Differing patterns of liver disease progression and hepatitis C virus (HCV) quasispecies evolution in children vertically coinfected with HCV and human immunodeficiency virus type 1

Abstract

Hepatitis C virus (HCV) quasispeciation was studied in two children vertically coinfected with HCV and human immunodeficiency virus type 1 (HIV-1). HCV quasispecies diversification and liver injury were more significant in patient C1, who was immunocompetent with anti-HIV therapy, than in patient C2, who was immunosuppressed, in consistency with modulation of HCV quasispeciation and liver injury by immunocompetence in coinfected children.

FIG. 1.

Clinical parameters measured in coinfected children. (A and D) CD4⁺ (open circles)- and CD8⁺ (filled circles)-T-lymphocyte counts were measured by flow cytometry. Introduction of and changes in antiretroviral therapy are indicated by arrowheads. Arrowhead 1, zidovudine-lamivudine-ritonavir; arrowhead 2, zidovudine monotherapy; arrowhead 3, zidovudine-lamivudine-saquinavir; arrowhead 4, amprenavir-didanosine-stavudine; arrowhead 5, lamivudine-stavudine-ritonavir-nelfinavir; arrowhead 6, didanosine-stavudine-lopinavir-ritonavir. (B and E) HIV-1 plasma RNA levels (open squares) were measured using the Quantiplex HIV RNA version 3.0 assay (Bayer, Pittsburgh, Pa.), with a sensitivity of 50 copies/ml; plasma HCV RNA levels (filled squares) were quantified using the COBAS Amplicor HCV Monitor assay version 2.0 (Roche Diagnostics, Montreal, Quebec, Canada), with a sensitivity of 100 IU/ml. (C and F) ALT aminotransferase (dashed lines) and AST aminotransferase (solid lines) levels were assayed on a Synchron LX20 system (Beckman Coulter, Palo Alto, Calif.). Sampling times are represented by vertical dashed lines.

FIG. 2.

Phylogenetic analysis of HCV HVR1 sequences derived from peripheral blood or liver samples obtained from coinfected children. (A) Case 1. A total of 115 independently derived HVR1 sequences (mean of 16.4 clones per time point; n = 13 to 20) were analyzed using the neighbor-joining method as described in the text. A transition/transversion ratio of 0.5 was used, and 500 bootstrap resamplings were performed. Letters correspond to the specific time point after birth at which sequences were isolated (A, 0.13 years; B, 0.14 years; C, 1.25 years; D, 1.56 years; E, 3.02 years; F, 4.88 years; G, 5.87 years). The asterisk indicates a sequence identical to that of a liver-derived variant. (B) Case 2. A total of 212 independently derived HVR1 sequences (mean of 21.2 clones per time point; n = 16 to 24) were analyzed as described above. Letters correspond to the specific time points after birth at which sequences were isolated (A, 0.21 years; B, 2.07 years; C, 2.30 years; D, 3.60 years; E, 3.83 years; F, 4.29 years; G, 7.07 years; H, 7.46 years; I, 7.76 years; J, 10.03 years; K, 12.65 years). Asterisks indicate the sequence of a liver-derived variant (b9) or sequences identical thereto. Predominant variants from the first time points were used as reciprocal outgroups in both analyses. The scale bars represent 0.1 nucleotide substitutions per site.

FIG. 3.

Longitudinal analysis of the distribution of HVR1 variants in two coinfected children. Each color corresponds to identical variants in within-group analysis but not in between-group analysis. (A) Case 1. A total of 115 independently derived clones were analyzed (mean of 16.4 clones per time point; n = 13 to 20). (B) Case 2. A total of 212 independently derived clones were analyzed (mean of 21.2 clones per time point; n = 16 to 24). Introduction of and changes in antiretroviral therapy are indicated by arrowheads. Arrowhead 1, zidovudine-lamivudine-ritonavir; arrowhead 2, zidovudine monotherapy; arrowhead 3, zidovudine-lamivudine-saquinavir; arrowhead 4, amprenavir-didanosine-stavudine; arrowhead 5, lamivudine-stavudine-ritonavir-nelfinavir; arrowhead 6, didanosine-stavudine-lopinavir-ritonavir.