In this research, we studied the therapeutic efficacy of a newly designed fusion protein containing Endoglin single-chain variable fragment and IP10 (Endoglin-scFv/IP10), together with our recently generated TRP2-specific CD8⁺ CD28⁺ CTLs (CD8⁺ CD28⁺ CTLs) in controlling melanoma growth in mice. The recombinant Endoglin-scFv/IP10 was expressed in E. coli, purified by affinity chromatography, and characterized in vitro for its chemotactic movement and immunoreactivity with endoglin-expressing cells. In vivo, melanoma xenografts were established in mice (C57BL/6) using B16F10 cells. After that, mice were treated with intravenous injections of vehicle (PBS), Endoglin-scFv/IP10 alone, CD8⁺ CD28⁺ CTLs alone, or Endoglin-scFv/IP10⁺ CD8⁺ CD28⁺ CTLs. The therapeutic efficacy was assessed by monitoring tumor growth, mouse survival and cellular biomarkers. Endoglin-scFv/IP10 fusion protein combined with CD8⁺ CD28⁺ CTLs observed a reduction in tumor growth, resulting in improved survival. On the cellular level, the combination treatment dramatically reduced the number of systemic and tumor associated myeloid-derived suppressor cells or regulatory T cells, increased tumor-responsive interferon-γ-producing lymphocytes and tumor-associated CD8⁺ CXCR3⁺ T cells, and inhibited proliferation and angiogenesis but stimulated apoptosis within melanoma tissue. This study demonstrates the therapeutic potential of Endoglin-scFv/IP10 fusion protein in combination with CD8⁺ CD28⁺ CTLs in melanoma treatment.

中文翻译：

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重组内皮糖蛋白单链可变片段/诱导蛋白10融合蛋白有效增强了过继转移的TRP2特异性CD8 + CD28 +细胞毒性T淋巴细胞在小鼠中的抗肿瘤功效。

在这项研究中，我们研究了一种新设计的包含Endoglin单链可变片段和IP10（Endoglin-scFv / IP10）融合蛋白，以及我们最近生成的TRP2特异性CD8 ⁺ CD28 ⁺ CTL（CD8 ⁺ CD28 ⁺ CTLs）来控制小鼠黑色素瘤的生长。重组Endoglin-scFv / IP10在大肠杆菌中表达，通过亲和层析纯化，并在体外表征其趋化运动和与表达Endglin的细胞的免疫反应性。体内，使用B16F10细胞在小鼠（C57BL / 6）中建立了黑色素瘤异种移植物。此后，对小鼠进行静脉内注射溶媒（PBS），单独使用Endoglin-scFv / IP10，单独使用CD8 ⁺ CD28 ⁺ CTL或使用Endoglin-scFv / IP10 ⁺ CD8 ⁺ CD28 ⁺ CTL治疗。通过监测肿瘤生长，小鼠存活率和细胞生物标志物评估治疗效果。Endoglin-scFv / IP10融合蛋白与CD8 ⁺ CD28 ^+结合CTLs观察到肿瘤生长减少，从而提高了生存率。在细胞水平上，联合治疗显着减少了全身和肿瘤相关的髓样抑制细胞或调节性T细胞的数量，增加了肿瘤反应性干扰素γ产生的淋巴细胞和肿瘤相关的CD8 ⁺ CXCR3 ⁺ T细胞的数量，并抑制了增殖和血管生成，但刺激黑色素瘤组织内的细胞凋亡。这项研究证明了内皮糖蛋白-scFv / IP10融合蛋白与CD8 ⁺ CD28 ⁺ CTL结合在黑素瘤治疗中的治疗潜力。