Department of Medicine, Stanford University, 269 Campus Drive, 94305 Stanford, CA, USA.
J Transl Med. 2013 Oct 2;11:242. doi: 10.1186/1479-5876-11-242.
Dendritic cells (DCs) are important mediators of anti-tumor immune responses. We hypothesized that an in-depth analysis of dendritic cells and their spatial relationships to each other as well as to other immune cells within tumor draining lymph nodes (TDLNs) could provide a better understanding of immune function and dysregulation in cancer.
We analyzed immune cells within TDLNs from 59 breast cancer patients with at least 5 years of clinical follow-up using immunohistochemical staining with a novel quantitative image analysis system. We developed algorithms to analyze spatial distribution patterns of immune cells in cancer versus healthy intra-mammary lymph nodes (HLNs) to derive information about possible mechanisms underlying immune-dysregulation in breast cancer. We used the non-parametric Mann-Whitney test for inter-group comparisons, Wilcoxon Matched-Pairs Signed Ranks test for intra-group comparisons and log-rank (Mantel-Cox) test for Kaplan Maier analyses.
Degree of clustering of DCs (in terms of spatial proximity of the cells to each other) was reduced in TDLNs compared to HLNs. While there were more numerous DC clusters in TDLNs compared to HLNs,DC clusters within TDLNs tended to have fewer member DCs and also consisted of fewer cells displaying the DC maturity marker CD83. The average number of T cells within a standardized radius of a clustered DC was increased compared to that of an unclustered DC, suggesting that DC clustering was associated with T cell interaction. Furthermore, the number of T cells within the radius of a clustered DC was reduced in tumor-positive TDLNs compared to HLNs. Importantly, clinical outcome analysis revealed that DC clustering in tumor-positive TDLNs correlated with the duration of disease-free survival in breast cancer patients.
These findings are the first to describe the spatial organization of DCs within TDLNs and their association with survival outcome. In addition, we characterized specific changes in number, size, maturity, and T cell co-localization of such clusters. Strategies to enhance DC function in-vivo, including maturation and clustering, may provide additional tools for developing more efficacious DC cancer vaccines.
树突状细胞（DCs）是抗肿瘤免疫反应的重要介质。我们假设，深入分析树突状细胞及其与肿瘤引流淋巴结（TDLNs）中其他免疫细胞之间的空间关系，可以更好地了解癌症中的免疫功能和失调。
我们使用一种新的定量图像分析系统，对 59 例至少有 5 年临床随访的乳腺癌患者的 TDLNs 中的免疫细胞进行免疫组织化学染色。我们开发了算法来分析癌症与健康乳腺内淋巴结（HLNs）中免疫细胞的空间分布模式，以获取有关乳腺癌中免疫失调潜在机制的信息。我们使用非参数 Mann-Whitney 检验进行组间比较，Wilcoxon 配对符号秩检验进行组内比较，对数秩（Mantel-Cox）检验进行 Kaplan-Meier 分析。
与 HLNs 相比，TDLNs 中的 DC （细胞之间空间接近程度）聚类程度降低。虽然 TDLNs 中的 DC 簇比 HLNs 中的多，但 TDLNs 中的 DC 簇中 DC 成员较少，并且显示 DC 成熟标志物 CD83 的细胞也较少。与未聚类的 DC 相比，在聚类 DC 的标准化半径内的 T 细胞数量增加，这表明 DC 聚类与 T 细胞相互作用有关。此外，在聚类 DC 的半径内的 T 细胞数量在肿瘤阳性 TDLNs 中比 HLNs 中减少。重要的是，临床结果分析表明，肿瘤阳性 TDLNs 中的 DC 聚类与乳腺癌患者无病生存时间相关。
这些发现是首次描述 TDLNs 中 DC 的空间组织及其与生存结果的关联。此外，我们还描述了这种簇的数量、大小、成熟度和 T 细胞共定位的特定变化。增强体内 DC 功能的策略，包括成熟和聚类，可能为开发更有效的 DC 癌症疫苗提供额外的工具。
