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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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Variation in the bases of 384 isolates. The 384 serum sample is used for the figure, with sequence changes found in the isolates indicated. The table lists each isolate and the base at each position in the sequence. The arrows indicate the limits of the reported sequences. The figure is adapted from Lyons et al. [29].
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Figure 8: Variation in the bases of 384 isolates. The 384 serum sample is used for the figure, with sequence changes found in the isolates indicated. The table lists each isolate and the base at each position in the sequence. The arrows indicate the limits of the reported sequences. The figure is adapted from Lyons et al. [29].

Mentions: We compared HCV RNA from patient 384 serum and four laboratory isolates to determine if changes were consistent with the current 2 D model of the 5'UTR RNA proposed by Honda et al. (2001). The 19 variant bases were either in regions that are not base paired, or where the changes would not affect base pairing (Figure 8). So although there were a number of changes to the sequence after culturing, these did not appear to cause any differences that would affect the overall 2 D structure of this model. All of the differences in the stems had either compensatory changes (e.g. GC to AU at bases 179 and 220) or could use alternative base pairing (e.g., GC to GU at bases 145 and 248). We are not sure how these changes impact the replication of HCV.


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)

Variation in the bases of 384 isolates. The 384 serum sample is used for the figure, with sequence changes found in the isolates indicated. The table lists each isolate and the base at each position in the sequence. The arrows indicate the limits of the reported sequences. The figure is adapted from Lyons et al. [29].
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 8: Variation in the bases of 384 isolates. The 384 serum sample is used for the figure, with sequence changes found in the isolates indicated. The table lists each isolate and the base at each position in the sequence. The arrows indicate the limits of the reported sequences. The figure is adapted from Lyons et al. [29].
Mentions: We compared HCV RNA from patient 384 serum and four laboratory isolates to determine if changes were consistent with the current 2 D model of the 5'UTR RNA proposed by Honda et al. (2001). The 19 variant bases were either in regions that are not base paired, or where the changes would not affect base pairing (Figure 8). So although there were a number of changes to the sequence after culturing, these did not appear to cause any differences that would affect the overall 2 D structure of this model. All of the differences in the stems had either compensatory changes (e.g. GC to AU at bases 179 and 220) or could use alternative base pairing (e.g., GC to GU at bases 145 and 248). We are not sure how these changes impact the replication of HCV.

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