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First experimental in vivo model of enhanced dengue disease severity through maternally acquired heterotypic dengue antibodies.

Ng JK, Zhang SL, Tan HC, Yan B, Martinez JM, Tan WY, Lam JH, Tan GK, Ooi EE, Alonso S - PLoS Pathog. (2014)

Bottom Line: Furthermore, a recent epidemiological study has challenged the influence of maternal antibodies in disease outcome.In this ADE model we demonstrated the role of TNF-α in DEN-induced vascular leakage.In vitro ELISA and ADE assays confirmed the cross-reactive and enhancing properties towards DENV2 of the serum from mice born to DENV1-immune mothers.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, Life Sciences Institute, National University of Singapore, Singapore; Immunology Programme, Yong Loo Lin School of Medicine, Life Sciences Institute, National University of Singapore, Singapore.

ABSTRACT
Dengue (DEN) represents the most serious arthropod-borne viral disease. DEN clinical manifestations range from mild febrile illness to life-threatening hemorrhage and vascular leakage. Early epidemiological observations reported that infants born to DEN-immune mothers were at greater risk to develop the severe forms of the disease upon infection with any serotype of dengue virus (DENV). From these observations emerged the hypothesis of antibody-dependent enhancement (ADE) of disease severity, whereby maternally acquired anti-DENV antibodies cross-react but fail to neutralize DENV particles, resulting in higher viremia that correlates with increased disease severity. Although in vitro and in vivo experimental set ups have indirectly supported the ADE hypothesis, direct experimental evidence has been missing. Furthermore, a recent epidemiological study has challenged the influence of maternal antibodies in disease outcome. Here we have developed a mouse model of ADE where DENV2 infection of young mice born to DENV1-immune mothers led to earlier death which correlated with higher viremia and increased vascular leakage compared to DENV2-infected mice born to dengue naïve mothers. In this ADE model we demonstrated the role of TNF-α in DEN-induced vascular leakage. Furthermore, upon infection with an attenuated DENV2 mutant strain, mice born to DENV1-immune mothers developed lethal disease accompanied by vascular leakage whereas infected mice born to dengue naïve mothers did no display any clinical manifestation. In vitro ELISA and ADE assays confirmed the cross-reactive and enhancing properties towards DENV2 of the serum from mice born to DENV1-immune mothers. Lastly, age-dependent susceptibility to disease enhancement was observed in mice born to DENV1-immune mothers, thus reproducing epidemiological observations. Overall, this work provides direct in vivo demonstration of the role of maternally acquired heterotypic dengue antibodies in the enhancement of dengue disease severity and offers a unique opportunity to further decipher the mechanisms involved.

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In vitro ELISA and ADE assays with the serum from mice born to DENV1 immune mothers.A) Total IgG antibodies against DENV1 or DENV2 were determined by ELISA in the serum from 5-week old mice born to DENV1 immune (black bar) or dengue naïve (white bar) mothers (n = 5 mice per group). B) Anti-DENV1 IgG antibody subclasses were analyzed by ELISA. Legend: serum from mice born to naïve mothers (white bar); serum from mice born to DENV1 immune mothers (black bar). C) Antibody-Dependent Enhancement (ADE) assay. Sera diluted 1/20 from 5-week old mice born to DENV1 immune or dengue naïve mothers (n = 5 mice per group) were mixed with DENV2 virus prior to infection of THP-1 macrophages. At 3 days p.i., the number of infectious particles produced in the supernatant was determined by plaque assay in BHK-21 cells. *, p<0.05.
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ppat-1004031-g008: In vitro ELISA and ADE assays with the serum from mice born to DENV1 immune mothers.A) Total IgG antibodies against DENV1 or DENV2 were determined by ELISA in the serum from 5-week old mice born to DENV1 immune (black bar) or dengue naïve (white bar) mothers (n = 5 mice per group). B) Anti-DENV1 IgG antibody subclasses were analyzed by ELISA. Legend: serum from mice born to naïve mothers (white bar); serum from mice born to DENV1 immune mothers (black bar). C) Antibody-Dependent Enhancement (ADE) assay. Sera diluted 1/20 from 5-week old mice born to DENV1 immune or dengue naïve mothers (n = 5 mice per group) were mixed with DENV2 virus prior to infection of THP-1 macrophages. At 3 days p.i., the number of infectious particles produced in the supernatant was determined by plaque assay in BHK-21 cells. *, p<0.05.

Mentions: To further characterize the nature and properties of the maternal DENV1 specific antibodies circulating in 5-week old mice born to DENV1 immune mothers, ELISA was carried out using UV-inactivated DENV1 or DENV2 particles as coating antigens. Expectedly, elevated absorbance readings were measured when probing for the presence of anti-DENV1 total IgG antibodies in the serum from 5-week old mice born to DENV1 immune mothers (Fig. 8A). In addition, serum from these mice cross-reacted significantly with DENV2 (Fig. 8A). Isotyping of the anti-DENV1 IgG antibodies revealed majority of IgG1 and IgG2a circulating in the serum of mice born to DENV1 immune mothers (Fig. 8B).


First experimental in vivo model of enhanced dengue disease severity through maternally acquired heterotypic dengue antibodies.

Ng JK, Zhang SL, Tan HC, Yan B, Martinez JM, Tan WY, Lam JH, Tan GK, Ooi EE, Alonso S - PLoS Pathog. (2014)

In vitro ELISA and ADE assays with the serum from mice born to DENV1 immune mothers.A) Total IgG antibodies against DENV1 or DENV2 were determined by ELISA in the serum from 5-week old mice born to DENV1 immune (black bar) or dengue naïve (white bar) mothers (n = 5 mice per group). B) Anti-DENV1 IgG antibody subclasses were analyzed by ELISA. Legend: serum from mice born to naïve mothers (white bar); serum from mice born to DENV1 immune mothers (black bar). C) Antibody-Dependent Enhancement (ADE) assay. Sera diluted 1/20 from 5-week old mice born to DENV1 immune or dengue naïve mothers (n = 5 mice per group) were mixed with DENV2 virus prior to infection of THP-1 macrophages. At 3 days p.i., the number of infectious particles produced in the supernatant was determined by plaque assay in BHK-21 cells. *, p<0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3974839&req=5

ppat-1004031-g008: In vitro ELISA and ADE assays with the serum from mice born to DENV1 immune mothers.A) Total IgG antibodies against DENV1 or DENV2 were determined by ELISA in the serum from 5-week old mice born to DENV1 immune (black bar) or dengue naïve (white bar) mothers (n = 5 mice per group). B) Anti-DENV1 IgG antibody subclasses were analyzed by ELISA. Legend: serum from mice born to naïve mothers (white bar); serum from mice born to DENV1 immune mothers (black bar). C) Antibody-Dependent Enhancement (ADE) assay. Sera diluted 1/20 from 5-week old mice born to DENV1 immune or dengue naïve mothers (n = 5 mice per group) were mixed with DENV2 virus prior to infection of THP-1 macrophages. At 3 days p.i., the number of infectious particles produced in the supernatant was determined by plaque assay in BHK-21 cells. *, p<0.05.
Mentions: To further characterize the nature and properties of the maternal DENV1 specific antibodies circulating in 5-week old mice born to DENV1 immune mothers, ELISA was carried out using UV-inactivated DENV1 or DENV2 particles as coating antigens. Expectedly, elevated absorbance readings were measured when probing for the presence of anti-DENV1 total IgG antibodies in the serum from 5-week old mice born to DENV1 immune mothers (Fig. 8A). In addition, serum from these mice cross-reacted significantly with DENV2 (Fig. 8A). Isotyping of the anti-DENV1 IgG antibodies revealed majority of IgG1 and IgG2a circulating in the serum of mice born to DENV1 immune mothers (Fig. 8B).

Bottom Line: Furthermore, a recent epidemiological study has challenged the influence of maternal antibodies in disease outcome.In this ADE model we demonstrated the role of TNF-α in DEN-induced vascular leakage.In vitro ELISA and ADE assays confirmed the cross-reactive and enhancing properties towards DENV2 of the serum from mice born to DENV1-immune mothers.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, Life Sciences Institute, National University of Singapore, Singapore; Immunology Programme, Yong Loo Lin School of Medicine, Life Sciences Institute, National University of Singapore, Singapore.

ABSTRACT
Dengue (DEN) represents the most serious arthropod-borne viral disease. DEN clinical manifestations range from mild febrile illness to life-threatening hemorrhage and vascular leakage. Early epidemiological observations reported that infants born to DEN-immune mothers were at greater risk to develop the severe forms of the disease upon infection with any serotype of dengue virus (DENV). From these observations emerged the hypothesis of antibody-dependent enhancement (ADE) of disease severity, whereby maternally acquired anti-DENV antibodies cross-react but fail to neutralize DENV particles, resulting in higher viremia that correlates with increased disease severity. Although in vitro and in vivo experimental set ups have indirectly supported the ADE hypothesis, direct experimental evidence has been missing. Furthermore, a recent epidemiological study has challenged the influence of maternal antibodies in disease outcome. Here we have developed a mouse model of ADE where DENV2 infection of young mice born to DENV1-immune mothers led to earlier death which correlated with higher viremia and increased vascular leakage compared to DENV2-infected mice born to dengue naïve mothers. In this ADE model we demonstrated the role of TNF-α in DEN-induced vascular leakage. Furthermore, upon infection with an attenuated DENV2 mutant strain, mice born to DENV1-immune mothers developed lethal disease accompanied by vascular leakage whereas infected mice born to dengue naïve mothers did no display any clinical manifestation. In vitro ELISA and ADE assays confirmed the cross-reactive and enhancing properties towards DENV2 of the serum from mice born to DENV1-immune mothers. Lastly, age-dependent susceptibility to disease enhancement was observed in mice born to DENV1-immune mothers, thus reproducing epidemiological observations. Overall, this work provides direct in vivo demonstration of the role of maternally acquired heterotypic dengue antibodies in the enhancement of dengue disease severity and offers a unique opportunity to further decipher the mechanisms involved.

Show MeSH
Related in: MedlinePlus