人β-防御素-2在喉癌组织中的表达及与CD1a~+树突状细胞浸润的相关性

目的 研究喉癌组织中人β-防御素-2(human beta-defensin-2,HBD-2)mRNA和蛋白的表达,探讨其在机体抗肿瘤免疫中的潜在价值.方法 采用链霉亲合素-生物素酶复合物法(strept actividin-biotin complex,SABC)检测34例喉鳞癌石蜡标本中HBD-2蛋白表达及CD1a~+树突状细胞(dendritic cell,DC)的浸润;收集18例喉鳞癌患者术中切除的新鲜肿瘤组织及其中5例距肿瘤边缘2 cm的癌旁上皮组织,应用互补DNA(cDNA)探针原位杂交检测HBD-2 mRNA的表达.分析HBD-2在不同的临床和病理状态下的表达及与CD1a~+树突状细胞浸润的关系.结果 ①免疫组化结果显示,在不同的病理分化组中HBD-2的表达差异有统计学意义(F=6.809,P＜0.05),高分化组HBD-2表达水平明显高于中分化和低分化组(P值均＜0.05).不同临床分型、T分期及有无淋巴转移组中HBD-2的表达水平差异无统计学意义.②原位杂交结果显示,18例喉鳞癌和5例癌旁上皮组织中HBD-2 mRNA的表达存在差异,喉癌低分化组明显弱于高分化组和癌旁上皮组织(F=16.391,P＜0.05).③连续切片染色结果显示,HBD-2蛋白表达水平与CD1a~+DC浸润程度呈正相关(r=0.343,P＜0.05),连续切片结果显示,图片染色的阳性区域相互对应,表示存在位置上的相关性.结论 HBD-2在喉鳞癌中的表达与癌细胞的分化程度有关,并受到转录水平和翻译水平上的调节;HBD-2可能通过趋化CD1a~+DC在肿瘤局部的浸润及功能的活化在机体获得性抗肿瘤免疫中发挥作用.     

树突状细胞表型抗原在喉咽癌颈部淋巴结中的表达

目的：探讨喉咽癌颈部淋巴结中树突状细胞（Dc）表型抗原的表达与喉咽癌转移及预后的关系。方法：对37例喉咽癌144枚淋巴结DC的表型抗原S-100、CD1a、CD83的表达用Envision HIS的方法检测,并观察CD45RO^＋T细胞与DC的关系。结果：S-100^＋DC、CD1a^＋DC、CD83^＋DC在转移组淋巴结中的数目较非转移组淋巴结中减少,差异有统计学意义。CD83^＋DC在转移淋巴结的癌巢中的数目较癌旁中的少,差异有统计学意义。在生存组淋巴结中的浸润数目较死亡组多,差异有统计学意义。CD45RO^＋T细胞主要分布在CD83^＋DC的周围。结论：表达不同表型抗原的DC具有不同的功能,CD83^＋DCs被认为是成熟DC分布在癌旁与其激活T细胞的抗原提呈功能有关,对于肿瘤免疫防御机制的建立起着重要作用。DC在淋巴结中的浸润程度及表型抗原的表达情况是反映宿主肿瘤免疫状况的重要指标,也是预测喉咽癌转移和预后的一项重要指标。     

CD45RO在喉鳞状细胞癌中的表达及临床意义

目的：探讨喉鳞状细胞癌中cD45RO的表达及其临床意义,为喉鳞状细胞癌的预后评估提供新的思路,及为其生物治疗提供理论依据。方法：2010—01—2012—12期间我院确诊并手术治疗的初发喉鳞状细胞癌标本50例。另取10例喉部正常黏膜作对照分析。所有标本均经10％甲醛固定,石蜡包埋、切片。采用免疫组织化学、苏木精伊红染色方法检测CD45RO的阳性表达,分析其与喉鳞状细胞癌临床病理特征之间的关系。结果：CD45RO在正常组织及喉鳞状细胞癌中的阳性表达率分别为30％、86％。在50例喉癌病例中,CD45RO与局部淋巴结转移呈负相关（r=-0．713,P〈0．05）,与肿瘤分区呈负相关（r=-0．750,P〈0．05）,与年龄、T分类、病理分化程度、临床分期无明显相关性。结论：①CD45RO在喉鳞状细胞癌组织中的表达高于正常喉黏膜组织中的表达;②CD45RO过表达可能减少区域淋巴结转移;③CD45RO过表达可能有利于喉鳞状细胞癌患者的预后;④传统UICC—TNM分期、组织学分级评估预后不同,通过对CD45RO的检测可以从另外一个角度评估喉鳞状细胞癌的预后情况。     

Dendritic cells in selected head and neck tumors

Specimens from 17 head and neck tumor patients were immunohistochemically stained with monoclonal antibodies against HLA-DR, CD1a, RFD1, LAG, CD3, CD4, CD8, CD45RO, CD68, and cytokeratin to identify the nature and distribution of dendritic cells (DCs), T cells, and macrophages. Small numbers of DCs were present in all but 2 specimens. They were located between the tumor cells and in the stroma, especially in areas of inflammatory cell infiltration. Variable numbers of T lymphocytes (cytotoxic and memory type) occurred in the same locations. Numerous macrophages were found in the epithelium, in the stroma, and in the vicinity of tumor cells. The presence of DCs in head and neck tumors indicates that the organism has activated the immune surveillance system and is trying to present tumor antigens. Considering the sparsity of DCs in the malignant tissues, the T cell response can be only limited.     

细胞外基质金属蛋白酶诱导因子在喉鳞状细胞癌组织中的表达

目的：了解细胞外基质金属蛋白酶诱导因子(EMMPRIN)在喉鳞状细胞癌(LSCC)及喉癌前病变的表达情况,探讨其与LSCC浸润和转移的关系。方法：用免疫组织化学SP法对42例LSCC、28例喉癌前病变和20例正常喉组织中的EMMPRIN表达情况进行检测。结果：EMMPRIN在LSCC和喉癌前病变的阳性表达率分别为88．1％和57．1％,明显高于正常喉组织的5．0％,Ⅲ～Ⅳ期LSCC组织的强阳性表达率为76．0％,高于Ⅰ～Ⅱ期的强阳性表达率17．6％。伴有颈淋巴结转移的LSCC组织的EMMPRIN强阳性表达率高于无淋巴结转移组。结论：EMMPRIN在LSCC有较高的阳性表达。EMMPRIN的表达与LSCC的分期和是否存在淋巴结转移有关。LSCC组织EMMPRIN的表达影响LSCC的浸润和转移。     

喉咽癌树突状细胞表型抗原表达

目的对喉咽癌中树突状细胞(Dendriticcells,DCs)的表型抗原的表达进行研究,探讨DC表型抗原的表达与喉咽癌各临床因素尤其是转移和预后的关系。方法对45例喉咽癌标本,用EnvisionHIS的方法检测DC的表型抗原S-100、CD1a、CD83的表达,并观察CD45RO T细胞与DC的关系。结果S-100 DCs在高分化组以及生存组中的数目明显多于中低分化组及死亡组,具有显著性差异。CD83 DCs的表达与喉咽癌的分化程度、远处转移、生存以及是否复发有关,差异具有显著性;CD83 DCs在癌巢中的数目较癌旁中的少,差异具有显著性。CD1a DCs的表达与喉咽癌的各临床因素间未发现明显相关性。CD45RO T细胞主要分布在CD83 DCs的周围。结论表达不同表型抗原的DC具有不同的功能,CD83 DCs被认为是成熟DC分布在癌旁与其激活T细胞的抗原提呈功能有关,对于肿瘤免疫防御机制的建立起着重要作用。DC在喉咽癌中的浸润程度及表型抗原的表达情况是反映宿主肿瘤免疫状况的重要指标,也是预测喉咽癌转移和预后的一项重要指标。     

喉癌颈淋巴结抗肿瘤免疫状态的研究

目的　通过对肿瘤引流淋巴结中增生性滤泡反应和树突状细胞 (dendriticcell,DC)浸润密度进行定量分析 ,探讨其在喉癌患者抗肿瘤免疫反应中的作用。方法　重新制备 4 7例行选择性颈淋巴结清扫术的喉癌患者的 15 7枚颈淋巴结石蜡切片 ,用抗S 10 0、CD4 5RO和CD2 0单克隆抗体免疫组化抗生物素 生物素过氧化物酶复合物 (avidin biotinperoxidase,ABC)法显示DC和增生性滤泡的分布并检测其密度。根据增生性滤泡反应的特点进行分型研究。结果　增生型滤泡反应的程度与类型和DC浸润密度有明显的相关性。术后生存 5年以上的患者淋巴结中滤泡计数和DC浸润密度明显多于生存期少于 5年者 (P <0 0 0 1)。无淋巴结转移组患者淋巴结中的滤泡计数和DC浸润密度较有转移组亦有明显增加 (P <0 0 0 1)。具有T细胞增生型滤泡反应的患者比T细胞非增生型患者有较高的 5年生存率 (P <0 0 1)和较低的淋巴结转移率 (P <0 0 5 )。结论　颈淋巴结中增生性滤泡反应和树突状细胞的浸润反映了机体免疫系统的抗肿瘤潜能。树突状细胞的浸润密度和增生性滤泡反应的程度及分型反映了该患者的抗肿瘤免疫状态 ,可以共同作为预后判定的重要指标。     

Relationship among lymphatic metastasis, pericancerous lymphocytic reaction and dendritic cell infiltration in laryngeal carcinoma cells]

OBJECTIVE: To study the role and the clinical significance of dendritic cell infiltration against tumor cells in patients with laryngeal squamous cell carcinoma. METHODS: Immunohistochemical method using S-100 protein antibody was employed to detect dendritic cells (DC) on paraffin-embedded tissue sections from 23 patients with laryngeal squamous cell carcinomas. The relationships among the density of dendritic cell infiltration in laryngeal carcinomas and cervical lymphatic metastasis, pericancerous lymphocytic reaction and other clinicopathologic parameters were observed. RESULTS: The numbers of dendritic cells infiltrating among laryngeal carcinoma cells in patients with no cervical lymph metastasis was significantly larger than those with cervical lymphatic metastasis (t = 4.889, P < 0.01). Significant increase DC infiltration among laryngeal carcinoma cells was found in the group with intensively positive reaction of pericancerous lymphocyte than in the medium and weakly positive groups. The number of infiltrating dendritic cells in patients who had survived more than 5 years was significantly larger than those survived less than 5 years (t = 4.423, P < 0.01). The numbers of the infiltrating dendritic cells in patients with well-differentiated squamous cell carcinoma and poorly differentiated squamous cells carcinoma were 2.2 +/- 1.07 and 14.6 +/- 7.14 respectively, which were significantly different (q = 4.532, P < 0.05). CONCLUSION: The study showed that DC infiltration among laryngeal squamous carcinoma cells played an important role in the host immune reaction against tumor. It indicated that when the density of infiltrating DC is higher, the patients had less chance of cervical lymph metastasis and may have a longer survival time. There was a coordinative effect between DC infiltration in tumor cells and pericancerous lymphocyte reaction. Thus, the DC infiltration among laryngeal squamous cells could be used as an index of prognosis.     

常年性持续性变应性鼻炎患者外周血单个核细胞CD45RA及CD45RO表达

目的 探讨常年性持续性变应性鼻炎(AR)患者外周血单个核细胞(peripheral blood mononuclear cells,PBMC)中CD45RA及CD45RO的表达及其与AR发病的关系.方法 应用荧光抗体标记、流式细胞仪检测30例AR患者PBMC中CD45RA及CD45RO的表达.结果 与对照组比较,①AR...     

Expression of CD68 CD45RO CD20 and proliferating cell nuclear antigen in nasal polyps]

OBJECTIVE: To study the cellular expression of CD45RO, CD20, CD68 and proliferating cell nuclear antigen (PCNA) in nasal polyps. METHODS: Nasal polyp tissues from 50 patients were evaluated for cellular expression of CD45RO, CD20, CD68 and PCNA using immunohistochemistry SP by counting the average number in 5 chosen high-power fields, Histopathological observations were combined with HE. Analyses were performed on SPSS10.0. RESULTS: CD68+ cells were expressed more in nasal polyps dominated by eosinophils than by neutrophils(P < 0.05). There was no difference between CD45RO and CD20, but both of them had negative correlation(P = 0.05). Significant correlation was found between CD68+ cells and eosinophils or PCNA positive cells on epithelium. PCNA positive cells on epithelium had significant correlation on fibroblast (P < 0.05). CONCLUSION: Inflammatory cell infiltration (eosinophilia CD45RO, CD20, CD68) and cell proliferating in epithelium cells, glandular cell and fibroblast are strongly correlated with formation of nasal polyps. The nasal polyps are not only characteristic of eosinophilia but also by lymphocytes dominated by CD45RO and CD68 positive cells. CD68 may be stem cell of nasal polyp.     

An immunohistochemical analysis of p53 protein expression in pre-malignant and malignant tissues of the oral cavity

This study looks at p53 protein expression in dysplastic and malignant lesions of the oral cavity using an immunohistochemical staining technique. Archival biopsy specimens of oral dysplasia of squamous cell carcinoma from 64 patients were analysed immunohistochemically. Sections from 90 oral biopsy specimens were examined in all. Positive immunohistochemical detection of the p53 protein, demonstrated by brown nuclear staining, was detectable in over 80% of mild, moderate and severe dysplastic tissues as well as carcinoma-in-situ and squamous carcinoma specimens. We concluded that p53 protein expression occurs frequently in both malignant and dysplastic lesions of the oral cavity, suggesting that abnormally detectable p53 protein is present at the very early stages of development of oral squamous carcinoma. Oral cancer may provide a good model for the study of multistage tumorigenesis in head and neck cancer as the lesions are frequently detected at the pre-invasive stage and are accessible to biopsy     

Clinicopathological features of Epstein-Barr virus-associated nasal T/NK cell lymphomas in southern Japan

OBJECTIVE: Nasal T/natural killer cell lymphomas (NTCL) are highly prevalent among Oriental populations. However the characteristic immunophenotype of NTCL is still controversial and it can be difficult to make a firm histologic diagnosis of malignancy. Therefore, 14 cases of NTCL in patients from southern Japan were evaluated for clinicopathological features and immunophenotypic status. Furthermore, the genetic variations in the latent membrane protein 1 (LMP-1) gene of Epstein-Barr virus (EBV)-related NTCL were evaluated. METHODS: Biopsy specimens were fixed in formalin and embedded in paraffin before examination using in situ hybridization for EBV-encoded RNA-1 (EBER-1) as well as immunohistochemical staining for CD3, CD3epsilon, CD4, CD8, CD43, CD45RA, and CD45RO. To detect genetic variations, single-strand conformation polymorphism (SSCP) analysis and DNA sequencing were applied. RESULTS: Clinically, 14 cases were divided into two groups. The first group consisting of eight patients showed good prognosis. The other group consisting of six patients showed poor prognosis. The vast majority of neoplastic cells in NTCL were EBER-1 positive. These cells did not express CD4, CD8, or CD45RA, but often expressed CD43 and CD45RO. In addition, they were negative for CD3 when stained with a mouse monoclonal antibody but stained for CD3epsilon when a rabbit polyclonal was used. The 3'-terminal of LMP-1 gene of seven cases were amplified and all of them have 30 base pair (bp) deletion. CONCLUSION: NTCL are a heterogeneous mix of cell types although EBV-associated NTCL in patients from southern Japan appear to originate from natural killer cells rather than T cells, and also prognosis is variable and not always poor. The ability to make a firm diagnosis can be enhanced through the combined use of in situ hybridization and immunohistochemistry. High prevalence of the 30-bp deletions of the LMP-1 gene in EBV-related NTCL may also reflect the prevalence of the deletion variant in the normal population in Japan.     

喉癌颈淋巴结抗肿瘤免疫状态的研究

OBJECTIVE: Densities of dendritic cells (DC) and hyperplastic follicular response in cervical lymph nodes were performed to prove their roles in immune responses against cancers. METHODS: Paraffin blocks were prepared for staining with monoclonal antibodies against CD45RO, CD20 and S-100 proteins,in 157 lymph nodes obtained from elective cervical lymphadenectomy in 47 cases of laryngeal squamous cell carcinomas. RESULTS: Quantitative analysis showed that the patients who survived longer than 5 years had significant higher number of follicles and higher extent of infiltration by DCs in the lymph nodes than those who less than 5 years (P < 0.001). According to negative or positive lymph node metastasis, there were statistically significant differences between two groups (P < 0.001). The patients who possessed T cell increase type follicular reaction had significant higher five-year survival rate ( P < 0.01) and lower lymph node metastasis rate (P < 0.05) than those who possessed T cell decrease type reaction. CONCLUSION: DCs and hyperplastic follicular response may be more directly involved in the host immune reaction against tumor. The classification of follicular reaction, the densities of DCs and follicular reaction, can serve as important indicators in assessing prognosis of laryngeal carcinomas.     

慢性扁桃体炎患者外周血T淋巴细胞亚群的分析与临床意义

目的：探讨慢性扁桃体炎患者外周血T淋巴细胞各亚群的变化特点及其与肾损害的关系。方法：采用流式细胞仪检测54例慢性扁桃体炎患者和52例健康体检者的外周血初始T细胞、功能T细胞、激活T细胞及凋亡T细胞亚群百分比,并结合血清半胱氨酸蛋白酶抑制剂（CystatinC）、ASO和ESR水平进行统计学分析。结果：慢性扁桃体炎患者外周血CD3^＋T细胞、CD3^＋CD8^＋T细胞、CD4^＋CD28^＋T细胞、CD8^＋CD28^＋T细胞、CD4^＋HLA—DR^＋T细胞、CD8^＋HLA—DR^＋T细胞、CD8^＋CD95^＋T细胞与健康对照组相应细胞亚群之间的差异无统计学意义（P〉0．05）。但CD4^＋T细胞、CD4^＋CD45RA^＋T细胞和CD4^＋CD25^＋T细胞减少,CD4^＋CD45RO^＋T细胞、CD8^＋CD38^＋T细胞和CD4^＋CD95^＋T细胞增加,CD4^＋／CD8^＋和CD4^＋CD45RA^＋／CD4^＋CD45RO^＋比值下降,与健康对照组相应细胞亚群之间的差异有统计学意义（P〈0．05）。慢性扁桃体炎患者的外周血CD4^＋CD45RA^＋T细胞百分比与其血清Cystatin C浓度负相关。结论：本研究通过对外周血各T细胞亚群变化特点的系统研究,证实了慢性扁桃体炎患者机体存在T细胞免疫平衡紊乱。CD4^＋CD45RA^＋T细胞的变化可以作为慢性扁桃体炎患者肾损害的相关参考指标。     

Immunohistochemical study of dysplasia and squamous cell carcinoma of the larynx

Formalin- or alcohol-fixed, paraffin-embedded laryngeal specimens were stained immunohistochemically with several kinds of monoclonal anti-Keratin antibody (KL-1, PKK-1, PKK-2, PKK-3), polyclonal anti-Keratin antibody, and epithelial membrane antigen. Immunohistochemical technique was used by Avidin-Biotin Complex method with the following subjects; normal epithelium 6 cases, dysplasia 15 cases, inflammation and polyp 12 cases, squamous cell carcinoma 47 cases. 6 specimens of squamous cell carcinoma were totally resected and alcohol-fixed, and other specimens were all biopsied and formalin-fixed. Comparing these two methods, the alcohol-fixed specimen gave superior result to those obtained with formalin-fixed in its staining. Non-staining area was recognized around basal layer by KL-1 stain in the normal epithelium. Enlargement of non-staining area was recognized in the dysplasia. Also, KL-1 staining showed positive staining with the keratinizing squamous cell carcinoma, and weak staining with the non-keratinizing squamous cell carcinoma. And, PKK-3 showed weak staining in the keratinizing squamous cell carcinoma. The results indicate that high molecular weight keratin (56 kilodalton) hardly localize around the basal layer in the squamous epithelium, and also in the non-keratinizing squamous cell carcinoma. The other way, this also suggests that low molecular weight keratin (45 kilodalton) hardly localize in the keratinizing squamous cell carcinoma. It is supposed that intracellular localization of keratin of dysplastic epithelium and squamous cell carcinoma varies from that in the normal squamous epithelium. Accordingly, monoclonal keratin stain is thought to be an useful method for the diagnosis of dysplasia and squamous cell carcinoma.     

巨噬细胞和淋巴细胞及增殖细胞核抗原在鼻息肉组织中的表达及病理学意义

目的　研究巨噬细胞 (CD68)、T细胞 (CD45RO)、B细胞 (CD2 0 )和增殖核细胞核抗原(proliferatingcellnuclearantigen ,PCNA)在鼻息肉组织中的表达。方法　应用免疫组化链霉菌抗生物素蛋白过氧化酶 (strept avidinoxidase,SP)法对 50例鼻息肉分别做CD2 0、CD45RO、CD68、PCNA的免疫组化染色 ,结合常规HE染色切片进行分析。结果　①CD68+ 细胞在嗜酸性粒细胞性鼻息肉较嗜中性粒细胞性鼻息肉表达率高 ,差异具有显著性意义 (P <0 0 5) ;②CD45RO、CD2 0在鼻息肉均有阳性表达 ,CD45RO与CD2 0呈负相关 (P =0 0 5) ;③CD68阳性细胞与嗜酸性粒细胞浸润及鼻息肉上皮PCNA阳性表达有相关性 (P <0 0 5)。鼻息肉上皮的PCNA阳性表达和成纤维细胞PCNA阳性表达有相关性 ,(P <0 0 5)。结论　鼻息肉的形成与炎性细胞浸润密切相关。鼻黏膜局部的细胞免疫与体液免疫异常导致上皮细胞、成纤维细胞增殖、腺体增生是鼻息肉发生的基础 ,CD68+ 细胞可能是鼻息肉中的炎性干细胞     

Immunohistochemical analysis of lymphocytic infiltration in the tumor microenvironment in patients operated on for laryngeal cancer

The aim of this study was to evaluate semiquantitative and qualitative analysis of lymphocytic infiltrations in a neoplasm microenvironment in patients with laryngeal cancers and the correlation analysis between the intensitivity degree and composition of lymphocytic infiltration in foreseeing a survival time and probability of the appearance of lymph node metastases. Postoperative specimens from 43 patients (Upper Silesia region) operated on for laryngeal cancer in the 2nd ENT Department, Silesian Medical University in Zabrze between 1985 and 1995 all had unfavorable courses due to tumor recurrences. The patients’ ages ranged from 39 to 79 years (mean 57 years). Tissue specimens were subjected to routine processing. The degree of pathological changes was ascertained and immunohistochemical preparations of laryngeal tissue were prepared according to generally accepted methods. The following primary monoclonal antibodies were used: CD 3, CD 20, CD 43, CD 45 RO, CD 56. The distribution analysis of the intensity of the phenotype CD 43 evaluated the lymphocytic infiltration in relation to differentiation of the whole study group. The intensity of CD 43 cell infiltration increased in the group of patients with lymph node metastases. In patients with stage IV disease, a relationship was found between survival time and intensity of cell infiltrations with CD 43 and CD 45 RO lymphocytes. The influence of these two lymphocyte phenotypes in the patient subgroups – one after total laryngectomy with confirmed lymph node metastases and the other group without lymph node metastases – showed their prognostic value. Our analysis of lymphocytic infiltration, mostly of CD 43 cells, in the neoplasm microenvironment indicated a prognostic value for determining a shorter survival time and the possibility of lymph node metastases in patients with recurrences of cancer. Received: 26 September 1998 / Accepted: 8 March 1999     

The Lublańska classification of precancerous lesions of the larynx

Laryngeal carcinomas are preceded by precancerous lesions in about 20% of cases. The macroscopical appearance of these lesions is not enough characteristic to define their malignant potential. The accurate identification of epithelial abnormalities of the laryngeal mucosa requires biopsy and microscopic evaluation. There are many histological classifications of laryngeal precancerous lesions used at present. Many of them are highly subjective and have low reproducibility. Moreover, the different grades of these classifications not always give distinct guidelines for clinician concerning the treatment modality. The Ljubljana classification seems to be easier, more readily applied and more reproducible. It uses the name "epithelial hyperplastic laryngeal lesion" (EHLL) which includes all alterations in laryngeal squamous epithelium. The four grades of EHLL are: 1. simple hyperplasia (thickening of epithelium due to augmentation of normal prickle cells), 2. abnormal hyperplasia (with increase of basal-like cells), 3. atypical, or risky hyperplasia (epithelium thickened by increase of basal-like cells with pronounced atypical features), and 4. carcinoma in situ (i.e. full thickness change with the features of malignancy but without stromal invasion). The criteria of Ljubljana classification are precise and gives a possibility to make a more clear-cut separation of cases with risk of developing carcinoma from those without it.     

A dendritic cell based hybrid cell vaccine generated by electrofusion for immunotherapy strategies in HNSCC

OBJECTIVE: Hybrid cells generated from dendritic cells (DC) and tumor cells provide tumor-associated antigens (TAA) in a polyvalent mode and therefore they have aroused interest in cancer immunotherapy. The present study was designed to investigate the hybrid cell generation and optimize its implementation for a TAA-target treatment of head and neck squamous cell carcinoma (HNSCC). METHODS: Hybrid cells from mature DC and laryngeal carcinoma cell line UTSCC-19A were generated by electrofusion. Fusion efficiency and viability were determined by flow cytometry, light and fluorescence microscopy analyses. RESULTS: The gradual electrofusion process constituted real human tumor and dendritic cell hybrids characterized by polynuclear cells and double staining as a result of overlay of red (HLA-DR:R-PE) and green (HEA:FITC) fluorescence. Furthermore, analyses have proven viability of fusion results, and factors influencing fusion yield were determined. CONCLUSION: Physical fusion of mature dendritic cells with laryngeal carcinoma cells provides a dendritic cell based hybrid cell vaccine as a quantitative prerequisite for anti-cancer vaccination. Specific cytotoxic T-lymphocytes need to be induced before hybrid cell application in clinical studies.     

Correlations between histopathological and biological findings in nasopharyngeal carcinoma and its prognostic significance

Forty-five consecutive cases of nasopharyngeal carcinoma were morphologically and immunocytochemically studied using monoclonal (anti-B and anti-T cell) and polyclonal (anti-S100 protein and antilysozyme) antibodies with the peroxidase-anti-peroxidase method to identify infiltrating lymphocytes (T and B cell) and histiocytes (monocytic/macrophagic and dendritic cells) in nasopharyngeal carcinoma. A variable density of dendritic cells was found within the tumor nests in 22 (49%) of 45 nasopharyngeal carcinomas examined; infiltrating macrophages were demonstrated in 15 (33%) specimens and around the tumor in almost all cases. Cases with moderate or marked density of dendritic cells (S100+) survived longer than those without such infiltration (mean 5-year survival rates of 31%, 55%, or 64% in patients with absent, moderate, or marked densities, respectively; P less than 0.05). A significant relationship between monocytic/macrophagic cells (lysozyme+) within the tumor and survival was also found (mean 5-year survival rate of 27% or 61% in patients with absent, moderate, or marked densities, respectively). However, lymphocytic infiltration was not statistically related to a better survival. Analyzing lymphocytic infiltration, we found a large prevalence of T cells in the neoplastic tissue without any prognostic significance. These data were correlated to different histological subtypes according to the principal histological classifications of nasopharyngeal carcinomas (Micheau, et al.; World Health Organization; Cologne University) to individualize the scheme which correlates best with prognosis and biological features of nasopharyngeal carcinomas. Our data suggest that, considering dendritic cells and macrophages within cancer nests, nasopharyngeal carcinoma histiotypes can be correlated to patient prognosis.