Comparison of P388D1 Mouse Macrophage Cell Line and Human Monocytes for Assay of Dengue-2 Infection-Enhancing Antibodies

Scott B. HalsteadDepartment of Tropical Medicine and Medical Microbiology, John A. Burns School of Medicine, University of Hawaii, Sir William Dunn School of Pathology, University of Oxford, Honolulu, Hawaii 96816, England

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Kay LarsenDepartment of Tropical Medicine and Medical Microbiology, John A. Burns School of Medicine, University of Hawaii, Sir William Dunn School of Pathology, University of Oxford, Honolulu, Hawaii 96816, England

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Srisakul KliksDepartment of Tropical Medicine and Medical Microbiology, John A. Burns School of Medicine, University of Hawaii, Sir William Dunn School of Pathology, University of Oxford, Honolulu, Hawaii 96816, England

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J. S. M. PeirisDepartment of Tropical Medicine and Medical Microbiology, John A. Burns School of Medicine, University of Hawaii, Sir William Dunn School of Pathology, University of Oxford, Honolulu, Hawaii 96816, England

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Jane CardosaDepartment of Tropical Medicine and Medical Microbiology, John A. Burns School of Medicine, University of Hawaii, Sir William Dunn School of Pathology, University of Oxford, Honolulu, Hawaii 96816, England

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James S. PorterfieldDepartment of Tropical Medicine and Medical Microbiology, John A. Burns School of Medicine, University of Hawaii, Sir William Dunn School of Pathology, University of Oxford, Honolulu, Hawaii 96816, England

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Tissue culture-adapted dengue 2 virus (DEN 2), strain 16681, exhibits antibody-dependent enhancement of infection (ADE) in P388D1 cells, a mouse macrophage-like cell line. ADE is dependent upon maintaining DEN 2 multiplicity of infection at between 0.1 and 0.001, and can be simply measured in multi-well plastic plates. The assay uses either trypsinized or non-trypsinized P388D1 cells at 5 × 105 cells per ml, an appropriate dilution of DEN 2 virus, and a source of antibody, and is most conveniently performed without further washing of stationary cultures, which are incubated in 5% CO2. Trypsinization of P388D1 cells prior to the addition of virus-serum mixtures reduced infection in control cultures thus increasing ADE. When cells were washed after incubation of virus-serum mixtures for 1 hour, a paradoxical increase of infection in cultures exposed to virus plus normal serum was noted, which reduced the sensitivity of the ADE assay. Using human cord blood sera, ADE titers measured in human monocytes and P388D1 cells were closely similar. This convenient and economical assay will facilitate large scale biological and epidemiological studies of dengue virus enhancing antibodies.

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