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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

Expression of HCV RNA positive and negative (replicative) strands in PBMC samples obtained at two collections approximately 5 years apart from individuals with self-resolved hepatitis C.(A) HCV RNA positive strand identification using 2 μg of total RNA extracted from PBMC treated ex vivo with PHA or C5 (*). (B) Detection of HCV RNA negative strand in HCV RNA positive strand reactive PBMC shown in A. Samples were obtained during the first (1) or the second (2) collection at follow-up time (years) indicated under panel A. As positive controls for HCV RNA positive strand detection, serial 10-fold dilutions of rHCV UTR-E2 carrying indicated copy numbers/reaction were used (panel A). For HCV RNA negative strand detection, synthetic HCV RNA positive strand (sHCV RNA pos) and HCV synthetic RNA negative strand (sHCV RNA neg) at 104 copies/reaction were used as positive and specificity controls, respectively (panel B). Water amplified in direct (DW) and nested (NW) reactions and a mock (M) extraction served as contamination controls. Positive signals demonstrated the expected 244-bp oligonucleotide fragments. Numbers under panels marked as relative DU represent relative densitometric units (DU) given by hybridization signals.
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pone.0140312.g002: Expression of HCV RNA positive and negative (replicative) strands in PBMC samples obtained at two collections approximately 5 years apart from individuals with self-resolved hepatitis C.(A) HCV RNA positive strand identification using 2 μg of total RNA extracted from PBMC treated ex vivo with PHA or C5 (*). (B) Detection of HCV RNA negative strand in HCV RNA positive strand reactive PBMC shown in A. Samples were obtained during the first (1) or the second (2) collection at follow-up time (years) indicated under panel A. As positive controls for HCV RNA positive strand detection, serial 10-fold dilutions of rHCV UTR-E2 carrying indicated copy numbers/reaction were used (panel A). For HCV RNA negative strand detection, synthetic HCV RNA positive strand (sHCV RNA pos) and HCV synthetic RNA negative strand (sHCV RNA neg) at 104 copies/reaction were used as positive and specificity controls, respectively (panel B). Water amplified in direct (DW) and nested (NW) reactions and a mock (M) extraction served as contamination controls. Positive signals demonstrated the expected 244-bp oligonucleotide fragments. Numbers under panels marked as relative DU represent relative densitometric units (DU) given by hybridization signals.

Mentions: PBMC found to be HCV RNA positive strand reactive were examined for expression of the HCV RNA negative (replicative) strand. This replication intermediate occurs at a lower copy number per cell than the positive strand and the assay detecting the negative strand is 10 to 100-fold less sensitive than that for the positive strand identification [3,20], so HCV RNA negative strand was tested only in PBMC found reactive for the positive strand. Thus, this replication intermediate was detected in 11 out of 16 (68.7%) PBMC investigated (Table 2 and Fig 2). Among the negative strand reactive PBMC samples, 7 (7/11; 63.6%) were obtained during the first collection (mean follow-up 5.1 years) and 4 (4/5; 80%) at the second (mean follow-up 14.5 years) (Table 2).


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)

Expression of HCV RNA positive and negative (replicative) strands in PBMC samples obtained at two collections approximately 5 years apart from individuals with self-resolved hepatitis C.(A) HCV RNA positive strand identification using 2 μg of total RNA extracted from PBMC treated ex vivo with PHA or C5 (*). (B) Detection of HCV RNA negative strand in HCV RNA positive strand reactive PBMC shown in A. Samples were obtained during the first (1) or the second (2) collection at follow-up time (years) indicated under panel A. As positive controls for HCV RNA positive strand detection, serial 10-fold dilutions of rHCV UTR-E2 carrying indicated copy numbers/reaction were used (panel A). For HCV RNA negative strand detection, synthetic HCV RNA positive strand (sHCV RNA pos) and HCV synthetic RNA negative strand (sHCV RNA neg) at 104 copies/reaction were used as positive and specificity controls, respectively (panel B). Water amplified in direct (DW) and nested (NW) reactions and a mock (M) extraction served as contamination controls. Positive signals demonstrated the expected 244-bp oligonucleotide fragments. Numbers under panels marked as relative DU represent relative densitometric units (DU) given by hybridization signals.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0140312.g002: Expression of HCV RNA positive and negative (replicative) strands in PBMC samples obtained at two collections approximately 5 years apart from individuals with self-resolved hepatitis C.(A) HCV RNA positive strand identification using 2 μg of total RNA extracted from PBMC treated ex vivo with PHA or C5 (*). (B) Detection of HCV RNA negative strand in HCV RNA positive strand reactive PBMC shown in A. Samples were obtained during the first (1) or the second (2) collection at follow-up time (years) indicated under panel A. As positive controls for HCV RNA positive strand detection, serial 10-fold dilutions of rHCV UTR-E2 carrying indicated copy numbers/reaction were used (panel A). For HCV RNA negative strand detection, synthetic HCV RNA positive strand (sHCV RNA pos) and HCV synthetic RNA negative strand (sHCV RNA neg) at 104 copies/reaction were used as positive and specificity controls, respectively (panel B). Water amplified in direct (DW) and nested (NW) reactions and a mock (M) extraction served as contamination controls. Positive signals demonstrated the expected 244-bp oligonucleotide fragments. Numbers under panels marked as relative DU represent relative densitometric units (DU) given by hybridization signals.
Mentions: PBMC found to be HCV RNA positive strand reactive were examined for expression of the HCV RNA negative (replicative) strand. This replication intermediate occurs at a lower copy number per cell than the positive strand and the assay detecting the negative strand is 10 to 100-fold less sensitive than that for the positive strand identification [3,20], so HCV RNA negative strand was tested only in PBMC found reactive for the positive strand. Thus, this replication intermediate was detected in 11 out of 16 (68.7%) PBMC investigated (Table 2 and Fig 2). Among the negative strand reactive PBMC samples, 7 (7/11; 63.6%) were obtained during the first collection (mean follow-up 5.1 years) and 4 (4/5; 80%) at the second (mean follow-up 14.5 years) (Table 2).

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