非小细胞肺癌组织中血管内皮生长因子表达与树突状细胞分布的关系

目的 探讨非小细胞肺癌组织中血管内皮生长因子（ＶＥＧＦ）表达与树突状细胞（ＤＣ）分布的关系。方法 ４６例原发性非小细胞肺癌（ＮＳＣＬＣ）冰冻组织切片经抗ＶＥＧＦ,抗ＣＤ８３抗体免疫组化染色,以观察ＮＳＣＬＣ组织中ＣＤ８３阳性之ＤＣ的密度及ＶＥＧＦ的表达强度,并分析两者的相关性。结果 ＶＥＧＦ免疫组化染色显示４６例肿瘤标本中阴性者１６例,１＋者１３例,２＋者５例,３＋者６例,４＋者６例;按每个高倍视     

食管癌与郎汉斯细胞和T细胞定量的相关性研究

探讨食管癌患者癌组织和癌旁间质中Ｌｃ、Ｔｃ的密度与组织学分类、生物学行为及与预后的关系。方法采用Ｓ－１００、ＣＤ３抗体和免疫组织化学ＡＢＣ方法,对２４９例原发食管癌分别标记,观察癌巢内和癌旁间质中Ｌｃ、Ｔｃ的形态、分布特点和密度。结果Ｔｃ阳性细胞阳性定位在细胞核和细胞质,不均质棕褐色颗粒。Ｔｃ阳性细胞阳性定位在胞膜呈棕黄色。癌巢内Ｌｃ数量与肿瘤分化程度、浸润深度、淋巴结转移至正相关,与患者生存期呈负相关。而Ｔｃ数量和癌旁间质Ｌｃ数量则相反。Ｌｃ、Ｔｃ分布数量与组织学分类无关。结论Ｌｃ、Ｔｃ在癌巢内和癌旁间质分布密度可作为判定食管癌预后的免疫学指标。     

Langerhans细胞,T—淋巴细胞浸润与宫颈癌患者临床,病理和预？…

为探讨中晚期宫颈癌组织中Ｌａｎｇｅｒｈａｎｓ细胞（ＬＣ），Ｔ－淋巴细胞（Ｔ－ｃｅｌｌ）的浸润与组织学分级及临床，预后的关系，用ＬＳＡＢ免疫组分方法，观察了１１３例宫颈组织，其中鳞癌９３例，腺癌１２例，正常宫颈组织８例，随访存活１０ａ以上５５列１ａ内死亡５０例，结果Ｓ－１００蛋白阳性的ＬＣ突起沿着癌组织之间的间隙伸入与癌组织紧密连接，宫颈鳞癌１，２级ＬＣ浸润阳性率４４％，Ⅲ级阳性率为２３．３％，（Ｐ     

食管癌与郎汉斯细胞和T细胞定理的相关性研究

目的 探讨食管癌患者癌组织和癌旁间质中Ｌｃ、Ｔｃ的密度与组织学分类、生物学行为及与预后的关系。方法 采用Ｓ－１００、ＣＤ３抗体和免疫组织化学ＡＢＣ方法，对２４９例原发食管癌分别标记观察癌巢内和癌旁质中Ｌｃ、ｃ的开矿、颁特点和密度。结果 Ｔｃ阳性细胞阳性定位在细胞核和细胞质，不均质褐色颗粒。Ｔｃ细胞性细胞阳性定位在胞膜呈棕黄色。癌巢内Ｌｃ数量与肿瘤分化程度、浸润深度淋巴结转转移呈正相关，与患者生存期     

非小细胞肺癌中P—gp,p53,bcl—2蛋白表达及其相关性研究

目的 研究非小细胞肺癌（ＮＳＣＬＣ）中ｐ５３、ｂｃｌ－２蛋白以及耐多药基因ＭＤＲ１产物Ｐ－糖蛋白（Ｐ－ｇｐ）的表达及相互关系,探讨它们在ＮＳＣＬＣ发展、预后及耐药中的作用。方法 采用免疫组化Ｓ－Ｐ法,对６０例ＮＳＣＬＣ及其癌旁正常肺组织中ｐ５３、ｂｃｌ－２蛋白和Ｐ－ｇｐ进行检测。结果 ｐ５３、ｂｃｌ－２蛋白及Ｐｇｐ在ＮＳＣＬＣ中表达明显高于癌旁正常组织（Ｐ〈０．０１）。两者都与患者预后显著负相关（     

大肠癌S－100~＋树突状细胞和预后的关系

用Ｓ－１００蛋白质抗体对７１例不同预后的大肠癌患者原位癌组织进行ＡＢＣ免疫组化染色。结果表明术后生存期≥５年者癌组织Ｓ－１００＋树突状细胞ＤＣ）数量明显高于术后生存期＜５年者（Ｐ＜０．００１）。在具有同一种病理因素的患者中，生存期不同者其癌组织ＤＣ数量差异有显著性。揭示癌组织ＤＣ数量可作为估计患者预后的指标之一。     

膀胱移行细胞癌Langerhans细胞的研究

膀胱移行细胞癌Ｌａｎｇｅｒｈａｎｓ细胞的研究图１正常膀味粘膜，ＬＣ位于粘膜下间质内，与移行细胞紧密相靠（ＳＩ。Ｏ２００Ｘ）图２移行细胞癌Ⅰ级，癌巢及周围淋巴组织中均可见到ＬＣ（Ｓ１０Ｏ２００×）图３移行细胞癌Ⅱ级，ＬＣ浸润于癌巢中（Ｓ１００２００×）...     

Langerhans细胞浸润与胃癌转移及分化程度关系的定量研究

Ｌａｎｇｅｒｈａｎｓ细胞（ＬＣ）是一种骨髓来源的具有免疫辅助功能的细胞，与肿瘤有一定的关系。本研究用免疫组织化学方法利用Ｓ－１００蛋白抗体对６５例胃癌组织中的ＬＣ进行了定量研究，以探讨肿瘤组织中ＬＣ的浸润与胃癌转移及分化程度的关系。结果表明，癌组织中ＬＣ浸润明显的患者，其局部淋巴结癌转移发生频率明显低于无ＬＣ浸润或ＬＣ浸润不明显者。３５例无淋巴结转移的患者癌组织中均可见ＬＣ浸润，而有淋巴结转移的３０例癌组织中只有９例可见ＬＣ浸润。另外，本研究还观察到分化程度高的癌组织中ＬＣ浸润较为明显，在１５例高分化癌组织中均可见ＬＣ浸润，１８例中分化的癌组织中有１４Ｎ可见ＬＣ浸润，而低分化的３２例癌组织中只有５例可见ＬＣ浸润。本研究结果提示ＬＣ浸润与胃癌转移和分化程度确有密切的关系。     

Langerhans细胞、T-淋巴细胞浸润与宫颈癌患者临床、病理和预后的关系

为探讨中晚期宫颈癌组织中Ｌａｎｇｅｒｈａｎｓ细胞（ＬＣ）、Ｔ－淋巴细胞（Ｔ－ｃｅｌ）的浸润与组织学分级及临床、预后的关系。用ＬＳＡＢ免疫组化方法，观察１１３例宫颈组织，其中鳞癌９３例，腺癌１２例，正常宫颈组织８例。随访存活１０ａ以上５５例，１ａ内死亡５０例。结果Ｓ－１００蛋白阳性的ＬＣ突起沿着癌组织之间的间隙伸入与癌组织紧密连接。宫颈鳞癌１、２级ＬＣ浸润阳性率４４％，Ⅲ级阳性率２３．３％，（Ｐ＜０．０５），Ｔ－ｃｅｌ与腺癌组织学分级关系不明显（Ｐ＜０．０５），Ｔ－ｃｅｌ浸润与腺癌组织学分级关系不明显（Ｐ＞０．０５）。患者存活１０ａ以上ＬＣ浸润阳性率４５．５％明显高于１ａ内死亡的２４％。（Ｐ＜０．０１）·Ｔ－淋巴细胞浸润存活１０ａ以上阳性率８５．５％高于１ａ内死亡６０％（Ｐ＜０．０１）。提示ＬＣ和Ｔ－ｃｅｌ浸润与宫颈癌组织分化程度及预后有关。     

ＰＴＥＮ、ＰＩ３Ｋ、Ｓｕｒｖｉｖｉｎ在非小细胞肺癌中的表达及相关性

目的　检测ＰＴＥＮ、ＰＩ３Ｋ和Ｓｕｒｖｉｖｉｎ蛋白在非小细胞肺癌（ＮＳＣＬＣ）组织中的表达,并对其表达水平与ＮＳＣＬＣ临床特征之间的关系进行相关性分析,探讨其与ＮＳＣＬＣ的浸润、转移和预后的关系。方法　免疫组织化学ＳＰ法检测６０例ＮＳＣＬＣ组织、１９例正常肺组织中ＰＴＥＮ、ＰＩ３Ｋ和Ｓｕｒｉｖｉｎ蛋白的表达水平,运用单因素（Ｌｏｇ-ｒａｎｋ）和多因素（Ｃｏｘ比例风险模型）生存分析比较三者与各临床特征及预后之间的关系。结果　ＰＩ３Ｋ和Ｓｕｒｖｉｖｉｎ在ＮＳＣＬＣ中的阳性表达率显著高于正常肺组织（Ｐ＜０.０５）,ＰＴＥＮ在ＮＳＣＬＣ中的阳性表达率显著低于正常肺组织（Ｐ＜０.０５）;ＰＴＥＮ与ＰＩ３Ｋ、Ｓｕｒｖｉｖｉｎ蛋白表达呈负相关;ＰＴＥＮ阴性组的总生存时间显著低于阳性组,而Ｓｕｒｖｉｖｉｎ和ＰＩ３Ｋ阳性组的总生存时间低于阴性组（Ｐ＜０.０５）;临床ＴＮＭ分期、ＰＴＥＮ、ＰＩ３Ｋ及Ｓｕｒｖｉｖｉｎ表达是影响ＮＳＣＬＣ患者预后的独立因素。结论　ＰＩ３Ｋ和Ｓｕｒｖｉｖｉｎ在ＮＳＣＬＣ中的表达上调,ＰＴＥＮ在ＮＳＣＬＣ中表达下调,ＰＴＥＮ与ＰＩ３Ｋ、Ｓｕｒｖｉｖｉｎ蛋白表达呈负相关,三者均为ＮＳＣＬＣ预后的独立因素,监测它们的表达对评估ＮＳＣＬＣ患者的预后具有一定的价值。     

肺癌细胞和蛙皮素抑制树突状细胞的产生和功能

目的：了解肺癌细胞株及蛙皮素对树突状细胞（DC）产生及功能的影响。方法：树突状细胞由健康人外周血单个核细胞CD14^ ，在完全细胞培养液中加入1000U／ml GM-CSF和1000U／ml IL-4,培养7天获得，肺癌细胞株CRL－5815，CRL－5826，Bombesin和Bombesin受体拮抗剂加入培养液中，Annxin V检测DC凋亡；流式细胞仪检测CD40，CD86，CD83，CD80，HLA－DR阳性表达。结果：培养7－10天后的DC前体细胞表达高水平的HLA－DR CD80 CD86 CD83 CD40。肺癌及蛙皮素可导致DC前体细胞凋亡，而蛙皮素受体拮抗剂可部分保护蛙皮素致DC凋亡的作用。加入肺癌细胞株或蛙皮素与DC共培养时明显抑制上述细胞表型的表达和DC刺激同种异体T细胞的增殖能力，当加入蛙皮素受体拮抗剂时，DC细胞表达HLA－DR CD80 CD86 CD83 CD40明显增加，接近DC正常对照组。结论：肺癌细胞株及蛙皮素可导致DC的凋亡，抑制树突状细胞的产生和功能。     

Dendritic cell and macrophage infiltration in microsatellite-unstable and microsatellite-stable colorectal cancer

High level microsatellite instability (MSI-H) is a hallmark of Lynch syndrome-associated colorectal cancer (CRC). MSI-H CRC express immunogenic tumour antigens as a consequence of DNA mismatch repair deficiency-induced frameshift mutations. Consequently, frameshift antigen-specific immune responses are commonly observed in patients with Lynch syndrome-associated MSI-H CRC. Dendritic cells (DC) and macrophages play a crucial role in the induction and modulation of immune responses. We here analysed DC and macrophage infiltration in MSI-H and microsatellite-stable CRC. Sixty-nine CRC (MSI-H, n = 33; microsatellite-stable, n = 36) were examined for the density of tumour-infiltrating DC, Foxp3-positive regulatory T cells, and CD163-positive macrophages. In MSI-H lesions, S100-positive and CD163-positive cell counts were significantly higher compared to microsatellite-stable lesions (S100: epithelium P = 0.018, stroma P = 0.042; CD163: epithelium P < 0.001, stroma P = 0.046). Additionally, numbers of CD208-positive mature DC were significantly elevated in the epithelial compartment of MSI-H CRC (P = 0.027). High numbers of tumour-infiltrating Foxp3-positive T cells were detected in tumours showing a low proportion of CD208-positive, mature DC among the total number of S100-positive cells. Our study demonstrates that infiltration with DC, mature DC, and macrophages is elevated in MSI-H compared to microsatellite-stable CRC. The positive correlation of Foxp3-positive Treg cell density with a low proportion of mature DC suggests that impaired DC maturation may contribute to local immune evasion in CRC. Our results demonstrate that DC and macrophages in the tumour environment likely play an important role in the induction of antigen-specific immune responses in Lynch syndrome. Moreover, impaired DC maturation might contribute to local immune evasion in CRC.     

S100A4蛋白在非小细胞肺癌中的表达与侵袭和转移的关系

背景与目的:实验证明S100A4在多种恶性肿瘤细胞中高表达,它可能在恶性肿瘤细胞的侵袭和转移等过程中发挥重要作用。本研究探讨S100A4在人非小细胞肺癌中的表达及其与侵袭和转移的关系。方法:采用免疫组化SP法检测41例非小细胞肺癌及6例正常肺组织中S100A4蛋白的表达水平。结果:S100A4在非小细胞肺癌中的阳性率(70.7%)显著高于正常肺组织(16.7%)(P<0.05);在肺腺癌中的阳性率(90.0%)明显高于肺鳞癌(52.4%)(P<0.01)。在TNM分期中,S100A4在Ⅲ Ⅳ期、Ⅱ期和Ⅰ期的阳性率分别为100.0%、66.7%和30.0%,Ⅲ Ⅳ期明显高于Ⅱ期和Ⅰ期(P<0.01),但是Ⅱ期和Ⅰ期间无显著性差异(P>0.05)。S100A4的阳性率在有淋巴结转移的非小细胞肺癌中(90.0%)明显高于无淋巴结转移组(52.4%)(P<0.01),且S100A4的表达与淋巴结转移密切相关(r=0.480,P=0.001)。肿瘤体积增大(<3cm,≥3cm),S100A4的阳性率明显升高(44.4%,91.3%)(P<0.001),且相关性显著(r=0.288,P=0.017)。S100A4的阳性率与非小细胞肺癌的病理分级无明显相关性(P>0.05)。结论:S100A4在非小细胞肺癌中的表达上调,且与淋巴结转移、TNM分期和肿瘤大小密切相关,提示S100A4与非小细胞肺癌的侵袭和转移有关。     

Restoration of CD44S in non-small cell lung cancer cells enhanced their susceptibility to the macrophage cytotoxicity

CD44 is a cell surface receptor for osteopontin (OPN) and hyaluronate. Transformation of normal tissue to a variety of cancers has been demonstrated to be associated with alterations of CD44 isoform expression. However, few reports have paid attention on differences in CD44S expression between non-small cell lung cancer (NSCLC) and adjacent normal lung tissue. In this study, we demonstrate that CD44S expression is down-regulated in NSCLC tissue when compared with paired normal lung tissue. To investigate the role of CD44S down-regulation in NSCLC cells, we reintroduced the CD44S back into the NSCLC cell line, H322 cells, which originally lack CD44S expression. The cytotoxicity by activated macrophage (RAW264.7 cells stimulated with lipopolysaccharide and interferon-gamma) against the H322 cells transfected with the CD44S gene (H322DeltaS) is more prominent than that against the H322 control transfectants (H322Deltaneo). The enhanced susceptibility of H322DeltaS cells to the activated macrophage cytotoxicity appears to be mediated by the interaction between CD44S expression on H322DeltaS cells and OPN produced by activated macrophages since it is completely blocked by either anti-OPN or anti-CD44 antibody. Moreover, H322DeltaS cells are attracted toward OPN produced by activated RAW264.7 cells to a much greater extent than H322Deltaneo cells. These findings suggest that CD44S down-regulation in NSCLC cells may confer a protective advantage of allowing escape from tumoricidal effector cells including activated macrophages of the host.     

结肠癌细胞中Ese-3对树突状细胞分化成熟的影响

目的:利用体外共培养实验,观察结肠癌细胞SW480中上皮特异性转录因子Ese-3表达水平的改变对树突状细胞（dendritic cell,DC）分化成熟的影响。方法:利用慢病毒系统获得过表达Ese-3的结肠癌细胞SW480（SW480 Ese-3）及其对照细胞株（SW480 NC）。利用磁珠分选方式,从人外周血单个核细胞（PBMC）中获得CD14+细胞,经rhGM-CSF、rhIL-4刺激获得不成熟的DC（iDC）。通过Transwell小室将SW480 Ese-3和SW480 NC细胞分别与iDC进行间接共培养。流式细胞术检测共培养后的DC细胞中HLA-DR、CD14、CD83、CD86 的表达。ELISA检测细胞培养上清中IL-12p70的水平。结果:SW480 Ese-3/DC共培养组DC细胞表面标志物HLA-DR、CD83、CD86水平,较SW480 NC/DC共培养组升高,CD14水平降低;SW480 Ese-3/DC共培养组上清中IL-12p70水平较对照组升高。结论:在体外共培养实验中,SW480细胞过表达Ese-3可促进DC细胞分化成熟。     

透毒复方对急性髓系白血病完全缓解患者CD+34细胞源树突状细胞生物学效应的影响

 目的　研究透毒复方青蒿鳖甲汤对急性髓系白血病完全缓解期（AML-CR）患者骨髓CD+34 细胞来源树突状细胞（DC）生物学效应的影响。方法　分离纯化骨髓CD+34 细胞,体外不同浓度透毒中药含药血清与细胞因子联合诱导分化为DC,观察DC形态特征,流式细胞术检测DC表面分子CD80、CD83、CD86的表达,将DC分别与自体、异体外周血T细胞共同培养,四甲基偶氮唑蓝（MTT）比色法检测激活的T细胞不同效靶比对人类白血病细胞株（K562细胞）的杀伤效应。结果　青蒿鳖甲汤联合细胞因子能促进AML-CR患者骨髓CD+34 分化为形态典型DC,不同剂量含药血清均能上调DC表面特征性分子及共刺激分子CD80、CD83、CD86比例,较单纯细胞因子组比较差异有统计学意义（P＜0.01）,且含药血清组较单纯细胞因子更能激发外周血T淋巴细胞对K562细胞的杀伤作用,两者间比较差异有统计学意义（P＜0.01）。结论　青蒿鳖甲汤能促进髓系微小残留白血病患者CD+34 细胞向DC转化,提高DC的生物学效应。     

负载肺癌干细胞膜微粒的 DC －CIK 对耐药肺癌细胞的靶向杀伤作用及对肺癌干细胞凋亡的影响

目的：探讨负载肺癌干细胞膜微粒的 DC －CIK 细胞对 EGFR －TKI 耐药肺癌细胞的杀伤作用及对肺癌干细胞凋亡的影响机制。方法：无血清悬浮细胞培养法富集 EGFR －TKI 耐药肺癌细胞 A549、H292干细胞样细胞,RT －PCR 检测干细胞标志物,裸鼠成瘤实验鉴定致瘤性。超滤和差速离心法获取肺癌干细胞膜微粒。流式细胞术分别测定共孵育组和常规培养组 DC 成熟标志 CD86和 CD83,测定两组 DC －CIK 细胞表型CD3＋、CD3＋CD8＋、CD3＋CD56＋、CD3＋CD4＋;形态观察及 MTT 法分别测定不同效靶比两组 DC －CIK 对A549、H292的杀伤效应;ELISA 法分别检测两组 DC －CIK 上清中 IL －2、IFN －γ、TNF －α分泌水平;流式细胞术分别测定两组 DC －CIK 对肺癌干细胞凋亡的影响。结果：富集培养获得的 EGFR －TKI 耐药肺癌干细胞样细胞高表达干细胞标志物 Sox2和 Oct4,并具较强裸鼠致瘤性。负载膜微粒的 DC 成熟标志 CD86和 CD83较常规 DC 表达显著升高;负载膜微粒的 DC －CIK 较常规 DC －CIK 细胞表型 CD3＋、CD3＋CD8＋、CD3＋CD56＋、CD3＋CD4＋升高,对 EGFR －TKI 耐药肺癌细胞杀伤效应高于常规 DC －CIK,并具有显著提高的靶向趋向性;负载膜微粒的 DC －CIK 分泌因子对 EGFR －TKI 耐药肺癌干细胞样细胞凋亡的影响与常规 DC －CIK 不同。结论：与常规培养 DC －CIK 相比,负载膜微粒的 DC －CIK 活性提高,对 EGFR －TKI 耐药肺癌细胞的体外特异靶向杀伤效应显著提高。细胞分泌因子可显著上调耐药肺癌干细胞的细胞凋亡率。     

High expression of PD-L1 in lung cancer may contribute to poor prognosis and tumor cells immune escape through suppressing tumor infiltrating dendritic cells maturation

The immunohistochemical analysis was used to evaluate the expression of PD-L1 in 109 non-small cell lung cancer (NSCLC) tissues and para-tumor tissues. Associations between expressed PD-L1 and tumor histological types, degree of differentiation, and lymph node metastasis were calculated, and overall survival was assessed. Meanwhile, immunohistochemistry and immunofluorescence double labeling technique were performed to detect the expressions of PD-L1, CD1α, and CD83 on TIDC of 20 lung cancer tissues, and the expression of PD-L1 in CD1α⁺DCs and CD83⁺DCs and their significances were also explored. We found that the expression rate of PD-L1 in NSCLC was associated with histological types and overall survival. Patients with either adenocarcinoma or survival time after surgery less than 3 years showed higher expression rate of PD-L1. Furthermore, Cox model analysis indicated that PD-L1 might be regarded as a poor prognostic factor. PD-L1 could be also detected in CD1α⁺ immature DC in NSCLC, indicating that as a class of key anti-tumor immunocyte in tumor microenvironment, DC expressing PD-L1 itself might play an important role in keeping its immature status and contributing to tumor cells immune escape and disease progression.     

Overexpression of S100A2 protein as a prognostic marker for patients with stage I non small cell lung cancer

S100A2, a calcium-binding protein, recently became of major interest because of its differential expression during transformation and metastasis in various tumors. The purpose of this study was to investigate the prognostic significance of S100A2 expression in the early-stage non small lung cancer (NSCLC). Immunohistochemical analysis to determine the percentage of cells staining positive for S100A2 was performed on 11 NSCLC tissue microarray slides containing samples from 113 patients with pathologic stage I NSCLC who had undergone curative surgery. S100A2 was expressed in samples from 79 patients (69.9%). Kaplan-Meier analysis showed that patients whose tumors had positive S100A2 expression had a significantly lower overall survival and disease-specific survival rate at 5 years after surgery than did patients with negative S100A2 expression (p < 0.001 and p < 0.001, respectively). Age at diagnosis, histologic type of cancer, degree of differentiation and smoking history did not have a statistically significant effect on survival. Multivariate analysis confirmed that S100A2 expression is a better predictor for disease-specific survival than were other clinical and histologic variables tested. Our results suggested that the expression of the S100A2 protein in stage I NSCLC indicates poor prognosis and may be used to identify patients with early-stage NSCLC who might benefit from adjuvant treatment.