电化学疗法对经典型卡波西肉瘤中人类疱疹病毒8动力学的影响。

Effect of electrochemotherapy on human herpesvirus 8 kinetics in classic Kaposi sarcoma.

作者信息

Starita Noemy, Di Monta Gianluca, Cerasuolo Andrea, Marone Ugo, Anniciello Anna Maria, Botti Gerardo, Buonaguro Luigi, Buonaguro Franco M, Tornesello Maria Lina

机构信息

Molecular Biology and Viral Oncology Unit, Istituto Nazionale Tumori IRCCS "Fond. G. Pascale", 80131 Naples, Italy.

Department of Surgery "Melanoma, Soft Tissues, Head and Neck, Skin Cancers", Istituto Nazionale Tumori IRCCS "Fond. G. Pascale", Naples, Italy.

出版信息

Infect Agent Cancer. 2017 Jun 19;12:35. doi: 10.1186/s13027-017-0147-4. eCollection 2017.

DOI:10.1186/s13027-017-0147-4

PMID:28649271

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5477158/

Abstract

BACKGROUND

Electrochemotherapy (ECT) has shown to be an effective treatment for cutaneous and subcutaneous Kaposi sarcoma (KS) lesions. However, no study has investigated the impact of ECT treatment on the kinetics of human herpesvirus type 8 (HHV8), which is considered the necessary causal agent of KS. We aimed to evaluate HHV8 viral load and expression levels in patients affected by classic KS who received one or more ECT treatments and have been followed semi annually for up to four years.

METHODS

A total of 27 classic KS patients were enrolled in this study. Tumour biopsies and blood samples were obtained before ECT treatment. Additional blood samples were collected at six month intervals for 12-48 months. HHV8 viral load and expression profiles of latent (ORF72 and ORF73) and lytic (K2, K8, K8.1, K10/K10.1, K10.5/K10.6 and ORF16) genes were assessed in all samples by real-time PCR. HHV8 ORF26 and K1 regions were amplified and subjected to direct nucleotide sequencing followed by phylogenetic analysis for variant identification.

RESULTS

All KS biopsies and 46.4% of peripheral blood mononuclear cells (PBMCs) collected before ECT treatment were positive for HHV8 DNA. Viral load ranged from 0.02 to 2.3 copies per cell in KS lesions and 3.0 × 10 to 6.9 × 10 copies per cell in PBMCs. Overall, latent ORF72 and ORF73 as well as lytic K2, K8 and K10/K10.1 were expressed in all KS biopsies. ORF16 mRNA was detected in 71.4% and both K8.1 and K10.5/K10.6 mRNAs in 57.1% of KS samples. The ORF72, ORF73 and K2 transcripts were amplified in 37.5%, 25% and 25% of PBMCs collected before ECT, respectively. After the first ECT session, complete response was achieved in 20 out of 27 (74.1%) patients and HHV8 DNA was detected in four out of 27 (14.8%) PBMC samples at six month follow up. Phylogenetic analysis of ORF26 amplimers showed that most viral variants belonged to A/C (82.3%), and few to C2 (5.9%) or C3 (11.8%) subtype. The K1/VR1 variants fell into A (33.3%) and C (66.7%) HHV8 clade. No correlation was found between HHV8 subtypes and ECT complete response.

CONCLUSIONS

ECT therapy has a significant effect on HHV8 kinetics in patients with classic KS. The complete remission of patients was accompanied by clearance of circulating virus.

摘要

背景

电化学疗法（ECT）已被证明是治疗皮肤和皮下卡波西肉瘤（KS）病变的有效方法。然而，尚无研究调查ECT治疗对人疱疹病毒8型（HHV8）动力学的影响，HHV8被认为是KS的必要致病因子。我们旨在评估接受一次或多次ECT治疗并每半年随访长达四年的经典KS患者的HHV8病毒载量和表达水平。

方法

本研究共纳入27例经典KS患者。在ECT治疗前获取肿瘤活检组织和血液样本。每隔6个月采集额外的血液样本，持续12 - 48个月。通过实时PCR评估所有样本中HHV8病毒载量以及潜伏（ORF72和ORF73）和裂解（K2、K8、K8.1、K10/K10.1、K10.5/K10.6和ORF16）基因的表达谱。对HHV8的ORF26和K1区域进行扩增并直接进行核苷酸测序，随后进行系统发育分析以鉴定变异体。

结果

ECT治疗前采集的所有KS活检组织和46.4%的外周血单个核细胞（PBMC）中HHV8 DNA呈阳性。KS病变中病毒载量范围为每细胞0.02至2.3拷贝，PBMC中为每细胞3.0×10至6.9×10拷贝。总体而言，所有KS活检组织中均表达潜伏的ORF72和ORF73以及裂解的K2、K8和K10/K10.1。71.4%的KS样本中检测到ORF16 mRNA，57.1%的KS样本中同时检测到K8.1和K10.5/K10.6 mRNA。ECT治疗前采集的PBMC中，分别有37.5%、25%和25%的样本扩增出ORF72、ORF73和K2转录本。第一次ECT治疗后，27例患者中有20例（74.1%）达到完全缓解，6个月随访时27例PBMC样本中有4例（14.8%）检测到HHV8 DNA。对ORF26扩增子的系统发育分析表明，大多数病毒变异体属于A/C（82.3%），少数属于C2（5.9%）或C3（11.8%）亚型。K1/VR1变异体属于HHV8的A（33.3%）和C（66.7%）分支。未发现HHV8亚型与ECT完全缓解之间存在相关性。