Ali S A, Rees R C, Anderson D Q, Reed M W, Goepel J R, Brown N J
Department of Life Sciences, Nottingham Trent University, Clifton Lane, Nottingham, NG11 8NS.
Br J Cancer. 2000 Oct;83(8):1061-8. doi: 10.1054/bjoc.2000.1403.
RENCA-IL-2 (Murine Renal Cell Carcinoma transfected with murine IL-2 gene) cells were rejected by immunocompetent (but not T-cell deficient) Balb/c mice, which developed 'immunity' to subsequent parental RENCA tumour cell challenge. Splenocytes adoptively transferred this immunity. CD4(+)and CD8(+)T-lymphocytes prepared from the spleens of 'tumour immune' mice were evaluated for their ability to traffic into the tumour environment using an in vivo model that enables visualization of events within the microvasculature. RENCA cells were implanted into the mouse cremaster muscle and the trafficking of syngeneic lymphocyte subpopulations, derived from naive and 'immune' animals, into both the RENCA tumour and the surrounding normal cremaster muscle microcirculation was measured by in vivo microscopy. Fluorescently labelled CD4(+)and CD8(+)T lymphocytes taken from the spleens of naive mice or mice previously immunized with RENCA-IL-2 were injected systemically into tumour-bearer mice. Naive effector cells migrated to, and flowed through both the tumour and the normal microcirculation, with negligible adhesion. However we observed the selective recruitment, localization and arrest of immune CD4(+)and CD8(+)T lymphocytes (P< 0.05) into the tumour microcirculation, and in some instances the subsequent extravasation of cells into the tumour interstitium. Lymphocyte rolling by 'immune' CD4(+)and CD8(+)T-cells in the tumour microcirculation was greatly reduced, suggesting impaired adhesion molecule expression on the tumour endothelium. This study clearly demonstrates, by direct in vivo microscopy assessment, the localization of effector cells, CD4(+)and CD8(+)lymphocytes into tumours.
RENCA-IL-2（转染了小鼠白细胞介素-2基因的小鼠肾细胞癌）细胞被具有免疫能力（但T细胞缺陷的小鼠除外）的Balb/c小鼠排斥，这些小鼠对随后的亲本RENCA肿瘤细胞攻击产生了“免疫力”。脾细胞可传递这种免疫力。使用能够可视化微脉管系统内事件的体内模型，评估了从“肿瘤免疫”小鼠脾脏制备的CD4(+)和CD8(+)T淋巴细胞进入肿瘤环境的能力。将RENCA细胞植入小鼠提睾肌，并通过体内显微镜测量来自未免疫和“免疫”动物的同基因淋巴细胞亚群进入RENCA肿瘤和周围正常提睾肌微循环的情况。将从未免疫小鼠或先前用RENCA-IL-2免疫的小鼠脾脏中获取的荧光标记CD4(+)和CD8(+)T淋巴细胞全身注射到荷瘤小鼠体内。未免疫的效应细胞迁移到肿瘤和正常微循环，并在其中流动，黏附作用可忽略不计。然而，我们观察到免疫CD4(+)和CD8(+)T淋巴细胞选择性募集、定位并停滞在肿瘤微循环中（P<0.05），在某些情况下，随后细胞外渗进入肿瘤间质。“免疫”CD4(+)和CD8(+)T细胞在肿瘤微循环中的滚动明显减少，提示肿瘤内皮细胞上黏附分子表达受损。这项研究通过直接的体内显微镜评估清楚地证明了效应细胞CD4(+)和CD8(+)淋巴细胞在肿瘤中的定位。
