Analysis of immunological response of regional lymph node lymphocytes from patients with gastric cancer after endoscopic local injection of biological response modifier (BRM)

Local injection of BRM (OK-432 or Lentinan) around the cancer lesions via endoscope was performed on 66 gastric cancer patients preoperatively, and the regional lymph node lymphocytes in the surgical specimens were examined on T-cell subset, production of IL-2, and responsiveness to IL-2. The results were as follows: 1) In the metastatic lymph nodes, proportions of T-lymphocyte subsets, production of IL-2 and responsiveness to IL-2 did not change after local administration of OK-432 or Lentinan. 2) In the non-metastatic lymph nodes, percentage of CD4+4B4+ cells were significantly increased and production of IL-2 was markedly augmented after local injection of OK-432. On the other hand, percentage of CD8+CD11- cells were significantly increased and production of IL-2 was markedly enhanced after the local injection of Lentinan. These data suggested that the antitumor response of the non-metastatic lymph node lymphocytes was augmented after the local injection of OK-432 or Lentinan.     

乳腺癌患者外周血中T细胞亚群与淋巴结转移和组织学分级的关系

目的:探讨乳腺癌患者外周血中T细胞亚群的变化及其与淋巴结转移和组织学分级的关系。方法:用流式细胞术检测86例乳腺癌患者以及20例乳腺腺病患者外周血T细胞亚群的百分率。结果:乳腺癌患者外周血总T细胞与CD4~+T细胞百分数与腺病患者无统计学差异(均P0.05),但CD8~+T细胞百分数低于腺病患者(P0.05)。乳腺癌患者中,淋巴结转移者CD4~+T细胞百分数高于无淋巴结转移者(P0.05);CD8~+T细胞百分数随组织学分级增加而升高(P0.05)。结论:乳腺癌患者存在细胞免疫功能紊乱,外周血中CD4~+T细胞、CD8~+T细胞比例的变化分别与淋巴结转移、组织学分级密切相关。监测外周血T细胞亚群的变化,有助于病情及预后的判断。     

Immunohistochemical study of the regional lymph nodes in gastric cancer

Immunohistochemical study with various monoclonal antibodies against the mononuclear cell surface antigen was performed on the regional lymph nodes of gastric cancer through Avidin-Biotin Complex (ABC) method. In the paracortical area (P.C.) of those lymph nodes without the metastasis of gastric cancer, T cells were dominant, and OKT3 positive (OKT3+) cells and OKT4+ cells were diffusely present, while OKT8+ cells were occasionally recognized. In the sinus, subsets of the above T cells, and OKT9+ cells and OKT10+ cells were observed. In the germinal center (G.C.), mantle zone (M.Z.) and primary follicle (P.F.), which were B cell regions, OKIa1+ cells and Leu12+ cells were diffusely present. OKB7+ cells, OKT9+ cells, OKT10+ cells and Leu7+ cells were also noted in G.C.. Leu8+ cells were observed in M.Z. and P.F.. OKIa1+ cells were occasionally noted in P.C. and sinus. In the lymph nodes with the metastasis, decrease of OKT4+ cells and increase of OKT8+ cells were noted in comparison to the lymph nodes without the metastasis. Using the tissue double fluorescence staining method, it was found that about half of the OKT4+ cells were helper T cells. The majority of OKT8+ cells were identified as cytotoxic T cells or their precursors. By preoperative endoscopic administration of OK-432 or PSK into the tumor, the IL-2 receptor+ cells, or OKM1+ cells and OKT4+ cells increased in the regional lymph nodes and the antitumor activity was intensified.     

Natural killer (NK) activity of peripheral blood and regional lymph nodes in patients with genitourinary malignant tumors. 2. NK activity and T-lymphocyte subsets in peripheral blood and regional lymph nodes

We investigated natural killer (NK) activity and T-lymphocyte subsets in peripheral blood and regional lymph nodes in order to examine the host's defence mechanisms against cancer. The materials were obtained from 26 patients with genitourinary cancer and 9 patients with benign diseases, who were used as controls. NK activity was measured by ATP-chemiluminescence (ATP) assay, a new method which is well correlated to the 51Cr-release assay, and T-lymphocyte subsets were stained with anti-Leu-2, -3, -4, -7, and -11 monoclonal antibody. The NK activity of regional lymph nodes was lower than that of the peripheral blood in all 35 cases, and the percentage of Leu 7+ and Leu 11+ cells was in agreement with that of the NK activity detected by ATP assay. In the peripheral blood, the percentage of Leu 3+ cells was lower in high stage patients than it was in low stage patients and controls, but in the regional lymph nodes it was paradoxically higher. The NK activity of regional lymph nodes against autologous tumor cells was low in 8 patients with bladder cancer. It appears from this study that the regional lymph nodes have no function as immunological barriers for metastasis and we therefore conclude that they should be dissected during surgery.     

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目的 研究乳腺癌腋窝淋巴结发生癌转移和未发生癌转移时的免疫功能。方法2004年8月至2005年7月采用流式细胞技术检测乳腺癌病人乳腺癌前哨淋巴结（SLN）与乳腺癌非前哨淋巴结（NSLN）中免疫细胞CD3^＋T、CD4^＋T、CD8^＋T、CD16^＋NK、CD^19^＋B的比例，并相互比较。结果淋巴结未发生癌转移时，SLN与NSLN的免疫细胞CD3^＋T、CD4^＋T、CD8^＋T、CD16^＋NK、CD^19^＋B的比例差异无统计学意义（P〉0．05）；而且当SLN与NSLN发生癌转移后，它们的免疫细胞的比例也无差异。但SLN发生癌转移与未发生癌转移时相比，其CD4^＋T、CD8^＋T、CD16^＋NK的比例发生显著改变（P〈0．05），CD3^＋T、CD19^＋B的比例改变无统计学意义（P〉0．05）。结论 当腋窝淋巴结未发生癌转移时，它仍有正常的免疫功能。当腋窝淋巴结发生癌转移后，它的免疫微环境发生了改变，免疫功能呈现一种抑制状态。     

A clinical study of immunological status in gastric cancer patients--with special reference to T-cell subsets in the lymphocytes of regional lymph nodes

Non-metastatic regional lymph node lymphocytes of 41 patients with gastric cancer were studied by using different monoclonal antibodies and flow cytometry. Used monoclonal antibodies were OKT3 (total T; CD3), OKT4 (helper/inducer T; CD4), OKT8 (suppressor/cytotoxic T; CD8) and Leu11 (NK/K cell; CD16). The results were as follows: 1. The percentage of CD3 cells and CD4 cells were about ten point fewer in lymph nodes than in peripheral blood. 2. CD8 cells were found to be one half or one third lesser in lymph nodes than in peripheral blood. 3. CD16 cells were found to be rare in lymph nodes. 4. The percentage of CD3, CD4 and CD8 cells were higher in distal lymph nodes than proximal ones. 5. The percentage of CD3, CD4 and CD8 cells were not different with progression of the cancer, whereas CD3 cells and CD8 cells were decreased in lymph nodes of stage IV. 6. The percentage of CD8 cells was higher in distal nodes of stage III. Regional lymph nodes are necessary to protect against cancer metastasis, and killer T cells and cytotoxic T cells were fewer in lymph nodes. These results suggested that killer activity and cytotoxicity of the lymph node lymphocytes are inactive and anergy.     

Studies on immunological parameters in patients with gastric carcinoma--with special reference to analyses with monoclonal antibodies of T-lymphocyte subsets and interleukin-2

We have investigated the distributions of T-lymphocyte subsets and interleukin-2 in peripheral blood of 62 patients with gastric carcinoma. The antibody (OKT3, OKT4, OKT8, OKT4/8, Leu11, and Leu7) treated cells were analyzed on a Spectrum III automated flow cytometer. Interleukin-2 was measured by the tritium-labelled thymidine CTLL assay on the supernatants of peripheral blood lymphocytes of gastric cancer patients after 24 hours stimulation with phytohemagglutinin. The number of OKT3, OKT4 cells and the ratio of OKT4/8 were decreased in patients with advanced gastric carcinoma compared with early gastric carcinoma. The numbers of Leu11 and Leu7 cells in patients with stage II and III were in higher levels than those with stage IV. Interleukin-2 production was depressed in patients in advanced stage compared with those of early stage. We concluded that the T-lymphocyte subsets and the interleukin-2 production were useful immunological parameters for gastric cancer patients.     

Immunohistochemical study using a double fluorescent staining on the regional lymph nodes of gastric cancers]

For the purpose to investigate the immunological anti-tumor function of lymphocytes in the regional lymph nodes of gastric cancer, a double staining procedure using several monoclonal antibodies against the surface membranes of T and NK cells with fluorescent antibody technique was conducted. The number of CD11b+ cells out of CD8+ cells was small with almost all being CD11b- belonging to cytotoxic T cells. Leu8+ cells and Leu8- cells out of CD4+ cells were recognized in equal numbers and were reciprocal. The ratio of CD4+ Leu8-/CD4+ demonstrated an increase in the group injected with OK-432. The ratio of OKT9+ or OKIa1+ occupied in CD8+ or CD4+ cells was only several percent, and few in number. An increase for the ratio of CD4+ OKT9+/CD4+ was observed in the group injected with IL-2. Similar increases for the ratio of CD4+ OKIa1+/CD4+ were obtained in the group injected with OK-432 and in the metastatic group, respectively. The Leu7+ CD16+ cells were not observed. The Leu7- CD16+ cells were observed in a part where metastases were focused. These results indicated that lymphocytes in regional lymphocyte of gastric cancer might hold the hidden anti-tumor effect, but did not display the full function without preoperative intratumor injection of BRM such as IL-2 or OK-432.     

Analysis of tissue lymphocytes by double fluorescent staining--gastric cancer tissue and regional lymph nodes

An immunohistochemical study was performed on human lymphocytes in the tissue of gastric cancer, and also in the regional lymph nodes, by double fluorescent staining, using monoclonal antibodies. Leu3a+8+ cells (inducer T cells) which consist about 30 per cent of Leu 3a+ cells were seen in the tissue surrounding the gastric cancer. The other 70 per cent of Leu 3a+ cells were Leu3a+8- cells (helper T cells). In the lymph nodes they were noted in T cell areas in almost the same proportions, while in germinal centers, only Leu3a+8- cells were found. On the other hand, OKT8+Leu15- cells (cytotoxic T cells) were noted in a large number, while OKT8+Leu15+ cells (suppressor T cells) were few. Further, an increase of OKT8+Leu15- cells was seen around gastric cancer or metastatic cancer in lymph nodes. These immunohistochemical findings suggest that cytotoxic T cells are the main component in the tissue of gastric cancers and the regional lymph nodes. Increases in inducer T cells and helper T cells are probably required to induce cytotoxic T cells around the cancer tissue.     

胃癌中CXCR4和CD133的表达及其对淋巴转移的影响

目的:探讨CXCR4和CD133在胃癌原发灶中的表达及其对淋巴转移的影响。方法:对50例原发性胃癌原发灶和癌旁胃黏膜组织行免疫组化染色法定位检测CXCR4和CD133蛋白;选用半定量RT-PCR及Western blot法测定CXCR4和CD133 mRNA与蛋白表达量,分析两者的相关性及其与淋巴管浸润和淋巴结转移的关系。结果:CXCR4和CD133分子均定位于肿瘤细胞膜表面,极少数CXCR4位于细胞核内。胃癌组织中CXCR4和CD133表达阳性率及mRNA和蛋白的表达量均明显高于癌旁胃黏膜组织(均P<0.05);CXCR4和CD133 mRNA相对灰度值在淋巴结转移组高于无淋巴结转移组(P=0.011,P=0.038);N1组CXCR4蛋白相对灰度值明显高于N0组(P=0.023),而低于N2+N3组(P=0.008),N1组和N2+N3组CD133蛋白灰度值明显高于N0组(P=0.04,P=0.01),但N1组与N2+N3组之间无明显差异;淋巴管浸润组中的CXCR4和CD133蛋白相对灰度值均高于淋巴管无浸润组(P<0.05);在淋巴转移患者中,CXCR4和CD133蛋白相对灰度值分别与转移淋巴结数(r=0.480,r=0.426)及转移性淋巴结比率(r=0.502,r=0.489)呈正相关。结论:CXCR4和CD133在胃癌原发灶中高表达,两者呈正相关,其联合表达与转移淋巴结比率和转移淋巴结数呈正相关,推测胃癌CD133阳性细胞亚群可能在CXCR4介导下更易导致淋巴管浸润和淋巴结转移。     

A comparative study of weight of regional lymph nodes in association with the presence of metastasis in primary lung cancer patients

The weight of regional lymph nodes was measured in 173 patients who underwent surgical resection of the primary lung cancer and lymph nodes from January, 1986 to January, 1989 in our hospital. Histological examination was also performed and correlation of metastasis and the weight of lymph nodes were studied. The average weight of metastatic lymph nodes was 2.34 g while that of non-metastatic ones was 0.83 g indicating a significant (p less than 0.05) increase of weight in metastatic lymph nodes. Although the percentage of metastasis increased as the weight of lymph nodes increased, 7.6% of lymph nodes weighing less than 0.5 g was positive for metastasis. On the other hand, 66.7% of the lymph nodes weighing more than 3.0 g in adenocarcinoma and 34.5% in squamous cell carcinoma were positive for metastasis indicating the difference of the metastatic tendency to the lymph nodes between the two histological types. The comparative study of the weight of each lymph node station according to the JJC criteria demonstrated the difference of average weight of non-metastatic lymph nodes among each lymph node station. The average weight of pretracheal (#3), subcarinal (#7), interlobar (#11), and segmental (#12) lymph nodes without tumor metastasis were more than 1.0 g, however those of anterior mediastinal (#3a), paraesophageal (#8), and pulmonary ligament (#9) were less than 0.5 g. The average weight of metastatic lymph nodes in each lymph node station was in proportion to those of non-metastatic ones.     

胃癌患者外周血中CD45^–CD44^+CD54^+细胞亚群含量及其临床意义

目的以胃癌肿瘤干细胞标志物CD44和CD54为靶点,利用流式细胞术检测胃癌患者外周血中CD45~–CD44^+CD54^+细胞亚群含量,并结合临床病理学特征分析其临床意义。方法纳入2016年12月至2017年9月期间四川大学华西医院胃肠外科38例胃癌患者作为研究对象,利用流式细胞术检测其外周血中CD45~–CD44^+CD54^+细胞亚群含量,并分析其临床意义。结果 38例患者的CD45~–CD44^+CD54^+细胞中位数为541.9个/mL(71.7~8 057.0个/mL),其中R0组为555.9个/mL(71.7~8 057.0)个/mL。CD45~–CD44^+CD54^+细胞中位数TNMⅠ–Ⅱ期患者为858.6个/mL(183.5~8 057.0个/mL),高于TNMⅢ–Ⅳ期患者的364.6个/mL(71.7~2 269.7个/mL),P=0.015;N0组为941.4个/mL(183.5~8 057.0个/mL,高于N^+组的379.3个/mL(71.7~2 269.7个/mL),P=0.002。T3–4期(P=0.025)、N^+期(P=0.009)和TNMⅢ–Ⅳ期(P=0.012)的患者中CD45~–CD44^+CD54^+高细胞量亚组所占比例较低。联合CD45~–CD44^+CD54^+细胞量和肿瘤大小可以对N分期和TNM分期做出更准确的判断,但其与其他临床病理学特征和预后无显著相关。结论 CD45~–CD44^+CD54^+细胞亚群数量与肿瘤进展之间有相关性,可能用于判断TNM分期和N分期,但本研究纳入样本较少,仍需扩大样本论证其在胃癌患者中的临床意义。     

Usefulness of inhibiting the lymph node metastasis in human gastric carcinoma by B7-1 gene transfection

INTRODUCTION: Lymph node metastasis is one of the crucial prognostic factors in gastric cancer. We have reported that ICAM-1 gene transfection was effective against lymph node metastases of gastric cancer. B7-1, one of the co-stimulatory factors, was reported to induce cytotoxic T lymphocytes when using melanoma and bladder cancer cell lines, as well as ICAM-1. In this study, we investigated the inhibitory effect of B7-1 on lymph node metastasis by B7-1 gene transfection into gastric cancer cells. MATERIALS AND METHODS: We transfected B7-1 genes into a gastric cancer cell line (OCUM-2MLN) and analyzed the effect of B7-1 transduction on lymph node metastasis, the in vitro adhesiveness and cytotoxicity assay of mononuclear lymphocytes to cancer cells and lymph node metastatic ability after orthotopic implantation of gastric cancer cells in vivo. RESULTS: We revealed that mononuclear lymphocytes showed significantly stronger adherence and cytotoxicity to B7-1 transfected cells (2MLN/B7) than its parent OCUM-2MLN cells. The tumor growth rate of 2MLN/B7 xenograft was significantly slower than OCUM-2MLN xenograft in nude mice. In orthotopic implantation experiments for nude mice, 2MLN/B7 cells in stomach developed significantly less lymph node metastasis than OCUM-2MLN cells. Histologic findings showed that leukocytes were intensively infiltrated in both the 2MLN/B7 tumors and its metastatic lesions, however, were scarcely observed in the lesions associated with 2MLN cells. CONCLUSION: B7-1 may play an important role in inhibiting lymph node metastasis by the mechanism of enhanced immunogenicity, and that B7-1 gene transduction might be effective against lymph node metastases of gastric cancer.     

Effect of lentinan on lymphocyte subsets of peripheral blood,lymph nodes,and tumor tissues in patients with gastric cancer

The effect of lentinan on lymphocyte subsets of peripheral blood, lymph nodes, and tumor tissues in gastric cancer patients was investigated. A 2-mg dose of lentinan was administered to 12 patients intravenously twice, the first at 3–9 days before and the second the day before surgery. The results were then compared with a control group without lentinan administration comprising 12 patients. Regarding peripheral blood and lymph nodes without metastasis, lymphocyte subsets defined with anti-CD3, anti-CD4, anti-CD8, anti-Leu7, and anti-Leu11 were analyzed by flowcytometry. As for tumor tissues, lymphocyte subsets defined with anti-CD3, anti-CD4, anti-CD8, anti-Leu11, and anti-M3 were analyzed after immunohistochemical staining. There were no significant changes in the lymphocyte subsets of peripheral blood between the two groups. In the lymph nodes, the CD4 cell ratios increased; otherwise in regard to tumor-infiltrating lymphocytes (TILs), the number of CD4, Leu11 and LeuM3 cells showed a prominent increase. Therefore, lentinan was observed to produce different effects in accordance with the subjective organs.     

曲马多对胃肠道肿瘤患者术中免疫功能的影响

目的 观察曲马多对胃肠道肿瘤患者术中T淋巴细胞亚群和NK细胞数量的影响.方法 30例胃肠道肿瘤行根治手术的患者随机均分为观察组和对照组.观察组麻醉前肌注曲马多1 mg/kg,对照组不使用.于麻醉前、手术1 h和术毕抽取外周静脉血,用流式细胞仪检测T淋巴细胞亚群(CD3+、CD3+CD4+、CD3+CD8+)、活化T细胞(CD3+HLA-DR+)和自然杀伤(NK)细胞(CD3-CDl6+CD56+)数量的变化.结果 两组手术1 h CD3+、CD3+CD4+、CD3+CD4+、CD3+CD8+、CD3+HLA-DR+和NK细胞数量均较麻醉前明显下降,对照组明显低于观察组(P＜0.05);术毕两组各指标有所回升,但对照组仍明显低于麻醉前水平和观察组(P＜0.05).结论 曲马多可减轻胃肠道肿瘤患者术中T淋巴亚群和NK细胞下降的程度,明显改善机体细胞免疫功能的抑制.     

3.0T MR扩散加权成像对直肠癌盆腔淋巴结转移的诊断价值

目的探讨3.0T MR扩散加权成像对直肠癌盆腔淋巴结转移的诊断价值。 方法回顾性分析经手术病理证实的53例直肠癌患者的临床资料、术前常规MRI及扩散加权成像(DWI)资料,以病理结果为参照,将淋巴结分为转移性和非转移性两组,分别测量定量参数ADC值,采用SPSS19.0软件包进行统计学处理,诊断效能采用ROC曲线分析,确定诊断转移性淋巴结的最佳临界点,计算诊断的敏感度、特异度和准确度。以P<0.05认为差异具有统计学意义。 结果116枚淋巴结中,转移性淋巴结47枚(40.5%),非转移性淋巴结69枚(59.5%)。转移性淋巴结的ADC值为(0.81±0.10)×10^－3mm²/s明显低于非转移性淋巴结ADC值(1.06±0.13)×10^－3mm²/s,差异具有显著统计学意义(t＝10.47, P<0.01);ROC曲线分析显示ADC值用于诊断转移性淋巴结的最佳临界点为0.95×10^－3mm²/s,其敏感度为91.5%,特异度为78.3%,准确度为83.6%。 结论3.0T MR扩散加权成像ADC值对直肠癌盆腔淋巴结转移具有较高的鉴别诊断价值,可为患者手术方案的选择和预后的评估提供更多的参考。     

胃癌患者外周血调节性T细胞的变化及其意义

目的 探讨胃癌患者外周血中CD4+CD25+T淋巴细胞、CD8+CD28-T淋巴细胞比例的变化及其临床意义.方法 采用流式细胞技术检测30例胃癌患者外周血中CD4+CD25+T淋巴细胞、CD8+CD28-T淋巴细胞玎分比,对照组为30例慢件胃炎患者.结果 胃癌患者外周血中CD4+CD25+调节性T细胞占T淋巴细胞的百分比为8.7%±1.3%,与胃炎患者相比差异无统计学意义.胃癌患者外周血中CD8+CD28-调节性T细胞占T淋巴细胞的百分比为30.3%±3.3%,明显高于胃炎患者的20.3%±2.7%.外周血中CD4+CD25+T淋巴细胞、CD8+CD28-T淋巴细胞百分比与胃癌淋巴结转移、病理分型无明显相关性.结论 外周血中调节性T细胞在胃癌发展中可能发挥一定作用.     

Presence of prostate cancer metastasis correlates with lower lymph node reactivity

BACKGROUND: Several reports suggest that the dissemination of neoplastic cells and cancer progression are associated with the generation of an immunosuppressive environment. METHODS: In this report, we investigated immunological effects of prostate cancer by comparing metastastic and non-metastatic pelvic lymph nodes (LNs) from 25 patients with carcinomatous involvement of LNs to the non-metastatic LNs from 26 control patients with no metastatic involvement by immunohistochemistry and histological analyses. RESULTS: Our results showed a decreased abundance of CD20+ B lymphocytes (P = 0.031), CD38+ activated lymphocytes (P = 0.038), and CD68+ macrophages (P < 0.001), and less evidence of follicular hyperplasia (P = 0.014), sinus hyperplasia (P < 0.001), and fibrosis (P=0.028) in metastatic LNs comparatively to control LNs. Finally, we observed that metastatic LNs were significantly smaller than control LNs (P = 0.005). CONCLUSIONS: Our results suggest that the development of prostate cancer LN metastasis is accompanied with smaller LN size and decreased LN reactivity suggesting the development of an immununosuppressive microenvironment.     

Foxp3+调节性T细胞与乳腺癌淋巴结转移及增殖的关系

目的:探讨Foxp3~+调节性T细胞(Tregs)与乳腺癌淋巴结转移及增殖的关系。方法:用免疫组化法检测168例女性乳腺癌患者癌组织中CD4和Foxp3(调节性T细胞标志)的表达,以42例女性乳腺良性病变乳腺组织为对照,分析Tregs与乳腺癌淋巴结转移及乳腺癌组织细胞核增殖相关抗原Ki-67表达的关系。结果:乳腺癌组织中CD4~+T细胞与Foxp3~+T细胞(Tregs)的数量均高于乳腺良性病变组织,差异均有统计学意义(均P0.05);有淋巴结转移的乳腺癌组织中CD4~+T细胞与Foxp3~+T细胞数均高于无淋巴结转移的乳腺癌组织,但仅后者差异有统计学意义(P0.05)。乳腺癌组织中Tregs的浸润数量与Ki-67表达之间无关(P0.05)。结论:乳腺癌组织微环境存在免疫抑制,Tregs浸润数量与淋巴结转移关系密切,但与肿瘤增殖无关,提示Tregs可作为判断乳腺癌患者有无淋巴结转移的新指标。     

CD44v6与胃癌淋巴结转移及预后的关系

目的 探讨CD44v6与胃癌淋巴结转移及预后的关系。方法 采用免疫组化二步法对100例进展期胃癌标本进行标记，分析CD44v6与临床病理及预后的关系。结果 胃癌原发灶CD44v6表达阳性率为64％(64／100)；CD44v6的表达随胃癌浸润深度、周围淋巴结转移而升高。结论 CD44v6在胃癌淋巴结转移中起着重要作用，可用于早期预测胃癌的转移潜能和预后。