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 × 10-7 to 6.9 × 10-4 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.
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【Effect of electrochemotherapy on human herpesvirus 8 kinetics in classic Kaposi sarcoma.].
【电化学疗法对经典卡波西肉瘤中人疱疹病毒8动力学的影响。】
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DOI：10.1186/s13027-017-0147-4文章类型：杂志文章
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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。但是，尚无研究调查ECT治疗对人类8型疱疹病毒（HHV8）动力学的影响，后者被认为是KS的必要病因。我们旨在评估经典KS患者的HHV8病毒载量和表达水平，这些患者接受一种或多种ECT治疗，并且每半年进行长达四年的随访。
方法：本研究共纳入27例经典KS患者。在ECT治疗之前获得了肿瘤活检和血液样本。在六个月的时间间隔内（12-48个月）收集更多的血液样本。通过实时评估所有样本中HHV8病毒载量和潜伏（ORF72和ORF73）和溶菌（K2，K8，K8.1，K10 / K10.1，K10.5 / K10.6和ORF16）基因的表达谱PCR。扩增HHV8 ORF26和K1区，并进行直接核苷酸测序，然后进行系统发育分析以鉴定变体。
结果：在ECT治疗前收集的所有KS活检和46.4％的外周血单个核细胞（PBMC）均为HHV8 DNA阳性。 KS病变中的病毒载量为每细胞0.02至2.3拷贝，PBMC中每细胞的病毒载量为3.0×10-7至6.9×10-4拷贝。总体而言，潜在的ORF72和ORF73以及溶解性K2，K8和K10 / K10.1在所有KS活检中均表达。在KS样本中，ORF16 mRNA的检出率为71.4％，K8.1和K10.5 / K10.6 mRNA的检出率为57.1％。 ORF72，ORF73和K2转录本分别在ECT前收集的PBMC中扩增了37.5％，25％和25％。在第一次ECT会议之后，在六个月的随访中，27例患者中有20例（74.1％）获得了完全缓解，27例PBMC中有4例（14.8％）检测到HHV8 DNA。系统发育分析的ORF26扩增子表明，大多数病毒变体属于A / C（82.3％），很少属于C2（5.9％）或C3（11.8％）亚型。 K1 / VR1变异分为A（33.3％）和C（66.7％）HHV8进化枝。在HHV8亚型和ECT完全应答之间未发现相关性。
结论：ECT治疗对经典KS患者的HHV8动力学有显着影响。患者的完全缓解伴随着循环病毒的清除。
方法：本研究共纳入27例经典KS患者。在ECT治疗之前获得了肿瘤活检和血液样本。在六个月的时间间隔内（12-48个月）收集更多的血液样本。通过实时评估所有样本中HHV8病毒载量和潜伏（ORF72和ORF73）和溶菌（K2，K8，K8.1，K10 / K10.1，K10.5 / K10.6和ORF16）基因的表达谱PCR。扩增HHV8 ORF26和K1区，并进行直接核苷酸测序，然后进行系统发育分析以鉴定变体。
结果：在ECT治疗前收集的所有KS活检和46.4％的外周血单个核细胞（PBMC）均为HHV8 DNA阳性。 KS病变中的病毒载量为每细胞0.02至2.3拷贝，PBMC中每细胞的病毒载量为3.0×10-7至6.9×10-4拷贝。总体而言，潜在的ORF72和ORF73以及溶解性K2，K8和K10 / K10.1在所有KS活检中均表达。在KS样本中，ORF16 mRNA的检出率为71.4％，K8.1和K10.5 / K10.6 mRNA的检出率为57.1％。 ORF72，ORF73和K2转录本分别在ECT前收集的PBMC中扩增了37.5％，25％和25％。在第一次ECT会议之后，在六个月的随访中，27例患者中有20例（74.1％）获得了完全缓解，27例PBMC中有4例（14.8％）检测到HHV8 DNA。系统发育分析的ORF26扩增子表明，大多数病毒变体属于A / C（82.3％），很少属于C2（5.9％）或C3（11.8％）亚型。 K1 / VR1变异分为A（33.3％）和C（66.7％）HHV8进化枝。在HHV8亚型和ECT完全应答之间未发现相关性。
结论：ECT治疗对经典KS患者的HHV8动力学有显着影响。患者的完全缓解伴随着循环病毒的清除。
