Human herpesvirus 8 (HHV-8) in familial Kaposi's sarcoma

Human herpesvirus 8 (HHV-8) has been detected in various epidemiological forms of Kaposi's sarcoma (KS). Since familial KS cases are exceedingly rare and the occurrence of familial KS in siblings has thought to depend rather on genetic factors than on a viral factor, familial KS has not been investigated for the presence of HHV-8. To investigate whether HHV-8 is present also in this rare form of KS, we examined tumor biopsies of 2 siblings with familial KS for the presence of HHV-8 specific DNA sequences by a nested PCR protocol. HHV-8 DNA sequences could be detected in KS specimens of both patients. Sequence analysis revealed an identical DNA sequence of HHV-8 in KS tissue of both siblings, but the sequence in our cases differs in one base pair at position 67 from the previously published HHV-8 KS330Bam fragment. The findings indicate that besides the yet poorly defined genetical factors involved in the pathogenesis of KS, HHV-8 may act as a cofactor also in familial KS. In addition, our data demonstrate that HHV-8 is found in all epidemiological forms of KS, including the rarely occurring familial KS. Familial KS may act as a further model to study the interaction of an oncogenic virus with genetic host factors in the context of a neoplastic disorder.     

Infection with Kaposi's sarcoma-associated herpesvirus among families of patients with classic Kaposi's sarcoma

BACKGROUND: Classic Kaposi's sarcoma (CKS) primarily affects elderly Mediterranean or Eastern European men. Incidence rates of CKS in Israel are among the world's highest. In practically all cases, antibodies against Kaposi's sarcoma-associated herpesvirus (KSHV) can be detected. A relatively high seroprevalence rate of KSHV in Israel generally correlates with the incidence of CKS. A sexual mode of virus transmission is recognized among homosexual men, whereas the precise transmission routes in the heterosexual population and those with CKS are still unclear. OBJECTIVE: To better assess the transmission routes of KSHV in Israeli patients with CKS and their first-degree relatives as compared with a control group. DESIGN: Serum was collected from all study participants and tested for KSHV antibodies by means of latent and lytic immunofluorescence assays. An open reading frame 65 (ORF65) Western blot assay was applied as a confirmatory tool. SETTING: Three dermatological departments in Israel. PATIENTS: Sixty-four Jewish patients with CKS, 143 of their first-degree relatives, and 186 hospital-based control subjects. RESULTS: Seropositivity to KSHV was detected in 62 (96.9%) of the patients with CKS, in 56 (39.2%) of their first-degree relatives, and in only 21 (11.3%) of the hospital controls (P<.001). The specific relationship with the index patient (spouse, offspring, or sibling) had no significant effect on the prevalence of serpositivity in the family members. CONCLUSION: Our serologic evidence of familial clustering of KSHV infection suggests a predominantly nonsexual horizontal transmission route of the virus.     

Expression of the CD34 antigen distinguishes Kaposi's sarcoma from pseudo-Kaposi's sarcoma (acroangiodermatitis)

Summary The differential diagnosis between Kaposi's sarcoma and the so-called 'pseudo-Kaposi's sarcoma' or acroangiodermatitis of the feet is often fraught with difficulty, not only on clinical but also on histological grounds. The aim of this study was to assess whether immunolabelling for the CD34 antigen, a marker of Kaposi's sarcoma cells, could be of value in the distinction between these two angioproliferative disorders. We comparatively examined 16 biopsy specimens from cases of Kaposi's sarcoma and seven biopsies from patients with pseudo-Kaposi's sarcoma, by a streptavidin-biotinperoxidase method, using a monoclonal antibody to the CD34 antigen. All cases of Kaposi's sarcoma showed CD34 labelling both on endothelial cells and on the characteristic spindle-shaped, perivascular cells. Biopsies of pseudo-Kaposi's sarcoma showed a strong labelling of endothelial cells of hyperplastic vessels. However, in sharp contrast with Kaposi's sarcoma, a complete absence of perivascular CD34 expression was noted. It seems therefore that immunolabelling for the CD34 antigen appears to be a valuable tool in the differential diagnosis between Kaposi's sarcoma and pseudo-Kaposi's sarcoma.     

HHV-8/KSHV during the development of Kaposi's sarcoma: evaluation by polymerase chain reaction and immunohistochemistry

The human gamma-herpes virus-8 (HHV-8) was first described in AIDS-related Kaposi's sarcoma (KS) tumour samples. In this study, we report comparative studies on paraffin-embedded biopsies of AIDS-related KS (AKS) and endemic KS (EKS) with regard to HHV-8 content as evaluated using polymerase chain reaction (PCR) and immunohistochemistry. DNA was extracted either using Chelex-100 or using Qia-gene kit and was evaluated with the help of a semiquantitative PCR assay. The PCR detection of HHV-8 was more sensitive to the Chelex method than to Qia-gene. The threshold for PCR test sensitivity with the help of serial dilution of DNA was at the level of five plasmid ORF-26 regions, and DNA from 25 body cavity-based lymphoma-1 cells. The results expressed as virus load/actin unit showed progressively higher HHV-8 levels in late (nodular) cases, compared to those in early (patch/plaque) stages. Evaluation of HHV-8 DNA levels in tumour tissues, thus, indicates a correlation between virus load and KS stage. Double immunostaining of spindle cells (SC) in KS biopsies for CD34 and HHV-8/latency-associated nuclear antigen (LANA) showed an increase in double-positive SC in the lesions of nodular AKS and EKS cases, compared to that in plaque and patch stages. However, 10-15% of CD34+/LANA- SC cells were observed during the development from patch to nodular cases of AKS and EKS. Our results indicate that PCR analysis is a simple and sensitive diagnostic method for HHV-8 evaluation in KS tissues, processed for conventional histopathology.     

Defective functions of circulating CD4+CD25+ and CD4+CD25- T cells in patients with chronic ordinary urticaria

BACKGROUND: Patients with chronic ordinary urticaria (CU) are divided into two groups: 30-50% have chronic autoimmune urticaria, and the remainder have chronic idiopathic urticaria. CD4(+)CD25(+) regulatory T (Treg) cells play critical roles in maintaining peripheral tolerance and preventing autoimmunity, but the characteristics of Treg cells have not yet been defined in CU. OBJECTIVE: To identify whether CD4(+) T cells play an important immunoregulatory role in the etiology of CU, we determined the frequencies and functions of circulating CD4(+)CD25(+) and CD4(+)CD25(-) T cells in CU patients and healthy control subjects, with special focus on the characteristics of CD4(+)CD25(+) T cells. METHODS: Peripheral blood mononuclear cells (PBMCs) were obtained from CU and healthy controls in this study. The frequency of CD4(+)CD25(+) T cells in PBMCs was detected by flow cytometry. The expression levels of forkhead box P3 (FOXP3) and transforming growth factor-beta (TGF-beta) in CD4(+)CD25(+) T cells were detected by real-time PCR. Furthermore, the suppressive function of CD4(+)CD25(+) T cells was analyzed. Additionally, the Th1/Th2 cytokine secretory profile in mitogen-stimulated CD4(+)CD25(-) T cells was measured by ELISA. RESULTS: An increased frequency of CD4(+)CD25(+) T cells was observed in CU patients (n=19) compared to control subjects (n=7). No significant difference was detected in the expression levels of FOXP3 or TGF-beta between CU patients (n=14) and control subjects (n=7). Strikingly, the suppressive capacity of CD4(+)CD25(+) Treg cells from 2 of 5 CU patients was partially defective. We also found that cytokine production from CD4(+)CD25(-) T cells was significantly reduced in CU patients (n=9) compared to healthy donors (n=11). CONCLUSIONS: Our data demonstrate that CD4(+)CD25(+) and CD4(+)CD25(-) T cells in PBMCs exhibit defective functions in CU patients.     

Detection of HHV-8 in pyogenic granuloma-like Kaposi sarcoma

Kaposi sarcoma (KS) is a low-grade vascular neoplasm associated with human herpesvirus-8 (HHV-8) infection. Clinically, lesions commence as blue-red macules that may develop into plaques and eventually into nodules. The histologic appearance spans a broad spectrum and varies with the stage of the lesion. At each stage, KS has significant morphologic overlap with other vasoproliferative lesions. Recently, we encountered 6 KS tumors that histologically mimicked pyogenic granuloma (PG), a common benign vascular tumor of the skin that usually does not figure in the histologic differential diagnosis of KS. We stained 6 PG-like KS and 28 PGs with a mouse monoclonal antibody (13B10) against HHV-8 latent nuclear antigen-1 (LNA-1) to determine the utility of immunoperoxidase staining in distinguishing KS from PG. All 6 PG-like KS demonstrated nuclear staining for HHV-8 LNA-1. No staining was identified in any of the 28 PGs. Histologic criteria often used to differentiate between these two entities were not helpful in our cases. The only distinguishing feature was the presence or absence of HHV-8 LNA-1 staining. The presence of HHV-8 LNA-1 nuclear staining seems to be a specific marker for KS when comparing PGs and PG-like KS. Immunoperoxidase staining for HHV-8 LNA-1 is a useful diagnostic tool in this setting.     

抗人疱疹病毒8型抗体效价在不同类型Kaposi肉瘤患者中的初步调查

研究表明,新疆不但是Kaposi肉瘤的高发区,亦是人疱疹病毒8型(HHV-8)的流行区(不同人群中感染率介于12.5%～48%)~([1-2]).Sims等[3]提出高效价抗HHV-8抗体的产生是导致HHV-8感染者罹患Kaposi肉瘤的绝对危险因素.而Newton等[4]的研究表明,抗LANA(潜伏态核抗原)高效价抗体与Kaposi肉瘤侵害程度无关.     

Disseminated Kaposi's sarcoma associated with idiopathic CD4+ lymphocytopenia and low dose steroid therapy

A case of disseminated Kaposi sarcoma in a 76-year-old woman treated with low-dose steroids for a seronegative polyarthritis is described. HIV serology was negative, whereas PCR for HHV-8 was positive. The patient presented with a profound reduction in the peripheral blood CD4+ cell count and a decrease in serum immunoglobulins and was diagnosed as having idiopathic CD4+ lymphocytopenia. The clinical picture was characterized by limited cutaneous lesions with typical features of KS associated with widespread visceral involvement and unusually aggressive behaviour.     

银屑病患者骨髓CD34~+细胞Hes-1基因的表达研究

目的:观察银屑病患者骨髓CD34~+细胞Hes-1基因的表达.方法:以β-肌动蛋白为内参照,采用反转录(RT)-PCR法半定量检测24例银屑病患者及18名正常人骨髓CD34~+细胞Hes-1 mRNA的表达水平.结果:与正常对照组相比,银屑病患者组Hes-1mRNA表达水平明显升高,二者差别有统计学意义.结论:造血细胞发育过程中重要的转录因子Hes-1的高表达可能与银屑病患者骨髓造血细胞的增殖活性异常有关.     

寻常型银屑病患者外周血CD4~+CD25~+及CD8~+CD25~+细胞的检测

目的研究进展期寻常型银屑病患者外周血CD4+CD25+和CD8+CD25+调节性T细胞的数量变化及其在银屑病免疫病理学发病机制中的作用。方法应用流式细胞术对进展期寻常型银屑病患者外周血CD4+CD25+和CD8+CD25+调节性T细胞进行检测。结果进展期寻常型银屑病外周血CD4+CD25+细胞及CD8+CD25+调节性T细胞数量与正常对照组相比,均显著降低(P<0.05,P<0.005),而CD4+CD25+/CD8+CD25+比值无显著性差异(P>0.05)。结论寻常型银屑病的发病与CD4+CD25+和CD8+CD25+调节性T细胞的同步降低有关,与二者的比值无关。     

Progressive HHV-8-positive classic Kaposi's sarcoma: rapid response to interferon α-2a but persistence of HHV-8 DNA sequences in lesional skin

The pathogenesis of Kaposi's sarcoma (KS) is often attributed to an infectious agent. In particular, the human herpesvirus 8 (HHV-8) was currently shown to be closely related to all known KS types, including HIV-associated KS, European classic KS, African endemic KS and iatrogenic KS. We report here on an HIV-negative, German patient of neither Jewish nor Mediterranean descent with disseminated classic KS showing unusual rapid progression into the tumour stage. After systemic administration of interferon α-2a over 4 weeks all tumour lesions cleared completely. Interestingly, HHV-8 DNA sequences detected by nested polymerase chain reaction in KS lesions before the onset of treatment were still present in lesional skin after complete remission of the tumour. No recurrence was seen after a follow-up period of 6 months.     

特应性皮炎患者外周血皮肤归巢的CD8+ T细胞研究

【摘要】 目的 探讨特应性皮炎（AD）患者外周血CD8+ T细胞皮肤归巢及杀伤功能相关蛋白的表达。 方法 15例AD患者和14例健康人,用流式细胞仪,检测外周血CD8+ T细胞、表达皮肤淋巴细胞相关抗原的CD8+ T细胞（CLA+CD8+ T细胞）的比例。 结果 CD8+ T细胞比例在AD组和对照组之间差异无统计学意义（P > 0.05）;CD8+ T细胞穿孔素、颗粒酶B和FasL的表达在两组间差异亦无统计学意义（均P > 0.05）。CLA+CD8+ T细胞比例在AD组（3.80% ± 1.46%）高于健康对照组（2.18% ± 0.85%）（t = 3.636,P < 0.01）,AD组CLA+CD8+ T细胞比例与SCORAD（SCORing of Atopic Dermatitis）评分呈正相关（r = 0.565,P < 0.05）;CCR4在CD8+ T细胞表达在AD组（13.86% ± 4.42%）高于健康对照组（9.50% ± 2.14%）（t = 3.738,P < 0.01）,而CCR10和CXCR6的表达在两组间差异均无统计学意义（P > 0.05）。CLA+CD8+ T细胞穿孔素的表达在AD组（74.27% ± 15.94%）高于健康对照组（57.20% ± 14.64%）（t = 2.998,P < 0.01）,颗粒酶B的表达在AD组（70.90% ± 13.85%）也高于健康对照组（56.41% ± 11.00%）（t = 3.104,P < 0.01）,而FasL的表达在两组间差异无统计学意义（P > 0.05）。CLA+CD8+ T细胞CCR4、CCR10和CXCR6的表达在AD组与对照组间差异无统计学意义（均P > 0.05）。 结论 AD患者外周血CLA+CD8+ T细胞数量增加,杀伤功能相关蛋白穿孔素、颗粒酶B的表达增强,可能参与了AD的发病。     

人疱疹病毒8型ORF75基因亚型与Kaposi肉瘤的相关性研究

目的 探讨人类疱疹病毒8型(HHV-8)ORF75基因亚型,与Kaposi肉瘤不同临床分型及侵袭性的相关性.方法 对25例新疆Kaposi肉瘤石蜡包埋组织进行HHV-8 DNA抽提、扩增及双向测序,使用Clustal W软件和PHYLIP软件包对测序结果进行发生学分析,从而确定HHV-8 ORF75基因哑型.结果 25例Kaposi肉瘤中,21例HHV-8阳性,阳性率为84%,其中7例AIDS相关型Kaposi肉瘤患者HHV-8均阳性.21例HHV-8阳性患者中,18例为HHV-8 ORF75 A亚型,3例为C亚型;不同亚型间Kaposi肉瘤患者有无黏膜损害及临床分型的分布差异均无统计学意义(P＞0.05).结论 新疆Kaposi肉瘤患者感染HHV-8 ORF75亚型属于A亚型和C亚型,HHV-8 ORF75不同亚型可能与新疆Kaposi肉瘤黏膜损害及临床分型无关.