The invention discloses a replication-defective recombinant lentivirus CAR-T transgenic vector targeting CD152 and a construction method of the replication-defective recombinant lentivirus CAR-T transgenic vector. The transgenic vector comprises a promoter for controlling a replication initiation site, a prokaryotic replicon for plasmid replication, a viral replicon for enhancing replication in eukaryotic cells, a resistance gene for detection, a regulatory element after viral transcription, a packaging cis element cis for lentiviral packaging and a chimeric antigen receptor CAR for recognition, transmission and initiation; the construction of the replication-defective recombinant lentiviral CAR-T transgenic vector comprises construction of a recombinant lentiviral plasmid, and packaging,concentration and purification of the recombinant lentiviral vector to obtain the replication-defective recombinant lentiviral CAR-T transgenic vector targeting CD152 with high purity. The recombinantlentivirus CAR-T transgenic vector provided by the invention can remarkably improve secretion of cytokines and specifically kill target cells CTLA-4, and provides a new treatment direction for inhibition of tumor microenvironment and attack of solid tumors.