Zika, chikungunya, and dengue viral infections in human peripheral blood mononuclear cells: cell susceptibility and gene expression

  • Ahmad Husein Alkaff Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Indonesia, Depok, Indonesia; Eijkman Institute for Molecular Biology, Ministry of Research and Technology/ National Agency for Research and Innovation, Jakarta, Indonesia https://orcid.org/0000-0001-8264-6494
  • Benediktus Yohan Eijkman Institute for Molecular Biology, Ministry of Research and Technology/ National Agency for Research and Innovation, Jakarta, Indonesia
  • Usman Sumo Friend Tambunan Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Indonesia, Depok, Indonesia https://orcid.org/0000-0001-6025-2195
  • R. Tedjo Sasmono Eijkman Institute for Molecular Biology, Ministry of Research and Technology/ National Agency for Research and Innovation, Jakarta, Indonesia http://orcid.org/0000-0003-0986-2590
Keywords: chemokines, chikungunya virus, cytokines, dengue virus, mononuclear leukocytes, Zika virus
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BACKGROUND Infections of Zika (ZIKV), dengue (DENV), and chikungunya viruses (CHIKV) are presented with similar clinical symptoms; these often lead to misdiagnosis. Viremia levels and host immune responses may contribute to disease severity. This study was aimed to characterize the ability of ZIKV, CHIKV, and DENV to infect human peripheral blood mononuclear cells (PBMCs) and assess the expression of tumor necrosis factor (TNF)-α, interleukin (IL)-10, and interferon gamma-induced protein (IP)- 10 genes in response to the viral infections. 

METHODS PBMCs were isolated from healthy donors using gradient centrifugation. Cells were infected with Indonesian isolates of ZIKV, CHIKV, and DENV for 48 hours. Plaque assays were performed to measure viable virus titers, while viral genomic RNA and the gene expression of TNF-α, IL-10, and IP-10 were determined using real-time quantitative reverse transcription-polymerase chain reaction. 

RESULTS The susceptibility of PBMCs to ZIKV, CHIKV, and DENV infection was observed, and the viable virus titer and viral genome quantity were found to be significantly higher in ZIKV and CHIKV. All viruses induced the expression of immune-related proteins. The TNF-α gene was upregulated by all viruses to relatively similar levels. IL-10 expression was highest in response to ZIKV, followed by CHIKV. In contrast, IP-10 expression was highly upregulated in DENV-infected cells and only moderately expressed in ZIKV- and CHIKV-infected cells. 

CONCLUSIONS ZIKV, CHIKV, and DENV clinical isolates infected PBMCs with different levels of virus infectivity. The gene expression of IL-10 was highly upregulated in ZIKV infection and IP-10 in DENV infection.


  1. Jones KE, Patel NG, Levy MA, Storeygard A, Balk D, Gittleman JL, et al. Global trends in emerging infectious diseases. Nature. 2008;451(7181):990-3. https://doi.org/10.1038/nature06536

  2. Paixão ES, Teixeira MG, Rodrigues LC. Zika, chikungunya and dengue: the causes and threats of new and re-emerging arboviral diseases. BMJ Glob Health. 2018;3(Suppl 1):e000530. https://doi.org/10.1136/bmjgh-2017-000530

  3. Haddow AD, Schuh AJ, Yasuda CY, Kasper MR, Heang V, Huy R, et al. Genetic characterization of Zika virus strains: geographic expansion of the Asian lineage. PLoS Negl Trop Dis. 2012;6(2):e1477. https://doi.org/10.1371/journal.pntd.0001477

  4. Perkasa A, Yudhaputri F, Haryanto S, Hayati RF, Ma'roef CN, Antonjaya U, et al. Isolation of Zika virus from febrile patient, Indonesia. Emerging Infect Dis. 2016;22(5):924-5. https://doi.org/10.3201/eid2205.151915

  5. Sasmono RT, Perkasa A, Yohan B, Haryanto S, Yudhaputri FA, Hayati RF, et al. Chikungunya detection during dengue outbreak in Sumatra, Indonesia: clinical manifestations and virological profile. Am J Trop Med Hyg. 2017;97(5):1393-8. https://doi.org/10.4269/ajtmh.16-0935

  6. Megawati D, Masyeni S, Yohan B, Lestarini A, Hayati RF, Meutiawati F, et al. Dengue in Bali: clinical characteristics and genetic diversity of circulating dengue viruses. PLoS Negl Trop Dis. 2017;11(5):e0005483. https://doi.org/10.1371/journal.pntd.0005483

  7. Banerjee A, Shukla S, Pandey AD, Goswami S, Bandyopadhyay B, Ramachandran V, et al. RNA-Seq analysis of peripheral blood mononuclear cells reveals unique transcriptional signatures associated with disease progression in dengue patients. Transl Res. 2017;186:62-78.e9. https://doi.org/10.1016/j.trsl.2017.06.007

  8. Luo H, Winkelmann ER, Fernandez-Salas I, Li L, Mayer SV, Danis-Lozano R, et al. Zika, dengue and yellow fever viruses induce differential anti-viral immune responses in human monocytic and first trimester trophoblast cells. Antiviral Res. 2018;151:55-62. https://doi.org/10.1016/j.antiviral.2018.01.003

  9. Colavita F, Bordoni V, Caglioti C, Biava M, Castilletti C, Bordi L, et al. ZIKV infection induces an inflammatory response but fails to activate types I, II, and III IFN response in human PBMC. Mediators Inflamm. 2018;2018:2450540. https://doi.org/10.1155/2018/2450540

  10. Chaaitanya IK, Muruganandam N, Sundaram SG, Kawalekar O, Sugunan AP, Manimunda SP, et al. Role of proinflammatory cytokines and chemokines in chronic arthropathy in CHIKV infection. Viral Immunol. 2011;24(4):265-71. https://doi.org/10.1089/vim.2010.0123

  11. Michlmayr D, Andrade P, Gonzalez K, Balmaseda A, Harris E. CD14+CD16+ monocytes are the main target of Zika virus infection in peripheral blood mononuclear cells in a paediatric study in Nicaragua. Nat Microbiol. 2017;2(11):1462-70. https://doi.org/10.1038/s41564-017-0035-0

  12. Ruiz Silva M, Aguilar Briseño JA, Upasani V, van der Ende-Metselaar H, Smit JM, Rodenhuis-Zybert IA. Suppression of chikungunya virus replication and differential innate responses of human peripheral blood mononuclear cells during co-infection with dengue virus. PLoS Negl Trop Dis. 2017;11(6):e0005712. https://doi.org/10.1371/journal.pntd.0005712

  13. Kelvin AA, Banner D, Silvi G, Moro ML, Spataro N, Gaibani P, et al. Inflammatory cytokine expression is associated with chikungunya virus resolution and symptom severity. PLoS Negl Trop Dis. 2011;5(8):e1279. https://doi.org/10.1371/journal.pntd.0001279

  14. Tappe D, Pérez-Girón JV, Zammarchi L, Rissland J, Ferreira DF, Jaenisch T, et al. Cytokine kinetics of Zika virus-infected patients from acute to reconvalescent phase. Med Microbiol Immunol. 2016;205(3):269-73. https://doi.org/10.1007/s00430-015-0445-7

  15. Rathakrishnan A, Wang SM, Hu Y, Khan AM, Ponnampalavanar S, Lum LC, et al. Cytokine expression profile of dengue patients at different phases of illness. PLoS One. 2012;7(12):e52215. https://doi.org/10.1371/journal.pone.0052215

  16. Rathakrishnan A, Klekamp B, Wang SM, Komarasamy TV, Natkunam SK, Sathar J, et al. Clinical and immunological markers of dengue progression in a study cohort from a hyperendemic area in Malaysia. PLoS One. 2014;9(3):e92021. https://doi.org/10.1371/journal.pone.0092021

  17. Kam YW, Leite JA, Lum FM, Tan JJL, Lee B, Judice CC, et al. Specific biomarkers associated with neurological complications and congenital central nervous system abnormalities from Zika virus-infected patients in Brazil. J Infect Dis. 2017;216(2):172-81. https://doi.org/10.1093/infdis/jix261

  18. Green S, Vaughn DW, Kalayanarooj S, Nimmannitya S, Suntayakorn S, Nisalak A, et al. Elevated plasma interleukin-10 levels in acute dengue correlate with disease severity. J Med Virol. 1999;59(3):329-34. https://doi.org/10.1002/(SICI)1096-9071(199911)59:3<329::AID-JMV12>3.0.CO;2-G

  19. Qin S, Rottman JB, Myers P, Kassam N, Weinblatt M, Loetscher M, et al. The chemokine receptors CXCR3 and CCR5 mark subsets of T cells associated with certain inflammatory reactions. J Clin Invest. 1998;101(4):746-54. https://doi.org/10.1172/JCI1422

  20. Hanaoka R, Kasama T, Muramatsu M, Yajima N, Shiozawa F, Miwa Y, et al. A novel mechanism for the regulation of IFN-γ inducible protein-10 expression in rheumatoid arthritis. Arthritis Res Ther. 2003;5(2):R74-81. https://doi.org/10.1186/ar616

  21. Kuan WP, Tam LS, Wong CK, Ko FW, Li T, Zhu T, et al. CXCL 9 and CXCL 10 as sensitive markers of disease activity in patients with rheumatoid arthritis. J Rheumatol. 2010;37(2):257-64. https://doi.org/10.3899/jrheum.090769

  22. Barros JBS, da Silva PAN, Koga CR, Gonzalez-Dias P, Carmo Filho JR, Nagib PRA, et al. Acute Zika virus infection in an endemic area shows modest proinflammatory systemic immunoactivation and cytokine-symptom associations. Front Immunol. 2018;9:821. https://doi.org/10.3389/fimmu.2018.00821

  23. Zhao L, Huang X, Hong W, Qiu S, Wang J, Yu L, et al. Slow resolution of inflammation in severe adult dengue patients. BMC Infect Dis. 2016;16:291. https://doi.org/10.1186/s12879-016-1596-x

How to Cite
Alkaff AH, Yohan B, Tambunan USF, Sasmono RT. Zika, chikungunya, and dengue viral infections in human peripheral blood mononuclear cells: cell susceptibility and gene expression. Med J Indones [Internet]. 2020Jun.30 [cited 2020Aug.7];29(2):129-35. Available from: https://mji.ui.ac.id/journal/index.php/mji/article/view/3548
Basic Medical Research