胸腺肽α1治疗重症肝炎的免疫调节研究

为了解日达仙的免疫调节作用,选取１８例下肝炎病人用日达仙（Ｔα１）治疗,并在治疗前后检测内毒素（ＬＳＰ）、肝瘤坏死因子（ＴＮＦ）、白细胞介素２受体（ＩＬ－２Ｒ）、白细胞介素４（ＩＬ－４）、白细胞介素６（ＩＬ－６）、及Ｔ细胞亚群（ＣＤ８＾＋、ＣＤ４＾＋）。结果治疗前ＬＳＰ、ＴＮＦ、ＩＬ－２Ｒ、ＩＬ－６、ＣＤ８＾＋均增高９Ｐ〈０．０１）,而ＩＬ－４及ＣＤ４＾＋降低。经Ｔα１治疗后ＬＳＰ、ＴＦ、ＩＬ－２     

芪黄冲剂对慢性乙型肝炎患者免疫功能的影响

目的：观察芪黄冲剂对慢性乙型病毒性肝炎免疫功能的影响。方法：治疗组以芪黄冲剂治疗,对照组以乙肝清热解毒冲剂合乙肝养阴活血冲剂治疗。两组患者采用间接免疫荧光法检测,治疗前后免疫球蛋白ＩｇＧ、ＩｇＭ、ＩｇＡ、Ｔ淋巴细胞亚群ＣＤ３＾＋、ＣＤ４＾＋、ＣＤ８＾＋、ＣＤ４＾＋／ＣＤ８＾＋、ＩＬ－２、ＩＬ－６、ＴＮＦ、ＮＫ。结果：芪黄冲剂具有益气活血,滋养肝肾兼以清热解毒作用,能升高ＣＤ４＾＋、ＮＫ细胞,提高ＣＤ４＾＋／ＣＤ８＾＋比值及ＩＬ－２水平,降低ＣＥ８＾＋,尤其对ＣＤ４＾＋、ＣＤ４＾＋／ＣＤ８＾＋、ＮＫ细胞以及ＩＬ０２的调整作用优于对照组（Ｐ〈０．０５￣０．０１）,能显著降低ＩＬ－６及ＴＮＦ水平（Ｐ〈０．０５￣０．０１）。同时还能降低ＩｇＧ、ＩｇＭ、ＩｇＳ,其中对ＩｇＡ的降低作用显著优于对照组。结论：芪黄冲剂不仅具有     

旋毛虫感染小鼠IgG,IL—2和T淋巴细胞亚群动态变化的 …

目的 了解小鼠感染旋毛虫后ＩｇＧ抗体水平、ＩＬ－２分泌量和Ｔ淋巴细胞亚群的动态变化。方法 分别于旋毛虫感染后第７、１４、２１、２８和３５天,采用ＥＬＩＳＡ方法检测血甭特异性ＩｇＧ抗体水平和ＩＬ－２含量,采用流式细胞仪检测ＣＤ４＾＋、ＣＤ８＾＋Ｔ细胞百分率。结果 ＩｇＧ抗体水平在感染后逐渐上升,感染后３５天达最高值;Ｔ淋巴细胞的变化表现为ＣＤ４＾＋细胞减少、ＣＤ８＾＋Ｔ细胞增多,ＣＤ４＾＋／ＣＤ８＾     

肝硬化和肝细胞癌患者血清IL-2、sIL-2R、IL-6、T细胞亚群的变化及相互关系

目的研究肝硬化和肝细胞癌患者ＩＬ－２、ｓＩＬ－２Ｒ、ＩＬ－６、Ｔ细胞亚群的变化和相互关系。方法分别采用双抗体夹心ＥＬＩＳＡ法、ＡＢＳ－ＥＬＩＳＡ法和红细胞花环实验,对４６例肝炎肝硬化和肝癌进行了ＩＬ－２、ＩＬ－６、ｓＩＬ－２Ｒ和Ｔ细胞亚群的测定,并与６６例健康献血员进行了对照。结果肝炎肝硬化（ＬＣ）、肝细胞癌（ＨＣＣ）患者血清ＩＬ－２、ＣＤ＋４／ＣＤ＋８水平明显低于正常对照（ＮＣ）组（Ｐ＜００１）,而ＩＬ－６、ｓＩＬ－２Ｒ水平明显升高,ＨＣＣ组的平均ＩＬ－６水平高出正常１０倍以上,较肝硬化组明显升高（Ｐ＜００１）。在肝炎肝硬化和肝癌患者均发现ＩＬ－２与ＣＤ＋４／ＣＤ＋８比值、ＩＬ－６与ｓＩＬ－２Ｒ正相关,ＩＬ－６与ＣＤ＋４／ＣＤ＋８比值显著负相关,而与ＩＬ－２无相关关系。结论肝炎肝硬化和肝细胞癌患者存在免疫功能紊乱和低下、淋巴因子网络失衡,这与其病理生理机制有关,也为临床上ＬＣ、ＨＣＣ应用ＩＬ－２治疗提供了理论依据。ＩＬ－６的显著增高有助于ＨＣＣ的早期诊断     

肝舒胶囊对慢性肝炎患者免疫功能的影响

为探讨肝舒胶囊对慢性肝炎患者免疫功能的调节作用，检测了肝舒胶囊治疗前后慢性肝炎患者的Ｔ淋巴细胞亚群，自然杀伤细胞及血清可溶性白细胞介素－２受体等指标，并以健康献血者为对照。结果肝炎组ＣＤ＾＋３，ＣＤ＾＋４，ＣＤ＾＋３／ＣＤ＾＋４，ＣＤ＾＋５７显著下降，ＣＤ＾＋８，ｓＩＬ－２Ｒ显著增高，经过肝舒胶囊治疗后，呈相对应负性改变。提示肝舒胶囊对免疫功能有显著改善和调节作用。     

瘤浸润淋巴细胞治疗肺癌的基础研究及临床应用

目的 探讨肺癌瘤浸润淋巴细胞（ＴＩＬ）的分离、培养增殖特性，杀伤活性，表型变化及回输病人后的临床毒副反应。方法 应用机械、酶消化法从６３例肺癌患者手术切除的实体瘤中分离得到ＴＩＬ，其中５１例经ＩＬ－２体外激活培养，１２例在培养前后应用＾３Ｈ－ＴｄＲ释放实验测定ＴＩＬ对自体瘤细胞和８０１－Ｄ细胞的杀伤作用，１３例用间接免疫荧光法测定淋巴细胞表型ＣＤ３＾＋、ＣＤ４＾＋、ＣＤ８＾＋比例的变化。１５例病人     

G—CSF配合大剂量CTX和VCR动员自体外周血干细胞的研究

本研究对１２例拟行ＡＰＢＳＣＴ的患者予ＶＣＲ１－２ｍｇ／ｍ２，体表面静注及ＣＴＸ７ｇ／＾２体表面积静滴，５－７天后予Ｇ－ＣＳＦ１００μｇ／ｍ＾２体表面积静注５－７天，白细胞升至３．０×１０＾９／Ｌ以上时，用ＣＳ３０００血细胞分离机单次收集单个核细胞ＭＮＣ〉２．８×１０＾８／ｋｇ，ＣＤ＾＋３４〉．６×１０＾６／ｋｇ，ＣＦＵ－ＧＭ〉３．９×１－０＾４／ｋｇ，ＡＰＢＳＣＴ后，全部患者于＋５天－＋１０天白     

未经治疗的原发性肝癌患者的免疫功能状态

对２８例未经任何治疗的中晚期原发性肝癌患者进行免疫功能的检测，并与正常比较，结果发现其ＩＬ－２系统中ＩＬ－２产生能力显著性降低，Ｔ淋巴细胞ｍＩＬ－２Ｒ表达非常显著性降低，而血清ｓＩＬ－２Ｒ水平非常显著性增加；外周血淋巴细胞绝对数及Ｔ淋巴细胞亚群分析表明前者明显降低，ＣＤ４细胞，ＣＤ４／ＣＤ８比值非常显著性降低，ＣＤ８细胞呈显著升高，相关性分析表明：ｓＩＬ－２Ｒ水平与ＣＤ４细胞，ＣＤ４／ＣＤ８比值呈     

乙型肝炎患者免疫促进剂治疗前后T淋巴细胞亚群及NK细胞的…

对６２例慢乙肝患者Ｔ淋巴细胞亚群及ＮＫ细胞检测，并观察分别经ｉＲＮＡ、猪苓多糖及ＬＡＫ细胞治疗前后的细胞数量变化。结果发现治疗前ＣＤ＾＋４细胞降低，ＣＤ＾＋８细胞增高。ＣＤ＾＋４／ＣＤ＾＋８比值下降。治疗后ＣＤ＾＋４细胞及ＮＫ细胞增高，ＣＤ＾＋４／ＣＤ＾＋８比值上升，其中以ＬＡＫ细胞治疗效果显著，与ＨＢＶＭ转阴呈正相关。说明提高机体免疫功能对清除病毒有一定作用。     

HBV特异性T细胞治疗慢性乙型肝炎的研究

为寻求一种治疗慢性乙型肝炎的新方法,应用ＨＢＶ特异性Ｔ细胞输注治疗慢性乙型肝炎９例,结果显示,在疗程结束后,患者的ＨＢｓＡｇ,ＨＢｅＡｇＨＢＶＤＮＡ的含量均有所下降,其中ＨＢｅＡｇ下降较明显,Ｐ〈０．０１,２例ＨＢｃＡｇ阳性患者,ＨＢｃＡｇ阴转,ＣＤ＾＋３,ＣＤ＾＋４,ＣＤ＾＋４／ＣＤ＾＋８ＮＫ活性上升,ＣＤ＾＋８,ｓＩＬ－２Ｒ较治疗前后所回复,表明特异性Ｔ细胞治疗慢性乙型肝炎有一定疗效,远期疗效     

Functional expression of β1 and β2 integrins on tumor infiltrating lymphocytes (TILs) in colorectal cancer

Integrins play an important role in various lymphocyte functions. In this study, tumor-infiltrating lymphocytes (TIL) were isolated from colorectal cancer tissues and the expression of β1 and β2 integrins on the TIL was quantitatively examined with two-color flow cytometry. In comparison with peripheral blood lymphocytes (PBL), TIL expressed a lower level of common β1 chain (CD29) in both CD4 and CD8 sub-populations. Among the associated α chains, the expressions of α1 (CD49a) and α2 (CD49b) were slightly higher in TIL than in PBL, whereas α4 (CD49d) and α6 (CD49f) were markedly downregulated in TIL. Both αL (CD11a) and β2 (CD18) were reduced in CD8(+) TIL but not in CD4(+) TIL. TIL with the CD8(+) cytotoxic phenotype showed significantly decreased binding to purified intracellular adhesion molecules (ICAM)-1, and vascular adhesion cell molecule (VCAM)-1, and HT29 colon cancer cells, compared with the in counterparts in PBL. The peculiar expression pattern and functional down regulation of these integrins may explain why TIL in colorectal cancer cannot eradicate the malignant cells. Received: August 24, 1998/Accepted: November 27, 1998     

Differentiation of anti-tumour cytotoxic T lymphocytes from autologous peripheral blood lymphocytes in non-Hodgkin's lymphomas

We have previously reported that specific anti-tumour cytotoxic T cells (CTL) can be differentiated from tumour-infiltrating lymphocytes (TIL) in non-Hodgkin's lymphoma. We found that the combination of interleukin (IL)-1, IL-2 and IL-12 was very efficient for expansion of CD8+ T-cell receptor (TCR)alphabeta+ T cells and for development of their ability to specifically lyse tumour cells. In this study, we investigated whether anti-tumour T cells could be generated from the peripheral blood of patients using the culture protocol developed for TIL. Autologous T cells and tumour B cells from five patients were included in this study. It was found that polyclonal anti-tumour cytotoxic effector cells were generated when cultured in the presence of IL-1beta, IL-2 and IL-12. Interestingly, tumour cells were lysed by perforin/granzyme-mediated cytolysis and not by CD95-mediated apoptosis. By performing inhibition experiments, it was observed that both CD8+ and CD4+ T cells were responsible for the cytotoxic effect and that they were able to recognize malignant B cells by either a major histocompatibility complex (MHC)-restricted or MHC-non-restricted mechanism. Intriguingly, in addition to interferon-gamma and tumour necrosis factor-alpha, IL-10 was secreted continuously during culture. The source of patient T cells used for the generation of anti-tumour CTL should be based on the results obtained with peripheral blood lymphocytes and TIL.     

Liver tumor infiltrating lymphocytes: Comparison of hepatocellular and cholangiolar carcinoma

AIM: To investigate the role of tumor infiltrating lymphocytes (TIL) in primary hepatocellular and cholangiolar carcinomas of the liver.METHODS: Immunohistochemical analysis was performed including antibodies to CD3, CD4, CD8, CD20, CD56 and TIA-1 in formalin-fixed and paraffin-embedded tissue of 35 liver resection specimens of hepatocellular or cholangiocellular carcinomas. Semiquantitative evaluation was performed with emphasis on the area of the tumor itself and of the tumor/liver interface.RESULTS: All hepatocellular carcinomas showed infiltration of lymphocytes predominantly around the tumor in the tumor/liver interface consisting mainly of CD3+ CD4+ T lymphocytes [164.3/10 high power fields (HPF)] and in the tumor itself of CD8+ cells (54.9/10 HPF). Cholangiocarcinomas contained a heterogeneous amount of TIL, composed mainly of CD3+ T cells with a predominance of CD8+ cells in the tumor tissue (52.6/10 HPF) and of CD4+ cells in the interface region (223.1/10 HPF). CD56+ cells of the innate immune system were scarce. There was no significant difference between hepatocellular or cholangiolar carcinoma. No correlation with the clinicopathological data was seen. CONCLUSION: Liver TIL consists of intratumoral CD8+ T cells and peritumoral CD4+ T cells independent of histogenetic origin. Different functions of lymphocytes in these regions seem possible.     

Liver tumor infiltrating lymphocytes： Comparison of hepatocellular and cholangiolar carcinoma

AIM: To investigate the role of tumor infiltrating lymphocytes (TIL) in primary hepatocellular and cholangiolar carcinomas of the liver.METHODS: Immunohistochemical analysis was performed including antibodies to CD3, CD4, CD8, CD20, CD56 and TIA-1 in formalin-fixed and paraffin-embedded tissue of 35 liver resection specimens of hepatocellular or cholangiocellular carcinomas. Semiquantitative evaluation was performed with emphasis on the area of the tumor itself and of the tumor/liver interface.RESULTS: All hepatocellular carcinomas showed infiltration of lymphocytes predominantly around the tumor in the tumor/liver interface consisting mainly of CD3+ CD4+ T lymphocytes [164.3/10 high power fields (HPF)] and in the tumor itself of CD8+ cells (54.9/10 HPF). Cholangiocarcinomas contained a heterogeneous amount of TIL, composed mainly of CD3+ T cells with a predominance of CD8+ cells in the tumor tissue (52.6/10 HPF) and of CD4+ cells in the interface region (223.1/10 HPF). CD56+ cells of the innate immune system were scarce. There was no significant difference between hepatocellular or cholangiolar carcinoma. No correlation with the clinicopathological data was seen. CONCLUSION: Liver TIL consists of intratumoral CD8+ T cells and peritumoral CD4+ T cells independent of histogenetic origin. Different functions of lymphocytes in these regions seem possible.     

Memory T-lymphocytes are the main population of tumor-infiltrating lymphocytes obtained from human primary liver tumors.

The phenotypic characteristics of freshly isolated tumor-infiltrating lymphocytes (TIL) obtained from human liver tumors were analyzed by two-color flow cytometry. TIL consisted of mainly CD3+ T-lymphocytes (70-80%). The ratio of CD4/CD8 in TIL from primary and metastatic liver tumors and autologous peripheral blood lymphocytes (A-PBL) was 1.3, 1.1 and 1.3, respectively. The majority of CD3+ T-lymphocytes (mean +/- SD; 95 +/- 11%) expressed T-cell antigen receptor (TCR) alpha/beta, and gamma/delta TCR positive T-cells were only 5 +/- 4.5% in TIL from both primary and metastatic liver tumors. TIL showed significantly higher percentages of transient activation markers, such as CD25 (Tac) and HLA-DR, than A-PBL. TIL also contained significantly more populations of CD3+ CD45RO+ T-lymphocytes, which are considered to be expressed on primed (memory) T-lymphocytes, than A-PBL. Furthermore, TIL from primary liver tumors demonstrated significantly higher percentages of CD3+ CD45RO+ T-cells than those from metastatic liver tumors. These data indicate that TIL from human liver tumors are an apparently distinct population from A-PBL, and that local immune responses against human primary and metastatic liver tumors might be different. Moreover, TIL from primary liver tumors consisted of mainly activated or primed (memory) T-cells, suggesting that they were sensitized and activated by autologous tumor cells in vivo. These observations may imply the possibility of adoptive immunotherapy using TIL against human primary liver tumors.     

Time course of changes in phenotypes of cultured tumor-infiltrating lymphocytes isolated from human colorectal cancer

A study was conducted to investigate the time course of changes in phenotypes of cultured tumor-infiltrating lymphocytes (TIL) isolated from 5 patients with colorectal cancer. Following initial incubation with protein phytohemagglutinin (PHA-P, final concentration 0.1 w/v%), one of the inducers of interleukin-2 (IL-2) receptor, and recombinant interleukin-2 (rIL-2, 1000U/ml) for 3 days, TIL was further cultured with rIL-2 alone for another 46 to 79 days. Phenotypes of cultured TIL were analyzed by a two color flow-cytometry. The following results were obtained: 1) Yields of TIL isolated from cancerous tissues were 1.0 x 10(6) to 1.6 x 10(6) cells/g wet weight, and TIL grew 31 to 3700 folds in the entire period of culture. These growth rates were significantly higher than those obtained by the conventional method of culture with rIL-2 alone (1 to 720 folds). 2) The population of cytotoxic T cells (CD8(+), CD11(-)) reached the maximum (92%, median value) at 5 weeks of culture, and thereafter gradually decreased to 61% at 9 and 10 weeks. In contrast, the population of activated natural killer cells (CD16(+) and Leu7 (+) or (-)) remained below 4% for the entire period of culture. 3) Maximum enhancement of specific cytotoxic activities of cultured TIL was observed during 3 to 4 weeks in culture, using K562 cells as well as Daudi cells as target cells. In conclusion, addition of PHA-P seems to beneficially affect the growth rate of TIL in culture.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of treatment of tumor-infiltrating lymphocytes on gastric cancer

AIM To study the effect of immune treatment on gastric cancer.METHODS Thirteen patients with advanced gastric cancer were given TIL adoptive immunotherapy in thisstudy. Histological findings showed that 13 patients had gastric adenocarcinoma. Patients received operationson their primary tumor, which could not be resected. Small tumor tissue and metastic lymph nodes were gotduring the operation for TIL preparation. Ten patients were treated as control group. During TILtreatment, the patients did not received any other treatment. Surgical speciments (metastatic lymph nodes )with pathological diagnosis were obtained from operating room. The lymph nodes were minced anddissociated in RPMI 1640 with 0.03% hyaluronidase type V (1500U/g), 0.05% collagenase type Ⅳ(200 U/g), and 0.008% deoxyribonuclease type I (100 U/g) (Sigma, USA) at 37℃ for 12 h. The cellmixture was then filtered through 4-layer gauge, washed twice in Hank's and separated on Ficoll-Hypague(Shanghai ist Chemical Reagent Factory) at 900×g for 20min. Finally, the cells were harvested andcounted. Cells suspension containing TIL and tumor cells were extensively washed and resuspended at a finalconcentration of 106 lymphocytes/mL in complete medium containing 15% human AB serum, 100 U/mLpenicillin and 100 μg/mL streptomycin in RPMI 1640 (Gibco). The final concentration of rIL-2 (Military Medical Institute, Nanjing ) was 500 U/mL. Cultured after 3 d-4 d, lymphoid cells were counted andculture was separated into more flasks when the concentration of lymmphoid cells reached or exceeded 2×106/mL until the total amount reached about 5×109/mL cells. Cytotoxic activities of TIL were tested by 6 h51Cr-release assay. Target cells (5×10s/mL) (human gastric adenocarcinoma) in 1 mL of culture mediumwere labelled with 100 μci of Na251CrO4(Beijing Atomic Energy Research Institute, Beijing), washed andadjusted to 105cells/mL. Labelled cells (E/T: 50: 1, 25: 1 and 12.5: 1) were seeded in round-bottom microtest plates (Corning, Japan) at 104cells/well. Isotope release was measured in a gamma counter (Packard,USA). The percentage of cytotoxicity was calculated according to the following formula: Cytotoxicity% = (Experimental- Spontaneous)/ (Maximum-Spontaneous)×100. Target cells without effective cells weremixed with 0.1 mL of culture media to obtain spontaneous release, and with 0.1 mL of 0.1 mol/L HCl toobtain maximum 51Cr release. TIL cells so induced were counted, washed twice, resupended in 100 mL 0.9%NaC1 solution and intravenouslly transferred. The number of total autologous TIL cells injected was morethan 5×109 cells for one patient and usually separated into 2 - 3 injection during the treatment, rIL-25000 U/d (Nanjing Military Medical institute, Nanjing) in 2 mL of 0.9% NaCl solution was intramuscularlyinjected starting from 5 days before TIL cells transfer to 5 days after transfer of TIL cells. All patients weregiven scheduled gastric roentgenograms. CT scanning, B type ultrasound, histological examination andimmune function were used to observe the changes before, during and after treatment. The curative effectswere judged by the standard of WHO. The methods of the assays of SIL-2R, NK cytotoxicity and CD4/CD8were carried out respectively according to the references.RESULTS The Nk cytotoxicity and CD4/CD8 were significantly increased (P<0.01) after 3- 6 monthstreatment. The soluble IL-2 receptor in sera of patients was significantly decreased (P<0.01) after 3- 6months treatment. There were no significantly differences in the test of CD4/CD8, the cytotoxicity of Nkcells and the soluble IL-2 receptor in serum between the group before treated by TIL and the control group(P >0.05). The NK cytotoxicity and CD4/CD8 in patients treated by TIL were significantly more than thosein the control group. On the contrary, the soluble IL-2 receptor in serum of patients treated by TIL wassignificantly less than those in serum of the control group. The patients of control group survived from 4.5months to 9 months (less than one year) after operation. However seven of the thirteen patients treated byTIL after operation survived over one year. Appetite was improved, sinew enhanced, weight increased andpain relieved in most of patients treated by TIL. On the contrary, the symptoms and signs of patients ofcontrol group were not improved. According to the standard of WHO, there were significantly differences ofPD(Disease Progress), MR (Minor remission), and PR(partial remission) between TIL group and controlgroup. The results indicated that tumor focus completely disapeared in 1(80%) of 13 patients, significantlydecreased in 4 (30%) of 13 patients and slightly decreased in 7 (53%) of 13 patients, suggesting that thetreatment of TIL in the patients with advanced cancer was effective. No side effects were found except fortransient fever in 2 patients.CONCLUSION TIL should be one of the fundamental therapies for the advanced gastric cancer, it canregulate the balance of immunity, relieve pain, improve symptoms and signs and prolong survival time.     

The in vitro effects of interleukin-12 upon tumor-infiltrating lymphocytes derived from renal cell carcinoma

Clinical trials utilising interleukin (IL)-2 with tumor-infiltrating lymphocytes (TIL) have demonstrated efficacy in the treatment of metastatic renal cell carcinoma (RCC). Several cytokines, as well as growth factors have demonstrated modulatory effects upon the biological properties of TIL from RCC, suggesting a potentially important role for cytokines other than IL-2 in the development of active and tumor-specific TIL. IL-12 was recently characterized as a natural-killer-cellstimulatory factor or cytotoxic-T-cell-maturation factor. These properties of IL-12 prompted us to investigate the impact of this cytokine upon the activation of TIL from human RCC. In an attempt to enhance the in vitro growth and activity of renal TIL, we have grown eight renal TIL cultures in varying concentrations of IL-2 (8, 40, 80, 400 U/ml) and IL-12 (200 U/ml). In addition, IL-12 (200 U/ml) was added to TIL cultures pre-activated with IL-2 (400 U/ml). Growth, cell expansion, and the ability of TIL to release certain cytokines upon tumor stimulation were determined. Proliferation assays, phenotypic analysis, and cytotoxicity assays were performed at an early and a late culture stage. IL-12, alone and when added to suboptimal concentrations of IL-2, failed to induce TIL growth. While the addition of IL-12 to optimal doses of IL-2 suppressed TIL culture expansion, sequential culture exposure first to IL-2 and then to IL-2+IL-12 increased the number of cells expressing CD3⁺/CD56⁺ and these cultures demonstrated enhanced in vitro lysis of autologous tumor. IL-12 clearly demonstrated a sequence-dependent impact of the biological behaviour of TIL from RCC. The optimal use of IL-12 in the in vitro expansion of renal TIL may result in cells with an enhanced specific anti-tumor effect.Abbreviations IL interleukin - TIL tumor-infiltrating lymphocytes - RCC renal cell carcinoma - NK natural killer - IFN interferon - TNF tumor necrosis factor - GM-CSF granulocyte/macrophage-colony-stimulating factor - LAK lymphokine-activated killer cells     

Human colorectal tumour infiltrating lymphocytes express activation markers and the CD45RO molecule, showing a primed population of lymphocytes in the tumour area.

This study investigated the phenotype of freshly isolated human tumour infiltrating lymphocytes (TIL) from 14 patients with colorectal tumours, and compared them with lymphocytes derived from the lamina propria of the unaffected mucosa and with lymphocytes derived from peripheral blood of the same patients. It was found that TIL expressed the activation markers CD25 and HLA-DR to a higher extent than the peripheral blood lymphocytes (p = 0.01), and that both lamina propria lymphocytes and TIL preferentially expressed the CD45RO + phenotype, associated with memory cells, in contrast with peripheral blood lymphocytes [corrected]. Both lamina propria lymphocytes and TIL contained few natural killer (NK) cells (CD3-CD56+) compared with peripheral blood lymphocytes (p = 0.001), and this was reflected in the cytotoxicity assays. After 1 to 2 weeks in culture with interleukin-2 100 U/ml, lymphocytes from all three compartments had a high cytolytic activity against all targets tested, consistent with the lymphokine activated killer cell phenomenon. No increase in the number of NK cells was noted after culture, but 20-30% of the T cells now coexpressed the CD56 molecule. This was most prominent in the CD8+ subset, but lymphokine activated killer cell activity was found in both CD4+ and CD8+ subsets. Possible tumour escape mechanisms are discussed.