分泌抗人角蛋白单抗杂交瘤细胞株的建立及其抗体的初步鉴定

角蛋白(Keratin)作为一组分子量为40,000～70,000的不溶于水的细胞结构蛋白,广泛存在于哺乳类动物的上皮细胞内。而各种非上皮细胞中该蛋白成份缺如,因而角蛋白被认为是上皮来源细胞的一种标志物,引起了细胞生物学及病理学工作者的兴趣,尤其在肿瘤病理诊断方面,如通过角蛋白特异性抗体对肿瘤细胞进行角蛋白的     

64例原发性肝癌角蛋白抗体标记观察

细胞角蛋白是上皮型细胞中丝蛋白的一种,它存在于绝大多数上皮细胞及其相应的肿瘤细胞中,因此用角蛋白制备的抗体可用于低分化恶性上皮性肿瘤的鉴别诊断.由于角蛋白种类多,不同种上皮所含     

乳腺上皮-肌上皮性肿瘤4例临床病理学分析

目的：探讨乳腺上皮-肌上皮性肿瘤(epithelial-myoepithelial tumor of breast)的临床病理学特点、免疫表型、诊断及鉴别诊断。方法：对4例乳腺上皮-肌上皮性肿瘤的临床特点、组织形态学及免疫组织化学结果进行分析,并复习相关文献。结果：患者：男性1例,女性3例,平均年龄51岁(27~63岁)。4例肿瘤直径1.5~3.0 cm(平均2.0 cm),无包膜,切面灰白色。显微镜下可见肿瘤由双相增生的肌上皮细胞和腺上皮细胞构成,肌上皮细胞环绕腺上皮细胞构成特征的套管结构。免疫组织化学染色,腺上皮细胞表达CK8/18、CK7,肌上皮细胞表达p63、Calponin、CK5/6。1例诊断为腺肌上皮瘤(adenomyoepithelioma,AME),3例诊断为伴有癌的腺肌上皮瘤(恶性腺肌上皮瘤, malignant adenomyoepithelioma,MAME)。结论：乳腺上皮–肌上皮性肿瘤是少见的肿瘤类型,需与导管内乳头状瘤、化生性癌等鉴别。     

成人食管上皮细胞的体外培养及生物学特性

目的 培养成人正常食管上皮细胞,建立能够体外长期培养的食管上皮细胞系,为上皮细胞的体外研究提供实验材料.方法 取食管癌患者正常食管上皮,用0.25%Dispase酶和0.25%胰蛋白酶/0.02?TA消化获取成人食管上皮细胞,使用无血清角化细胞培养液培养,通过细胞形态学观察和角蛋白、上皮膜抗原(EMA)免疫组织化学染色鉴定细胞.结果 原代培养8d后,细胞汇合成片呈铺路石样生长,细胞角蛋白、上皮膜抗原表达阳性,可连续传代.结论 为体外分离培养成人正常食管上皮细胞建立了方便可行的方法.     

糖蛋白非转移性黑色素瘤蛋白B在肾嗜酸性肿瘤鉴别诊断中的意义

目的探讨糖蛋白非转移性黑色素瘤蛋白B(glycoprotein nonmetastatic melanoma protein B, GPNMB)在各种肾嗜酸性肿瘤中的表达情况, 并比较GPNMB与细胞角蛋白(CK)20、CK7、CD117在肾嗜酸性肿瘤鉴别诊断中的价值。方法收集南京大学医学院附属鼓楼医院病理科2017年1月至2022年3月传统类型肾肿瘤嗜酸细胞亚型, 包括22例肾透明细胞癌嗜酸细胞亚型(e-ccRCC)、19例肾乳头状细胞癌嗜酸细胞亚型(e-papRCC)、17例肾嫌色细胞癌嗜酸细胞亚型(e-chRCC)、12例肾嗜酸细胞瘤(RO)和新兴的具有嗜酸细胞特征的肾肿瘤类型, 包括3例嗜酸性实性囊性肾细胞癌(ESC RCC)、3例肾低级别嗜酸细胞肿瘤(LOT)、4例延胡索酸水合酶缺陷型肾细胞癌(FH-dRCC)以及5例肾脏上皮样血管平滑肌脂肪瘤(E-AML), 采用免疫组织化学EnVision法染色检测GPNMB与CK20、CK7、CD117在上述肿瘤中的表达情况, 并统计分析其在以上各种肾肿瘤鉴别诊断中的意义。结果 GPNMB在ESC RCC、LOT、FH-dRCC这3种新...     

兔角膜缘上皮细胞羊膜种植及生物学特性研究

研究兔角膜缘上皮细胞(含干细胞)在完整羊膜和去上皮羊膜上种植及生物学特征,探讨组织工程技术体外重建角膜上皮良好的方式和方法.将兔角膜缘上皮细胞分别种植于去上皮羊膜基底膜和完整羊膜上进行体外培养扩增.从倒置显微镜下的生长特性、光镜下的组织结构、超微结构、免疫组织化学检测等方面对培养获得的复合组织进行观察.结果表明兔角膜缘上皮细胞在完整羊膜上不易贴壁、生长缓慢、很难汇合成片.而在去上皮羊膜上细胞能黏附生长并增殖形成由羊膜基底膜和4～5层上皮细胞组成的复合组织,基底层细胞与羊膜之间有半桥粒连接,细胞之间可见桥粒连接,细胞表达角蛋白3.该复合物可作为组织工程化角膜上皮组织.     

人巨细胞病毒感染对腮腺导管上皮细胞角蛋白K8和K18表达的影响

目的 研究人巨细胞病毒(HCMV)感染对腮腺导管上皮细胞角蛋白K8和K18表达的影响.方法 用免疫组化方法研究腮腺巨细胞包涵体病(PCID)石蜡包埋组织中巨细胞病毒立即早期抗原和角蛋白K8、K18的表达.结果 PCID腮腺组织导管上皮出现包涵体巨细胞;包涵体巨细胞HCMV抗原DDG9/CCH2阳性;包涵体巨细胞角蛋白K8表达呈阴性,而K18表达呈强阳性.结论 HCMV感染腮腺导管上皮细胞诱发角蛋白K8降解,K18表达上调是一种反应性改变;单层上皮角蛋白网具有维持上皮细胞机械力学完整性的功能.     

BCL-2、CK20、CD10、AR表达在基底细胞癌和毛母细胞瘤鉴别诊断中的意义

目的 观察BCL-2、CK20、CD10、AR在基底细胞癌(basal cell carcinoma,BCC)和毛母细胞瘤(trichoblastoma,TB)中表达的不同,以提高它们在两者诊断和鉴别诊断中的作用.方法 对确诊的32例BCC和8例TB石蜡包埋标本行BCL-2、CK20、CD10、AR免疫组化染色,观察它们在肿瘤细胞及其周围间质中的表达.结果 32例BCC中BCL-2肿瘤上皮弥漫阳性表达30例,阴性2例;CK20均为阴性;CD10肿瘤上皮部分表达23例,阴性9例,肿瘤间质阳性4例,阴性28例;AR阳性表达7例.8例TB中BCL-2上皮部分均阳性表达,CK20散在阳性6例,阴性2例,CD10部分上皮阳性4例,阴性4例,间质阳性3例,阴性5例;AR均为阴性.统计结果,BCL-2、CK20表达差异有显著性意义(P＜0.01),AR、CD10差异无显著性意义.结论 BCL-2弥漫强阳性、CK20阴性支持诊断BCC,反之支持诊断TB.     

近端型上皮样肉瘤临床病理研究

目的 探讨近端型上皮样肉瘤(PES)的临床病理特点、诊断及鉴别诊断依据.方法 收集5例PES患者资料,免疫组织化学EnVision法染色.第一抗体选用细胞角蛋白(CK)、波形蛋白、上皮细胞膜抗原(EMA)、CD34、β-catenin、S-100蛋白、平滑肌肌动蛋白(SMA)、肌调节蛋白(MyoD1)、结蛋白、HMB45、CK7及CK20,观察和分析其临床病理学形态及免疫表型特征.结果 5例PES中女1例,男4例;发病年龄19-46岁,发生部位分别为:会阴部2例,下腹部、髂前上棘和臀部各1例,均表现为进行性增大的无痛性单发肿块.光镜下肿瘤细胞呈结节状排列,浸润性生长,瘤细胞大部分由相对独特的上皮样细胞组成,胞质丰富,嗜酸性;核卵圆形,肿瘤中心常见坏死.免疫组织化学染色示5例瘤细胞均表达波形蛋白,4例表达CK、EMA,3例表达β-catenin、CD34,1例表达S-100蛋白;而SMA、MyoD1、结蛋白、HMB-45、CK7及CK20均阴性.结论 根据PES组织学和病理学特征,结合免疫组织化学染色结果可以做出明确诊断.     

北京中山生物技术有限公司推荐新抗体

1 鼠抗ＣＫ5 / 6 (Ｄ5 / 16Ｂ4)单抗 :鼠抗ＣＫ5 / 6能与人角蛋白 5和 6起反应 ,并与角蛋白 4有微弱的交叉反应 ,但它与角蛋白 1、7、8、10、13、14、18和 19没反应。在正常鳞状上皮中该抗体能使基底细胞及部分棘细胞显色 ,也能显示部分前列腺上皮的基层细胞 ,一般来说它不能使其他单层上皮的腺体着色 ,因此该抗体可以用来鉴别鳞癌和腺癌。此外 ,ＣＫ5 / 6还可用做上皮和间皮分化的标志物。在正常乳腺中大多的肌上皮细胞和末梢导管、小叶单位的腺上皮细胞为主要表达部位。在良性导管增生性疾病 ,包括非典型性导管上皮增生则没有ＣＫ5 / 6的…     

Marker profile of different phases in the transition of normal human ovarian epithelium to ovarian carcinomas.

To investigate whether early changes in the transformation of normal ovarian epithelial cells into tumor cells can be detected with monoclonal antibodies, a comparative immunohistochemical study was performed on normal human ovarian mesothelial cells, cystomas, cystadenomas, ovarian carcinomas, as well as granulosa cell tumor. Using monoclonal antibodies against different keratin subtypes, it was shown that mesothelial cells, ovarian cysts, cystadenomas, and carcinomas all reacted positively with broad-spectrum anti-keratin monoclonal antibodies (MAbs), as well as with MAbs to keratins 7, 8, 18, and 19. Keratins 4 and 13 were not found in mesothelial cells, but positive groups of cells were identified in several cystomas, adenomas, and carcinomas. While mesothelial cells did not react with the pan-epithelial marker BW495/36, invaginating metaplastic mesothelial cells, inclusion cysts, cystomas, adenomas, and carcinomas showed an increasing reactivity with BW495/36, with an increasing degree of malignancy. The reactivity of MAbs against ovarian carcinoma-associated antigens (OV-TL 3, OC 125, MOv 18, and OV-TL 10) was limited to weak staining reaction in some mesothelial cells but were found to be positive on more than 50% of the ovarian cystadenomas and more than 90% of the ovarian carcinomas. Thecal and granulosa cells of primordial, primary, and secondary follicles all reacted positively with antibodies to the broad-spectrum keratins OV-TL 12/5 and RCK 102, and to keratins 8 and 18, but not with keratins 4, 7, 13, and 19. These keratins decreased or disappeared in granulosa cells of mature follicles (Graafian follicles), whereas granulosa cell tumors did not react with anti-keratin antibodies. The reactivity of BW 495/36 was negative or limited to traces in some granulosa cells. Ovarian carcinoma-associated antigens were not expressed in granulosa cells or granulosa cell tumors. The data indicate that mesothelial cells undergoing metaplastic changes finally resulting in ovarian cystadenomas (and carcinomas) initiate the synthesis of a 200-kd glycoprotein recognized by MAb (BW 495/36), the production of ovarian carcinoma associated antigens, in addition to focal production of keratin 4 and/or 13, as seen in several samples. The granulosa cell tumors decrease or switch off their keratin production and remain negative for the 200-kd glycoprotein and the ovarian carcinoma-associated antigens.     

The distribution of vimentin and keratin in epithelial and nonepithelial neoplasms. A comprehensive immunohistochemical study on formalin- and alcohol-fixed tumors

Vimentin has been regarded as the intermediate filament characteristic of normal and neoplastic mesenchymal cells and keratins as typical of epithelial cells and the neoplasms derived from them. However, many epithelial cells in tissue culture or in effusion, as well as cells in some solid epithelial neoplasms such as renal, endometrial, ovarian, pulmonary, and thyroid adenocarcinomas, have been shown to coexpress vimentin and keratin. The recent availability of monoclonal antibodies that work reasonably well in paraffin-embedded tissue led us to carry out a comprehensive immunohistochemical study on formalin- and alcohol-fixed specimens of neoplasms in which we used monoclonal antibodies against vimentin. These results were compared with our previous study in which the same tumor tissues were investigated using antibodies against keratin. The antigenicity of vimentin was found to be preserved in all alcohol-fixed specimens and in 63% of formalin-fixed tissues. Vimentin was the sole intermediate filament present in virtually all sarcomas, meningiomas, schwannomas, and melanomas. In addition, variable percentages (10-57%) of carcinomas, neuroendocrine carcinomas, neuroblastomas, thymomas, and mesotheliomas were positive for vimentin, which, except in the neuroblastomas, was coexpressed with keratins. Among the adenocarcinomas, more than 50% of papillary carcinomas of the thyroid, as well as renal, endometrial, ovarian, and lung carcinomas, coexpressed keratins and vimentin. A distinctive paranuclear and basal localization of vimentin was observed in the cells of many of these tumors, in contrast to the predominantly apical distribution of the keratins. The authors conclude that coexpression of vimentin and keratin is more widespread than previously reported and that antibodies to vimentin, by themselves, are of limited value for the differentiation of epithelial from mesenchymal neoplasms.     

Epithelial cell heterogeneity in the guinea pig thymus: immunohistochemical characterization of four thymic epithelial subsets defined by monoclonal anti-keratin antibodies

Keratins are a family of related polypeptides constitutive of the cytoskeleton of epithelial cells and are never found in nonepithelial tissues. Thymic epithelial cells (TEC), known to induce T cell differentiation, are the keratin-containing cells within the thymus. Using four monoclonal anti-keratin antibodies (KL1, KL4, AE2, AE3) directed against keratins of different molecular weight, we have investigated the guinea pig thymic epithelium. The immunohistochemical analysis of thymic cryostatic sections revealed that the keratin expression of TEC varied according to their location in the thymic lobula; the thymic cortex was specifically stained by AE3 whereas the thymic medulla and the subcapsular cortex were recognized by KL4. In addition, KL1 and AE2 exclusively labeled Hassall's corpuscles. The biochemical analysis of keratins extracted from the thymus showed that each TEC subset was characterized by an unique pattern of keratin polypeptides. This study extends the concept of thymic epithelium heterogeneity and suggests that anti-keratin antibodies which allow the typing of TEC subsets may be valuable tools for studying the differentiation of thymic epithelium and its in vitro function on T lymphocytes.     

Expression of a neural type of intermediate filament as a distinguishing feature between oat cell carcinoma and other lung cancers.

Expression of intermediate filaments was studied immunohistologically in oat-cell (6 cases), epidermoid (9 cases), adeno- (7 cases), large-cell anaplastic (3 cases), and bronchioalveolar carcinoma (3 cases), of lung. Affinity-purified antibodies against epithelial (anti-keratin), neural (anti-neurofilament), muscle (anti-desmin) and mesenchymal (anti-vimentin) intermediate filaments were used. In indirect immunofluorescence microscopy of oat-cell carcinomas a positive cytoplasmic fluorescence was seen only with antibodies against neural intermediate filaments, neurofilaments, while no staining of the tumor cells was observed with antibodies against other types of intermediate filaments. On the other hand, all the epidermoid, adeno, anaplastic, and bronchioalveolar carcinomas showed constantly a strong reaction with anti-keratin antibodies in a varying number of cells but no decoration with anti-neurofilament antibodies. The results show that expression of neural intermediate filaments is a major distinguishing feature between oat-cell carcinomas and other lung cancers and suggest that anti-neurofilament antibodies can be used as a diagnostic aid in the surgical pathologic study of pulmonary neoplasms.     

The limited difference between keratin patterns of squamous cell carcinomas and adenocarcinomas is explicable by both cell lineage and state of differentiation of tumour cells.

AIM: To study the differentiation of epithelial tissues within their histological context, and to identify hypothetically, on the basis of keratin pattern, the putative tissue origin of a (metastatic) carcinoma. METHODS: Using well characterised monoclonal antibodies against individual keratins 7, 8, 18, and 19, which are predominantly found in columnar epithelia, and keratins 4, 10, 13, and 14, predominantly expressed in (non)-keratinising squamous epithelia, the keratin patterns for a series of 45 squamous cell carcinomas and 44 adenocarcinomas originating from various epithelial tissues were characterised. RESULTS: The predominant keratins in all adenocarcinomas proved to be 8, 18, and 19. In addition, these keratins were also abundantly present in squamous cell carcinomas of the lung, cervix, and rectum and, to a lesser extent, of the larynx, oesophagus, and tongue, but not in those of the vulva and skin. Keratins 4, 10, 13, and 14 were present in almost all squamous cell carcinomas, but also focally in some of the adenocarcinomas studied. CONCLUSIONS: There is a limited differential expression of distinctive keratin filaments between squamous cell carcinomas and adenocarcinomas. Apparently, squamous cell carcinomas that originate from columnar epithelium by squamous metaplasia gain the keratins of squamous cells but retain the keratins of columnar epithelial cells. However, the simultaneous expression of two of three squamous keratins (4, 10, and 13) identifies a squamous cell carcinoma, and thus might be useful in solving differential diagnostic problems.     

Mapping of the keratin polypeptides in meningiomas of different types: an immunohistochemical analysis of 463 cases

Keratins are known to be expressed in some meningiomas, and they have been consistently reported in secretory meningiomas. However, the expression of individual keratin polypeptides has not yet been systematically explored, and such a study could provide important information for the differential diagnosis of meningioma and other tumors, particularly metastatic carcinomas. In this study we analyzed the keratin polypeptide patterns of 463 meningiomas of different types using well-characterized monoclonal antibodies specific to 11 individual keratins and 3 additional antibodies that recognize more than 1 keratin. Archnoid tissues from autopsies were examined for comparison. Secretory meningiomas showed consistent positivity for all simple epithelial keratins K7, K8, K18, and K19, usually limited to the slender spindle cells lining the gland-like lumina. Other keratins variably detected in a minority of gland-like structures were K5/6, K14, K16, and K17. Among other meningiomas, keratin 18 was commonly present in a significant number of tumor cells in different subtypes (30% to 80% of cases), often extensively. The K18 positivity of meningiomas paralleled that observed in normal arachnoids, which were negative for K7, K8, K19, and AE1. Only rare meningiomas, other than the secretory ones, had significant positivity for K7, K8, K19, and other keratins than K18. The concentric spindle cells around psammoma bodies and few other spindle cell foci were often focally positive for K7 and to lesser degree for K8, K14, and K19. The complex pattern of keratins in meningiomas has to be considered in the differential diagnosis of meningioma and metastatic carcinoma. In this context, antibodies to K7, K8, and K19 and the antibodies AE1 and AE3 are useful, because they only rarely label significant numbers of meningioma cells but are positive in a great majority of carcinomas. Careful histologic analysis is necessary to differentiate anaplastic meningiomas and metastatic carcinomas, which have overlapping patterns of several keratins except K20, which was never present in any type of meningioma.     

Immunolocalization of keratin proteins in sweat gland tumours by the use of monoclonal antibody

A total of 34 cases (eccrine poroma: 2, cylindroma: 2, eccrine spiradenoma: 4, syringocystadenoma papilliferum: 1, hydroadenoma papilliferum: 1, clear cell hydroadenoma: 7, mixed tumour: 16) of sweat gland tumours of the skin were described in terms of immunohistochemical distribution of keratins using polyclonal anti-keratin antiserum (TK, detecting 41-56 KDa keratins) and monoclonal antibodies (KL1, 55-57 KDa; PKK1, 40, 45, 52.5 KDa). Keratin expression in eccrine poroma, spiradenoma and syringocystadenoma was similar to that in the ductal segment of normal sweat glands. Cylindroma showed usually slight staining for kertins. Tumour cells of hydroadenomas showed not so prominent staining for any of the keratins; however, histologically, tumour cells indicated marked variation, and the degree of keratin proteins also was different among these histological variants. Mixed tumours of the skin were strongly decorated with anti-keratin antibodies along the luminal surface cells of typical structures, while no staining occurred in outer side cells. Luminal tumour cells may be differentiated from secretory coil cells, whereas outer side cells may have a myoepithelial origin, as outer layer cells found in pleomorphic adenoma of salivary glands.     

Prognostic value of keratin subtyping in transitional cell carcinoma of the upper urinary tract

To investigate the prognostic value of keratin subtyping in invasive transitional cell carcinomas (TCCs), we performed a systematic study applying 15 different monoclonal keratin antibodies on 53 upper urinary tract TCCs using a tissue microarray technique. Immunoreactivity was correlated with pT stages and tumour grades using the Fishers exact test. Impact on disease-free survival was analysed using the Kaplan-Meier method and compared by the log-rank test. Immunoreactivity for keratins 5/6, 6, 7, 8, 13, 14, 17, 18, 19, 20, low molecular weight (LMW) keratins (8, 18) and high molecular weight (HMW) keratins (1, 5, 10, 14) was detected in varying quantities. Regarding semi-quantitative assessment, a prognostic impact was found for keratins 5/6, 7, 8, 13, 17, 20 and HMW keratins, with reduced expression or loss of immunoreactivity being significantly associated with disease progression. With respect to analysis of staining patterns, the retention of a basally accentuated labelling for keratin 5/6 and HMW keratin as well as a superficially accentuated labelling for keratin 20 was significantly associated with a favourable outcome. In conclusion, this investigation is the first to demonstrate a possible prognostic value for keratin subtyping in invasive (upper urinary tract) TCCs with respect to metastasis-free survival. Further studies, however, are needed to substantiate our results.     

An immunoperoxidase study of renal cell carcinomas: correlation with nuclear grade, cell type, and histologic pattern

We applied a panel of antibodies to formalin-fixed, paraffin-embedded sections of 55 renal cell carcinomas using a three-stage immunoperoxidase technique. The antibody panel included two anti-keratins, AE1 and CAM5.2, anti-epithelial membrane antigen (EMA), anti-vimentin, anti-S100 protein, and the anti-leukocyte marker PD7/26. Forty-eight of 55 renal cell carcinomas expressed keratins. CAM5.2 stained 46 tumors (84%) and AE1 stained 37 neoplasms (67%). AE1 reacted with two CAM5.2-negative tumors. EMA was expressed by 35 carcinomas (64%), including three of the CAM5.2-negative neoplasms. Therefore, using all three antibodies, 50 neoplasms (91%) expressed antigens of epithelial differentiation. Anti-EMA and AE1 were complementary to each other; the combination stained 46 of the carcinomas, comparable with CAM5.2 alone. Vimentin was expressed by 26 tumors (47%), and S100 was expressed by one. PD7/26 did not stain any of the cases. Vimentin expression correlated with nuclear grade; low nuclear grade neoplasms infrequently expressed vimentin, while the converse was true for high nuclear grade tumors. Keratin expression was related to tumor cell type and histologic pattern, as fewer neoplasms of clear cell type and with a solid pattern expressed keratins. In contrast, all papillary and eight of nine (89%) spindled carcinomas expressed keratins.     

A series of 14 new monoclonal antibodies to keratins: characterization and value in diagnostic histopathology.

A series of 14 new mouse monoclonal antibodies (MAbs) to keratins is described and the data suggesting their potential value in the differential diagnosis of human tumours are reported. The specificities of individual MAbs of the 'C-series' presented here range from monospecificity for keratin No. 7 (MAbs C-18, C-35, C-62, and C-68), keratin No. 8 (MAbs C-15, C-43, and C-15), and keratin No. 18 (MAbs C-04 and C-08) up to the broadly reacting 'pan-keratin' MAb C-11, with the target epitopes of the remaining four MAbs being shared by different pairs of keratin polypeptides. The results of the biochemical characterization of the MAbs, together with their immunohistochemical staining patterns on frozen as well as on paraffin sections of normal human tissues, suggest that they represent a significant contribution to the growing list of anti-keratin MAbs applicable in both research and routine diagnostic pathology. The immunohistochemical examination of a wide range of human neoplasms with the new MAbs not only confirmed their value in making distinctions between carcinomas, on the one hand, and lymphomas, and gliomas, on the other, but also verified the possibility of more subtle subdivisions within the group of adenocarcinomas and their metastases. Furthermore, the identification of small subsets of breast carcinomas with decreased levels or apparent loss of the keratin No. 7 polypeptide and some cases of stomach carcinoma with apparently induced expression of this keratin suggests that such 'exceptions' must be considered when using keratin spectra as one of the criteria in differential diagnosis.