The role of human herpesvirus 8 and Epstein-Barr virus in the pathogenesis of giant lymph node hyperplasia (Castleman's disease)
The role of human herpesvirus 8 and Epstein-Barr virus in the pathogenesis of giant lymph node hyperplasia (Castleman's disease)
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DOI：
10.1093/clinids/22.6.1120
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发表时间：
1996-06-01
影响因子：
11.8
通讯作者：
Parravicini, C
中科院分区：
文献类型：
--
作者：
Corbellino, M;Poirel, L;Parravicini, C
图1.从Castleman病的浆细胞型（病例I和4）和透明血管型（病例5）患者中提取的DNA的有限稀释半定量PCR分析结果。注意，在浆细胞Castleman病的两种情况下，人类疱疹病毒8（HHV-8）均呈强阳性，而EB病毒（EBV）仅存在于病例4中。在PCR分析之前，将DNA从500 ng连续稀释10倍至0.05ng。用人的特异性引物进行扩增。α-珠蛋白、EBV [8]和HHV-8 [3]，如前所述。在1.5%琼脂糖凝胶上分析扩增物，转移到尼龙膜上，并与32 P标记的寡聚体探针杂交。淋巴结炎作为对照包括在研究中。来自HHV-8阳性KS病变的DNA和不含DNA的试管（9）分别用作阳性和阴性内部对照。没有患者，或随后有，KS或淋巴瘤在一年后活检。淋巴结样本和对照组进行平行处理，根据35个循环的PCR扩增的协议，从500 ng的DNA蛋白酶K消化，酚氯仿提取，乙醇沉淀。在整个过程中采取严格措施，以监测假阳性结果的发生[10]。浆细胞型CD患者的活检标本均经PCR扩增出HHV-8 DNA序列，而透明血管型CD患者的活检标本和对照淋巴结活检标本均为阴性。相比之下，EBV被发现在只有两个活检标本francial浆细胞CD和两个活检标本HIV感染患者持续全身淋巴结病，所有其他活检标本均为阴性。通过有限稀释，半定量PCR分析显示浆细胞CD患者中HHV-8和EBV载量存在显著差异。特别是，
Figure 1. Results of limiting dilution semiquantitative PCR analysis of DNA extracted from patients with the plasma cell type (cases I and 4) and the hyaline vascular type (case 5) ofCastleman's disease. Note the strong positivity for human herpesvirus 8 (HHV-8) in both cases of plasma cell Castleman's disease, as opposed to Epstein-Barr virus (EBV), which is present in only case 4. Before PCR analysis, DNA was serialIy diluted 10-fold from 500 ng to 0.05 ng. Amplifications were performed with specific primers for human. a-globin, EBV [8], and HHV-8 [3], as described previously. Amplimers were analyzed on 1.5% agarose gels, transferred onto nylon membranes, and hybridized with 32P-labeled oligomer probes. lymphadenitis were included in the study as controls. DNA from an HHV-8-positive KS lesion and tubes without DNA (9] were used as positive and negative internal controls, respectively. None of the patients had, or subsequently had, KS or lymphoma in the year following biopsy.Lymph node samples and controls were processed in parallel according to the protocol of a 35-cycle PCR amplification starting with 500 ng of DNA obtained by proteinase K digestion, phenolchloroform extraction, and ethanol precipitation. Strict measures were followed throughout the entire procedure to monitor the occurrence of false-positive results [10]. DNA sequences of HHV-8 were detected by PCR analysis in all biopsy specimens from patients with plasma cell CD, while both biopsy specimens from patients with hyaline vascular CD and control lymph node biopsy specimens were negative. By contrast, EBV was found in only two biopsy specimens frOID patients with plasma cell CD and two biopsy specimens from HIV-infected patients with persistent generalized lymphadenopathy; all other biopsy specimens were negative. By limiting dilution, semiquantitative PCR analysis revealed striking differences in the HHV-8 and EBV loads in patients with plasma cell CD. In particular,