Division of Infectious Diseases, Los Angeles Biomedical Research Institute, Harbor-University of California at Los Angeles (UCLA) Medical Center, 1124 West Carson Street, Torrance, CA 90502, USA.
J Infect Dis. 2010 Jun 1;201(11):1708-17. doi: 10.1086/652496.
Invasive fungal infections cause considerable morbidity and mortality in neutropenic patients. White blood cell transfusions are a promising treatment for such infections, but technical barriers have prevented their widespread use.
To recapitulate white blood cell transfusions, we are developing a cell-based immunotherapy using a phagocytic cell line, HL-60. We sought to stably transfect HL-60 cells with a suicide trap (herpes simplex virus thymidine kinase), to enable purging of the cells when desired, and a bioluminescence marker, to track the cells in vivo in mice.
Transfection was stable despite 20 months of continuous culture or storage in liquid nitrogen. Activation of these transfected cells with retinoic acid and dimethyl sulfamethoxazole enhanced their microbicidal effects. Activated transfected killer (ATAK) cells were completely eliminated after exposure to ganciclovir, confirming function of the suicide trap. ATAK cells improved the survival of neutropenic mice with lethal disseminated candidiasis and inhalational aspergillosis. Bioluminescence and histopathologic analysis confirmed that the cells were purged from surviving mice after ganciclovir treatment. Comprehensive necropsy, histopathology, and metabolomic analysis revealed no toxicity of the cells.
These results lay the groundwork for continued translational development of this promising, novel technology for the treatment of refractory infections in neutropenic hosts.
侵袭性真菌感染会导致中性粒细胞减少症患者产生相当大的发病率和死亡率。白细胞输注是治疗此类感染的一种很有前途的方法，但技术障碍阻止了它们的广泛应用。
为了重现白细胞输注，我们正在使用吞噬细胞系 HL-60 开发一种基于细胞的免疫疗法。我们试图用自杀陷阱（单纯疱疹病毒胸苷激酶）稳定转染 HL-60 细胞，以便在需要时清除细胞，并用生物发光标记物在小鼠体内跟踪细胞。
尽管经过 20 个月的连续培养或在液氮中储存，转染仍很稳定。用维甲酸和二甲亚砜激活这些转染细胞增强了它们的杀菌作用。用更昔洛韦处理后，激活的转染杀伤（ATAK）细胞完全被清除，证实了自杀陷阱的功能。ATAK 细胞改善了患有致死性播散性念珠菌病和吸入性曲霉菌病的中性粒细胞减少症小鼠的存活率。生物发光和组织病理学分析证实，在用更昔洛韦治疗后，细胞从存活的小鼠中被清除。全面尸检、组织病理学和代谢组学分析显示细胞没有毒性。
这些结果为继续开发这种有前途的新型技术用于治疗中性粒细胞减少症宿主的难治性感染奠定了基础。
