Volume 80, Issue 1
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645


We describe microneutralization assays that used automated 96-well enzyme-linked immunospot (ELI-SPOT) readout instrumentation to measure human anti-dengue virus (DENV) antibodies in CV-1 cells that were stably transfected to express human FcγRIIA (CD32) using conventional Vero cells as a comparator. Classic plaque reduction neutralization test (PRNT) end-point titers were determined by probit analysis. Neutralization titers against DENV measured in CV-1 transfectants were expressed in terms of both conventional 50% to 90% PRNT end-point titers and differential infectivity of antibody-treated virus in control and CD32-expressing CV-1 cells. Significantly reduced PRNT titers and strikingly heightened infectivity (up to 100-fold) of antibody-treated DENV was observed in CV-1 CD32 transfectants compared with that observed in control CV-1 or Vero cells. Because DENVs may preferentially replicate in CD32-expressing monocytes/macrophages and dendritic cells, , it is possible that CD32 introduced into a conventional DENV neutralization assay might provide results that better correlate with protection.


Article metrics loading...

Loading full text...

Full text loading...



  1. Vorndam V, Beltran M, 2002. Enzyme-linked immunosorbent assay-format microneutralization test for dengue viruses. Am J Trop Med Hyg 66 : 208–212.
  2. Thacker WL, Lewis VJ, Baer GM, Sather GE, 1978. A rapid fluorescent focus-inhibition test for determining dengue neutralizing antibody and for identifying prototype dengue viruses. Can J Microbiol 24 : 1553–1556.
  3. Roehrig JTHJ, Barrett ADT, 2008. Guidelines for plaque-reduction neutralization testing of human antibodies to dengue viruses. Viral Immunol 21 : 123–132.
  4. Durbin AP, Vargas MJ, Wanionek K, Hammond SN, Gordon A, Rocha C, Balmaseda A, Harris E, 2008. Phenotyping of peripheral blood mononuclear cells during acute dengue illness demonstrates infection and increased activation of monocytes in severe cases compared to classic dengue fever. Virology 376 : 429–435.
  5. Wu SJ, Grouard-Vogel G, Sun W, Mascola JR, Brachtel E, Putvatana R, Louder MK, Filgueira L, Marovich MA, Wong HK, Blauvelt A, Murphy GS, Robb ML, Innes BL, Birx DL, Hayes CG, Frankel SS, 2000. Human skin Langerhans cells are targets of dengue virus infection. Nat Med 6 : 816–820.
  6. Halstead SB, 1988. Pathogenesis of dengue: challenges to molecular biology. Science 239 : 476–481.
  7. Blackley S, Kou Z, Chen H, Quinn M, Rose RC, Schlesinger JJ, Coppage M, Jin X, 2007. Primary human splenic macrophages, but not T or B cells, are the principal target cells for dengue virus infection in vitro. J Virol 81 : 13325–13334.
  8. Schlesinger JJ, Brandriss MW, Walsh EE, 1987. Protection of mice against dengue 2 virus encephalitis by immunization with the dengue 2 virus non-structural glycoprotein NS1. J Gen Virol 68 : 853–857.
  9. Russell PK, Nisalak A, Sukhavachana P, Vivona S, 1967. A plaque reduction test for dengue virus neutralizing antibodies. J Immunol 99 : 285–290.
  10. Rodrigo WW, Jin X, Blackley SD, Rose RC, Schlesinger JJ, 2006. Differential enhancement of dengue virus immune complex infectivity mediated by signaling-competent and signaling-incompetent human Fcgamma RIA (CD64) or Fcgamma RIIA (CD32). J Virol 80 : 10128–10138.
  11. Morens DM, Rigau-Perez JG, Lopez-Correa RH, Moore CG, Ruiz-Tiben EE, Sather GE, Chiriboga J, Eliason DA, Casta-Velez A, Woodall JP, 1986. Dengue in Puerto Rico, 1977: public health response to characterize and control an epidemic of multiple serotypes. Am J Trop Med Hyg 35 : 197–211.
  12. Messer WB, Vitarana UT, Sivananthan K, Elvtigala J, Preethimala LD, Ramesh R, Withana N, Gubler DJ, De Silva AM, 2002. Epidemiology of dengue in Sri Lanka before and after the emergence of epidemic dengue hemorrhagic fever. Am J Trop Med Hyg 66 : 765–773.
  13. Parren PW, Burton DR, 2001. The antiviral activity of antibodies in vitro and in vivo. Adv Immunol 77 : 195–262.
  14. Endy TP, Nisalak A, Chunsuttitwat S, Vaughn DW, Green S, Ennis FA, Rothman AL, Libraty DH, 2004. Relationship of preexisting dengue virus (DV) neutralizing antibody levels to viremia and severity of disease in a prospective cohort study of DV infection in Thailand. J Infect Dis 189 : 990–1000.
  15. Simmons CP, Chau TN, Thuy TT, Tuan NM, Hoang DM, Thien NT, Lien le B, Quy NT, Hieu NT, Hien TT, McElnea C, Young P, Whitehead S, Hung NT, Farrar J, 2007. Maternal antibody and viral factors in the pathogenesis of dengue virus in infants. J Infect Dis 196 : 416–424.
  16. Halstead SB, 1989. Antibody, macrophages, dengue virus infection, shock, and hemorrhage: a pathogenetic cascade. Rev Infect Dis 11 (Suppl 4): S830–S839.

Data & Media loading...

  • Received : 25 Jun 2008
  • Accepted : 19 Sep 2008

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error