Role of NS1 antibodies in the pathogenesis of acute secondary dengue infection

Jayathilaka, Deshni; Gomes, Laksiri; Jeewandara, Chandima; Jayarathna, Geethal. S. Bandara; Herath, Dhanushka; Perera, Pathum Asela; Fernando, Samitha; Wijewickrama, Ananda; Hardman, Clare S.; Ogg, Graham S.; Malavige, Gathsaurie Neelika

doi:10.1038/s41467-018-07667-z

Cited by 77 publications

(71 citation statements)

References 49 publications

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“…In order to measure NS1 antibody levels in patient serum samples, an in-house indirect ELISA was carried out as previously described by Jayathilaka, D. et.al [18]. Serum samples diluted at 1:5000 were added to 96-well plates coated with DENV-2 NS1 protein (Native antigen, USA) and blocked with blocking buffer (PBS containing 0.05% Tween 20 and 1% Bovine serum albumin (BSA).…”

Section: Determining Ns1 Antibody Levels In Patient Seramentioning

confidence: 99%

“…The Tfh cells were also shown to be able to produce IL-21 and IFNγ upon stimulation with DENV peptides, suggesting that they were capable of providing B cell help, which is largely dependent on production of IL-21 [17]. Therefore, increase in frequency and activation of Tfh cells could lead to higher DENV specific antibody levels which may lead to protection, or possibly disease enhancement [18,19]. In order to further understand the pathogenesis of acute dengue and the constituents of a protective immune response, it would be important to study how the functionality and phenotype of Tfh cells associate with DENV-specific antibody responses and clinical disease severity.…”

Section: Introductionmentioning

confidence: 99%

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Phenotype and functionality of follicular helper T cells in patients with acute dengue infection

et al. 2020

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Background: The association of functionality and phenotype of follicular helper T cells (Tfh) with dengue virus (DENV) specific antibody responses and clinical disease severity has not been well studied. Methods: We investigated the phenotype and functionality of Tfh cells and plasmablasts in adult patients (DF = 18, DHF = 22) with acute dengue (day 4 to 8 since onset of fever) of varying severity using multiparametric flowcytometry. The properties of Tfh cells were correlated with viraemia, disease severity, plasmablast responses and DENV-specific serum antibody responses. We further evaluated the kinetics of neutralizing antibodies (Neut50) throughout the course of illness in order to evaluate their association with clinical disease severity and viraemia. Results: Tfh cells (especially those producing IL-21 and co-expressing PD-1 and ICOS) were found to be significantly expanded (p < 0.0001) and highly activated in patients with DHF compared to those with DF. The frequency of Tfh cells significantly correlated with DENV-specific IgG, NS1-specific antibodies and Neut50 antibody titres in patients with DHF but not in those with DF. Although the Neut50 titres increased during the course of acute secondary DENV infection, they showed differences based on serotype. For instance, the Neut50 titres were significantly higher during the latter part of illness in patients with DF compared to DHF in DENV1 infection, while in DENV2, patients with DHF had significantly higher titres. The viral loads during early illness did not correlate with the subsequent rise in the Neut50 antibody titres during any time point of illness. Conclusions: The expansion of Tfh cells is associated with DHF and DENV-specific IgG, NS1-specific and neutralizing antibodies. Neut50 titres did not associate with disease severity or viraemia at the point of first presentation during the febrile phase, but later titres do show differential association with severity in patients with DENV1 compared to DENV2.

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Section: Determining Ns1 Antibody Levels In Patient Seramentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Phenotype and functionality of follicular helper T cells in patients with acute dengue infection

et al. 2020

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“…Although the majority of DENV specific antibodies are specific to the DENV envelope protein, 27% of the DENV antibody repertoire has shown to be specific to DENV NS1 protein [29]. Therefore, we measured DENV2 specific NS1 antibodies using an in-house ELISA as described before [18]. As observed earlier, NS1-antibody levels were significantly higher in patients with DHF (median 1.7, IQR 0.76 to 2.3 OD values) compared to those with DF (median 0.56, IQR 0.23 to 1.4 OD values).…”

Section: Resultsmentioning

confidence: 99%

“…In order to measure NS1 antibody levels in patient serum samples, an inhouse indirect ELISA was carried out as previously described by Jayathilaka, D. et.al [18]. Serum samples diluted at 1:5000 were added to 96-well plates coated with DENV-2 NS1 protein (Native antigen, USA) and blocked with blocking buffer (PBS containing 0.05% Tween 20 and 1 % Bovine serum albumin (BSA).…”

Section: Methodsmentioning

confidence: 99%

Phenotype and functionality of follicular helper T cells in patients with acute dengue infection

Wijesinghe

Gamage

Goonewardana

et al. 2019

Preprint

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AbstractBackgroundThe association of functionality and phenotype of follicular helper T cells (Tfh) with dengue virus (DENV) specific antibody responses and clinical disease severity is has not been well studied.Methodology/Principal findingsWe investigated the phenotype and functionality of Tfhs in adult patients (DF = 18, DHF = 22) with acute dengue of varying severity using multiparametric flowcytometry. We determined if the properties of Tfhs were correlated with viraemia, disease severity, plasmablast responses and DENV-specific serum antibody responses. We further evaluated the changes in neutralizing antibodies (Neut50) with viraemia and clinical disease severity in a different cohort of patients with acute secondary DENV1 (DF=12, DHF=10) and DENV2 infection (DF=8, DHF=9).Tfhs (especially those producing IL-21 and co-expressing PD-1 and ICOS) were found to be significantly expanded (p<0.0001) and highly activated in patients with DHF compared to those with DF. The frequency of Tfh cells significantly correlated with DENV-specific IgG, NS1-specific antibodies and Neut50 antibody titres, which were also significantly higher in patients with DHF. Although the Neut50 titres increased during the course of acute secondary DENV infection, they showed differences based on serotype. For instance, the Neut50 titres were significantly higher during the latter part of illness in patients with DF compared DHF in DENV1 infection, while in DENV2, patients with DHF had significantly higher titres. The viral loads during early illness did not correlate with the subsequent rise in the Neut50 antibody titres during time point of illness.Conclusions/SignificanceThe expansion of Tfhs is associated with DHF and DENV-specific IgG, NS1-specific and neutralising antibodies. Neut50 titres did not associate with disease severity or viraemia at the point of first presentation during the febrile phase, but later titres do show differential association with severity in patients with DENV1 compared to DENV2.Author summaryFollicular helper T cells (Tfh cells) are a subset of T cells which are important in activation of germinal centre T cells and induction of long-lasting virus specific antibodies. The association of the functional status and phenotype of Tfh cells in relation to dengue virus (DENV) specific antibodies, clinical disease severity and the degree of viraemia in patients with acute dengue has not been extensively studied. Here we show that the Tfh cells are significantly expanded in patients with severe clinical disease and associate with levels of serum DENV envelop specific IgG, NS1-specific antibodies and neutralising antibodies (Neu50). Although DENV Neu50 antibody titres are thought to associate with protection, the Neut50 antibody titres were similar in patients who proceeded to develop mild or severe clinical disease and also the increase in their titres showed significant variation based on infecting DENV serotype and clinical disease severity. Therefore, although Tfh cells are expanded in patients with more severe forms of disease, and associate with DENV-specific antibody responses, they do not appear to have a protective role.

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“…Antibodies against the viral non-structural protein NS1 have also been shown to bind to endothelial cells in secondary infections, causing their apoptosis (7-9). In addition, NS1 protein is abundantly secreted into patient serum and has been shown to directly induce endothelial cell hyperpermeability (10, 11). Serum NS1 concentrations in dengue patients vary widely.…”

Section: Introductionmentioning

confidence: 99%

A Critical Role for Perivascular Cells in Amplifying Viral Haemorrhage Induced by Dengue Virus Non-Structural Protein 1

Cheung

Mastrullo

Maselli

et al. 2020

Preprint

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23Dengue is the most prevalent arthropod-borne viral disease affecting humans, with 24 severe dengue typified by potentially fatal microvascular leakage and hypovolaemic shock. 25Blood vessels of the microvasculature are composed of a tubular structure of endothelial cells 26 ensheathed by perivascular cells (pericytes). Pericytes support endothelial cell barrier 27 formation and maintenance through paracrine and contact-mediated signalling, and are critical 28 to microvascular integrity. Pericyte dysfunction has been linked to vascular leakage in 29 noncommunicable pathologies such as diabetic retinopathy, but has never been linked to 30 infection-related vascular leakage. Dengue vascular leakage has been shown to result in part 31 from the direct action of the secreted dengue virus (DENV) non-structural protein NS1 on 32 endothelial cells. Using primary human vascular cells, we show here that NS1 also causes 33 pericyte dysfunction, and that NS1-induced endothelial hyperpermeability is more pronounced 34 in the presence of pericytes. Notably, NS1 specifically disrupted the ability of pericytes to 35 support endothelial cell function in a 3D microvascular assay, with no effect on pericyte viability 36 or physiology. These effects are mediated at least in part through contact-independent 37 paracrine signals involved in endothelial barrier maintenance by pericytes. We therefore 38 identify a role for pericytes in amplifying NS1-induced microvascular hyperpermeability in 39 severe dengue, and thus show that pericytes can play a critical role in the aetiology of an 40 infectious vascular leakage syndrome. These findings open new avenues of research for the 41 development of drugs and diagnostic assays for combating infection-induced vascular 42 leakage, such as severe dengue.43 44 45 46 47 3 SIGNIFICANCE STATEMENT 48 The World Health Organisation considers dengue one of the top ten global public 49 health problems. There is no specific antiviral therapy to treat dengue virus and no way of 50 predicting which patients will develop potentially fatal severe dengue, typified by vascular 51 leakage and circulatory shock. We show here that perivascular cells (pericytes) amplify the 52 vascular leakage-inducing effects of the dengue viral protein NS1 through contact-53 independent signalling to endothelial cells. While pericytes are known to contribute to 54 noncommunicable vascular leakage, this is the first time these cells have been implicated in 55 the vascular effects of an infectious disease. Our findings could pave the way for new 56 therapies and diagnostics to combat dengue, and potentially other infectious vascular leakage 57 syndromes. 58 59 60 61 62 63 64 65 66 67 68 69 355 MR/R010315/1 to KM and a

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Role of NS1 antibodies in the pathogenesis of acute secondary dengue infection

Cited by 77 publications

References 49 publications

Phenotype and functionality of follicular helper T cells in patients with acute dengue infection

Phenotype and functionality of follicular helper T cells in patients with acute dengue infection

Phenotype and functionality of follicular helper T cells in patients with acute dengue infection

A Critical Role for Perivascular Cells in Amplifying Viral Haemorrhage Induced by Dengue Virus Non-Structural Protein 1

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