Department of Virology, LG-Schreiber, Bernhard Nocht Institute for Tropical Medicine, Bernhard Nocht Str. 74, 20359 Hamburg, Germany.
Department of Pathology and Neuropathology, Asklepios Kliniken Hamburg GmbH, Asklepios Klinik Nord, Standort Heidberg, 22417 Hamburg, Germany.
Int J Mol Sci. 2023 Feb 24;24(5):4467. doi: 10.3390/ijms24054467.
This study aimed to isolate cells from grade 4 glioblastoma multiforme tumors for infection experiments with Zika virus (ZIKV) prME or ME enveloped HIV-1 pseudotypes. The cells obtained from tumor tissue were successfully cultured in human cerebrospinal fluid (hCSF) or a mixture of hCSF/DMEM in cell culture flasks with polar and hydrophilic surfaces. The isolated tumor cells as well as the U87, U138, and U343 cells tested positive for ZIKV receptors Axl and Integrin αvβ5. Pseudotype entry was detected by the expression of firefly luciferase or green fluorescent protein (gfp). In prME and ME pseudotype infections, luciferase expression in U-cell lines was 2.5 to 3.5 logarithms above the background, but still two logarithms lower than in the VSV-G pseudotype control. Infection of single cells was successfully detected in U-cell lines and isolated tumor cells by gfp detection. Even though prME and ME pseudotypes had low infection rates, pseudotypes with ZIKV envelopes are promising candidates for the treatment of glioblastoma.
本研究旨在从 4 级多形性胶质母细胞瘤肿瘤中分离细胞,用于感染寨卡病毒(ZIKV)prME 或 ME 包膜 HIV-1 假型。从肿瘤组织中获得的细胞在含有极性和亲水表面的人脑脊液(hCSF)或 hCSF/DMEM 混合物中在细胞培养瓶中成功培养。分离的肿瘤细胞以及 U87、U138 和 U343 细胞均对 ZIKV 受体 Axl 和整合素 αvβ5 呈阳性。假型进入通过萤火虫荧光素酶或绿色荧光蛋白(gfp)的表达来检测。在 prME 和 ME 假型感染中,U 细胞系中的荧光素酶表达比背景高 2.5 至 3.5 对数,但仍比 VSV-G 假型对照低 2 对数。通过 gfp 检测,成功检测到 U 细胞系和分离的肿瘤细胞中的单细胞感染。尽管 prME 和 ME 假型的感染率较低,但带有 ZIKV 包膜的假型是治疗胶质母细胞瘤的有前途的候选物。