Persistence of circulating memory B cell clones with potential for dengue virus disease enhancement for decades following infection - PubMed (original) (raw)

Persistence of circulating memory B cell clones with potential for dengue virus disease enhancement for decades following infection

Scott A Smith et al. J Virol. 2012 Mar.

Abstract

Symptomatic dengue virus infection ranges in disease severity from an influenza-like illness to life-threatening shock. One model of the mechanism underlying severe disease proposes that weakly neutralizing, dengue serotype cross-reactive antibodies induced during a primary infection facilitate virus entry into Fc receptor-bearing cells during a subsequent secondary infection, increasing viral replication and the release of cytokines and vasoactive mediators, culminating in shock. This process has been termed antibody-dependent enhancement of infection and has significantly hindered vaccine development. Much of our understanding of this process has come from studies using mouse monoclonal antibodies (MAbs); however, antibody responses in mice typically exhibit less complexity than those in humans. A better understanding of the humoral immune response to natural dengue virus infection in humans is sorely needed. Using a high-efficiency human hybridoma technology, we isolated 37 hybridomas secreting human MAbs to dengue viruses from 12 subjects years or even decades following primary or secondary infection. The majority of the human antibodies recovered were broadly cross-reactive, directed against either envelope or premembrane proteins, and capable of enhancement of infection in vitro; few exhibited serotype-specific binding or potent neutralizing activity. Memory B cells encoding enhancing antibodies predominated in the circulation, even two or more decades following infection. Mapping the epitopes and activity of naturally occurring dengue antibodies should prove valuable in determining whether the enhancing and neutralizing activity of antibodies can be separated. Such principles could be used in the rational design of vaccines that enhance the induction of neutralizing antibodies, while lowering the risk of dengue shock syndrome.

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Figures

Fig 1

Serotype-specific strongly neutralizing E protein binding human MAbs. (A) The ability of purified human MAb 5J7 to neutralize the four serotypes of DENV is shown over a concentration range. (B) The ability of purified human MAb 2D22 to neutralize the four serotypes of dengue virus is shown over a concentration range. (C) Purified human MAb 5J7 neutralization of serotype 3 virus over a detailed, broader range of halving dilutions. (D) Purified human MAb 2D22 neutralization of serotype 2 virus over a detailed range of halving dilutions. Flow cytometric neutralization assays were performed using U937-DC-SIGN cells.

Fig 2

Cross-reactive poorly neutralizing E protein binding human MAbs. (A) The ability of purified human MAb 2A15, 2A10, or 1L5 to neutralize or enhance DENV 4 is shown over a concentration range. These MAbs represent a class of fully cross-reactive, weakly neutralizing, and moderately enhancing antibodies. (B) The ability of purified human MAb 3F13, 1A15, or 3H9 to neutralize or enhance DENV 4 is shown over a concentration range. These MAbs represent a class of fully cross-reactive, non-neutralizing, and nonenhancing antibodies. Flow cytometric neutralization assays were performed using U937-DC-SIGN cells. ADE assays were performed using U937 Fc receptor expressing cells. In the absence of antibody, the percentage of U937 cells infected in the ADE assay with DENV 4 was 2.09%.

Fig 3

Cross-reactive poorly neutralizing prM protein binding human MAbs. The ability of purified human MAb 1G6, 4E9, 4F8, or 5L20 to neutralize or enhance DENV1 (A), DENV2 (B), DENV3 (C), or DENV4 (D) is shown over a concentration range. Flow cytometric neutralization assays were performed with U937-DC-SIGN cells. ADE assays were performed using U937 Fc receptor expressing cells. In the absence of antibody, the percentages of U937 cells infected in the ADE assay with DENV1, DENV2, DENV3, or DENV4 were 0.06, 0.17, 0.31, and 1.31%, respectively.

References

1. Anonymous 2010. Locally acquired dengue–Key West, Florida, 2009-2010. MMWR Morb. Mortal. Wkly. Rep. 59:577–581 - PubMed
1. Balsitis SJ, et al. 2010. Lethal antibody enhancement of dengue disease in mice is prevented by Fc modification. PLoS Pathog. 6:e1000790. - PMC - PubMed
1. Beltramello M, et al. 2010. The human immune response to dengue virus is dominated by highly cross-reactive antibodies endowed with neutralizing and enhancing activity. Cell Host Microbe 8:271–283 - PMC - PubMed
1. Bhardwaj S, Holbrook M, Shope RE, Barrett AD, Watowich SJ. 2001. Biophysical characterization and vector-specific antagonist activity of domain III of the tick-borne flavivirus envelope protein. J. Virol. 75:4002–4007 - PMC - PubMed
1. Crill WD, Hughes HR, Delorey MJ, Chang GJ. 2009. Humoral immune responses of dengue fever patients using epitope-specific serotype-2 virus-like particle antigens. PLoS One 4:e4991. - PMC - PubMed

Persistence of circulating memory B cell clones with potential for dengue virus disease enhancement for decades following infection - PubMed (original) (raw)