Propagation and characterization of human herpesvirus-7 (HHV-7) isolates in a continuous T-lymphoblastoid cell line (SupT1)
| Autor: | J.E. Whitman, Dharam V. Ablashi, M Handy, L.G Chatlynne, Anthony L. Komaroff, W. Lapps, Z N Berneman, J Bernbaum, B. Kramarsky |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 1998 |
| Předmět: |
Adult
Virus Cultivation viruses T-Lymphocytes Fluorescent Antibody Technique Enzyme-Linked Immunosorbent Assay Herpesvirus 7 Human medicine.disease_cause Antibodies Viral Lymphocyte Activation Peripheral blood mononuclear cell Polymerase Chain Reaction Sensitivity and Specificity Virus Herpesviridae Cell Line Antigen Cytopathogenic Effect Viral Virology medicine Morphogenesis Humans Child Antigens Viral Infectivity Immunodiagnostics Fatigue Syndrome Chronic biology virus diseases Microscopy Electron Cell culture DNA Viral biology.protein Antibody |
| Zdroj: | Journal of virological methods |
| ISSN: | 0166-0934 |
| Popis: | After initial culture of HHV-7 in PHA-stimulated human cord blood mononuclear cells (HCBMC), six HHV-7 isolates were propagated successfully in an immature continuous T-lymphoblastoid cell line SupT1. All six isolates infected efficiently the SupT1 cells, and the infected cells became grossly enlarged and multinucleated 7–21 days post-infection. Various stages of HHV-7 morphogenesis were detected. Cell-free supernatants from HHV-7-infected SupT1 cells were infectious to HCBMC as well as to SupT1 cells. The HHV-7-infected SupT1 and HCBMC cell lysates contained more infectious virus than the centrifuged cell culture fluid supernates from the same culture. The HHV-7 isolates H7-2, H7-3, JHC, and JB, concentrated 500 times, had average infectivity titers of 103.0 TCID50/ml while strains H7-4 and KHR titered approximately 1–2 logs higher. When all six HHV-7 isolates were propagated in SupT1 and culture fluid supernatants were examined 14–21 days post-infection by negative stain electron microscopy they contained an average of 1.9×109 virus particles/liter. IFA and ELISA, using HHV-7/SupT1 cell lysate as an antigen, seem to correlate well in detecting high and low HHV-7 antibody in sera from chronic fatigue patients and healthy donors as controls. HHV-7 from SupT1 cell culture was free of HHV-6 and other human herpesviruses as tested by PCR, and the HHV-7 PCR signal was still strong when the viral preparation was diluted to 4.82×102 genome copies. Since HCBMC are expensive to obtain and available in only small amounts, it is difficult to obtain large quantities of HHV-7 antigen. On the other hand, the SupT1 cell is an excellent source to produce consistently sufficient quantities of HHV-7 for purification studies, development of immunodiagnostics, in vivo infectivity studies, evaluation of antiviral drugs, and molecular biological studies. |
| Databáze: | OpenAIRE |
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