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Circulating T follicular helper (cTfh) cells have been identified as counterparts of germinal center Tfh (GC Tfh) cells in humans and can support T-dependent B cell maturation and antibody production  in vitro . However, the role of cTfh cells in neutralizing antibody (nAb) responses in HCV infection remains unclear. Here, we characterized the phenotype and function of cTfh cells and demonstrated the associations of cTfh cells and their subsets with nAb responses in HCV infection. A total of 38 HCV-infected individuals and 28 healthy controls were enrolled from a pool of injection drug users. The frequency and function of blood Tfh cells were analyzed by flow cytometry. The titers and breadths of serum nAbs were measured using HCV pseudo-particle neutralization assays. Herein, we report several key observations. First, HCV infection skewed cTfh toward CXCR3 +  cTfh cell differentiation. Second, the frequency of CXCR3 +  cTfh cells positively correlated with HCV nAb titers and breadths. Third, CXCR3 +  cTfh cells showed higher expression of Tfh-associated molecules (PD-1, ICOS, IL-21, Bcl-6) compared with CXCR3 −  cTfh cells from individuals with HCV infection. Coculture of cTfh cells and autologous memory B cells  in vitro  indicated that CXCR3 +  cTfh cells show a superior ability to support HCV E2-specific B cell expansion compared with CXCR3 −  cTfh cells from individuals with HCV infection. HCV infection skews cTfh cells toward CXCR3-biased Tfh cell differentiation, which positively correlates with the magnitude and breadth of the HCV nAb response. It is our hope that these findings will provide insights for the rational design of a nAb-based HCV vaccine.

Circulating CXCR3+ Tfh cells positively correlate with neutralizing antibody responses in HCV-infected patients