桥本甲状腺炎合并甲状腺癌患者肿瘤内抗肿瘤免疫临床意义

目的 观察桥本甲状腺炎合并甲状腺癌患者肿瘤内抗肿瘤免疫并探讨其临床意义.方法 应用免疫组织化学二步法检测桥本甲状腺炎合并甲状腺乳头状癌患者肿瘤组织内S-100蛋白呈阳性的树突状细胞及HLA-DR抗原呈阳性表达的细胞并探讨其临床意义.结果 桥本甲状腺炎合并甲状腺乳头状癌患者肿瘤组织内存在大量树突状细胞(8/8),数量明显多于不伴桥本甲状腺炎的甲状腺癌患者(1/20),树突状细胞的胞浆突起插入癌细胞之间并与之接触.癌细胞呈不同程度HLA-DR抗原阳性表达.在树突状细胞及HLA-DR抗原阳性表达的癌细胞间可见大量淋巴细胞围绕.癌细胞存在不同程度坏死、崩解、脱落.结论 桥本甲状腺炎合并甲状腺乳头状癌患者肿瘤组织内存在由树突状细胞、淋巴细胞及HLA-DR抗原阳性表达的癌细胞引起的特异性抗肿瘤免疫,与患者预后可能密切相关.     

树突状细胞肿瘤免疫基础及临床研究进展

树突状细胞（dendritic cells,DCs）是目前已知的功能最强的专职抗原提呈细胞(antigen presenting cells,APCs),其功能主要是能诱导肿瘤免疫耐受和抗肿瘤免疫。肿瘤的发生发展与机体免疫密切相关,肿瘤微环境中存在大量的免疫抑制性因子,可抑制DC免疫功能;而通过调节微环境中的免疫因子可增强DC抗肿瘤效应,逐渐优化DC免疫治疗。同时,放射治疗杀伤照射野内的肿瘤细胞,也能激活机体免疫系统,诱导细胞因子的表达和树突状细胞的功能变化,两者之间存在复杂的作用关系。本文针对树突状细胞免疫功能及其治疗作一综述。     

食管癌组织中肿瘤浸润性树突状细胞表型改变及功能分析

目的:探讨食管癌患者肿瘤组织中肿瘤浸润性树突状细胞表型改变及功能。方法:收集2017年01月至2018年09月我院收治的食管癌患者92例作为观察组,选择同时期我院收治的92例食管良性肿物患者作为对照组。术中分别取食管癌标本和良性肿物标本。检测相应组织中肿瘤浸润性树突状细胞表达水平及表型,同时检测T细胞亚群表达情况,分析肿瘤浸润性树突状细胞表达情况和食管癌患者临床特征的相关性。结果:与对照组比较,观察组肿瘤浸润性树突状细胞密度、MHC-Ⅱ阳性树突状细胞和CD54阳性树突状细胞百分比均显著降低(P＜0.05);观察组CD4+T细胞增高［(24.81±3.72）% vs （20.77±3.63）%,P=0.000］;CD8+T细胞降低［(20.90±4.12）% vs （23.08±4.42）%,P=0.001］。食管癌组织中肿瘤浸润性树突状细胞密度、MHC-Ⅱ阳性树突状细胞和CD54阳性树突状细胞百分比与肿瘤直径、TNM分期和淋巴结转移有关(P＜0.05)。食管癌组织中肿瘤浸润性树突状细胞密度、MHC-Ⅱ阳性树突状细胞和CD54阳性树突状细胞与CD4+T细胞显著负相关,与CD8+T细胞显著正相关(P＜0.05)。结论:肿瘤浸润性树突状细胞在食管癌组织中低表达,功能低下,与T细胞亚群失衡和预后不良有关。     

抗原负载树突状细胞介导肝癌患者肿瘤杀伤细胞功能状态研究

目的:探讨肝癌细胞抗原负载树突状细胞激活肝癌患者外周血细胞毒性T细胞(CTL)、肿瘤浸润淋巴细胞(TI1)的抗肿瘤作用.方法:以细胞计数、间接荧光表型测定分析肝癌患者DC功能状态;MTT法测定肝癌细胞抗原负载DC介导肝癌患者外周血CTL、TIL对肝癌细胞的杀伤活性.结果:肝癌患者DC表达CD1a、CD80、CD83和HLA-DR等分子水平,DC介导的CTL、TIL对肝癌细胞的杀伤活性明显低于健康对照组(P＜0.05,P＜0.01).结论:肝癌患者存在DC功能缺陷,致使其介导的CTL、TIL对肝癌细胞的杀伤作用明显降低.     

H22细胞全细胞抗原负载的树突状细胞激活肿瘤浸润性淋巴细胞抗小鼠肝癌的实验

目的探讨H22小鼠肝癌细胞（H22细胞）全细胞抗原致敏的树突状细胞激活肿瘤浸润淋巴细胞抗小鼠肝癌细胞活性。方法取得小鼠骨髓细胞并诱导生成树突状细胞,由冻融法制备的H22细胞全细胞抗原致敏,然后用已致敏的树突状细胞激活肿瘤浸润性淋巴细胞,测定致敏前后的DC表面抗原CD11c、CD80、CD86、CD40、MHCⅡ,并评估激活前后的TIL对H22细胞的杀伤活性,同时脾淋巴细胞作为杀伤对照。结果CD11c阳性细胞中CD80、CD86、CD40、MHCⅡ阳性细胞所占比例在致敏后的DC表现为明显上调。经致敏后成熟DC激活的TIL对H22细胞杀伤活性明显高于未激活的TIL,并高于激活或未激活的小鼠脾脏淋巴细胞。结论在H22细胞全抗原致敏后,小鼠成熟DC中CD80、CD86、CD40、MHCⅡ的表达率明显高于未成熟DC。经H22细胞全细胞抗原致敏的DC能诱导活化TIL,明显提高其在体外对H22细胞的杀伤活性。     

自体树突状细胞疫苗治疗雌、孕激素受体双阴性乳腺癌的疗效观察

目的 评价自体肿瘤冻融抗原致敏的自体树突状细胞（ADC）疫苗免疫治疗,对处于Ⅱ、ⅢA期雌、孕激素受体双阴性表达乳腺癌患者的疾病进展及生存情况的影响。方法利用自体肿瘤冻融抗原致敏的CD14+前体细胞产生树突状细胞,在细胞因子的作用下开始成熟,制备ADC疫苗。选取符合入组标准的63例患者,根据治疗方案分为实验组和对照组,实验组共接受4次皮下ADC回输。测定实验组疫苗接种前后IFN-γ+／CD8+T细胞含量及两组针对抗肿瘤裂解物发生的Ⅳ型变态反应情况,随访两组的疾病进展情况。结果 疫苗接种后提高了IFN-γ+／CD8+T细胞数量,与疫苗接种前比较差异有统计学意义[（23.4±4.1）％ vs.（14.3±2.0）％, P＜0.05];实验组18例（58.1％）为Ⅳ型变态反应阳性,而对照组均为阴性,差异有统计学意义（P＜0.05）。实验组的3年无进展生存率高于对照组,差异有统计学意义（76.9％ vs. 31.0％, P＜0.05）。实验组未发生严重不良反应。结论ADC疫苗可能通过触发乳腺癌患者的免疫应答机制来治疗乳腺癌,具有较好的效果,并延长无进展生存期。     

树突状细胞及其肿瘤疫苗

树突状细胞（DC）是功能强大的抗原提呈细胞，在抗肿瘤免疫中处于中心地位。近年来DC疫苗的研究取得较大进展，现就DC的培养、肿瘤患者DC的特点及DC疫苗的构建等作一综述。     

肿瘤诱导树突状细胞凋亡研究进展

树突状细胞作为最重要的抗原递呈细胞,在抗肿瘤免疫中起重要作用.恶性肿瘤可以通过分泌细胞因子、蛋白质、神经酰胺和神经节苷脂引起树突状细胞功能缺陷及数量减少,还可以诱导树突状细胞发生凋亡.因此,肿瘤相关抗原不能有效传递给T细胞,激活抗肿瘤免疫应答.利用某些蛋白和细胞因子对树突状细胞进行预处理,可以减少肿瘤诱导凋亡的发生,基因技术可以增强树突状细胞耐受凋亡的能力.现综述恶性肿瘤诱导树突状细胞凋亡的研究进展.     

树突状细胞与热休克处理抗原联合诱导抗肿瘤免疫的研究

目的建立体外培养与扩增脐血树突状细胞(DC)的方法,并利用DC与热休克处理的肿瘤抗原联合诱导产生一种高效特异的抗肿瘤免疫.方法用GM-CSF和IL-4联合刺激诱导脐血单个核细胞分化为DC,再用加热处理的肿瘤抗原负载,与同源的淋巴因子活化的杀伤(LAK)细胞共培养.用电镜观察DC形态,CD1a和HLA-DR试剂盒检测DC细胞表面CD1a和HLA-DR的表达情况,MTT法测定细胞杀伤活性.结果在体外,DC-LAK对同源的SPC-A-1细胞具有相对特异的杀伤作用,LAK和DC-L的作用不具有明显特异性,DC-LAK的细胞毒作用最强;加热抗原组作用明显优于未加热处理组.结论将DC与热休克处理的肿瘤抗原联合,可以诱导产生一种高效并具有相对特异性的抗肿瘤免疫活性细胞.     

树突状细胞的肿瘤抗原负载

树突状细胞（DC）是体内功能最强的专职抗原提呈细胞，在诱导特异性抗肿瘤免疫反应中起着至关重要的作用。通过体外负载肿瘤抗原致敏DC可以有效地增强其抗原提呈能力，所制备的肿瘤抗原DC疫苗回输体内后可促进机体产生特异性细胞毒T淋巴细胞（CTL）和其他抗肿瘤免疫应答机制，有效地杀伤肿瘤细胞。目前已发展了多种针对DC负载肿瘤抗原的方法，有些方法已被应用到了人体肿瘤治疗。     

HIFU治疗后肿瘤抗原对树突状细胞的活化及其抗肿瘤效应

目的：探讨HIFU治疗小鼠H22抑制性肝癌后产生的肿瘤抗原对机体抗肿瘤免疫功能增强的机制。方法：正常小鼠骨髓中提取骨髓细胞，在rmIL-4、rmGM-CSF奈件下培养7d，制备小鼠骨髓树突状细胞，用HIFU治疗小鼠移植性肝癌后产生的肿瘤抗原活化树突状细胞，再用活化后的树突状细胞激活T淋巴细胞为细胞毒性T细胞，用MTT法检测CTL在体外特异性杀伤肿瘤靶细胞的能力。结果：B16肿瘤HSP70-肽复合物组和H22肿瘤HSP70-肽复合物组的脾淋巴细胞的增殖率均高于阴性对照组、H22肿瘤粗提物组和HIFU后H22肿瘤粗提物组，P〈0．001；但两组之间差异无统计学意义，P〉0．05。H22肿瘤HSP70-肽复合物组CTL对H22肿瘤细胞的杀伤率为70．0％，明显高于阴性对照组、H22肿瘤粗提物组和HIFU后H22肿瘤粗提物组（P〈0．001），但对非靶细胞B16肿瘤的杀伤率与上述各组的差异无统计学意义；B16肿瘤HSP70-肽复合物组CTL对B16肿瘤细胞的杀伤率为78．5％，对H22细胞的杀伤率为21．4％，表明CTL对肿瘤细胞的杀伤作用具有特异性。结论：HIFU治疗后坏死肿瘤组织中的HSP70-肽复合物作为肿瘤疫苗，通过活化DC和刺激T淋巴细胞增殖为CTL，发挥特异性抗肿瘤免疫功能。     

膀胱移行细胞癌树突状细胞浸润和PCNA表达的临床病理研究

目的　探讨膀胱移行细胞癌组织中树突状细胞 (DCs)浸润和PCNA的表达与临床病理特征的关系。方法　采用免疫组织化学SP法检测 3 1例膀胱移行细胞癌术后组织标本中增殖细胞核抗原 (PCNA )以及DCs标志蛋白S 10 0的表达。结果　不同病理分级及临床分期以及单发癌灶与多发癌灶的膀胱移行细胞癌 ,其DCs数量相互比较 ,均有显著性差异 (P <0 .0 5 ) ;原发与复发膀胱癌的DCs浸润数无显著性差异 (P >0 .0 5 )。不同病理分级膀胱移行细胞癌PCNA阳性表达细胞数比较有显著性差异 (P<0 .0 5 )。结论　膀胱癌DCs浸润数量随病理分级、临床分级的升高而降低 ,而PCNA随病理分级的升高而增高。DC可以作为预测膀胱癌生物行为的指标之一     

胃癌及癌前病变中树突状细胞浸润、HLA-DR抗原表达的临床意义

 目的　探讨S 10 0 + DC浸润、HLA DR抗原的表达在胃癌发生、发展中的作用及临床意义。方法　应用免疫组织化学方法对 4 3例胃癌、2 4例癌前病变组织中的S 10 0 + DC、HLA DR进行检测。结果　S 10 0 + DC、HLA DR在胃癌组、癌前病变组中的表达均显著高于正常胃粘膜组 (P <0 .0 5 )。在低分化、淋巴结转移和远处转移胃癌患者中S 10 0 + DC数目明显减少 (P <0 .0 1)。HLA DR在低分化胃癌中的阳性表达率显著降低 (P <0 .0 5 )。S 10 0 + DC与HLA DR的表达有正相关性。结论　胃粘膜中S 10 0 + DC与HLA DR的变化反映了胃癌局部免疫状态 ,二者在胃癌发生、发展中起重要作用。     

The number of intratumoral dendritic cells and zeta-chain expression in T cells as prognostic and survival biomarkers in patients with oral carcinoma

BACKGROUND: Dendritic cells (DCs) are antigen-presenting cells with a unique ability to cross prime T cells and generate strong antitumor responses. This study evaluates the presence and prognostic significance of DCs as well as functional T cells, which accumulate in the microenvironment in patients with oral squamous cell carcinoma (OSCC). METHODS: Immunohistochemistry for S-100 positive or p55 positive DCs and for T-cell receptor (TcR)-associated zeta-chain expression in tumor-infiltrating lymphocytes (TILs) was performed in 132 paraffin embedded specimens from patients with primary OSCC. The median clinical follow-up for the patients was 50 months. The numbers of intratumoral DCs or TILs expressing the zeta chain were determined microscopically and compared with clinical and pathohistologic prognostic parameters, including disease stage, T stage or tumor grade, lymph node involvement, as well as disease free survival and overall survival. RESULTS: Immunostaining identified DCs in the epithelial compartment of the tumors (S-100 positive) as well as interdigitating reticular DCs (p55 positive) in peritumoral areas. Based on S-100 staining, intratumoral DC infiltrates were low (<10 DCs per high-power field [HPF]) in 20% of OSCC specimens, intermediate (10-20 DCs per HPF) in 42% of OSCC specimens, and high (>20 DCs per HPF) in 37% of OSCC specimens. The number of S-100 positive DCs was positively and significantly correlated with that of p55 and of TILs with normal zeta-chain expression. A low number of infiltrating S-100 positive DCs was more predictive of poor survival (hazard ratio, 7.95) than lymph node involvement (hazard ratio, 3.36) or late T stage (hazard ratio, 2.92). A significant but weaker association of p55 positive DC infiltration with survival was observed. Low density of DCs and low or absent expression of the zeta chain in TILs correlated with each other and predicted the poorest survival and the greatest risk. CONCLUSIONS: The number of DCs infiltrating the tumor is a highly significant prognostic parameter in patients with OSCC. Furthermore, the absence or paucity of DCs is strongly linked to abnormalities in the TcR-associated zeta chain in TILs. The two biomarkers, zeta-chain expression in TILs and the number of S-100+ DCs in the tumor, independently predict overall survival, disease free survival, and time to disease recurrence in patients with OSCC.     

Tumor infiltrating dendritic cells predict treatment response to immmunotherapy in patients with metastatic renal cell carcinoma

BACKGROUND: Although renal cell carcinoma (RCC) is considered to be an immunogenic tumor, the role of immunogenicity in this tumor for predicting treatment response has been little investigated. PATIENTS AND METHODS: Resected RCC specimens from 25 patients who received cytokine treatment for metastases were investigated using immunohistochemistry for CD83+ or S100+ dendritic cells (DCs), CD8+ T-cells, HLA-DR+ tumor cells, CD68+ tumor associated macrophages, microvascular density and vascular endotherial growth factor. RESULTS: Among the examined parameters, DCs status showed predictive value, that is, higher numbers of CD83+ or S100+ cells in tumors were associated with favorable treatment response. However, only higher CD83 status, which indicates mature and activated DCs, contributed to better survival (p = 0.0339). CONCLUSION: Increased tumor infiltration of mature DCs would be a predictor of treatment response and outcome in metastatic RCC patients, who receive immunotherapy.     

肿瘤抗原致敏的树突状细胞联合IL-2活化的淋巴细胞胸腹腔输注治疗晚期癌性胸腹水

目的：观察肿瘤抗原致敏的树突状细胞联合白细胞介素2活化的淋巴细胞胸膜腹腔内输液治疗晚期癌性胸腹水的效果,为肿瘤的生物治疗提供新的思路。方法：分离晚期癌症患者外周血中单个核细胞,制备单核细胞来源的DC（MoDC)和IL－2活性的淋巴细胞。用患者胸腹腔积液中分离的肿瘤细胞制备冻融抗原致敏MoDC后,联合IL－2活化的淋巴细胞给患者胸腹腔输注,X线或B超监测免疫治疗后胸腹腔积液的变化,FACS分析治疗前后胸腹水中淋巴细胞表面细胞因子受体的表达情况,结果：经肿瘤抗原MoDC联合IL－2活化的淋巴细胞胸瘤腔输液后,胸腹水中表达IL－2R的淋巴细胞数目明显增多,表达IL－10R的淋巴细胞数目明显减少,光镜下可见较多的DC对淋巴细胞黏附及淋巴细胞对肿瘤细胞的杀伤,免疫治疗组完全缓解率46．9％,部分结解率53．1％,有效率100．0％,明显高于DDP对照组（P＜0．05）,结论：肿瘤抗原致敏的MoDC联合IL－2活化的淋巴细胞胸腹腔内输注能显著地促进胸腹水中肿瘤浸润性淋巴细胞的增殖和活化。提高了癌症患者的免疫功能,有效地治疗癌性胸腹水,其机制与肿瘤抗原致敏的MoDC对肿瘤抗原的有效提呈和IL－2活化的淋巴细胞对肿瘤细胞的直接杀伤作用有关。     

Immunohistochemical tracking of an immune response in mammary Paget's disease

Dendritic cells (DC) are the most potent antigen-presenting cells of the organism. They are specialized to capture, process, and present antigen via the MHC class II as well as the MHC class I pathways to CD4+ and CD8+ T cells, respectively. This results in T cell-mediated immune responses that are likely to counteract the generation and propagation of tumors in vivo. Therefore, we studied the distribution of dendritic cells in mammary Paget's disease. Paraffin-embedded samples of Paget's disease of the breast (n=27) and of disease-free epidermis of the nipple (n=10) were investigated immunohistochemically for the presence of dendritic cells, in particular of Langerhans cells, using antibodies against S-100, CD1a, and HLA-DR, as well as novel reagents against Langerin/CD207, DC-LAMP/CD208 and p55 (Fascin), the latter two being specific for mature dendritic cells. Paget samples presented a decrease of CD1a+, S-100+, and Langerin+ intraepidermal Langerhans cells in almost all cases. This was paralleled by a concentration of immature dendritic cells in the tumor-infiltrated tissue itself. Similar to infiltrating breast carcinoma we observed a marked increase of DC-LAMP+ and p55+ mature dendritic cells in the corial tissue beneath the tumor. These cells were almost always found in ribbon-like or nodular lymphocytic infiltrates. Moreover, rare mature dendritic cells were also found in the Paget cell-infiltrated epidermis of the nipple, i.e. in the tumorous lesion itself. These findings may indicate an effective ongoing anti-tumor immune response in this part of spreading breast cancer.     

树突状细胞局部免疫抗瘤作用的初步研究

目的：探讨树突状细胞（ＤＣ）瘤内注射的局部免疫方式对小鼠Ｈ２２肿瘤的治疗效果。方法：体外诱导生成树突状细胞,经Ｈ２２肿瘤裂解抗原致敏,实验分为对照组、ＤＣ组和ＤＣ＋ＣｐＧ-ＯＤＮ组,分别与Ｔ细胞共培养,收获的Ｔ细胞与肿瘤细胞共培养,观察其对肿瘤细胞的杀伤。体内试验：ＢＡＬＢ／ｃ小鼠皮下接种Ｈ２２肿瘤细胞制作成荷瘤鼠,第４天时进行局部瘤内免疫治疗,分为３组：生理盐水组、ＤＣ组和ＤＣ＋ＣｐＧ-ＯＤＮ组,免疫治疗１０天后处死小鼠,观察其治疗小鼠Ｈ２２肿瘤的效果。结果：体外实验中,对照组、ＤＣ组和ＤＣ＋ＣｐＧ-ＯＤＮ组的肿瘤杀伤率分别为（１０.８０±３.２７）％、（３８.２６±５.６０）％和（４２.６６±９.００）％,后两组杀伤率均高于对照组（Ｐ＜０.０１）;体内实验中,生理盐水组、ＤＣ组和ＤＣ＋ＣｐＧ-ＯＤＮ组的平均瘤重（ｇ）分别为１.８０４±０.４２２、１.２１６±０.３３５和０.７３３±０.１９１（Ｐ＜０.０１）。结论：ＤＣ瘤苗局部瘤内注射可抑制小鼠Ｈ２２肿瘤的生长,联合应用非甲基化ＣｐＧ-ＯＤＮ可以明显提高抑瘤效果。     

Anti-tumor Efficacy of a Hepatocellular Carcinoma Vaccine Based on Dendritic Cells Combined with Tumor-derived Autophagosomes in Murine Models

The majority of hepatocellular carcinoma (HCC) patients have a poor prognosis with current therapies, andnew approaches are urgently needed. We have developed a novel therapeutic cancer vaccine platform based ontumor cell derived autophagosomes (DRibbles) for cancer immunotherapy. We here evaluated the effectivenessof DRibbles-pulsed dendritic cell (DC) immunization to induce anti-tumor immunity in BALB/c mouse HCCand humanized HCC mouse models generated by transplantation of human HCC cells (HepG2) into BALB/c-numice. DRibbles were enriched from H22 or BNL cells, BALB/c-derived HCC cell lines, by inducing autophagyand blocking protein degradation. DRibbles-pulsed DC immunization induced a specific T cell response againstHCC and resulted in significant inhibition of tumor growth compared to mice treated with DCs alone. Antitumorefficacy of the DCs-DRibbles vaccine was also demonstrated in a humanized HCC mouse model. Theresults indicated that HCC/DRibbles-pulsed DCs immunotherapy might be useful for suppressing the growthof residual tumors after primary therapy of human HCC.