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Suppression of EBV release from irradiated B lymphoblastoid cell-lines: superior activity of ganciclovir compared with acyclovir. Cytotherapy 2003;5(4):323-35

Date

08/29/2003

Pubmed ID

12944239

DOI

10.1080/14653240310002243

Scopus ID

2-s2.0-0142031139   14 Citations

Abstract

BACKGROUND: B lymphoblastoid cell-lines (BLCL), generated by exposure of PBMC to a laboratory strain of EBV, are commonly utilized in the preparation of T cells used for immunotherapy. Although most B cells are latently infected, BLCL contain a subset of cells that harbor infectious virus, which could be released into the infusion product during preparation. To reduce this known risk, laboratories have pretreated BLCL for > or = 14 days with 100 microM acyclovir (ACV), an inhibitor of viral DNA polymerase, prior to use. We tested the effectiveness of ACV in preventing the release of infectious virus from irradiated fresh and previously frozen BLCL, and compared its effects with those of ganciclovir (GCV).

METHODS: BLCL were grown for 14 days in medium containing various doses of ACV or GCV, washed, irradiated, and tested for the presence of infectious virus in co-culture assays with cord blood mononuclear cells(CBMC) (21 CBMC to BLCL). B-cell transformation was assessed at 3-4 weeks of culture.

RESULTS: Both fresh and previously frozen BLCL released infectious virus, which transformed nearly all (92%) of CBMC co-cultures (n = 52). Transformation was not prevented by treatment with 100 microM ACV (88%, n = 52). Increasing the ACV dose to 200 microM (or 50 microg/mL) still allowed transformation in 4/9 (44%) cultures, while this and higher doses severely reduced the proliferation rate of the BLCL during ACV exposure. Infectious virus release was detectable within 1 day of ACV removal and BLCL irradiation. In contrast, GCV was able to prevent infectious virus release in 12/12 co-cultures at a concentration (15 microM) that only modestly reduced BLCL growth.

DISCUSSION: These results indicate that GCV is more effective at preventing release of infectious EBV from irradiated BLCL than ACV at concentrations that do not severely inhibit B-cell growth.

Author List

Keever-Taylor CA, Behn B, Konings S, Orentas R, Davies B, Margolis D

Author

David A. Margolis MD Interim Chair, Professor in the Pediatrics department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Acyclovir
Antigens, CD20
B-Lymphocytes
CD3 Complex
CD56 Antigen
Cell Division
Cell Line, Transformed
Cell Transformation, Viral
Coculture Techniques
Cytomegalovirus
Cytotoxicity Tests, Immunologic
Dose-Response Relationship, Drug
Fetal Blood
Fibroblasts
Flow Cytometry
Freezing
Ganciclovir
HLA-DR Antigens
Herpesvirus 4, Human
Humans
Immunoglobulin G
Kinetics
Leukocyte Common Antigens
Leukocytes, Mononuclear
T-Lymphocytes, Cytotoxic
Viral Load
Viral Plaque Assay