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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

Detection of HCV RNA in plasma obtained at two separate collections from individuals with a past episode of spontaneously resolved hepatitis C.Total RNA extracted from 250 μl or a pellet recovered after ultracentrifugation of ~5 ml plasma (*) acquired during the first (1) and the second (2) sample collections were amplified with 5'-UTR specific primers and amplicon specificity verified by NAH. Serial 10-fold dilutions of recombinant HCV 5'-UTR-E2 (rHCV UTR-E2) fragment carrying indicated copy numbers/reaction were used as positive and specificity controls. Water amplified in direct (DW) and nested (NW) reactions and a mock (M) extraction served as contamination controls. Positive signals showed the expected 244-bp oligonucleotide fragments. Numbers under the panels represent follow-up time in years after resolution of hepatitis C (upper line) and relative densitometric units (DU) given by hybridization signals (lower line).
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pone.0140312.g001: Detection of HCV RNA in plasma obtained at two separate collections from individuals with a past episode of spontaneously resolved hepatitis C.Total RNA extracted from 250 μl or a pellet recovered after ultracentrifugation of ~5 ml plasma (*) acquired during the first (1) and the second (2) sample collections were amplified with 5'-UTR specific primers and amplicon specificity verified by NAH. Serial 10-fold dilutions of recombinant HCV 5'-UTR-E2 (rHCV UTR-E2) fragment carrying indicated copy numbers/reaction were used as positive and specificity controls. Water amplified in direct (DW) and nested (NW) reactions and a mock (M) extraction served as contamination controls. Positive signals showed the expected 244-bp oligonucleotide fragments. Numbers under the panels represent follow-up time in years after resolution of hepatitis C (upper line) and relative densitometric units (DU) given by hybridization signals (lower line).

Mentions: Plasma samples obtained during the first (n = 24) or the second (n = 9) collections were uniformly HCV RNA negative by standard clinical testing (Table 1). However, when the same samples were re-analyzed using RT-PCR/NAH assay, 58.3% (14/24) and 44.4% (4/9) were HCV RNA reactive at mean follow-up 6.4 and 12.7 years (ranges 0.3 to 14.4 and 5.5 to 20.7 respectively) (Table 2 and Fig 1). The majority of HCV RNA reactive samples from the first collection (11/24; 45.8%) were positive when RNA from 250 μl plasma was analyzed. The remaining (3/14; 21.4%) were HCV RNA positive when pellets from ~5 ml of plasma were tested (Table 2). Inversely, 3 of 4 (75%) plasma samples from the second collection were HCV RNA positive after ultracentrifugation (Table 2). Overall, 17 of 24 (70.8%) individuals carried HCV in either one or two plasma samples collected.


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)

Detection of HCV RNA in plasma obtained at two separate collections from individuals with a past episode of spontaneously resolved hepatitis C.Total RNA extracted from 250 μl or a pellet recovered after ultracentrifugation of ~5 ml plasma (*) acquired during the first (1) and the second (2) sample collections were amplified with 5'-UTR specific primers and amplicon specificity verified by NAH. Serial 10-fold dilutions of recombinant HCV 5'-UTR-E2 (rHCV UTR-E2) fragment carrying indicated copy numbers/reaction were used as positive and specificity controls. Water amplified in direct (DW) and nested (NW) reactions and a mock (M) extraction served as contamination controls. Positive signals showed the expected 244-bp oligonucleotide fragments. Numbers under the panels represent follow-up time in years after resolution of hepatitis C (upper line) and relative densitometric units (DU) given by hybridization signals (lower line).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0140312.g001: Detection of HCV RNA in plasma obtained at two separate collections from individuals with a past episode of spontaneously resolved hepatitis C.Total RNA extracted from 250 μl or a pellet recovered after ultracentrifugation of ~5 ml plasma (*) acquired during the first (1) and the second (2) sample collections were amplified with 5'-UTR specific primers and amplicon specificity verified by NAH. Serial 10-fold dilutions of recombinant HCV 5'-UTR-E2 (rHCV UTR-E2) fragment carrying indicated copy numbers/reaction were used as positive and specificity controls. Water amplified in direct (DW) and nested (NW) reactions and a mock (M) extraction served as contamination controls. Positive signals showed the expected 244-bp oligonucleotide fragments. Numbers under the panels represent follow-up time in years after resolution of hepatitis C (upper line) and relative densitometric units (DU) given by hybridization signals (lower line).
Mentions: Plasma samples obtained during the first (n = 24) or the second (n = 9) collections were uniformly HCV RNA negative by standard clinical testing (Table 1). However, when the same samples were re-analyzed using RT-PCR/NAH assay, 58.3% (14/24) and 44.4% (4/9) were HCV RNA reactive at mean follow-up 6.4 and 12.7 years (ranges 0.3 to 14.4 and 5.5 to 20.7 respectively) (Table 2 and Fig 1). The majority of HCV RNA reactive samples from the first collection (11/24; 45.8%) were positive when RNA from 250 μl plasma was analyzed. The remaining (3/14; 21.4%) were HCV RNA positive when pellets from ~5 ml of plasma were tested (Table 2). Inversely, 3 of 4 (75%) plasma samples from the second collection were HCV RNA positive after ultracentrifugation (Table 2). Overall, 17 of 24 (70.8%) individuals carried HCV in either one or two plasma samples collected.

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