Department of Immunology, School of Basic Medicine, Fourth Military Medical University, Xi'an, China.
Military Medical Innovation Center, Fourth Military Medical University, Xi'an, China.
Int Immunopharmacol. 2022 Feb;103:108505. doi: 10.1016/j.intimp.2021.108505. Epub 2022 Jan 5.
Taxol-resistance gene 1 (TXR1) is closely correlated with the paclitaxel resistance in the cancer chemotherapy. However, due to the lack of monoclonal antibodies (mAbs) with strong specificity and high sensitivity, little information is found about TXR1 target-related tumor therapy.
We developed an TXR1 recombinant DNA vaccine by inserting TXR1 DNA sequence into lysosome-associated membrane protein 1 (LAMP1). Adaptive immune responses were assessed by indirect enzyme-linked immunosorbent assay (ELISA), Enzyme-linked immunospot test (ELISpot), and cytotoxic T-lymphocyte (CTL) cytotoxicity.
The pGEX4T-1-TXR1 reconstructed prokaryotic expression plasmid was constructed for producing high-purity TXR1 protein. Subsequently, a total of four mAbs for TXR1 and two PcAbs were successfully constructed and identified. We further found that TXR1 was highly expressed in breast cancer tissue than normal controls. Therefore, we constructed four tumor vectors, pVAX1-LAMP/TXR1, pVAX1-LAMP, pVAX1/TXR1 and pVAX1, for immunization. After three times of immunization, ELISpot data showed that single peptide 6,9,11 could stimulate T cells secreting IFN-γ in pVAX1-LAMP/TXR1 group. Moreover, the number of specific T cells and immune response effects significantly increased comparing to the pVAX1-LAMP control group. In addition, cytotoxicity showed that when the effect to target ratio was 40:l the killing effect of pVAX1-LAMP/TXR1 group was significantly higher than the pVAX1-TXR1 group.
Our results provides new evidence for the TXR1 related tumor immunology and aids the early prevention of cancer.
紫杉醇耐药基因 1（TXR1）与癌症化疗中的紫杉醇耐药密切相关。然而，由于缺乏特异性强、灵敏度高的单克隆抗体（mAbs），关于 TXR1 靶相关肿瘤治疗的信息很少。
我们通过将 TXR1 DNA 序列插入溶酶体相关膜蛋白 1（LAMP1）构建了 TXR1 重组 DNA 疫苗。通过间接酶联免疫吸附试验（ELISA）、酶联免疫斑点试验（ELISpot）和细胞毒性 T 淋巴细胞（CTL）杀伤活性评估适应性免疫反应。
构建了 pGEX4T-1-TXR1 重组原核表达质粒，用于生产高纯度的 TXR1 蛋白。随后，成功构建并鉴定了针对 TXR1 的 4 种 mAbs 和 2 种 PcAbs。我们进一步发现，TXR1 在乳腺癌组织中的表达明显高于正常对照。因此，我们构建了四个肿瘤载体，pVAX1-LAMP/TXR1、pVAX1-LAMP、pVAX1/TXR1 和 pVAX1，用于免疫。三次免疫后，ELISpot 数据显示，单肽 6、9、11 可刺激 pVAX1-LAMP/TXR1 组 T 细胞分泌 IFN-γ。此外，与 pVAX1-LAMP 对照组相比，特异性 T 细胞数量和免疫反应效果显著增加。此外，细胞毒性显示当效靶比为 40:1 时，pVAX1-LAMP/TXR1 组的杀伤效果明显高于 pVAX1-TXR1 组。
我们的结果为 TXR1 相关肿瘤免疫学提供了新的证据，并有助于癌症的早期预防。
