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Analysis of the 5'UTR of HCV genotype 3 grown in vitro in human B cells, T cells, and macrophages.

Revie D, Alberti MO, Prichard JG, Kelley AS, Salahuddin SZ - Virol. J. (2010)

Bottom Line: Results revealed a number of sequence changes as compared to the serum RNA.The HCV RNA produced efficiently by infected macrophages, B-cells, and T-cells had sequences similar to HCV-1, which suggests that selection of the variants was performed at the level of macrophages.Therefore, in our opinion, HCV-3 does not replicate efficiently in macrophages, and patients infected with HCV-3 may contain a population of HCV-1 in their blood.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biology, California Lutheran University, Thousand Oaks, California, USA.

ABSTRACT

Background: Previously, we have reported the isolation and molecular characterization of human Hepatitis C virus genotype 1 (HCV-1) from infected patients. We are now reporting an analysis of HCV obtained from patients infected with HCV genotype 3 (HCV-3) as diagnosed by clinical laboratories.

Results: HCV was cultured in vitro using our system. HCV RNA was isolated from patients' blood and from HCV cultured in various cell types for up to three months. The 5'UTR of these isolates were used for comparisons. Results revealed a number of sequence changes as compared to the serum RNA. The HCV RNA produced efficiently by infected macrophages, B-cells, and T-cells had sequences similar to HCV-1, which suggests that selection of the variants was performed at the level of macrophages. Virus with sequences similar to HCV-1 replicated better in macrophages than HCV having a 5'UTR similar to HCV-3.

Conclusions: Although HCV-3 replicates in cell types such as B-cells, T-cells, and macrophages, it may require a different primary cell type for the same purpose. Therefore, in our opinion, HCV-3 does not replicate efficiently in macrophages, and patients infected with HCV-3 may contain a population of HCV-1 in their blood.

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Related in: MedlinePlus

Comparisons of 5'UTR consensus sequences between patient 314 serum and isolates of HCV. H77, GenBank accession number NC_004102, is shown for comparison purposes and was not used to determine the consensus.
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Figure 3: Comparisons of 5'UTR consensus sequences between patient 314 serum and isolates of HCV. H77, GenBank accession number NC_004102, is shown for comparison purposes and was not used to determine the consensus.

Mentions: The first sample of HCV-3 analyzed was from patient 314. Few or no changes in the sequences of the 5'UTR were observed for cultured HCV-1, but 19 changes in the sequences for patient 314 RNA were observed (Figure 3). The sequences for 314 T1 and 314 T2 were the same, showing that consecutive transfers of HCV into the same cell type do not affect the sequence. The 314 T1 and T2 sequences were almost identical to genotype 1 H77, therefore the isolation system for HCV-3 replicated virus which had a 5'UTR similar to HCV-1. This was unexpected, but suggested that there were some HCV-1 in the blood of patient 314 that preferentially replicated in macrophages.


Analysis of the 5'UTR of HCV genotype 3 grown in vitro in human B cells, T cells, and macrophages.

Revie D, Alberti MO, Prichard JG, Kelley AS, Salahuddin SZ - Virol. J. (2010)

Comparisons of 5'UTR consensus sequences between patient 314 serum and isolates of HCV. H77, GenBank accession number NC_004102, is shown for comparison purposes and was not used to determine the consensus.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2913957&req=5

Figure 3: Comparisons of 5'UTR consensus sequences between patient 314 serum and isolates of HCV. H77, GenBank accession number NC_004102, is shown for comparison purposes and was not used to determine the consensus.
Mentions: The first sample of HCV-3 analyzed was from patient 314. Few or no changes in the sequences of the 5'UTR were observed for cultured HCV-1, but 19 changes in the sequences for patient 314 RNA were observed (Figure 3). The sequences for 314 T1 and 314 T2 were the same, showing that consecutive transfers of HCV into the same cell type do not affect the sequence. The 314 T1 and T2 sequences were almost identical to genotype 1 H77, therefore the isolation system for HCV-3 replicated virus which had a 5'UTR similar to HCV-1. This was unexpected, but suggested that there were some HCV-1 in the blood of patient 314 that preferentially replicated in macrophages.

Bottom Line: Results revealed a number of sequence changes as compared to the serum RNA.The HCV RNA produced efficiently by infected macrophages, B-cells, and T-cells had sequences similar to HCV-1, which suggests that selection of the variants was performed at the level of macrophages.Therefore, in our opinion, HCV-3 does not replicate efficiently in macrophages, and patients infected with HCV-3 may contain a population of HCV-1 in their blood.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biology, California Lutheran University, Thousand Oaks, California, USA.

ABSTRACT

Background: Previously, we have reported the isolation and molecular characterization of human Hepatitis C virus genotype 1 (HCV-1) from infected patients. We are now reporting an analysis of HCV obtained from patients infected with HCV genotype 3 (HCV-3) as diagnosed by clinical laboratories.

Results: HCV was cultured in vitro using our system. HCV RNA was isolated from patients' blood and from HCV cultured in various cell types for up to three months. The 5'UTR of these isolates were used for comparisons. Results revealed a number of sequence changes as compared to the serum RNA. The HCV RNA produced efficiently by infected macrophages, B-cells, and T-cells had sequences similar to HCV-1, which suggests that selection of the variants was performed at the level of macrophages. Virus with sequences similar to HCV-1 replicated better in macrophages than HCV having a 5'UTR similar to HCV-3.

Conclusions: Although HCV-3 replicates in cell types such as B-cells, T-cells, and macrophages, it may require a different primary cell type for the same purpose. Therefore, in our opinion, HCV-3 does not replicate efficiently in macrophages, and patients infected with HCV-3 may contain a population of HCV-1 in their blood.

Show MeSH
Related in: MedlinePlus