Although immune checkpoint inhibitors (ICIs) show durable responses in various cancers, relapse remains common. The efficacy of retreatment with ICIs is controversial. We conducted a multicenter, single-arm, phase 2 trial (NCT05824468) including 30 advanced cervical cancer patients that progressed on or after prior ICI therapy. Participants received a combined regimen of zimberelimab and lenvatinib (immune rechallenge). Single-cell multi-omics analysis of sequential biopsies of relapsed tumors and blood samples showed immune rechallenge induced a more cytotoxic phenotype in CD8+T cells in responders, while an NK-like CD8+T and progenitor-exhausted CD8+T phenotype in the blood of non-responders. In tumor, responders showed more effector memory CD8+T cells and reduced exhausted CD8+T cell post-treatment. A population of CD45+CD3+Lyz+dyad cells, composed of T cells and myeloid cells, was correlated with clinical benefit. Our findings proved immune rechallenge could be an effective treatment for patients with advanced cervical cancer who progressed on or after prior ICIs therapy.