A major goal of this proposal is to elucidate processes by which innate immune sensing mechanisms program B lineage cells to produce protective anti-West Nile virus (WNV) IgM and IgG antibodies. Humoral immunity to WNV is both necessary and sufficient to protect hosts from disease, however, the early events that coordinate the initial innate and subsequence adaptive antibody (Ab) responses to WNV are not well understood. We will elucidate how dendritic cells (DCs) and B cells respond to WNV and how rapid IgM and IgG anti-WNV neutralizing Abs develop. We hypothesize that initial innate programming of DC subsets dictate both the quality and the duration of protective Ab responses to WNV. To investigate this hypothesis we will conduct the following Specific Aims:
Aim 1 : We will compare the course of pathogenic WNV Texas (WN-TX) and attenuated WNV Madagascar (WN-MAD) strains early after in vivo infection. We propose that high and low pathogenic WNVs differ in how they infect, alter and program DCs. We will quantify changes early after infection in DC subset numbers, survival, cytokine production and infection. With Project 1, we will determine what innate immune factors regulate initial DC responses and susceptibility to WNV infection by comparing DC responses and infection in wildtype (WT) mice vs. C3 and IPS-1 KO mice Using mixed bone marrow (BM) chimeras, in which DCs have been ablated, we will determine how DC ablation or the absence in DCs of IPS-1, TLR-3 or other innate immune factors affects early events after WNV infection.
Aim 2 : To determine the molecular and cellular requirements for the development of rapid IgM neutralizing antibody responses to WNV. To determine the role of marginal zone (MZ) B cells in protective IgM immunity to WNV, we will assess IgM responses and susceptibility of MZ B cell-deficient CD22 KO and Notch2fl/fl conditional KO mice. DCs play a pivotal role in rapid IgM responses to encapsulated bacteria;using mixed BM chimeras, in which DCs have been ablated, we will determine how DC ablation or the absence in DCs of IPS-1, BAFF or other innate immune factors affects the development of neutralizing IgM Ab responses.
Aim 3 : We will investigate if MZ B cells are required for immune IgM to promote protective IgG immunity. Delivery of Ag to MZ-associated DCs can lead to a strong IgG antibody response. We will examine if delivery of WNV E protein to MZ DCs can induce IgM and IgG neutralizing Ab responses, and if so whether this form of Ag delivery can be used to induce protective, sterilizing immunity to WNV. These studies will lead to new insights into how best to design vaccines to WNV and other flaviviruses.

Public Health Relevance

WNV is an NIAID Category B infectious agent that has recently emerged in the Western hemisphere. It continues to be a serious public health threat to both humans and animals, and yet relatively little is known about how best to induce sustained humoral immunity to WNV or other flaviviruses. These studies will lead to new insights into how best to design effective vaccines that induce neutralizing antibodies to WNV and other flaviviruses.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Program--Cooperative Agreements (U19)
Project #
Application #
Study Section
Special Emphasis Panel (ZAI1-BDP-I)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Washington
United States
Zip Code
Bekerman, Elena; Neveu, Gregory; Shulla, Ana et al. (2017) Anticancer kinase inhibitors impair intracellular viral trafficking and exert broad-spectrum antiviral effects. J Clin Invest 127:1338-1352
Daniels, Brian P; Jujjavarapu, Harsha; Durrant, Douglas M et al. (2017) Regional astrocyte IFN signaling restricts pathogenesis during neurotropic viral infection. J Clin Invest 127:843-856
Da Costa, Andreia; Garza, Esteban; Graham, Jessica B et al. (2017) Extrinsic MAVS signaling is critical for Treg maintenance of Foxp3 expression following acute flavivirus infection. Sci Rep 7:40720
Gorman, Jacquelyn A; Hundhausen, Christian; Errett, John S et al. (2017) The A946T variant of the RNA sensor IFIH1 mediates an interferon program that limits viral infection but increases the risk for autoimmunity. Nat Immunol 18:744-752
Jagger, Brett W; Miner, Jonathan J; Cao, Bin et al. (2017) Gestational Stage and IFN-? Signaling Regulate ZIKV Infection In Utero. Cell Host Microbe 22:366-376.e3
Daniels, Brian P; Snyder, Annelise G; Olsen, Tayla M et al. (2017) RIPK3 Restricts Viral Pathogenesis via Cell Death-Independent Neuroinflammation. Cell 169:301-313.e11
Li, Junwei; Baird, Madison A; Davis, Michael A et al. (2017) Dramatic enhancement of the detection limits of bioassays via ultrafast deposition of polydopamine. Nat Biomed Eng 1:
Luo, Huanle; Winkelmann, Evandro; Xie, Guorui et al. (2017) MAVS Is Essential for Primary CD4+ T Cell Immunity but Not for Recall T Cell Responses following an Attenuated West Nile Virus Infection. J Virol 91:
Tajfirouz, Deena; West, Devin M; Yin, Xiao-Tang et al. (2017) CXCL9 compensates for the absence of CXCL10 during recurrent Herpetic stromal keratitis. Virology 506:7-13
Green, Richard; Wilkins, Courtney; Thomas, Sunil et al. (2017) Oas1b-dependent Immune Transcriptional Profiles of West Nile Virus Infection in the Collaborative Cross. G3 (Bethesda) 7:1665-1682

Showing the most recent 10 out of 120 publications