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

Abstract

BackgroundThe 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.\n\nMethodology/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).\n\nTfhs (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.\n\nConclusions/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.\n\nAuthor 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.