人疱疹病毒6型对EB病毒复制影响的研究

目的探讨人疱疹病毒６型（ＨＨＶ６）对ＥＢ病毒（ＥＢＶ）复制的影响。方法用ＨＨＶ６感染Ｒａｊｉ细胞和ＥＢＶ转化的淋巴细胞（ＬＣＬ），并用抗ＨＨＶ６单克隆抗体（ＭＡｂ）作免疫萤光分析（ＩＦＡ）确定ＨＨＶ６已感染这二种细胞。用抗ＥＢＶ人血清作ＩＦＡ观察ＥＢＶ抗原表达。结果ＨＨＶ６感染Ｒａｊｉ细胞后可见细胞病变效应（ＣＰＥ）；而ＬＣＬ未见ＣＰＥ。这二种细胞株用抗ＨＨＶ６的ＭＡｂ作ＩＦＡ均检出ＨＨＶ６，而对照组未检出ＨＨＶ６；用抗ＥＢＶ人血清作ＩＦＡ，感染ＨＨＶ６的二种细胞中均检出ＥＢＶ抗原。结论ＨＨＶ６促进了ＥＢＶ抗原表达，并提示在鼻咽癌的发生中ＨＨＶ６和ＥＢＶ可能起协同致病作用。     

血管内皮生长因子及其受体对喉癌细胞生长的影响

为探讨血管内皮生长因子（ＶＥＧＦ）在喉癌中的作用及可能的作用方式，用抗ＶＥＧＦ抗体及其受体ｆｌｔ抗体通过微波处理暴露抗原后，进行标准化的ＥｌｉｔｅＡＢＣ染色；将不同浓度的抗ＶＥＧＦ及抗ｆｌｔ抗体分别加入ＨＥＰ－２细胞的培养液中，培养５ｄ后测ＨＥＰ－２细胞的活性（ＭＴＴ法）。结果：免疫组化染色显示ＶＥＧＦ及其受体ｆｌｔ主要在喉癌细胞和血管内皮细胞表达；不同浓度的ＶＥＧＦ抗体及抗ｆｌｔ抗体对ＨＥＰ－２     

鼻咽泡状核细胞癌和低分化鳞癌的临床生物学行为的对比研究

目的：探讨鼻咽泡状核细胞癌（ＶＮＣＣ）和低分化鳞癌（ＰＤＳＣＣ）的临床生物学行为的差异性。方法：用原位杂交和免疫组化染色（ＳＰ法）技术,检测ＶＮＣＣ和ＰＤＳＣＣ癌组织中ＣｅｒｂＢ２、表皮生长因子受体（ＥＧＦＲ）和增殖细胞核抗原（ＰＣＮＡ）表达,用原位末端标记（ＩＳＬＥ）方法检测癌细胞凋亡,并对其预后和临床特征进行对比分析。结果：①ＶＮＣＣ的ＩＳＥＬ和ＰＣＮＡ染色强度指数（ＳＩ）明显高于ＰＤＳＣＣ;②ＶＮＣＣ的ＣｅｒｂＢ２、ＥＧＦＲｍＲＮＡ和ＣｅｒｂＢ２蛋白表达与ＰＤＳＣＣ无明显差别;③ＶＮＣＣ的５年生存率比ＰＤＳＣＣ高。结论：与ＰＤＳＣＣ比较,ＶＮＣＣ是一种生长增殖活跃、分化程度更低,但有较好预后趋向的一种癌型。     

γ—干扰素对喉癌细胞株HEP—2增殖活性的影响

用单克隆抗体Ｋｉ－６７（抗ＰＣＮＡ），以链霉菌素－生物素技术（ＬＳＡＢ）检测喉癌细胞株ＨＥＰ－２增殖细胞核抗原（ＰＣＮＡ）表达。人重组γ－干扰素（ｒｈｕ－ＩＦＮ－γ）抑制ＰＣＮＡ表达（即抗增殖活性）强弱与其剂量有关；雌激素对ｒｈｕ－ＩＦＮ－γ抗增殖作用无影响。提示：ｒｈｕ－ＩＦＮ－γ对喉癌细胞株ＨＥＰ－２有抗增殖作用，因而在喉癌的治疗中具有应用价值的。     

头颈部癌化疗药敏试验研究

为指导头颈肿瘤患者化疗的用药选择对２７例新鲜头颈部Ⅲ、Ⅳ期癌组织行四唑蓝（Ｍｅｔｈｙｌｔｈｉａｚｏｌｙｌｔｅｔｒａｚｏｌｉｕｍ）比色法（简称ＭＴＴ法）８种药物敏感性测试，成功率为９２．６％。药敏阳性率与对应药物文献报道的单药化疗有效率极为相似，药敏顺序为：ＰＹＭ（平阳霉素，Ｐｉｎｇｙａｎｇｍｙｃｉｎ）＞ＨＨＡ（高三尖杉酯碱，Ｈｏｍｏｈａｒｒｉｎｇｔｏｎｉｎｅ）＞ＭＴＸ（甲氨喋呤，Ｍｅｔｈｏｔｒｅｘａｔｅ）＞ＣＢＤＣＡ（卡铂Ｃａｒｂｏｐｌａｔｉｎ）＞ＭＭＣ（丝裂霉素Ｃ，ＭｉｔｏｍｙｃｉｎＣ）＞５－Ｆｕ（氟尿嘧啶，５－Ｆｌｕｏｒｏｕｒａｃｉｌ）＞ＶＣＲ（长春新碱，Ｖｉｎｃｒｉｓｔｉｎｅ）＞ＶＰ１６（足叶乙甙，Ｅｔｏｐｏｓｉｄ）。认为ＭＴＴ法为头颈部癌药敏测试较为理想的方法。头颈癌对ＨＨＡ有较高敏感性，可将其用于头颈癌药敏测试和化疗中。头颈癌原发部位、鳞癌分化程度及雌激素受体（ｅｓｔｒｏｇｅｎｒｅｃｅｐｔｏｒ，以下简称为ＥＲ）表达不同，其化疗敏感性无差异。鳞癌的化疗敏感性显著高于腺样囊性癌。异倍体癌肿的化疗敏感性显著高于二倍体癌肿。     

EB病毒感染P53及P16表达与鼻腔咽淋巴环非霍奇金淋巴瘤 …

目的 观察ＥＢ病毒（Ｅｐｓｔｉｎ－Ｂａｒｒ ｖｉｒｕｓ,ＥＢＶ）、抑癌基因Ｐ５３、Ｐ１６在鼻、鼻咽和扁桃体非霍奇金淋巴瘤（ｎｏｎ－Ｈｏｄｇｋｉｎ’ｓ ｌｙｍｐｈｏｍａ,ＮＨＬ）中的表达,并分析其相互间关系。方法 用原位分子杂交检测３６例病变组织的ＥＢＶ－ＤＮＡ、Ｐ１６ｍＲＮＡ,用免疫组化技术检测ＬＭＰ１、Ｐ５３及Ｐ１６蛋白,５例原位杂交检测为ＥＢＶ阴性的病例用原位ＰＣＲ检测ＥＢＶ－ＤＮＡ。结果 ①     

鼻咽癌活检组织中HLA,EBNA及间质中T淋巴细胞亚群的…

本实验对人类白细胞抗原（ＨＬＡ）、爱波斯坦－巴尔病毒核抗原（ＥＢＮＡ）、Ｔ细胞亚群的表达及分布进行综合分析，探讨其在癌变过程中的作用。用单克隆抗体Ｗ６／３２（抗ＨＬＡ－Ａ，Ｂ，Ｃ）、ＣＲ３／４３（抗ＨＬＡ－ＤＲ，ＤＰ，ＤＱ）、和Ｔ４、Ｔ８、Ｔ１１，以碱性磷酸酶抗碱性磷酸酶法（ＡＰＡＡＰ）和用鼻咽癌患者血清以抗补体Ｃ３免疫荧光法，检测鼻咽癌（２５例）及慢性鼻咽炎（１０例）活检组织中ＨＬＡ、Ｔ细胞亚群     

人类中耳渗出液中的血小板活化因子-乙酰水解酶

血小板活化因子（ＰＡＦ）是一种有效的脂质中间物,在中耳渗出液（ＭＥＥ）中起着重要的作用。该项研究旨在观察慢性分泌性中耳炎（ＣＯＭＥ）血清和粘液型中耳液中的血小板活化因子－乙酰水解酶（ＰＡＦ－ＡＨ）的活性以揭示慢性中耳炎渗出液的生物化学特性。ＭＥＥ从ＣＯＭＥ鼓膜切开或鼓室植入管中收集,每种标本立即在－２０℃下保存。使用时ＭＥＥ用０．２５ｍｏｌ／Ｌ蔗糖等容稀释１０次。人类血浆取自ＣＯＭＥ患者和正常人,以ＥＤＴＡ作为抗凝剂,每种标本于－８０℃下保存,使用时血浆用０．２５ｍｏｌ／Ｌ蔗糖等容稀释１０次。细…     

白三烯C_4和D_4能增加离体人类上呼吸道粘膜纤毛摆动频率

抗原刺激能够影响人类上呼吸道粘膜纤毛摆动频率（ＣＢＦ）,在变态反应过程中上呼吸道白三烯Ｃ４（ＬＴＣ４）和Ｄ４（ＬＴＤ４）水平均有升高。由于腺样体上皮和其它上呼吸道粘膜在组织学上的相同性,作者研究了ＬＴＣ４和ＬＴＤ４在５小时内对离休人类腺样体ＣＢＦ的作用。腺样体上皮标本培养于ＭＥＭ培养基中。ＬＴＣ４和ＬＴＤ４的使用浓度分别为１０－６和１０－８Ｍ,白三烯受体特异阻断剂ＬＹ－１７７８８３的浓度为１０－６Ｍ。ＣＢＦ用相差显微技术和显微成像系统测定。结果发现,加白三烯之前ＣＢＦ均值为５．５１±０．５８Ｈｚ…     

鼻咽癌活检组织中HLA、EBNA及间质中T淋巴细胞亚群的分布

本实验对人类白细胞抗原（ＨＬＡ）、爱波斯坦－巴尔病毒核抗原（ＥＢＮＡ）、Ｔ细胞亚群的表达及分布进行综合分析，探讨其在癌变过程中的作用。用单克隆抗体Ｗ＿６／３２（抗ＨＬＡ－Ａ，Ｂ，Ｃ）、ＣＲ＿３／４３（抗ＨＬＡ－ＤＲ，ＤＰ，ＤＱ）、和Ｔ＿４、Ｔ＿８、Ｔ＿（１１），以碱性磷酸酶抗碱性磷酸酶法（ＡＰＡＡＰ）和用鼻咽癌患者血清以抗补体Ｃ＿３免疫荧光法，检测鼻咽癌（２５例）及慢性鼻咽炎（１０例）活检组织中ＨＬＡ、Ｔ细胞亚群分型和ＥＢＮＡ的表达。结果显示，鼻咽癌癌细胞多有ＨＬＡ－Ⅰ和Ⅱ型抗原表达，而以ＨＬＡ－Ⅱ型抗原表达为明显；慢性鼻咽炎上皮细胞表达ＨＬＡ－Ⅰ和Ⅱ型抗原较弱，与癌细胞比较，差异有显著性（Ｐ＜０．０５）。提示在癌变过程中，存在着ＨＬＡ表达的异常，体内ＥＢ病毒特异性Ｔ细胞可能不能识别带ＨＬＡ－Ⅱ型抗原的癌细胞，因而使其逃脱了机体的免疫监视。鼻咽癌组织中，癌细胞ＥＢＮＡ呈阳性和强阳性；癌间质中Ｔ细胞总数、Ｔ辅助、Ｔ杀伤细胞数量均减少，证明鼻咽癌局部细胞免疫功能降低。     

Cell fusion by nasopharyngeal carcinoma-derived Epstein-Barr virus

Cell fusion experiments using Epstein-Barr virus (EBV) obtained from nasopharyngeal carcinoma (NPC) hybrid cells (NPC-KT) were carried out to examine whether human adenoid epithelial cells (Ad-AH) could be fused with lymphoblastoid cells (BJAB, Raji, and A2L) superinfected or infected by EBV, and if Ad-AH nuclei of the fused cells could express EBV-associated nuclear antigen. The presence of NPC EBV could induce EBV early antigen in all three lymphoblastoid cell lines. By autoradiography, we found that NPC EBV could induce cell fusion between Ad-AH cells and tritiated thymidine-labeled lymphoblastoid cells. In addition, we found that Ad-AH nuclei of the fused cells expressed EBV-associated nuclear antigen four days after NPC EBV-mediated cell fusion of Ad-AH cells with lymphoblastoid cells.     

Suppressive effect of CDDP on induction of the Epstein-Barr virus antigen synthesis by n-butyrate

n-Butyrate has been shown to induce Epstein-Barr virus (EBV) antigen synthesis in certain EBV genome-carrying lymphoblastoid cell lines. We have studied the effect of cis-dichloro-diamine-platinum (CDDP) on induction of EBV antigen synthesis not only by n-butyrate treatment of the EBV producer, P3HR-1 cells, but also by superinfection of EBV to the EBV non-producer, Raji cells. Simultaneous treatment of the cells with n-butyrate and CDDP or MMC blocked VCA and EA induction, but no suppressive effect was observed by the treatment of the cells with n-butyrate and Ara-C. The induction of EBV antigens was also observed in EBV-superinfected Raji cells, even in the presence of DNA synthesis inhibitor, CDDP or MMC. From these results, we discussed the mechanisms of the EBV antigens synthesis in EBV latently infected cells.     

Primary EBV infection of human umbilical cord lymphocytes and EBV genome-negative lymphoblastoid cell lines (BJAB and Ramos)

The appearance of Epstein-Barr virus (EBV)-associated nuclear antigen (EBNA) and induction of EBV-induced early antigen (EA) in human umbilical cord blood lymphocytes (HUCLs) and two EBV genome-negative Burkitt's lymphoma (BL) lines (BJAB and Ramos) were studied by infection with EBVs prepared from three different cell lines: marmoset cell line (B95-8) derived from infections mononucleosis, BL-derived cell line (P3HR-1) and human epithelial hybrid cell line (NPC-KT) derived from nasopharyngeal carcinoma. B95-8 virus can transform HUCLs but cannot superinfect Raji cells. P3HR-1 virus can transform HUCLs cells but cannot transform HUCLs. NPC-KT virus can transform HUCLs and can superinfect Raji cells. We have examined the time sequence of EBNA appearance and EA induction in HUCLs, BJAB cells and Ramos cells, in order to determine if three different strains of EBV differ in their abilities to infect their cells. We found that all three strains of EBV can induce EBNA in HUCLs, BJAB cells and Ramos cells. On the other hand, we found that P3HR-1 virus and NPC-KT virus can induce EA in BJAB cells and Ramos cells, but B95-8 virus cannot induce EA in their cells.     

Interactions between Epstein-Barr virus (EBV) and human cell lines: growth and EBV induction

Differences in in vitro growth characteristics between Epstein-Barr virus (EBV) genome-negative and genome-positive lymphoblastoid cell lines and between EBV genome-negative and genome-positive epithelial cell lines were investigated. All EBV genome-positive cells were able to grow in medium containing only 0.75% fetal calf serum (FCS), whereas no EBV genome-negative cells were able to grow at this low concentration. However, there was no clear difference between the growth of the two EBV genome-negative lymphoblastoid cell lines (BJAB and Ramos) and two EBV genome-positive BJAB and Ramos sublines (BJAB/B95-8 and Ramos/B95-8) in medium containing more than 2.5% FCS. On the other hand, an EBV genome-positive clone of A2L/AH epithelial hybrid cells (cl-1) grew in both concentrations of FCS better than an EBV genome-negative clone of A2L/AH cells (cl-654). In addition, there was a difference in EBV induction between EBV genome-positive lymphoblastoid (BJAB/B95-8 and Ramos/B95-8) and epithelial (cl-1) cells after treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA). Treatment of BJAB/B95-8 and Ramos/B95-8 cells with TPA induced a low percentage of EBV-induced early antigen but not viral capsid antigen, whereas treatment of cl-1 cells with TPA induced both early antigen and viral capsid antigen.     

Epstein-Barr virus (EBV) receptors on epithelial hybrid cells derived from nasopharyngeal carcinoma

We previously established an Epstein-Barr virus (EBV) genome-positive epithelial hybrid cell line, designated NPC-KT, which was prepared by fusing primary epithelial cells derived from nasopharyngeal carcinoma (NPC) with an epithelial Ad-AH cell line. In this study, we measured the presence of the EBV receptors using radiolabelled EBV. It was determined that similar amounts of Burkitt's lymphoma (BL)-derived P3HR-1 or NPC-KT-derived EBV can attach BL-derived Raji cells, but that only P3HR-1 virus can attach to NPC-KT cells. In addition, the superinfection of NPC-KT cells with P3HR-1 virus could not be inhibited by pretreatment with a monoclonal antibody against C3d (OKB7). Raji cells adsorbed with OKB7 became much less susceptible to superinfection with P3HR-1 or NPC virus. The data suggest that EBV receptors unrelated to C3d receptor exist.     

Role of cytomegalovirus, Epstein-Barr virus, and human herpes virus-8 in benign lymphoepithelial cysts of the parotid gland

OBJECTIVE: To provide background and evaluate the role of herpesviruses in benign lymphoepithelial cysts (BLC) of the parotid gland. STUDY DESIGN: Case series derived from review of pathology specimens. METHODS: Radiolabeled polymerase chain reaction (PCR) analysis was used to detect for the presence of cytomegalovirus (CMV), Epstein-Barr virus (EBV), and human herpes virus 8 (HHV-8) DNA sequences in 14 paraffin embedded specimens and 1 freshly aspirated BLC specimen. Thirteen normal parotid tissue specimens obtained from paraffin embedded blocks were used as a control group. RESULTS: CMV was detected with nearly equal frequency between the two groups (23% of normal vs. 20% in BLC). HHV-8 was found in 13% of the BLC group and in none of the normal group (P =.4841). There was significant difference in EBV detection between the normal (0%) and the BLC (33%) groups (P =.0437). CONCLUSION: CMV and HHV-8 does not appear to be associated with BLCs. Although EBV is found more frequently in BLC than in normal parotid controls, further studies are needed to elucidate the role of this virus in BLC pathogenesis.     

Infection of herpes simplex virus (HSV) and Epstein-Barr virus (EBV) in acute tonsillitis--histopathological assessment by optical and electron microscopic observation of biopsy specimens of tonsils]

Infection with HSV or EBV was studied by measuring serum antiviral antibody titers in adults with acute tonsillitis, and results were compared to light and electron microscopy findings of tonsil biopsy specimens. The clinical and laboratory features of acute tonsillitis caused by HSV or EBV were also studied. Subjiects were 42 patients with acute tonsillitis treated at the Department of Otorhinolaryngology at Tokyo Women's Medical University Daini Hospital between August 1997 and March 2000. They had failed to respond to antimicrobial agents prescribed by a physician, and had severe oropharyngeal mucosal lesions, liver dysfunction, skin eruptions, or cervical lymphadenopathy, with hospitalization required because of impaired food intake due to sore throat or deterioration in general condition. Subjects were 24 men (mean age: 30.8 years) and 18 women (mean age: 28.3 years) aged 16 to 78 years (mean: 29.8 years). A underwent, bacteriological and hematology tests and palatine tonsil biopsy specimens were obtained to examine tissue changes by light microscopy and electron microscopy due to detect HSV antigen by immunohistochemistry and EBV nucleic acids by EBV-encoded small nuclear RNA 1 and 2 (EBER) in situ hybridization (ISH). Among patients, the serum antiviral antibody profile indicated that 4 (9.5%) had acute tonsillitis due to primary HSV infection and 5 (11.9%) had acute tonsillitis due to primary EBV infection. The findings characteristic of acute tonsillitis due to primary HSV infection included stomatitis, skin eruptions, atypical lymphocytes, and liver dysfunction. Findings characteristic of acute tonsillitis due to primary EBV infection included petechiae of the soft palate, an increase of lymphocytes, atypical lymphocytes, and liver dysfunction. At the initial test, serum anti-HSV antibody was positive in 14 patients (33.3%), and more than half had no history of prior infection. Anti-EBNA antibody was positive in 32 (76.2%), and many had been infected previously. It should be noted that a decrease in positive HSV antibody means that acute tonsillitis due to primary HSV infection is not uncommon in adults and is expected to increase steadily. Light microscopy revealed histological changes in 2 patients. HSV antigen was positive in 2 (50%) with acute tonsillitis due to primary HSV infection, while EBER cells were positive in 5 (100%) with acute tonsillitis due to primary EBV infection, so special staining of the tissues was found to be useful. Electron microscopy failed to detect viral particles in ultrathin sections and no differences were seen in morphological changes or tissue damage between patients with positivity for HSV antigen and with EBER-positive cells. Detection of HSV antigen and EBV nucleic acids in pathological specimens from patients with acute tonsillitis requires careful judgment, but is considered useful for making an early diagnosis and for making a diagnosis in patients without an increase of the antiviral antibody titer and in those with reinfection or reactivation. Pathological examination (including special staining) and careful observation of clinical features may help to identify HSV or EBV infection and allow decisions to be made with regard to the therapeutic strategy and prevention of complications.     

Epstein-Barr virus infection of epithelial cells derived from primary cultures of adenoidal tissue

Summary Epithelial cells derived from primary cultures of adenoidal tissue were exposed to Epstein-Barr virus (EBV) from the throat washings of a patient with infectious mononucleosis (IM) and P3HR-1 and B95-8 cell strains. They were then examined for EBV-specific antigens by immunofluorescence. EBV from both the P3HR-1 cell strains and the throat washings of the IM patient infected the epithelial cells at the 6th and 9th days, respectively. Although the EBV-antigen-positive cells did not increase in number and disappeared at the 12th day after infection, EBV antigens of the virus's replication cycle could still be detected in the epithelial cells shed from cell culture layer at this time. However, the virus obtained from the cell-free 12th day's culture medium was incapable of transforming cord blood lymphocytes.Offprint requests to: M. Furukawa     

Marmoset lymphoblastoid cells transformed by NPC-KT-derived Epstein-Barr virus which possesses transforming and superinfecting characteristics

New world monkey (cotton-top marmoset; CTM) lymphocytes were transformed by two different strains of Epstein-Barr virus (EBV) derived from human nasopharyngeal epithelial/hybrid cells (NPC-KT and A2L/AH). M-KT cells were a CTM lymphoblastoid cell line which was transformed by EBV derived from NPC-KT cells (NPC-KT EBV). M-BA2L cells were a CTM lymphoblastoid cell line which was transformed by EBV derived from A2L/AH cells (A2L/AH EBV). EBV obtained from M-KT cells, like parental NPC-KT EBV, could transform human cord blood lymphocytes (HCBLs) and superinfect Raji cells. EBV obtained from M-BA2L cells, like parental A2L/AH EBV, could transform HCBLs but could not superinfect Raji cells. We have compared the EBV-DNA associated with M-KT cells to NPC-KT EBV-DNA. The results obtained using a DNA restriction enzyme (Hind III) showed that virus DNA prepared from NPC-KT cells is different from EBV-DNA prepared from CTM lymphocytes transformed by EBV derived from infectious mononucleosis (B95-8), but that virus DNA prepared from M-KT cells is identical with parental NPC-KT EBV. These results indicate the possibility that a single NPC-KT EBV is associated with two biological activities.     

Search for Herpesviruses in cerebrospinal fluid of facial palsy patients by PCR

CONCLUSIONS: Herpes simplex virus 1 (HSV-1) and varicella-zoster virus (VZV) DNA were not detected in the cerebrospinal fluid (CSF) of patients with acute idiopathic peripheral facial palsy (Bell's palsy). Our results indicate either the absence of these viruses or the presence of technical shortcomings. The role of human herpesvirus 6 (HHV-6) in this disorder and the significance of a positive HHV-6 DNA finding in the central nervous system need further investigation. OBJECTIVE: Our goal was to determine whether DNA of HSV-1, VZV, or HHV-6 can be found by polymerase chain reaction (PCR) in the CSF of peripheral facial palsy patients. MATERIALS AND METHODS: We used PCR to detect the presence of HSV-1, VZV, and HHV-6 DNA in CSF. This was a retrospective case control study with 33 peripheral facial palsy patients (34 CSF samples) in the study group (26 with Bell's palsy, 5 with simultaneously diagnosed herpesvirus infection, 1 with puerperal facial palsy, 1 with Melkersson-Rosenthal syndrome). The control group included 36 patients, most with diagnosed or suspected Borreliosis and facial palsy or sudden deafness. RESULTS: One patient with Bell's palsy had HHV-6 DNA in CSF. Neither HSV-1 nor VZV DNA was detected in patients or controls.