Tsujihashi H, Matsuda H, Uejima S, Akiyama T, Kurita T
Department of Urology, Kinki University School of Medicine, Osaka, Japan.
J Urol. 1988 Oct;140(4):890-4. doi: 10.1016/s0022-5347(17)41851-9.
Tissue infiltrating lymphocytes (TIL) in bladder tumors have been assumed to be an expression of local host resistance against the tumor. We investigated the functional activity of TIL compared to peripheral blood lymphocytes (PBL). Isolation of TIL was performed using the enzyme cocktail treatment with Ficoll-Hypaque discontinuous gradient centrifugation. Analysis of lymphocyte subsets by flow cytometry demonstrated Leu 4, 43.6% (T cells); Leu 10, 10.5% (B cells) and Leu 7, 13.1% (natural killer (NK) cells) in TIL. The cytotoxic activity of TIL and PBL was tested in a four hour 51Cr-release assay. Myeloid K562 cells (NK sensitive), HT 1197 (bladder tumor) and fresh bladder tumors were used as target cells. The spontaneous NK cell activity of PBL was 23.7%, whereas that of TIL was only 3.5%. However, in vitro culture with IL2 induced a significant augmentation of NK activity in TIL as well as in PBL. On the other hand, the spontaneous lymphokine activated killer cell (LAK) activity of PBL and TIL was very low. IL2-cultured PBL and TIL exhibited the highest levels of lysis against fresh bladder tumors. Unlike PBL, IL2-induced cytotoxicity of TIL against autologous bladder tumors was higher than that against allogenic bladder tumors. Immunomodulators OK432 and Il2 were injected intratumorally during endoscopy. Analysis of the lymphocyte subsets in TIL showed an increase of T and NK cells following immunomodulator injection. Endoscopic injection of immunomodulators into bladder tumors augmented NK cell functional activity in TIL as well as PBL. These findings suggest that local immunosurveillance is directed against bladder tumors. Further studies are required to understand more fully the local and systemic host immune responses in cancer.
膀胱肿瘤中的组织浸润淋巴细胞（TIL）被认为是局部宿主对肿瘤抵抗力的一种表现。我们研究了TIL与外周血淋巴细胞（PBL）相比的功能活性。采用酶混合物处理并结合Ficoll-泛影葡胺不连续梯度离心法分离TIL。通过流式细胞术分析淋巴细胞亚群，结果显示TIL中Leu 4阳性细胞占43.6%（T细胞）；Leu 10阳性细胞占10.5%（B细胞）；Leu 7阳性细胞占13.1%（自然杀伤（NK）细胞）。在4小时的51Cr释放试验中检测了TIL和PBL的细胞毒性活性。髓系K562细胞（NK敏感细胞）、HT 1197细胞（膀胱肿瘤细胞）和新鲜膀胱肿瘤组织用作靶细胞。PBL的自发NK细胞活性为23.7%，而TIL的仅为3.5%。然而，用白细胞介素2（IL2）进行体外培养可显著增强TIL以及PBL中的NK活性。另一方面，PBL和TIL的自发淋巴因子激活杀伤细胞（LAK）活性非常低。经IL2培养的PBL和TIL对新鲜膀胱肿瘤的裂解水平最高。与PBL不同，IL2诱导的TIL对自体膀胱肿瘤的细胞毒性高于对同种异体膀胱肿瘤的细胞毒性。在内镜检查期间将免疫调节剂OK432和IL2瘤内注射。对TIL中的淋巴细胞亚群分析显示，注射免疫调节剂后T细胞和NK细胞增加。向膀胱肿瘤内内镜注射免疫调节剂可增强TIL以及PBL中的NK细胞功能活性。这些发现表明局部免疫监视针对膀胱肿瘤。需要进一步研究以更全面地了解癌症中局部和全身宿主免疫反应。
