In summary, our findings firstly revealed that TLR4 played a vital role in sensing HSV-2 infection in human genital epithelial cells, and TLR4-MyD88/TRIF-AP-1 pathway is essential for HSV-2-induced up-regulation of TLR4 expression, which implicated that TLR4 could be as a virus sensor for herpes infection. Here, TLR4 is linked to Herpesviridae infectious disease.