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Objective To investigate the levels of different follicular helper T cells in peripheral blood of syphilis patients and discuss its clinical significance. Methods Eighty-four syphilis patients and 37 healthy controls were enrolled from Dermatology Hospital of Southern Medical University from May 2015 to March 2016. Flow cytometry was used to measure CD4+CXCR5+ICOS+、CD4+CXCR5+、CD4+ICOS+Tfh cells in the peripheral blood. Correlation of TRUST titers with different Tfh cells in syphilis patients, including untreated, treated, cured or sero-fast, was analyzed. Results The proportion of CD4+CXCR5+ICOS+Tfh cells in CD4+ T cells was significantly higher in the pre-treatment group than in healthy controls (P<0.001). The proportions of CD4+CXCR5+ICOS+Tfh cells in CD4+ T cells in the post-treatment, cured and sero-fast syphilis groups were significantly lower than that in the pre-treatment group (P=0.039, P<0.001, P=0.009). The proportion of CD4+CXCR5+ICOS+Tfh cells in CD4+ T cells was positively correlated with TRUST titers (r=0.30, P<0.01). However, the proportion of neither CD4+CXCR5+ Tfh cells nor CD4+ICOS+ Tfh cells in CD4+ T cells was correlated with TRUST titers (P=0.519, P=0.118). Conclusion Tfh cells in peripheral blood may be involved in the onset and the development of syphilis.