1. Volume 134, Issue 3, Pages 812-822 (March 2008)
Hepatitis C Virus Replicates in the Same Immune Cell Subsets in Chronic Hepatitis C and Occult Infection 
Tram N.Q. Pham, Dawn King, Sonya A. MacParland, Jerry S. McGrath, S. Bharati Reddy, Ford R. Bursey, Tomasz I. Michalak 
Gastroenterology 
Volume 134, Issue 3, Pages (March 2008)
DOI: /j.gastro
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2. Figure 1
Expression HCV-RNA–positive and HCV-RNA–negative strands in PBMC and immune cell subsets from patients with CHC. PBMCs and purified CD4+ and CD8+ T lymphocytes from patients with CHC (cases 2 and 3a) were left untreated (ut) or were treated with PHA/IL-2 (t), whereas B cells and monocytes (Mo) were tested as untreated. RNA extracted from 2–4 × 106 cells was analyzed by RT-PCR/NAH for (A) HCV-RNA–positive strand using HCV synthetic RNA (sRNA) as a positive control and (B) HCV-RNA–negative strand using HCV-sRNA–negative strand (neg) as a positive control and HCV-sRNA–positive strand (pos) as a specificity control. Contamination controls included water added instead of cDNA and amplified by direct (dw) and nested (nw) PCR and a mock (m) treated as test RNA. β-actin amplified in parallel served as a loading control. Positive signals showed the expected 346-bp (direct) and 244-bp (nested) 5′-UTR fragments, and the 239-bp amplicon for β-actin.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2008 AGA Institute Terms and Conditions

3. Figure 2
Detection of HCV in PBMCs and immune cell subsets from individuals with occult HCV infection. PBMCs and purified CD4+ and CD8+ T cells, B cells, and monocytes (Mo) from cases 10a, 12a, and 13a were analyzed by RT-PCR/NAH for (A) HCV-RNA–positive and (B) HCV-RNA–negative strands, as outlined in the legend to Figure 1.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2008 AGA Institute Terms and Conditions

4. Figure 3
Expression of HCV NS5A protein in different immune cell subsets from a patient with CHC. Purified CD4+ and CD8+ T lymphocytes, B cells, and monocytes (Mo) from case 3b were double-stained with anti-HCV NS5A and antitubulin mAb, and analyzed by confocal microscopy. Huh7 cells, either naive or expressing HCV AB12-A2 replicon, were stained with the same mAbs and used as negative and positive controls, respectively. HCV-NS5A–positive cells on images depicting staining with antitubulin are marked with arrows. Images were captured at 60× magnification.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2008 AGA Institute Terms and Conditions

5. Figure 4
HCV NS5A protein detection in PBMCs and immune cell subtypes from individuals with occult HCV infection. PBMCs and purified CD4+ and CD8+ T cells, B cells, and monocytes (Mo) from (A) case 14 and (B) cases 10b, 12b, and 13b were stained for HCV NS5A protein and analyzed as outlined in the legend to Figure 3. Examples of representative staining patterns are shown. aCells treated with PHA/IL-2 before staining. HCV-NS5A–positive cells on images depicting staining with antitubulin are marked with arrows. Images were captured at 60× magnification.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2008 AGA Institute Terms and Conditions

6. Figure 5
HCV-IRES sequences detected in PBMCs, purified immune cell subtypes, and plasma from patients with CHC and occult HCV infection. HCV 5′-UTR sequences from immune cells from cases 4, 12a, and 13a were cloned and 9–10 randomly selected clones were sequenced bidirectionally. HCV sequence detected in plasma collected at the time of PBMC acquisition was used as a reference, except in case 13a in which HCV from plasma obtained during CHC served as the reference. Representative sequences are shown and the number at the end of the sequence depicts the number of clones in which a given variant was found. Nucleotides identical to those in the reference sequence are shown as dots, different as letters, and deleted as dashes.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2008 AGA Institute Terms and Conditions