Lymphocyte motility is governed by a complex array of mechanisms, and highly dependent on external microenvironmental cues. Tertiary lymphoid sites in particular have unique physical structure such as collagen fiber alignment, due to matrix deposition and remodeling. Three dimensional studies of human lymphocytes in such environments are lacking. We hypothesized that aligned collagenous environment modulates CD8+ T cells motility. We encapsulated activated CD8+ T cells in collagen hydrogels of distinct fiber alignment, a characteristic of tumor microenvironments. We found that human CD8+ T cells move faster and more persistently in aligned collagen fibers compared with nonaligned collagen fibers. Moreover, CD8+ T cells move along the axis of collagen alignment. We showed that myosin light chain kinase (MLCK) inhibition could nullify the effect of aligned collagen on CD8+ T cell motility patterns by decreasing T cell turning in unaligned collagen fiber gels. Finally, as an example of a tertiary lymphoid site, we found that xenograft prostate tumors exhibit highly aligned collagen fibers. We observed CD8+ T cells alongside aligned collagen fibers, and found that they are mostly concentrated in the periphery of tumors. Overall, using an in vitro controlled hydrogel system, we show that collagen fiber organization modulates CD8+ T cells movement via MLCK activation thus providing basis for future studies into relevant therapeutics.

中文翻译：

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胶原纤维结构引导细胞毒性 T 淋巴细胞的 3D 运动。

淋巴细胞运动受到一系列复杂机制的控制，并且高度依赖于外部微环境线索。由于基质沉积和重塑，第三淋巴部位尤其具有独特的物理结构，例如胶原纤维排列。缺乏对此类环境中人类淋巴细胞的三维研究。我们假设对齐的胶原环境调节 CD8+ T 细胞的运动性。我们将活化的 CD8+ T 细胞封装在具有不同纤维排列的胶原水凝胶中，这是肿瘤微环境的一个特征。我们发现，与未排列的胶原纤维相比，人类 CD8+ T 细胞在排列的胶原纤维中移动得更快、更持久。此外，CD8+ T 细胞沿着胶原排列轴移动。我们发现，肌球蛋白轻链激酶 (MLCK) 抑制可以通过减少未排列的胶原纤维凝胶中 T 细胞的转动来消除排列的胶原蛋白对 CD8+ T 细胞运动模式的影响。最后，作为三级淋巴部位的一个例子，我们发现异种移植前列腺肿瘤表现出高度排列的胶原纤维。我们观察到 CD8+ T 细胞与排列的胶原纤维一起，发现它们主要集中在肿瘤的周围。总体而言，使用体外控制的水凝胶系统，我们表明胶原纤维组织通过 MLCK 激活调节 CD8+ T 细胞的运动，从而为未来相关治疗的研究提供基础。