Tumor-infiltrating　CD8+T　cells　and　programmed　death-1　(PD1)　levels　are　critical　indicators　for　tumor　immunophenotyping　and　therapeutic　decision-making.　Noninvasive　optical　imaging　in　the　second　near　infrared　window　(NIR-II)　is　particularly　well-suited　for　investigating　the　biological　processes　within　tumors　in　live　mammals,　thanks　to　its　deep-tissue　penetration　and　superior　spatiotemporal　resolution.　However,　in　vivo　NIR-II　imaging　has　primarily　been　restricted　to　a　single　probe　at　a　time.　Methods:　Herein,　we　developed　a　two-plex　NIR-II　molecular　imaging　method　utilizing　the　non-overlapping　fluorescence　emission　of　indocyanine　green　(ICG)　in　the　NIR-IIa　window　(1000-1200　nm)　and　PbS/CdS　core-shell　quantum　dots　(QDs)　in　the　NIR-IIb　window　(1500-1700　nm).　By　integrating　PD1　aptamer-labeled　ICG　(ICG-Apt-PD1,　targeting　PD1)　and　CD8　aptamer-labeled　QDs　(QDs@Apt-CD8,　targeting　CD8+　T　cells),　our　two-plex　NIR-II　molecular　imaging　enabled　simultaneous　and　noninvasive　monitoring　of　the　number　of　CD8+　T　cells　and　PD1　levels　in　tumors.　Results:　QDs@Apt-CD8　demonstrated　the　excellent　ability　for　in　vivo　imaging　of　tumor　infiltrating　CD8+　T　cells,　owing　to　its　strong　NIR-IIb　luminescence　and　the　high　selectivity　and　specificity.　This　two-plex　in　vivo　molecular　imaging　allowed　for　dynamic　monitoring　for　PD1　levels　and　the　number　of　CD8+　T　cells　in　tumors.　We　observed　the　heterogeneous　bio-distributions　of　PD1　and　CD8+　T　cells　across　different　tumor　types　and　revealed　the　tumor　immunophenotypes.　Moreover,　our　findings　indicated　that　the　low　PD1　and　high　CD8+　T　cells　levels　in　tumors　predicted　a　better　anti-tumor　effect.　Conclusions:　Such　in　vivo　noninvasive　NIR-II　molecular　imaging　would　complement　ex　vivo　biopsy-based　diagnostic　techniques,　and　it　could　contribute　to　developing　an　in　vivo　tumor　immune-scoring　algorithm　to　offer　a　more　precise　prediction　for　immunotherapeutic　response.