Continuous release of hepatitis C virus (HCV) by peripheral blood mononuclear cells and B-lymphoblastoid cell-line cultures derived from HCV-infected patients

Abstract

In order to investigate hepatitis C virus (HCV) persistence and replication in peripheral blood mononuclear cells (PBMC) from a group of haemophilic individuals, HCV production and release to PBMC culture supernatants (SNs) from HCV singly infected patients and HIV/HCV co-infected patients was studied. HCV RNA + SNs were found more frequently from HIV/HCV co-infected individuals (89?5 %) with poor reconstitution of their immune status than from singly HCV-infected patients (57 %) or from HIV/HCV co-infected individuals with a good response to highly active anti-retroviral therapy (50 %). The presence of the HCV genome in culture SNs was associated with lower CD4 + T-cell counts and with a more severe clinical picture of HIV infection. In spite of prolonged negative HCV viraemia, PBMC from HIV/HCV co-infected patients released the HCV genome after culture. HCV permissive PBMC allowed generation of HCV productive B cell lines with continuous HCV replication. These findings add further weight to the involvement of PBMCs in persistence of HCV infection and emphasize the role of B lymphocytes as HCV reservoirs.

Extrahepatic replication of HCV: Insights into clinical manifestations and biological consequences

“…20 Although there is limited data to suggest that HCV replication occurs in B lymphocytes, [21][22][23] chronic antigenic stimulation by the virus may trigger B cell proliferation resulting in a wide spectrum of pathology ranging from minor expansion of B cell populations to an aggressive high-grade lymphoma. A high prevalence of HCV in patients with B cell non-Hodgkin's lymphoma (NHL) has been reported in some, but not all studies.…”

“…Several complementary strategies to investigate extrahepatic replication of HCV include DNA-based expression systems, 41,[49][50][51][52][53] lymphocyte cell lines that constitutively express HCV, 21,23,54 peripheral blood mononuclear cell (PBMC) cultures from HCV-infected persons, 21,55,56 direct viral infection of extrahepatic cell types with HCV-infected sera, 35,[57][58][59][60] in situ hybridization and immunohistochemistry, 38,[61][62][63][64] and laser capture microdissection. 37,65 The relevance of each of these strategies to extrahepatic replication of HCV is not currently known and can only be determined through extensive validation and cross-comparison amongst all available methodologies.…”

Clearance of hepatitis C virus RNA from the peripheral blood mononuclear cells of blood donors who spontaneously or therapeutically control their plasma viremia

“…Numerous publications have analyzed extrahepatic compartments of viral replication that could potentially contribute to plasma viremia, most frequently peripheral blood mononuclear cells (PBMC). [7][8][9][10][11][12][13][14] The contribution of PBMC-based HCV replication to total viremia remains unclear, however, as the level of negative-strand HCV RNA in PBMC is very low with respect to that in liver. [15][16][17][18] The continued presence of viral RNA in the PBMC of subjects who had either spontaneously cleared their plasma viremia or cleared viremia following antiviral therapy has recently been reported, raising concerns that PBMC may serve as a long-lived HCV reservoir capable of rekindling systemic infection.…”

Mixed Genotypes in Hepatitis C Virus Infection

“…In our first report that was carried out studying only one culture per patient, HCV+ results were observed in 12 of 21 (57%) patients for the HCV monoinfected group and 23 of 31 (74%) of the coinfected group had HCV positive cultures (Baré et al, 2005). The difference between the populations did not reach statistical differences.…”

“…Using a cell culture system that allows the detection of the HCV genome during prolonged time periods (Baré et al, 2005), we investigated the presence of HCV in peripheral blood mononuclear cell (PBMC) cultures and the HCV genotypes associated to the lymphoid cells. Although peripheral blood mononuclear cells (PBMC) are not the primary site of HCV replication, previous reports emphasize their role as viral reservoirs (Radkowski et al, 2005;Pham et al, 2004).…”