Chimeric antigen receptor (CAR)-T cell therapies for T-cell malignancies face significant challenges, including cannibalism among CAR-T cells and product contamination from cell explosion. Allogeneic CAR-T cells generated from healthy donor T cells can provide ready-to-use, blast-free therapeutic products, but their application may be complicated by graft-versus-host disease (GvHD) and host rejection. Here, we developed CD7-targeting CAR-T cells (RD13-01) from healthy donors that were genetically modified to resist cannibalism, GvHD, and allorejection and to enhance antitumor function . A phase I clinical trial (NCT04538599) enrolled 12 patients (11 with T-cell leukemia/lymphoma and 1 with CD7-expressing acute myeloid leukemia). All patients achieved the prespecified endpoint, and 11 patients were evaluated for efficacy. No dose-limiting toxicities, GvHD, immune effector cell-related neurotoxicity, or severe cytokine release syndrome (≥Grade 3) were observed. After 28 days of infusion, 81.8% (9/11) of patients experienced objective response, with a complete response rate of 63.6% (7/11, including AML patients). Three responding patients received allogeneic hematopoietic stem cell transplantation. At a median follow-up of 10.5 months, 4 patients remained in complete response. Reactivation of cytomegalovirus (CMV) and/or Epstein-Barr virus (EBV) was observed in several patients, one of whom died from EBV-related diffuse large B-cell lymphoma (DLBCL). Expansion of CD7-negative normal T cells was detected after infusion. In summary, we present the first report of a phase I clinical trial using healthy donor-derived CD7-targeting allogeneic CAR-T cells to treat CD7+ hematological malignancies. Our results demonstrate the encouraging safety and efficacy of RD13-01 allogeneic CAR-T cells for the treatment of CD7+ tumors.