Frequency of Chromosomally-Integrated Human Herpesvirus 6 in Children with Acute Lymphoblastic Leukemia

Gravel, Annie; Sinnett, Daniel; Flamand, Louis

doi:10.1371/journal.pone.0084322

Cited by 22 publications

(10 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Four microliters (approximately 50 ng) of the DNA was mixed with primers and probes in a final volume of 10 l, mixed with an equal volume of 2ϫ Rotor-Gene multiplex PCR kit (Qiagen), and subjected to 40 cycles of amplification. The conditions for detection of HHV-6A/B were described previously (6,35). Primers and probe used are listed in Table 4.…”

Section: Methodsmentioning

confidence: 99%

Cell Culture Systems To Study Human Herpesvirus 6A/B Chromosomal Integration

Gravel¹,

Dubuc²,

Wallaschek

et al. 2017

J Virol

Self Cite

View full text Add to dashboard Cite

Human herpesviruses 6A/B (HHV-6A/B) can integrate their viral genomes in the telomeres of human chromosomes. The viral and cellular factors contributing to HHV-6A/B integration remain largely unknown, mostly due to the lack of efficient and reproducible cell culture models to study HHV-6A/B integration. In this study, we characterized the HHV-6A/B integration efficiencies in several human cell lines using two different approaches. First, after a short-term infection (5 h), cells were processed for single-cell cloning and analyzed for chromosomally integrated HHV-6A/B (ciHHV-6A/B). Second, cells were infected with HHV-6A/B and allowed to grow in bulk for 4 weeks or longer and then analyzed for the presence of ciHHV-6. Using quantitative PCR (qPCR), droplet digital PCR, and fluorescent in situ hybridization, we could demonstrate that HHV-6A/B integrated in most human cell lines tested, including telomerase-positive (HeLa, MCF-7, HCT-116, and HEK293T) and telomerase-negative cell lines (U2OS and GM847). Our results also indicate that inhibition of DNA replication, using phosphonoacetic acid, did not affect HHV-6A/B integration. Certain clones harboring ciHHV-6A/B spontaneously express viral genes and proteins. Treatment of cells with phorbol ester or histone deacetylase inhibitors triggered the expression of many viral genes, including U39, U90, and U100, without the production of infectious virus, suggesting that the tested stimuli were not sufficient to trigger full reactivation. In summary, both integration models yielded comparable results and should enable the identification of viral and cellular factors contributing to HHV-6A/B integration and the screening of drugs influencing viral gene expression, as well as the release of infectious HHV-6A/B from the integrated state.IMPORTANCE The analysis and understanding of HHV-6A/B genome integration into host DNA is currently limited due to the lack of reproducible and efficient viral integration systems. In the present study, we describe two quantitative cell culture viral integration systems. These systems can be used to define cellular and viral factors that play a role in HHV-6A/B integration. Furthermore, these systems will allow us to decipher the conditions resulting in virus gene expression and excision of the integrated viral genome resulting in reactivation.KEYWORDS chromosomal integration, ddPCR, telomere

show abstract

Section: Methodsmentioning

confidence: 99%

Cell Culture Systems To Study Human Herpesvirus 6A/B Chromosomal Integration

Gravel¹,

Dubuc²,

Wallaschek

et al. 2017

J Virol

Self Cite

View full text Add to dashboard Cite

show abstract

“…Consequently, individuals with such inherited chromosomally integrated HHV-6 (iciHHV-6) carry a copy of the viral genome in every cell of their body. Diagnosis of iciHHV-6 is easily made by measuring and comparing the relative number of HHV-6 DNA copy per cell either using quantitative PCR (qPCR) and/or digital droplet PCR (9,10). iciHHV-6+ subjects have approximately one million copies per million cells, whereas those that have acquired HHV-6 naturally typically have between 60 and 80 viral DNA copies per million cells (11).…”

mentioning

confidence: 99%

“…Genomic DNA extracted from blood-spotted filters was used for HHV-6 qPCR and droplet digital PCR (ddPCR) analyses. Because iciHHV-6 individuals carry one viral genome copy per cell, identification of subjects with iciHHV-6 is easily made (9,34). The overall prevalence of iciHHV-6 in the CaG cohort was determined, and disease prevalence in subjects with iciHHV-6 was compared with that of subjects without iciHHV-6.…”

mentioning

confidence: 99%

Inherited chromosomally integrated human herpesvirus 6 as a predisposing risk factor for the development of angina pectoris

Gravel

Dubuc

Morissette

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

View full text Add to dashboard Cite

Inherited chromosomally integrated human herpesvirus-6 (iciHHV-6) results in the germ-line transmission of the HHV-6 genome. Every somatic cell of iciHHV-6+ individuals contains the HHV-6 genome integrated in the telomere of chromosomes. Whether having iciHHV-6 predisposes humans to diseases remains undefined. DNA from 19,597 participants between 40 and 69 years of age were analyzed by quantitative PCR (qPCR) for the presence of iciHHV-6. Telomere lengths were determined by qPCR. Medical records, hematological, biochemical, and anthropometric measurements and telomere lengths were compared between iciHHV-6+ and iciHHV-6− subjects. The prevalence of iciHHV-6 was 0.58%. Two-way ANOVA with a Holm-Bonferroni correction was used to determine the effects of iciHHV6, sex, and their interaction on continuous outcomes. Two-way logistic regression with a Holm-Bonferroni correction was used to determine the effects of iciHHV6, sex, and their interaction on disease prevalence. Of 50 diseases monitored, a single one, angina pectoris, is significantly elevated (3.3×) in iciHHV-6+ individuals relative to iciHHV-6− subjects (P = 0.017; 95% CI, 1.73-6.35). When adjusted for potential confounding factors (age, body mass index, percent body fat, and systolic blood pressure), the prevalence of angina remained three times greater in iciHHV-6+ subjects (P = 0.015; 95%CI, 1.23-7.15). Analyses of telomere lengths between iciHHV-6− without angina, iciHHV-6− with angina, and iciHHV-6+ with angina indicate that iciHHV-6+ with angina have shorter telomeres than age-matched iciHHV-6− subjects (P = 0.006). Our study represents, to our knowledge, the first largescale analysis of disease association with iciHHV-6. Our results are consistent with iciHHV-6 representing a risk factor for the development of angina.HHV-6 | telomere | angina | chromosomal integration | ciHHV-6

show abstract

“…Thus, the incidence of ciHHV‐6 in Japan appears to be low in comparison with previous studies in Western countries . Previous studies revealed a similar frequency of ciHHV‐6 between healthy controls and children with acute lymphoblastic leukemia, patients with classical Hodgkin lymphoma, and patients with breast cancer . However, ciHHV‐6 was associated with angina pectoris based on a large‐genome cohort study .…”

Section: Discussionmentioning

confidence: 65%

Chromosomally integrated human herpesvirus 6 in the Japanese population

Miura

Kawamura

Hattori

et al. 2018

Journal of Medical Virology

View full text Add to dashboard Cite

The objectives of the work are to elucidate the incidence and virological findings of chromosomally integrated human herpesvirus 6 (ciHHV-6) in Japanese population and to analyze an association between ciHHV-6 and the clinical manifestation of exanthema subitum (ES). Real-time polymerase chain reaction was performed to determine HHV-6 DNA loads in 2347 cord blood samples from healthy neonates (cohort A), febrile children less than 5 years old (cohort B), and hematopoietic cell transplant recipients (cohort C). CiHHV-6 was confirmed by detection of high copy numbers of viral DNA in somatic cells. The integration site was determined by fluorescent in situ hybridization analysis. In the ciHHV-6 subjects of cohorts A and B, HHV-6 antibody titers were measured, the history of ES was obtained, and the incidence of ES was compared with non-ciHHV-6 children without primary HHV-6B infection in the cohort B. CiHHV-6 was detected in 14 (0.60%) of the 2347 samples: A (6/1006, 0.60%), B (6/790, 0.76%), and C (2/551, 0.36%). The integration sites were on chromosome 22q in seven cases, Yp in two cases, and 17q and Xp in one case. No past history of ES was observed in 11 of the 12 subjects. Nine children with ciHHV-6 underwent serological analysis and were found to be positive for HHV-6 IgG antibodies. Incidence of ES was statistically higher in the control subjects than the ciHHV-6 subjects (P = 0.0039). In Japan, the frequency of ciHHV-6 was 0.60%. A high incidence of ciHHV-6A, specifically in chromosome 22, is a characteristic finding among the Japanese. CiHHV-6 may interfere with the clinical symptoms of primary HHV-6B infection.

show abstract

Frequency of Chromosomally-Integrated Human Herpesvirus 6 in Children with Acute Lymphoblastic Leukemia

Cited by 22 publications

References 42 publications

Cell Culture Systems To Study Human Herpesvirus 6A/B Chromosomal Integration

Cell Culture Systems To Study Human Herpesvirus 6A/B Chromosomal Integration

Inherited chromosomally integrated human herpesvirus 6 as a predisposing risk factor for the development of angina pectoris

Chromosomally integrated human herpesvirus 6 in the Japanese population

Contact Info

Product

Resources

About