been identified, which serve as the basis for the development of anti-tumor DNA vaccines. The immunogenicity and efficacy of these drugs, however, remain low. The bottlenecks in using DNA vaccines to treat cancer are considered to be imperfect design of polyepitope constructs, as well as inefficient delivery of therapeutic molecules directly to the target cells. A partial solution to these problems may be represented by the use of oncolytic viruses as vectors for the delivery of artificial immunogens. The recombinant vaccinia virus was obtained by transient dominant selection. The cytolytic activity of the obtained virus was tested using the MTT assay. The oncolytic activity of the virus was assessed in the mouse xenograft model obtained using malignant SK-Mel-28 cells. This paper reports the production of a recombinant L-IVP_oncoM virus, the oncolytic virus for the delivery of anticancer therapeutic genes into the cells, on the basis of the vaccinia virus strain L-IVP. Toward this end, the gene encoding the Granulocyte-macrophage colony-stimulating factor (GM-CSF) and the artificial gene encoding a polyepitope immunogen containing the epitopes of the antigens over-expressed in melanoma cells were inserted into the virus genome. These insertions were located in the proximity of the genes encoding thymidine kinase (J2R) and viral growth factor (C11L), respectively. The properties of L-IVP_oncoM were studied in in vitro experiments using cell cultures of various origin and in the in vivo experiments using the mouse xenograft model. The basic experiments to assess the biological properties of the obtained L-IVP_oncoM, which are necessary to characterize the oncolytic virus, have been carried out.

中文翻译：

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有望用于治疗黑色素瘤的重组痘苗病毒

摘要 黑色素瘤是产生黑色素的色素细胞（黑色素细胞）恶性转化而形成的肿瘤。它是最具侵袭性的皮肤癌形式，其特征是对化疗药物具有高度耐药性，因此需要探索该疾病治疗的替代方法。目前，正在大力开发新的癌症治疗方法，溶瘤免疫疗法就是其中之一。这种方法包括使用病毒作为能够刺激肿瘤特异性和非特异性免疫反应的靶向肿瘤特异性溶细胞剂。目前，大量研究旨在通过将编码免疫调节蛋白或肿瘤特异性抗原决定簇的基因插入病毒基因组来改善病毒的免疫刺激特性。对于黑色素瘤，已鉴定出数量最多的肿瘤相关抗原 (TAA)，这是开发抗肿瘤 DNA 疫苗的基础。然而，这些药物的免疫原性和功效仍然很低。使用 DNA 疫苗治疗癌症的瓶颈被认为是多表位构建体的设计不完善，以及将治疗分子直接传递到靶细胞的效率低下。这些问题的部分解决方案可以通过使用溶瘤病毒作为递送人工免疫原的载体来表示。通过瞬时显性选择获得重组痘苗病毒。使用MTT测定法测试获得的病毒的细胞溶解活性。在使用恶性 SK-Mel-28 细胞获得的小鼠异种移植模型中评估病毒的溶瘤活性。本文报道了在痘苗病毒株 L-IVP 的基础上生产重组 L-IVP_oncoM 病毒，该病毒用于将抗癌治疗基因传递到细胞中。为此，将编码粒细胞-巨噬细胞集落刺激因子 (GM-CSF) 的基因和编码包含在黑色素瘤细胞中过度表达的抗原表位的多表位免疫原的人工基因插入到病毒基因组中。这些插入分别位于编码胸苷激酶 (J2R) 和病毒生长因子 (C11L) 的基因附近。在使用各种来源的细胞培养物的体外实验和使用小鼠异种移植模型的体内实验中研究了 L-IVP_oncoM 的特性。