树突状细胞激活的肿瘤浸润性淋巴细胞体外对自体肝癌细胞杀伤活性的研究

目的探讨树突状细胞(DC)激活的肿瘤浸润性淋巴细胞(TIL)体外对自体肝癌细胞杀伤活性.方法从肝癌患者外周血获取DC,应用粒/巨噬细胞集落刺激因子(GM-CSF)、白细胞介素-4(IL-4)和肿瘤抗原激活DC,然后用DC来激活TIL,观察TIL在体外对自体肝癌细胞的杀伤活性.结果DC激活的TIL具有很高的对自体肝癌细胞杀伤活性,杀伤率为(89.39±3.05)%,明显高于未经DC激活的TIL、CD激活的T淋巴细胞和未经DC激活的T淋巴细胞对自体肝癌细胞的杀伤率.结论肝癌患者外周血DC能诱导TIL产生高效而特异的抗肝癌免疫.     

肺癌TIL的体外培养及其表型分析

应用APAAP法对15例肺癌TIL部分膜抗原的表达动态进行了免疫组化分析。结果显示:未经激活的TIL细胞,其膜抗原(CD~+3,CD~+4,CD~+8)的表达动态基本恒定,经IL-2激活的TIL细胞体外培养一段时期后,其CD~+3细胞数明显上升,同时CD~+8的表达逐渐减少,而CD~+4的表达在培养后期明显增多。     

人体恶性实体瘤TIL增殖力、表型和杀伤力研究

在建立高活力肿瘤浸润淋巴细胞（ＴＩＬ）分离培养方法的基础上，对８３例恶性实体瘤ＴＩＬ的细胞增殖动力学、部份ＴＩＬ表型与杀伤力等作了较为系统的研究。结果提示，本实验室所建立分离培养的ＴＩＬ增殖能力较强且稳定，有６５％的增殖倍数大于１０００倍；３Ｈ－ＴｄＲ掺入高峰在培养的第４５～７５天；对其自身的原代肿瘤细胞杀伤力可以维持至５６天以上；对５６份经ｒＩＬ２诱导的ＴＩＬ（培养时间１７±５．２天）表型分析：ＣＤ３８０±２１％，ＣＤ４３７±２１％，ＣＤ８４４±１８％，ＨＬＡＤＲ６９±２４％，与ｒＩＬ２诱导前比较，ＣＤ３、ＣＤ４、ＣＤ８和ＨＬＡＤＲ均增多，尤以ＣＤ３与ＣＤ８表型增多明显。     

不同来源的TIL临床应用研究

以５１例晚期肿瘤患者作为研究对象，对其不同来源的肿瘤浸润淋巴细胞（ＴＩＬ）进行治疗研究，发现肿瘤组织来源的ＴＩＬ其有效率及一年生存率明显高于胸腹水及转移淋巴结的ＴＡＬ；其中肿瘤组织的ＴＩＬ，胸腹水、转移淋巴结的ＴＡＬ有效率分别为８０．９％、４０．０％、４６．６％；三者治疗肿瘤患者的一年生存率分别为８１．０％、４６．７％、６０．０％；另外发现胸腹水ＴＡＬ对实质性肿块治疗疗效较低，为４０％，其中静脉滴     

MHC-Ⅰ类分子在肿瘤浸润性淋巴细胞识别人黑色素瘤细胞中的作用

应用人白细胞抗原－Ａ２阳性（ＨＬＡ－Ａ＋２）黑色素瘤病人的瘤块中分离得到的肿瘤浸润性淋巴细胞（ＴＩＬ），进行ＨＬＡ－Ａ２限制性ＴＩＬ抗人黑色素瘤细胞作用的研究。在实验中发现，ＴＩＬ对自身和同种异体的ＨＬＡ－Ａ＋２黑色素瘤细胞具有杀伤作用，而对ＨＬＡ－Ａ－２黑色素瘤细胞及ＨＬＡ－Ａ＋２非黑色素瘤细胞无作用。选择重组白细胞介素４（ｒＩＬ－４）＋肿瘤坏死因子α（ＴＮＦα）能促进黑色素瘤细胞ＨＬＡ－Ａ２表达量，并增强ＴＩＬ对瘤细胞的杀伤活性。抗ＣＤ３、抗ＨＬＡ－ＡＢＣ和抗ＨＬＡ－Ａ２单抗具有明显抑制ＴＩＬ的抗瘤细胞活性。结果表明ＴＩＬ杀伤黑色素瘤细胞依赖于Ｔ细胞受体（ＴＣＲ）对瘤细胞共同抗原的识别，并有主要组织相容性复合体－Ⅰ类分子参与。     

Type 1 and type 2 tumor infiltrating effector cell subpopulations in progressive breast cancer

Effector T cells fall into two subpopulations based on cytokine-secretion. Type 1 cells secrete IFN-gamma, whereas type 2 cells secrete IL-4, IL-10, and GM-CSF. NKT cells represent a third subpopulation that secretes similar cytokines and have been associated with immunoregulation. Using the TS/A adenocarcinoma, we assessed the phenotype and kinetics of tumor-infiltrating lymphocytes (TIL) in mice challenged subcutaneously in the mammary region. Flow cytometric analysis shows that T cells do not infiltrate the primary tumor site until days 7-14 following tumor challenge. Both CD4 and CD8 TILs were predominantly CD44(High) and expressed CD25, CD69, and CD95 cell surface activation markers. Activated CD4/CD44(High) TIL numbers reached peak levels at day 21 that precipitously decreased by day 28 whereas corresponding CD8 cell numbers progressively increased, however, at lower levels and with later kinetics. Intracellular cytokine staining showed that greater numbers of IL-4-producing Th2 cells were elicited and with earlier kinetics than that of IFN-gamma-producing Th1 cells. T cells co-expressing DX5 (CD3(+)/DX5(+)) emerged (>21 days), suggesting a recruitment of NK-like T cells at later stages of tumor progression. Moreover, tumors selectively up-regulated TGF-beta, MIF, and IP-10 gene expression at times as early as day 4, with peak levels at day 7 in vivo. Such gene expression remained elevated and correlated with a continued progression in tumor growth suggesting that preferential effector cell recruitment and production of select factors during different stages of tumor maturation may aid in regulating effective endogenous antitumor responses in progressive breast cancer.     

TIL治疗肿瘤研究进展

肿瘤组织中有淋巴细胞浸润现象,证明浸润的淋巴细胞反映了机体对肿瘤的特异性免疫反应.1986年Rosenberg等首先将小鼠肿瘤中分离的浸润淋巴细胞在体外培养,用IL-2扩增达到一定数目后回输给荷瘤小鼠,能有效地控制肝和肺中转移灶的生长,这种淋巴细胞称为肿瘤浸润的淋巴细胞(tumor-infiltrating lymphocytes,TIL).     

卵巢癌患者术后化疗联合应用S—TIL细胞与T淋巴细胞亚群的变化

目的 :观察卵巢癌患者术后化疗辅加冷冻卵巢癌瘤苗激活的肿瘤浸润淋巴细胞 (S- TIL)治疗前后 T淋巴细胞亚群的变化。方法 :将 76例卵巢癌术后行化疗的患者分为 S- TIL 治疗组及对照组。 2组分别于化疗前及治疗后第 7,14,2 1天采外周静脉血检查 T淋巴细胞亚群 ,并进行比较。结果 :治疗组 CD3 +、CD4+、CD8+、CD4+/ CD8+比值在 14d左右即上升 ,与对照组比较有显著性差异 (P <0 .0 5 )。结论 :卵巢癌术后化疗联合应用 S- TIL 治疗可提高机体细胞免疫功能     

Expression of BCL-2 oncoprotein on tumor cells and tumor-infiltrating lymphocytes (TIL) in meningiomas

The Expression of the antiapoptotic oncoprotein BCL-2 and its correlation to tumor grade in 62 meningiomas (48 classic, 9 atypical, and 5 anaplastic) using single and double immunohistochemistry was investigated. BCL-2 expression was found in two different cell populations identified as lymphocytes (BCL-2+CD3+) and tumor cells (BCL+/CD3–). Tumor-infiltrating lymphocytes (TIL) (CD3+) were found within classic (9.5% of cells), atypical (2.4% of cells), and anaplastic (1.8% of cells) meningiomas. In classic meningiomas, 66.5% of TIL were BCL-2-positive, in atypical meningiomas 79.2%, and in anaplastic meningiomas 37.9%. In 33 (68.8%) of the classic meningiomas, medium to high counts of BCL-2+ tumor cells were detected. Atypical meningiomas showed nearly equal percentages of high (two patients), medium (five patients), and low (two patients) BCL-2+ tumor cell counts, whereas anaplastic meningiomas showed only medium (two patients) and low (three patients) BCL-2 tumor cell counts or were BCL-2-negative (one patient). In summary, a significant inverse correlation between the number of BCL-2-positive tumor cells and tumor grade in meningiomas was found. These findings support the hypothesis of cell survival prolongation by the antiapoptotic ability of BCL-2 proto- oncogenes and demonstrate the prognostic relevance of BCL-2 immunoreactivity in meningiomas. Received: 10 June 1998 / Accepted: 23 February 1999     

肿瘤患者T细胞克隆性增殖与预后的分析

 目的　通过分析肿瘤患者肿瘤浸润性淋巴细胞 (tumor infiltratinglymphocyte ,TIL)和外周血淋巴细胞 ( peripheralbloodlymphocytes,PBL)中克隆性增殖T细胞的TCRβ基因谱型的分子特征 ,研究T细胞免疫在肿瘤预后中的作用。方法　采用RT PCR及变性聚丙烯酰胺测序凝胶电泳方法扩增并分离TCRβ基因V J重排片断。通过PCR产物直接测序的方法 ,判断T细胞克隆的特征 ,得到CDR3区氨基酸序列。结果　在结直肠癌、肺癌TIL和术前PBL中均发现TCRβ基因的寡克隆性扩增 ,它们在术后的PBL中均消失。具有术前PBL的克隆性增殖T细胞的患者还可能影响肿瘤的预后。结论　肿瘤患者的肿瘤组织和术前外周血中都存在肿瘤肽特异性激活的T细胞克隆 ,T细胞免疫系统的激活在肿瘤的预后中可能起着重要作用