A novel approach to analyze the immunological responses in human colorectal carcinoma

1：大肠癌组织巨噬细胞、树突状细胞和淋巴细胞的流式细胞分析(1)我们通过酶切和Ficoll-Paque分离从大肠癌肿瘤和正常大肠癌组织中分离出单个核细胞。(2)流式细胞术分析显示CRC浸润缘的cd14阳性巨噬细胞多于正常粘膜组织。(3)与外周血淋巴细胞（PBL）相比，来自结直肠癌的T细胞以cd4为主，其中90%以上为记忆T淋巴细胞。(4)与PBL相比，结直肠癌中分离的T细胞以th1为主，且ⅰ型趋化因子CXCR3和CCR5阳性。(5)混合淋巴细胞反应实验表明，位于CRC的cd14阳性巨噬细胞具有刺激T细胞反应的能力。2：免疫组化分析(1)肿瘤单个核细胞表面抗原的免疫组化表达与流式细胞分析数据一致。(2) CRC浸润缘巨噬细胞Fas配体（FasL）阳性。fasl阳性巨噬细胞周围凋亡癌细胞的数量与fasl阳性巨噬细胞的数量相关。(3) CD4+ T细胞和CD8+ T细胞CXCR3和CCR5阳性。CD8+ T淋巴细胞表达RANITES，癌细胞和巨噬细胞表达IP- 10。

1: Flowcytometric analyses of macrophages, dendritic cells, and lymphocytes isolated from colorectal cancer(CRC) tissue(1) We isolated mononuclear cells from colorectal tumors and normal colorectal tissues by enzymatic digestion and Ficoll-Paque separation.(2) Flowcytometric analyses showed more CD14-positive macrophages in the invasive margin of CRC than in normal mucosal fissue.(3) T cells derived from CRC were CD4-dominant compared to peripheral blood lymphocytes (PBL) and more than 90 % of those were memory T lymphocytes.(4) The T cells isolated from CRC were Th 1-dominant compared to PBL and positive for type l chemokines, CXCR3 and CCR5.(5) A mixed lymphocyte reaction experiment demonstrated that CD14-positive macrophages located in CRC possessed the ability to stimulate T cell responses.2: lmmunohistochemlcal analyses(1) lmmunohistochemical expression of the surface antigens in mononuclear cells in tumor was consistent with the data of the flowcytometric analyses.(2) Macrophages along the invasive margin of CRC were Fas ligand (FasL)-positive. The number of apoptotic cancer cells around the FasL-positive macrophages was correlated with the number of the FasL-positive macrophages.(3) CD4+ T cells and CD8+ T cells were positive for CXCR3 and CCR5. CD8+ T lymphocytes expressed RANITES, and cancer cells and macrophages expressed IP- 10.