Efficient infection of a human B cell line with cell-free Kaposi's sarcoma-associated herpesvirus.

[primary effusion lymphoma]

Kaposi 's sarcoma -associated herpesvirus (KSHV ) is causatively linked to two B cell lymphoproliferative disorders , multicentric Castleman 's disease and primary effusion lymphoma . Latently infected B cells are a major KSHV reservoir , and virus activation from tonsillar B cells can result in salivary shedding and virus transmission . Paradoxically , human B cells (primary and continuous ) are notoriously refractory to infection , thus posing a major obstacle to the study of KSHV in this cell type . By performing a strategic search of human B cell lymphoma lines , we found that MC 116 cells were efficiently infected by cell-free KSHV . Upon exposure to recombinant KSHV .219 , enhanced green fluorescent protein reporter expression was detected in 17 to 20 % of MC 116 cells . Latent-phase transcription and protein synthesis were detected by reverse transcription-PCR and detection of latency-associated nuclear antigen expression , respectively , in cell lysates and individual cells . Selection based on the puromycin resistance gene in KSHV .219 yielded cultures with all cells infected . After repeated passaging of the selected KSHV-infected cells without puromycin , latent KSHV was maintained in a small fraction of cells . Infected MC 116 cells could be induced into lytic phase with histone deacetylase inhibitors , as is known for latently infected non-B cell lines , and also selectively by the B cell-specific pathway involving B cell receptor cross-linking . Lytic -phase transition was documented by red fluorescent protein reporter expression , late structural glycoprotein (K 8 .1 A , gH ) detection , and infectious KSHV production . MC 116 cells were CD 27 (-)/CD 10 (+), characteristic of transitional B cells . These findings represent an important step in the establishment of an efficient continuous B cell line model to study the biologically relevant steps of KSHV infection . Kaposi 's sarcoma -associated herpesvirus (KSHV ) causes two serious pathologies of B cells , the antibody-producing cells of the immune system . B cells are a major reservoir for KSHV persistence in the body . Paradoxically , in the laboratory , B cells are extremely difficult to infect with KSHV ; this problem greatly hinders scientific analysis of B cell infection . We describe our search for and successful identification of a stable human B cell line that can be efficiently infected by KSHV . Upon infection of these cells , the virus goes into a quiet latent phase , a characteristic feature of many herpesvirus infections . The virus can be triggered to enter an active lytic phase by treatments known to stimulate normal B cell functions . These findings suggest that the new B cell line will be a valuable model in which to study KSHV infection of this major target cell type .

Diseases
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Diseases presenting "latently infected non-b" symptom

primary effusion lymphoma

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