Research ArticleBasic Science Investigation

Noninvasive Monitoring of Immunotherapy in Lung Cancer by Lymphocyte Activation Gene 3 PET Imaging of Tumor-Infiltrating Lymphocytes

Zhen Quan, Zhaoguo Han, Yang Yang, Jiannan Wang, Hui Wang, Lili Yang, Rong A, Xinxin Hu, Jing Wang, Xiaona Li, Xiaoqian Li, Hong Yu, Wei Chen, Kai Wang and Xilin Sun
Journal of Nuclear Medicine November 2023, jnumed.123.266002; DOI: https://doi.org/10.2967/jnumed.123.266002

Article Figures & Data

Figures

  • Figure
  • FIGURE 1.
    FIGURE 1.

    (A) Radiochemical synthesis of [68Ga]Ga-NOTA-C25. (B) Flow cytometry and quantitative analysis of CD8+ LAG-3+ in murine immune cells that were activated or resting. (C) Cell uptake of [68Ga]Ga-NOTA-C25 (6.788 × 10−9 μM) in murine immune cells with blocking assays by nonradiolabeled C25 (9.613 × 10−6 μM) and αLAG-3 (1.333 × 10−7 μM). n = 3. **P < 0.01. ****P < 0.0001.

  • FIGURE 2.
    FIGURE 2.

    Representative PET/CT images and quantitative analysis in KLN-205, CMT-167, and LLC tumor–bearing mice at 30, 60, and 120 min after injection of [68Ga]Ga-NOTA-C25 with blocking assays by nonradiolabeled C25 (100 μg in 100 μL; 0.0961 μmol). Yellow arrows indicate tumor. n ≥ 3. *P < 0.05. **P < 0.01. ***P < 0.001. ****P < 0.0001.

  • FIGURE 3.
    FIGURE 3.

    (A) Time line of immunotherapy and PET imaging in LLC tumor–bearing mice. (B) Representative [68Ga]Ga-NOTA-C25 PET/CT and quantitative analysis in LLC models treated with PBS, αPD-1, MSA-2, and αPD-1 + MSA-2 at days 4 and 13. Yellow arrows indicate tumor. Red arrows indicate spleen. (C) Biodistribution in tumor and main immune organs excised from LLC models at days 4 and 13. n ≥ 6. *P < 0.05. **P < 0.01. ***P < 0.001. ****P < 0.0001.

  • FIGURE 4.
    FIGURE 4.

    Flow cytometry detection of CD8+LAG-3+ T cells and quantitative analysis in tumor, spleen, and TDLN excised from LLC tumor models at day 13. n ≥ 6. *P < 0.05. **P < 0.01. ***P < 0.001. ****P < 0.0001.

  • FIGURE 5.
    FIGURE 5.

    Immunofluorescence images and quantitative analysis of LAG-3 and CD3 staining in tissue sections from LLC models at day 13. Blue = DAPI. Green = CD3. Red = LAG-3. Scale bar, 50 μm. n = 3. *P < 0.05. **P < 0.01. ***P < 0.001. ****P < 0.0001.

  • FIGURE 6.
    FIGURE 6.

    (A) Tumor growth curves of LLC tumor–bearing mice treated with PBS, αPD-1, MSA-2, and αPD-1 + MSA-2. (B) Survival analysis of LLC models for different groups. n ≥ 6. *P < 0.05. **P < 0.01. ****P < 0.0001.

  • FIGURE 7.
    FIGURE 7.

    (A) Quantitative analysis of flow cytometry for hCD8+hLAG-3+ in hPBMCs. (B) Cell uptake and blocking assays (nonradiolabeled C25, 9.613 × 10−6 μM) of [68Ga]Ga-NOTA-C25 in hPBMCS. (C) Representative PET/CT images and quantitative analysis in H460 and H460-HIS tumor models at 60 min after injection of [68Ga]Ga-NOTA-C25. Yellow arrows indicate tumor. n = 3. *P < 0.05. **P < 0.01.

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