College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, Zhejiang, China.
PLoS One. 2013 Sep 3;8(9):e74363. doi: 10.1371/journal.pone.0074363. eCollection 2013.
Pinelliapedatisecta agglutinin (PPA) has previously been used in labeling fractions of myeloid leukemia cells in our laboratory. We report here that a bacterial expressed recombinant PPA domain b tagged with soluble coxsackie and adenovirus receptor (sCAR-PPAb) preferentially recognized drug resistant cancer cells K562/ADR and H460/5Fu, as compared to their parental cell lines. Pretreatment of K562/ADR cells with sCAR-PPAb significantly enhanced phagocytosis of K562/ADR by macrophages in vivo. Meanwhile, in a K562/ADR xenograft model, intratumoral injection of sCAR-PPAb induced macrophage infiltration and phagocytosis. Furthermore, immunoprecipitation, mass spectrometry and Western blot identified the membrane target of PPA on K562/ADR as sarcolemmal membrane associated protein (SLMAP). An antibody against SLMAP significantly promoted the phagocytosis of K562/ADR by macrophages in vitro. These findings suggest that PPA not only could be developed into a novel agent that can detect drug resistant cancer cells and predict chemotherapy outcome, but also it has potential value in immunotherapy against drug resistant cancer cells through inducing the tumoricidal activity of macrophages.
被孢霉凝集素(PPA)先前已被用于标记本实验室髓系白血病细胞的馏分。我们在此报告,一种带有可溶性柯萨奇病毒和腺病毒受体(sCAR-PPAb)标签的细菌表达的重组 PPA 结构域 b 优先识别耐药性癌细胞 K562/ADR 和 H460/5Fu,与它们的亲本细胞系相比。用 sCAR-PPAb 预处理 K562/ADR 细胞,可显著增强巨噬细胞在体内对 K562/ADR 的吞噬作用。同时,在 K562/ADR 异种移植模型中,肿瘤内注射 sCAR-PPAb 诱导巨噬细胞浸润和吞噬作用。此外,免疫沉淀、质谱和 Western blot 鉴定出 K562/ADR 上 PPA 的膜靶标为肌膜相关蛋白(SLMAP)。针对 SLLMAP 的抗体显著促进了巨噬细胞对 K562/ADR 的吞噬作用。这些发现表明,PPA 不仅可以开发成一种新型试剂,用于检测耐药性癌细胞并预测化疗结果,而且通过诱导巨噬细胞的杀瘤活性,它在针对耐药性癌细胞的免疫治疗中具有潜在价值。