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Comparative Study
. 2002 Dec;76(23):12242-9.
doi: 10.1128/jvi.76.23.12242-12249.2002.

Virus replication and cytokine production in dengue virus-infected human B lymphocytes

Yu-Wen Lin  1 , Kuan-Ju Wang, Huan-Yao Lei, Yee-Shin Lin, Trai-Ming Yeh, Hsiao-Sheng Liu, Ching-Chuan Liu, Shun-Hua Chen
Affiliations
Comparative Study

Virus replication and cytokine production in dengue virus-infected human B lymphocytes

Yu-Wen Lin et al. J Virol. 2002 Dec.

Abstract

Dengue virus (DV) replication, antibody-enhanced viral infection, and cytokine responses of human primary B lymphocytes (cells) were characterized and compared with those of monocytes. The presence of a replication template (negative-strand RNA intermediate), viral antigens including core and nonstructural proteins, and increasing amounts of virus with time postinfection indicated that DV actively replicated in B cells. Virus infection also induced B cells to produce interleukin-6 and tumor necrosis factor alpha, which have been previously implicated in virus pathogenesis. In addition, a heterologous antibody was able to enhance both virus and cytokine production in B cells. Furthermore, the levels of virus replication, antibody-enhanced virus replication, and cytokine responses observed in B cells were not statistically different from those in monocytes. These results suggest that B cells may play an important role in DV pathogenesis.

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Figures

FIG. 1.
FIG. 1.
Active DV replication in human B cells. (A) B cells were infected with DV2 (solid diamonds) or DV3 (open triangles) and monocytes were infected with DV2 (open diamonds) at an MOI of 10. Culture supernatants were collected at the indicated times to determine virus titers. Numbers adjacent to symbols designate individual donors, and bars represent the mean values for each group. (B) After infection or mock infection of B cells of donor 4 with DV2 at an MOI of 10 in the absence or presence of control or DV3 immune serum, total RNA was isolated from B cells at 24 h p.i. Portions of each sample were hybridized with β2 and D1 primers (top) and the D2 primer (middle) to analyze negative- and positive-strand RNA genomes. Afterward, half of each sample was incubated with (+) or without (−) RT, amplified by PCR, and run in adjacent lanes. β-Actin (bottom) served as an internal control. M, DNA marker. (C) Staining of DV2-infected B cells. After mock treatment (1) or DV2 infection (2 to 4) at an MOI of 10, B cells of donor 6 were collected 48 h later and stained for intracellular viral core protein (1 and 2) and cell surface CD19 molecules (3). Panel 4 is the compiled image of panels 2 and 3.
FIG. 1.
FIG. 1.
Active DV replication in human B cells. (A) B cells were infected with DV2 (solid diamonds) or DV3 (open triangles) and monocytes were infected with DV2 (open diamonds) at an MOI of 10. Culture supernatants were collected at the indicated times to determine virus titers. Numbers adjacent to symbols designate individual donors, and bars represent the mean values for each group. (B) After infection or mock infection of B cells of donor 4 with DV2 at an MOI of 10 in the absence or presence of control or DV3 immune serum, total RNA was isolated from B cells at 24 h p.i. Portions of each sample were hybridized with β2 and D1 primers (top) and the D2 primer (middle) to analyze negative- and positive-strand RNA genomes. Afterward, half of each sample was incubated with (+) or without (−) RT, amplified by PCR, and run in adjacent lanes. β-Actin (bottom) served as an internal control. M, DNA marker. (C) Staining of DV2-infected B cells. After mock treatment (1) or DV2 infection (2 to 4) at an MOI of 10, B cells of donor 6 were collected 48 h later and stained for intracellular viral core protein (1 and 2) and cell surface CD19 molecules (3). Panel 4 is the compiled image of panels 2 and 3.
FIG. 2.
FIG. 2.
Antibody-enhanced DV infection of B-cell line Raji. Viruses were incubated with control or DV3 immune serum diluted as indicated before infecting cells at an MOI of 10. Culture supernatants were collected at the indicated times, and virus titers were determined. The means ± SEM of duplicate samples are shown. ∗, P < 0.05 when immune serum-treated samples are compared to control serum-treated samples.
FIG. 3.
FIG. 3.
Antibody-enhanced DV infection of primary mononuclear cells. (A) Viruses were incubated with a 1:60,000 dilution of control serum or DV3 immune serum before infecting B cells of donor 4 at MOIs of 1 and 10. Culture supernatants were collected at the indicated times, and virus titers were determined. The means ± SEM of duplicate samples are shown. (B) B cells (solid diamonds) and monocytes (open diamonds) of donors were infected with DV2 at an MOI of 10 in the presence of control serum or DV3 serum. Culture supernatants were collected at 48 h p.i., and virus titers were determined. The fold increase of virus titer due to ADE was calculated as follows: fold increase = virus titer of DV3 immune serum sample/virus titer of control serum sample. Numbers adjacent to symbols designate individual donors, and bars represent the mean values for each group.
FIG. 4.
FIG. 4.
Antibody-enhanced NS1 expression in infected B cells. DV2 was incubated with normal human serum or DV3 immune serum before infecting B cells of donors 4, 5, and 8 at an MOI of 1. At 48 h p.i., infected cells were collected and subjected to flow-cytometric analysis of the viral NS1 protein. The results are expressed as the mean percentages of cells that express the NS1 protein of DV ± SEM.
FIG. 5.
FIG. 5.
Cytokine production in DV-infected human mononuclear cells. The same 48-h supernatant samples used for virus titration in Fig. 1A and their corresponding mock-treated samples of B cells (solid diamonds) and monocytes (open diamonds) were assayed for IL-6 (A) and TNF-α (C) by using ELISA kits. Numbers adjacent to symbols represent individual donors, and bars represent mean values for each group. (B and D) Correlation of IL-6 and virus production (B) and correlation of TNF-α and virus production (D) in infected B cells (top) and monocytes (bottom). The amounts of the indicated cytokines produced in the infected samples were plotted against the amounts of virus produced in the same samples. The best-fit lines were generated by linear regression of log10 values. Star, virus production in B cells for donor 8.
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References

    1. Anderson, R., S. Wang, C. Osiowy, and A. C. Issekutz. 1997. Activation of endothelial cells via antibody-enhanced dengue virus infection of peripheral blood monocytes. J. Virol. 71:4226-4232. - PMC - PubMed
    1. Bethell, D. B., K. Flobbe, X. T. Cao, N. P. Day, T. P. Pham, W. A. Buurman, M. J. Cardosa, N. J. White, and D. Kwiatkowski. 1998. Pathological and progonostic role of cytokines in dengue hemorrhagic fever. J. Infect. Dis. 177:778-782. - PubMed
    1. Boehme, M. W., Y. Deng, U. Raeth, A. Bierhaus, R. Ziegler, W. Stremmel, and P. P. Nawroth. 1996. Release of thrombomodulin from endothelial cells by concerted action of TNF-α and neutrophils: in vivo and in vitro studies. Immunology 87:134-140. - PMC - PubMed
    1. Boonpucknavig, S., N. Bhamarapravati, S. Nimmannitya, A. Phalavadhtana, and J. Siripont. 1976. Immunofluorescent staining of the surfaces of lymphocytes in suspension from patients with dengue hemorrhagic fever. Am. J. Pathol. 85:37-48. - PMC - PubMed
    1. Brandt, W. E., J. M. McCown, F. H. J. Top, W. H. Bancroft, and P. K. Russell. 1979. Effect of passage history on dengue-2 virus replication in subpopulations of human leukocytes. Infect. Immun. 26:534-541. - PMC - PubMed

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