Departments of Sugery, University of Pittsburgh, Pittsburgh, PA 15213 USA.
Department of Urology, University of Pittsburgh, Pittsburgh, PA 15213 USA.
J Immunother Cancer. 2015 Mar 24;3:6. doi: 10.1186/s40425-015-0050-8. eCollection 2015.
BCG is a prototypal cancer immunotherapeutic factor currently approved of bladder cancer. In attempt to further enhance the effectiveness of immunotherapy of bladder cancer and, potentially, other malignancies, we evaluated the impact of BCG on local production of chemokines attracting the desirable effector CD8(+) T cells (CTLs) and undesirable myeloid-derived suppressor cell (MDSCs) and regulatory T(reg) cells, and the ability of bladder cancer tissues to attract CTLs.
Freshly resected bladder cancer tissues were either analyzed immediately or cultured ex vivo in the absence or presence of the tested factors. The expression of chemokine genes, secretion of chemokines and their local sources in freshly harvested and ex vivo-treated tumor explants were analyzed by quantitative PCR (Taqman), ELISAs and immunofluorescence/confocal microscopy. Migration of CTLs was evaluated ex vivo, using 24-transwell plates. Spearman correlation was used for correlative analysis, while paired Students T test or Wilcoxon was used for statistical analysis of the data.
Bladder cancer tissues spontaneously expressed high levels of the granulocyte/MDSC-attractant CXCL8 and Treg-attractant CCL22, but only marginal levels of the CTL-attracting chemokines: CCL5, CXCL9 and CXCL10. Baseline CXCL10 showed strong correlation with local expression of CTL markers. Unexpectedly, BCG selectively induced only the undesirable chemokines, CCL22 and CXCL8, but had only marginal impact on CXCL10 production. In sharp contrast, the combination of IFNα and a TLR3 ligand, poly-I:C (but not the combinations of BCG with IFNα or BCG with poly-I:C), induced high levels of intra-tumoral production of CXCL10 and promoted CTL attraction. The combination of BCG with IFNα + poly-I:C regimen did not show additional advantage.
The current data indicate that suboptimal ability of BCG to reprogram cancer-associated chemokine environment may be a factor limiting its therapeutic activity. Our observations that the combination of BCG with (or replacement by) IFNα and poly-I:C allows to reprogram bladder cancer tissues for enhanced CTL entry may provide for new methods of improving the effectiveness of immunotherapy of bladder cancer, helping to extend BCG applications to its more advanced forms, and, potentially, other diseases.
卡介苗（BCG）是一种典型的癌症免疫治疗因子，目前已被批准用于膀胱癌的治疗。为了进一步提高膀胱癌免疫治疗的效果，并可能提高其他恶性肿瘤的治疗效果，我们评估了卡介苗对吸引理想效应 CD8(+)T 细胞（CTL）和不理想的髓系来源抑制细胞（MDSC）和调节性 T 细胞（Treg）的局部产生的化学趋化因子的影响，以及膀胱癌组织吸引 CTL 的能力。
新鲜切除的膀胱癌组织立即进行分析，或在不存在或存在测试因子的情况下进行离体培养。通过定量 PCR（Taqman）、ELISA 和免疫荧光/共聚焦显微镜分析新鲜采集和离体处理的肿瘤标本中趋化因子基因的表达、趋化因子的分泌及其局部来源。使用 24 孔迁移小室体外评估 CTL 的迁移。采用 Spearman 相关分析进行相关性分析，采用配对学生 t 检验或 Wilcoxon 检验进行数据的统计学分析。
膀胱癌组织自发表达高水平的粒细胞/MDSC 趋化因子 CXCL8 和 Treg 趋化因子 CCL22，但仅表达低水平的 CTL 趋化因子：CCL5、CXCL9 和 CXCL10。基线 CXCL10 与局部 CTL 标志物的表达呈强相关性。出乎意料的是，BCG 仅选择性地诱导不理想的趋化因子 CCL22 和 CXCL8，但对 CXCL10 的产生只有轻微影响。相比之下，IFNα 和 TLR3 配体聚肌苷酸（poly-I:C）的组合（而不是 BCG 与 IFNα 或 BCG 与 poly-I:C 的组合）诱导了高水平的肿瘤内 CXCL10 产生，并促进了 CTL 的趋化。BCG 与 IFNα+poly-I:C 方案联合使用并未显示出额外的优势。
目前的数据表明，BCG 重新编程癌症相关趋化因子环境的能力不足可能是限制其治疗活性的一个因素。我们的观察结果表明，BCG 与（或替代）IFNα 和 poly-I:C 的组合可重新编程膀胱癌组织以增强 CTL 进入，这可能为提高膀胱癌免疫治疗效果提供新的方法，有助于将 BCG 的应用扩展到其更高级的形式，并可能扩展到其他疾病。
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