Juliana M Sousa-Canavez, Flavio C Canavez, Kátia R M Leite, Luiz H Camara-Lopes
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RNA extracted from prostate tumor lineage (LNCAP) was introduced into these cells by electroporation and co-incubation. Transfection success was measured by immunocytochemistry of the PSA expression level in DCs.</p><p><strong>Results: </strong>Cell surface markers, including CD14, CD80, CD86, CCR7, CD11c, and CD1a, confirmed mature and immature DC phenotypes. Both cell maturation stages were successfully used for RNA introduction as shown by PSA characterization.</p><p><strong>Conclusion: </strong>Our data support the use of mature and immature DCs for vaccine preparation with either RNA electroporation or RNA co-incubation. The highest efficiency, however, was observed when RNA was delivered by electroporation into mature DCs. Due to in vitro RNA transcription, this method allows small tumors to be used for DC vaccine preparation; it is therefore a promising approach for the treatment of metastatic prostate cancer.</p>","PeriodicalId":12596,"journal":{"name":"Genetic Vaccines and Therapy","volume":" ","pages":"2"},"PeriodicalIF":0.0000,"publicationDate":"2008-01-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2259348/pdf/","citationCount":"0","resultStr":"{\"title\":\"Therapeutic dendritic cell vaccine preparation using tumor RNA transfection: a promising approach for the treatment of prostate cancer.\",\"authors\":\"Juliana M Sousa-Canavez, Flavio C Canavez, Kátia R M Leite, Luiz H Camara-Lopes\",\"doi\":\"10.1186/1479-0556-6-2\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Early prostate adenocarcinoma can be diagnosed through seric prostate-specific antigen (PSA) screenings. 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引用次数: 0
摘要
背景:早期前列腺腺癌可通过血清前列腺特异性抗原(PSA)筛查确诊。然而,一部分患者会发展为无法治愈的转移性疾病。因此,治疗这些患者的新型疗法极为可取。以携带肿瘤抗原的树突状细胞(DC)为基础的治疗性疫苗已成为启动针对肿瘤细胞的免疫反应的一种有前途的策略。这些疫苗可采用不同的方法制备,如本研究中所述的肿瘤 mRNA 应用:方法:通过向单核细胞培养物中添加特定细胞因子,在体外获得成熟和不成熟的 DC。通过电穿孔和共孵育将从前列腺肿瘤(LNCAP)中提取的 RNA 导入这些细胞。转染成功与否通过免疫细胞化学法检测 DC 中 PSA 的表达水平来衡量:结果:细胞表面标志物(包括 CD14、CD80、CD86、CCR7、CD11c 和 CD1a)证实了成熟和未成熟 DC 的表型。正如 PSA 表征所示,这两个细胞成熟阶段都能成功用于 RNA 导入:我们的数据支持使用成熟和未成熟的直流细胞通过 RNA 电穿孔或 RNA 共孵育制备疫苗。然而,通过电穿孔将 RNA 送入成熟的 DC 时效率最高。由于RNA是体外转录的,这种方法可将小肿瘤用于制备DC疫苗;因此,它是治疗转移性前列腺癌的一种很有前途的方法。
Therapeutic dendritic cell vaccine preparation using tumor RNA transfection: a promising approach for the treatment of prostate cancer.
Background: Early prostate adenocarcinoma can be diagnosed through seric prostate-specific antigen (PSA) screenings. However, a fraction of patients progress to an incurable metastatic disease. Therefore, novel therapies for treating these patients are extremely desirable. Therapeutic vaccines based on Dendritic Cells (DCs) carrying tumor antigens have emerged as a promising strategy to initiate an immune response against tumor cells. These vaccines can be prepared using different methodologies, such as the application of tumor mRNA described in this work.
Methods: Mature and immature DCs were obtained in vitro by adding specific cytokines to monocyte cell cultures. RNA extracted from prostate tumor lineage (LNCAP) was introduced into these cells by electroporation and co-incubation. Transfection success was measured by immunocytochemistry of the PSA expression level in DCs.
Results: Cell surface markers, including CD14, CD80, CD86, CCR7, CD11c, and CD1a, confirmed mature and immature DC phenotypes. Both cell maturation stages were successfully used for RNA introduction as shown by PSA characterization.
Conclusion: Our data support the use of mature and immature DCs for vaccine preparation with either RNA electroporation or RNA co-incubation. The highest efficiency, however, was observed when RNA was delivered by electroporation into mature DCs. Due to in vitro RNA transcription, this method allows small tumors to be used for DC vaccine preparation; it is therefore a promising approach for the treatment of metastatic prostate cancer.