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Persistence of Hepatitis C Virus Traces after Spontaneous Resolution of Hepatitis C.

Chen AY, Hoare M, Shankar AN, Allison M, Alexander GJ, Michalak TI - PLoS ONE (2015)

Bottom Line: Telaprevir entirely eliminated HCV replication in the PBMC examined.An apparently effective host immune response curtailing hepatitis appears insufficient to completely eliminate the virus.The long-term morbidity of asymptomatic HCV carriage should be examined even in individuals who achieve undetectable HCV by standard testing and their need for treatment should be assessed.

View Article: PubMed Central - PubMed

Affiliation: Molecular Virology and Hepatology Research Group, Faculty of Medicine, Memorial University, St. John's, Newfoundland and Labrador, Canada.

ABSTRACT
Hepatitis C virus (HCV) frequently causes chronic hepatitis, while spontaneous recovery from infection is infrequent. Persistence of HCV after self-limited (spontaneous) resolution of hepatitis C was rarely investigated. The current study aimed to assess incidence and robustness of HCV persistence after self-resolved hepatitis C in individuals with normal liver enzymes and undetectable virus by conventional tests. Applying high sensitivity HCV RNA detection approaches, we analyzed plasma and peripheral blood mononuclear cells (PBMC) from individuals with previous hepatitis C infection. Parallel plasma and PBMC from 24 such non-viraemic individuals followed for 0.3-14.4 (mean 6.4) years were examined. Additional samples from 9 of them were obtained 4.5-7.2 (mean 5.9) years later. RNA was extracted from 250 μl plasma and, if HCV negative, from ~5 ml after ultracentrifugation, and from ex vivo stimulated PBMC. PBMC with evidence of HCV replication from 4 individuals were treated with HCV protease inhibitor, telaprevir. HCV RNA was detected in 14/24 (58.3%) plasma and 11/23 (47.8%) PBMC obtained during the first collection. HCV RNA replicative strand was evident in 7/11 (63.6%) PBMC. Overall, 17/24 (70.8%) individuals carried HCV RNA at mean follow-up of 5.9 years. Samples collected 4.5-7.2 years later revealed HCV in 4/9 (44.4%) plasma and 5/9 (55.5%) PBMC, while 4 (80%) of these 5 PBMC demonstrated virus replicative strand. Overall, 6/9 (66.7%) individuals remained viraemic for up to 20.7 (mean 12.7) years. Telaprevir entirely eliminated HCV replication in the PBMC examined. In conclusion, our results indicate that HCV can persist long after spontaneous resolution of hepatitis C at levels undetectable by current testing. An apparently effective host immune response curtailing hepatitis appears insufficient to completely eliminate the virus. The long-term morbidity of asymptomatic HCV carriage should be examined even in individuals who achieve undetectable HCV by standard testing and their need for treatment should be assessed.

No MeSH data available.


Related in: MedlinePlus

Inhibition of HCV infection in PBMC expressing initially both positive and negative strand of HCV RNA from individuals with a past spontaneous resolution of hepatitis C.PBMC naturally infected with HCV derived from asymptomatic persons followed for 7.4 (14-47/F), 16.1 (15-46/F) or 20.6 (20-49/F) years were treated (T) with 4 μM of TLV or left untreated (UT) in culture for 72 h. Except 20-49/F PBMC, the experiment was performed in duplicate. (A) HCV RNA positive strand was detected by RT-PCR with 5’-UTR-specific primers and amplicon specificity verified by NAH. (B) Virus negative (replicative) strand was identified by the strand-specific RT-PCR/NAH in which synthetic HCV RNA positive (pos) and negative (neg) strands at 104 copies/reaction were used as specificity controls. Other controls were as described in the legend to Fig 2.
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pone.0140312.g005: Inhibition of HCV infection in PBMC expressing initially both positive and negative strand of HCV RNA from individuals with a past spontaneous resolution of hepatitis C.PBMC naturally infected with HCV derived from asymptomatic persons followed for 7.4 (14-47/F), 16.1 (15-46/F) or 20.6 (20-49/F) years were treated (T) with 4 μM of TLV or left untreated (UT) in culture for 72 h. Except 20-49/F PBMC, the experiment was performed in duplicate. (A) HCV RNA positive strand was detected by RT-PCR with 5’-UTR-specific primers and amplicon specificity verified by NAH. (B) Virus negative (replicative) strand was identified by the strand-specific RT-PCR/NAH in which synthetic HCV RNA positive (pos) and negative (neg) strands at 104 copies/reaction were used as specificity controls. Other controls were as described in the legend to Fig 2.

Mentions: Treatment of PBMC reactive for HCV RNA positive and negative strands with TLP completely inhibited HCV expression in PBMC (Fig 5). Thus, PBMC from 14-47/F, 15-46/F and 20-49/F acquired during the second sample collection at 7.4, 16.1 and 20.7 years of follow-up, respectively, totally aborted expression of HCV RNA negative and positive strands after exposure to TLP, while both virus RNA strands remained detectable in control cells treated with 0.5% DMSO alone.


Persistence of Hepatitis C Virus Traces after Spontaneous Resolution of Hepatitis C.

Chen AY, Hoare M, Shankar AN, Allison M, Alexander GJ, Michalak TI - PLoS ONE (2015)

Inhibition of HCV infection in PBMC expressing initially both positive and negative strand of HCV RNA from individuals with a past spontaneous resolution of hepatitis C.PBMC naturally infected with HCV derived from asymptomatic persons followed for 7.4 (14-47/F), 16.1 (15-46/F) or 20.6 (20-49/F) years were treated (T) with 4 μM of TLV or left untreated (UT) in culture for 72 h. Except 20-49/F PBMC, the experiment was performed in duplicate. (A) HCV RNA positive strand was detected by RT-PCR with 5’-UTR-specific primers and amplicon specificity verified by NAH. (B) Virus negative (replicative) strand was identified by the strand-specific RT-PCR/NAH in which synthetic HCV RNA positive (pos) and negative (neg) strands at 104 copies/reaction were used as specificity controls. Other controls were as described in the legend to Fig 2.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4608821&req=5

pone.0140312.g005: Inhibition of HCV infection in PBMC expressing initially both positive and negative strand of HCV RNA from individuals with a past spontaneous resolution of hepatitis C.PBMC naturally infected with HCV derived from asymptomatic persons followed for 7.4 (14-47/F), 16.1 (15-46/F) or 20.6 (20-49/F) years were treated (T) with 4 μM of TLV or left untreated (UT) in culture for 72 h. Except 20-49/F PBMC, the experiment was performed in duplicate. (A) HCV RNA positive strand was detected by RT-PCR with 5’-UTR-specific primers and amplicon specificity verified by NAH. (B) Virus negative (replicative) strand was identified by the strand-specific RT-PCR/NAH in which synthetic HCV RNA positive (pos) and negative (neg) strands at 104 copies/reaction were used as specificity controls. Other controls were as described in the legend to Fig 2.
Mentions: Treatment of PBMC reactive for HCV RNA positive and negative strands with TLP completely inhibited HCV expression in PBMC (Fig 5). Thus, PBMC from 14-47/F, 15-46/F and 20-49/F acquired during the second sample collection at 7.4, 16.1 and 20.7 years of follow-up, respectively, totally aborted expression of HCV RNA negative and positive strands after exposure to TLP, while both virus RNA strands remained detectable in control cells treated with 0.5% DMSO alone.

Bottom Line: Telaprevir entirely eliminated HCV replication in the PBMC examined.An apparently effective host immune response curtailing hepatitis appears insufficient to completely eliminate the virus.The long-term morbidity of asymptomatic HCV carriage should be examined even in individuals who achieve undetectable HCV by standard testing and their need for treatment should be assessed.

View Article: PubMed Central - PubMed

Affiliation: Molecular Virology and Hepatology Research Group, Faculty of Medicine, Memorial University, St. John's, Newfoundland and Labrador, Canada.

ABSTRACT
Hepatitis C virus (HCV) frequently causes chronic hepatitis, while spontaneous recovery from infection is infrequent. Persistence of HCV after self-limited (spontaneous) resolution of hepatitis C was rarely investigated. The current study aimed to assess incidence and robustness of HCV persistence after self-resolved hepatitis C in individuals with normal liver enzymes and undetectable virus by conventional tests. Applying high sensitivity HCV RNA detection approaches, we analyzed plasma and peripheral blood mononuclear cells (PBMC) from individuals with previous hepatitis C infection. Parallel plasma and PBMC from 24 such non-viraemic individuals followed for 0.3-14.4 (mean 6.4) years were examined. Additional samples from 9 of them were obtained 4.5-7.2 (mean 5.9) years later. RNA was extracted from 250 μl plasma and, if HCV negative, from ~5 ml after ultracentrifugation, and from ex vivo stimulated PBMC. PBMC with evidence of HCV replication from 4 individuals were treated with HCV protease inhibitor, telaprevir. HCV RNA was detected in 14/24 (58.3%) plasma and 11/23 (47.8%) PBMC obtained during the first collection. HCV RNA replicative strand was evident in 7/11 (63.6%) PBMC. Overall, 17/24 (70.8%) individuals carried HCV RNA at mean follow-up of 5.9 years. Samples collected 4.5-7.2 years later revealed HCV in 4/9 (44.4%) plasma and 5/9 (55.5%) PBMC, while 4 (80%) of these 5 PBMC demonstrated virus replicative strand. Overall, 6/9 (66.7%) individuals remained viraemic for up to 20.7 (mean 12.7) years. Telaprevir entirely eliminated HCV replication in the PBMC examined. In conclusion, our results indicate that HCV can persist long after spontaneous resolution of hepatitis C at levels undetectable by current testing. An apparently effective host immune response curtailing hepatitis appears insufficient to completely eliminate the virus. The long-term morbidity of asymptomatic HCV carriage should be examined even in individuals who achieve undetectable HCV by standard testing and their need for treatment should be assessed.

No MeSH data available.


Related in: MedlinePlus