Chaudhuri Suhnrita, Singh Manoj K, Bhattacharya Debanjan, Datta Ankur, Hazra Iman, Mondal Somnath, Faruk Sk Md Omar, Ronsard Larance, Ghosh Tushar K, Chaudhuri Swapna
Department of Laboratory Medicine, Cellular and Molecular Immunology Lab, School of Tropical Medicine, Kolkata, West Bengal 700073, India.
Department of Physiology, University of Calcutta, Kolkata, West Bengal 700009, India.
J Cell Physiol. 2018 Feb;233(2):759-770. doi: 10.1002/jcp.26047. Epub 2017 Jul 11.
Malignant glioma is the most fatal of astrocytic lineage tumors despite therapeutic advances. Onset and progression of gliomas is accompanied by severe debilitation of T-cell defense and T-cell survival. One of the chief contributors to T-cell survival downstream of activation is the PI3K-AKT pathway. Our prior studies showed that the novel immunotherapeutic molecule T11-target structure (T11TS) blocks T-cell apoptosis in glioma. We also showed activation of immunological synapse components and calcineurin-NFAT pathway following T11TS immunotherapy of glioma-bearing rats. This lead to investigations whether such T-cell activation upon T11TS therapy translates into activation of downstream PI3K/AKT signals which may be related to observed blockade of T-cell apoptosis. For the purpose, we assessed by flowcytometry and immunoblotting, expressions of PI3K, PDK1, AKT, p-AKT, and PTEN in splenic T-cells of normal, experimentally-induced glioma-bearing rats and glioma-bearing rats receiving first, second and third doses of T11TS. We also determined comparative nuclear translocation of NF-κB across groups. We found significant increases in T-cell expressions of PDK1, PI3K, and p-AKT in T11TS-treated animal groups compared to sharp downregulations in glioma. AKT levels remained unchanged across groups. PTEN levels declined sharply after T11TS immunotherapy. T11TS also caused enhanced NF-κB translocation to the T-cell nucleus compared to glioma group. Results showed heightened activation of the PI3K-AKT pathway in glioma-bearing rats following T11TS immunotherapy. These results illustrate the novel role of T11TS immunotherapy in ameliorating the PI3K pathway in T-cells in glioma-bearing animals to enhance T-cell survival, according greater defense against glioma. The study thus has far-reaching clinical outcomes.
尽管治疗取得了进展，但恶性胶质瘤仍是星形细胞谱系肿瘤中最致命的。胶质瘤的发生和进展伴随着T细胞防御和T细胞存活的严重衰弱。激活下游T细胞存活的主要因素之一是PI3K-AKT通路。我们之前的研究表明，新型免疫治疗分子T11-靶结构（T11TS）可阻断胶质瘤中的T细胞凋亡。我们还显示，对荷瘤大鼠进行T11TS免疫治疗后，免疫突触成分和钙调神经磷酸酶-NFAT通路被激活。这引发了对于T11TS治疗后这种T细胞激活是否转化为下游PI3K/AKT信号激活的研究，而这可能与观察到的T细胞凋亡阻断有关。为此，我们通过流式细胞术和免疫印迹法评估了正常大鼠、实验性诱导的荷瘤大鼠以及接受第一、第二和第三剂T11TS的荷瘤大鼠脾T细胞中PI3K、PDK1、AKT、p-AKT和PTEN的表达。我们还确定了各组间NF-κB的相对核转位情况。我们发现，与胶质瘤组中这些蛋白的急剧下调相比，T11TS治疗的动物组中T细胞PDK1、PI3K和p-AKT的表达显著增加。各组间AKT水平保持不变。T11TS免疫治疗后PTEN水平急剧下降。与胶质瘤组相比，T11TS还导致NF-κB向T细胞核的转位增强。结果显示，T11TS免疫治疗后荷瘤大鼠的PI3K-AKT通路激活增强。这些结果说明了T11TS免疫治疗在改善荷瘤动物T细胞中PI3K通路以提高T细胞存活方面的新作用，从而增强对胶质瘤的防御能力。因此，该研究具有深远的临床意义。
