Multifunctional T cell response in convalescent patients two years after ZIKV infection

Abstract

Zika is an important emerging infectious disease in which the role of T cells remains elusive. This study aimed to evaluate the phenotype of multifunctional T cells in individuals 2 yr after exposure to Zika virus (ZIKV). We used a library of 671 synthetic peptides covering the whole polyprotein of ZIKV in pools corresponding to each viral protein (i.e., capsid, membrane precursor or prM, envelope, NS1 [nonstructural protein], NS2A + NS2B, NS3, NS4A + NS4B, and NS5) to stimulate PBMCs from individuals previously exposed to ZIKV. We observed an increased frequency of ZIKV-specific IFN , IL-17A, TNF, and IL-10 production by T cell populations. IFN and TNF production were especially stimulated by prM, capsid, or NS1 in CD8+ T cells and by capsid or prM in CD4+ T cells. In addition, there was an increase in the frequency of IL-10+ CD8+ T cells after stimulation with prM, capsid, NS1, NS3, or NS5. Multifunctional properties were observed in ZIKVspecific T cells responding especially to prM, capsid, NS1 or, to a smaller extent, NS3 antigens. For example, we found a consistent IFN + TNF+ CD8+ T cell population in response to most virus antigens and CD4+ and CD8+ T cells that were IFN + IL-17A+ and IL-17A+IL-10+, which could also produce TNF, in response to capsid, prM, NS1, or NS3 stimulation. Interestingly, CD8+ T cells were more prone to a multifunctional phenotype than CD4+ T cells, and multifunctional T cells were more efficient at producing cytokines than single-function cells. This work provides relevant insights into the quality of ZIKV-specific T cell responses and ZIKV immunity. K E Y W O R D S immunity, multifunctional T cells, Zika, ZIKV 1 INTRODUCTION Zika virus (ZIKV) is an arbovirus of the Flaviviridae family and Flavivirus genus that comprises more than 70 viruses distributed worldwide, among which approximately 40 viruses are described as causing diseases in humans. 1 Among the Flavivirus viruses of particular interest, Abbreviations: CZS, Congenital Zika syndrome; DENV, Dengue virus; MFI, mean fluorescence intensity; NS, nonstructural protein; prM, membrane precursor; Tc, CD8+ cytotoxic T cell; YFV, Yellow fever virus; ZIKV, Zika virus. the most prominent members are dengue virus (DENV), responsible for an annual incidence of 96 million infections with symptomatic dengue fever; yellow fever virus (YFV); West Nile virus (WNV); Japanese encephalitis virus (JEV); and ZIKV, which are classified as emerging infectious viruses. 1 It is estimated that only 20-25% of ZIKV-infected individuals develop symptoms, with nonspecific clinical manifestations common to other arboviruses, 2 such as rash, low-grade fever, nonpurulent conjunctivitis, myalgia, and arthritis/arthralgia, which are usually benign and self-limiting. ZIKV infection has also been associated with

Involvement of Th1Th17 Cell Subpopulations in the Immune Responses of Mothers Who Gave Birth to Children with Congenital Zika Syndrome (CZS)

“…They stimulated the PBMCs from these individuals with a wide range of synthetic peptides from the entire ZIKV polyprotein, divided into pools according to each of the viral protein's region. They showed that IFN-γ+ IL-17A+ and IL-17A+ IL-10+T cells could also produce TNF in response to stimulation with antigens of the capsid, prM, NS1 or NS3 regions [78].…”

Evaluation of the Expression of CCR5 and CX3CR1 Receptors and Correlation with the Functionality of T Cells in Women infected with ZIKV during Pregnancy

“…Multifunctional properties were observed in ZIKV-specific T cells responding especially to prM, capsid, NS1, or, to a smaller extent, NS3 antigens. In addition, CD8+ T cells were more prone to a multifunctional phenotype than were CD4+ T cells [ 51 ].…”

Peculiarities of Zika Immunity and Vaccine Development: Lessons from Dengue and the Contribution from Controlled Human Infection Model

“…Experimental models suggest that an effective immune response against ZIKV is driven by multifunctional CD4+ and CD8+ T cells [ 59 , 63 ]. We have found that ZIKV-specific T cells display a dominant multifunctional profile secreting multiple cytokines, especially IFNg, IL-17A, TNF and IL-10, simultaneously (quadruple-producing T cells) or its combinations [ 64 ] ( Figure 1 ). Interestingly, we observed that CD8+ T cells displayed higher frequencies of multifunctional lymphocytes when compared to CD4+ T cells [ 64 ].…”

“…We have found that ZIKV-specific T cells display a dominant multifunctional profile secreting multiple cytokines, especially IFNg, IL-17A, TNF and IL-10, simultaneously (quadruple-producing T cells) or its combinations [ 64 ] ( Figure 1 ). Interestingly, we observed that CD8+ T cells displayed higher frequencies of multifunctional lymphocytes when compared to CD4+ T cells [ 64 ]. Indeed, multifunctional T cell responses and cellular immunity have been considered important mechanisms for host resistance against other flaviviruses like DENV [ 51 , 65 ] and immunity to YFV [ 66 , 67 , 68 ].…”