Detection of HCV RNA in EBV+ cell lymphoblastoid cell lines (B-LCLs) derived from HIV/HCV co-infected patients months after establishment in vitro.

BARÉ P; MASSUD I; PICCHIO G; BELMONTE L; CORTI M; PÉREZ BIANCO R; DE TEZANOS PINTO; DE BRACCO MM; RUIBAL ARES B

Keystone, Denver Colorado, Estados Unidos

Simposio; Keystone Symposia, HIV Pathogenesis; 2001

HCV replicates in vitro in PBMC from some HCV infected individuals. In contrast, HCV replication is readily detectable in PBMC from a large proportion of HIV/HCV co-infected patients. The enhanced  HCV replication observed in the presence of HIV suggests that interactions between these two viruses probably exist. We have previously described an in vitro culture system that favors HIV replication in macrophages/monocytes, often leading to the generation of EBV-transformed B-LCLs. Using this method, we established 14 B-LCLs from 9 HIV/HCV co-infected patients. Quantitative measurements of HCV RNA and HIV p24Ag were made in culture supernatants (SN) at different times during and after the establishment of these cell lines. The mean number of HCV RNA copies/ml in the SN of 11 B-LCLs was 18,197 (4.26log). HCV RNA was detected in the SN of B-LCLs  that had been in culture for up to 7 months, excluding the possibility that the detected HCV RNA was derived from plasma virions adsorbed to the surface of PBMC. The establishment of B-LCLs positive for HCV RNA was preceded by the appearance in PBMC culture SN of HIV p24 Ag in concentrations ranging between 27-2300 pg/ml. In contrast, B-LCL negative for HCV RNA (n=3) were preceded by low (<20 pg/ml) or undetectable p24 Ag levels in culture SN. A positive correlation between the preceding (before day 35) HIV p24 Ag and HCV RNA concentrations in culture SN was observed. Our results suggest that HIV replication in PBMC from HIV/HCV co-infected patients plays a role in the establishment of  HCV infection in the resulting B-LCLs. In addition, HCV+ B-LCLs may constitute a useful model for studying HCV replication in vitro.