Institute of Translational Medicine, Department of Molecular and Clinical Cancer Medicine, University of Liverpool, 2nd Floor Sherrington Building, Ashton Street, Liverpool, L69 3GE, UK.
National Research Centre, 12662, Dokki, Giza, Egypt.
Sci Rep. 2020 Jan 15;10(1):342. doi: 10.1038/s41598-019-57293-y.
In carcinomas, the nature of CD40 ligand shapes the outcome of CD40 ligation. To date, the consequences of membrane-bound CD40L (mCD40L) on its immune-stimulatory function are unknown. Here, we examined the impact of mCD40L versus soluble CD40L (sCD40L) on T24 bladder carcinoma gene expression profiling. Of 410 differentially expressed genes, 286 were upregulated and 124 downregulated by mCD40L versus sCD40L. Gene ontology enrichment analysis revealed immune-stimulatory function as the most significant enriched biological process affected by upregulated transcripts, while those downregulated were critical for cell growth and division. Furthermore, immature dendritic cells (iDC) responded to mCD40L with enhanced maturation and activation over sCD40L evidenced by higher expression levels of CD83, CD86, HLA-DR and CD54, increased secretion of IL12 and IL10 and higher tumour-antigen (TA) uptake capacity. Furthermore, autologus CD3+ T cells responded to TA-loaded mCD40L-activated DC with increased proliferation and cytotoxic response (CD107a and IFN-γ-producing CD3+ CD8+ T cells) to the tumour-loaded autologous PBMCs compared to sCD40L. Thus, these data indicate that mCD40L enhances the immunostimulatory capacity over sCD40L. Furthermore, the ability of mCD40L to also directly induce cell death in CD40-expressing carcinomas, subsequently releasing tumour-specific antigens into the tumour microenvironment highlights the potential for mCD40L as a multi-faceted anti-cancer immunotherapeutic.
在癌中，CD40 配体的性质决定了 CD40 交联的结果。迄今为止，膜结合 CD40L（mCD40L）对其免疫刺激功能的后果尚不清楚。在这里，我们研究了 mCD40L 与可溶性 CD40L（sCD40L）对 T24 膀胱癌基因表达谱的影响。在 410 个差异表达基因中，mCD40L 相对于 sCD40L 上调了 286 个，下调了 124 个。基因本体富集分析显示，免疫刺激功能是受上调转录物影响的最显著的富集生物学过程，而下调的转录物对细胞生长和分裂至关重要。此外，未成熟树突状细胞（iDC）对 mCD40L 的反应是通过增强成熟和激活来实现的，这一点可以从 CD83、CD86、HLA-DR 和 CD54 的更高表达水平、IL12 和 IL10 的更高分泌以及更高的肿瘤抗原（TA）摄取能力得到证明。此外，自体 CD3+T 细胞对负载 TA 的 mCD40L 激活的 DC 的反应是增加增殖和细胞毒性反应（CD107a 和产生 IFN-γ的 CD3+CD8+T 细胞）对负载肿瘤的自体 PBMCs，与 sCD40L 相比。因此，这些数据表明 mCD40L 增强了相对于 sCD40L 的免疫刺激能力。此外，mCD40L 还能直接诱导表达 CD40 的癌死亡，随后将肿瘤特异性抗原释放到肿瘤微环境中，这突出了 mCD40L 作为一种多方面的抗癌免疫治疗的潜力。
