Bacillus Calmette-Guerin cell-wall skeleton enhances the killing activity of cytotoxic lymphocyte-activated human dendritic cells transduced with the prostate-specific antigen gene
Bacillus Calmette-Guerin cell-wall skeleton enhances the killing activity of cytotoxic lymphocyte-activated human dendritic cells transduced with the prostate-specific antigen gene
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DOI：
10.1111/j.1464-410x.2009.08703.x
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发表时间：
2009-12-01
影响因子：
4.5
通讯作者：
Hara, Isao
中科院分区：
文献类型：
--
作者：
Fujii, Reona;Iwahashi, Makoto;Hara, Isao
目的探讨前列腺特异性抗原（PSA）基因转导的树突状细胞（DC）能否诱导PSA特异性细胞毒性淋巴细胞（CTL）杀伤前列腺癌细胞，以及卡介苗（BCG）细胞壁骨架（CWS）是否能促进DC-PSA的成熟和随后诱导的PSA特异性CTL的杀伤活性。（AxCA-PSA），使用粘粒末端蛋白复合物方法。用AxCA-PSA离心法感染DC。使用51 Cr-释放测定评估CTL裂解表达PSA的靶细胞（即PSA阳性前列腺癌细胞系LNCap和表达PSA的PSA转导的自体植物血凝素（PHA）母细胞）的能力。流式细胞术检测BCG CWS刺激DC PSA成熟的情况。用~（51）Cr释放法检测BCG-CWS对LNCaP的杀伤活性，结果表明BCG-CWS诱导的PSA特异性CTL对LNCaP的杀伤活性为85%（效靶比E：T = 50：1）。然而，对PSA阴性细胞的细胞毒活性非常低。抗CD 8和抗主要组织相容性（MHC）I类抗体阻断PSA特异性细胞毒性。PSA特异性杀伤对表达PSA的自体PHA母细胞是可重复的，独立于人白细胞抗原单倍型。DC-PSA与BCG-CWS联合应用可显著增强PSA对表达PSA的PHA细胞的特异性杀伤作用（E：T = 50：1时，增强15-30%）。结论DC-PSA可诱导MHC I类限制性PSA特异性CD 8 + CTL反应，BCG-CWS成熟的DC-PSA可增强PSA特异性细胞毒作用。DC-PSA联合BCG-CWS治疗晚期前列腺癌可能是一种有效的方法。
OBJECTIVETo determine whether dendritic cells (DC) transduced with the prostate-specific antigen (PSA) gene can induce PSA-specific cytotoxic lymphocytes (CTL) against prostate cancer cells, and whether bacillus Calmette-Guerin (BCG) cell-wall skeleton (CWS) can enhance the maturation of DC-PSA and the killing activity of subsequently induced PSA-specific CTL.MATERIALS AND METHODSWe generated an adenovirus encoding the PSA gene (AxCA-PSA) using the cosmid-terminal protein complex method. DC were infected with AxCA-PSA using the centrifugal method. The ability of CTL to lyse target cells expressing PSA, i.e the PSA-positive prostate cancer cell line, LNCap, and PSA-transduced autologous phytohaemagglutinin (PHA) blasts expressing PSA, was assessed using the 51Cr-release assay. The maturation of DC-PSA stimulated by BCG-CWS was assayed by flow cytometry. The cytotoxic activity enhanced by BCG-CWS was assessed by the 51Cr-release assay.RESULTSDC-PSA induced PSA-specific CTL with 85% cytotoxic activity against LNCaP (effector: target ratio, E:T, of 50:1). However, the cytotoxic activity against PSA-negative cells was very low. Anti-CD8 and anti-major histocompatibility (MHC) class I antibodies blocked PSA-specific cytotoxicity. The PSA-specific killing was reproducible against autologous PHA blast cells expressing PSA, independently of human leukocyte antigen haplotype. Furthermore, the combination of DC-PSA with BCG-CWS remarkably enhanced the PSA-specific cytotoxicity against PHA blasts expressing PSA (15-30% at an E:T ratio of 50:1).CONCLUSIONThese findings suggest that DC-PSA can induce MHC class I-restricted PSA-specific CD8+ CTL responses and that DC-PSA matured by BCG-CWS enhance PSA-specific cytotoxicity. The combination of DC-PSA with BCG-CWS might be a useful approach for treating advanced prostate cancer.