PMC8339907_01
The timeline of the patient's treatment course.
fonc-11-674328-g001.jpg
PMC8339907_01_fonc-11-674328-g001.jpg
f1
The clinical course and examination are shown in  Figure 1 .
Changes in clinical indexes before and after neoadjuvant immunochemotherapy. (A) Representative CT images during neoadjuvant immunochemotherapy. (B) Carcinoembryonic antigen test results during neoadjuvant immunochemotherapy. (C) Histological results of the pretreatment tumor biopsy (hematoxylin and eosin (HE) staining, magnification x200) and surgically resected tissue (HE, magnification x100).
fonc-11-674328-g002.jpg
PMC8339907_01_fonc-11-674328-g002.jpg
f2
Tumor cells accounted for 90% of the cells in the pretreatment tumor biopsy ( Figure 2A ). After neoadjuvant PD-1 blockade and chemotherapy, radiological images showed that the tumor was significantly reduced ( Figure 2A ). Serum tumor marker levels (CEA and CA125) were decreased compared with pretreatment levels ( Figure 2B ). Pathological assessment showed that the residual viable tumor cells accounted for 60% of the resection sample ( Figure 2C ).
Changes in the immune microenvironment before and after neoadjuvant immunochemotherapy. (A) Biopsy before neoadjuvant immunochemotherapy (PD-L1, IHC). (B) Surgically resected tissue after neoadjuvant immunochemotherapy and surgery (PD-L1, IHC). (C) Representative images of the pretreatment biopsy (multiplex immunofluorescence staining, magnification, x200). (D) Representative images of surgically resected tissue (multiplex immunofluorescence staining, magnification, x200). With this staining technique, visible structures include CD8+ T cells (green), CD68+ macrophages (cyan), CD57+ cells (red), PD-1+ cells (magenta), and PD-L1+ cells (orange). (E, F) Quantitative multiplex immunohistochemistry results of pretreatment and posttreatment samples in the tumor and stroma regions.
fonc-11-674328-g003.jpg
PMC8339907_01_fonc-11-674328-g003.jpg
f3
In the pretreatment tumor biopsy, the immunohistochemical results showed that tumor cell PD-L1 expression was negative ( Figure 3A ). NK cells, CD68+ macrophages, and CD68+ PD-L1+ macrophages infiltrated the tumor ( Figure 3B ). Next-generation sequencing (NGS) showed wild-type EGFR, ALK, ROS, NTRK, and STK11 genes and mutant In the post-treatment resection specimen, PD-L1 immunohistochemistry (IHC) assays showed that PD-L1 expression was 50% in the resected tumor ( Figure 3C ). The mIHC results revealed more CD8+ and CD8+ PD-1+ T cell infiltration posttreatment relative to pretreatment ( Figures 3D-F ).
Changes in related indexes during adjuvant immunochemotherapy and irAEs. (A) Pulmonary CTA indicated multiple cords, plaques, and flocculent high-density shadows in both lungs, bilateral pleural effusion, and a clinical diagnosis of immune-associated pneumonia (CTCAE level 4). (B) Bronchoscopy revealed a fistula at the anastomosis of the right bronchus and the formation of a bloody abscess cavity. (C) Multiplexed bead immunoassay results during the entire treatment period. (D) Representative CRP levels during adjuvant immunochemotherapy and irAEs. (E-K) Representative multiplexed bead immunoassay results during the entire treatment period.
fonc-11-674328-g004.jpg
PMC8339907_01_fonc-11-674328-g004.jpg
f4
During adjuvant therapy, immune-related lung injury (grade 4) occurred ( Figure 4A ). On October 14th, bronchoscopy showed hemorrhagic vomica in the bronchial anastomosis ( Figure 4B ). Serum cytokine changes during the neoadjuvant phase and adjuvant PD-1 blockade combined with chemotherapy therapy were detected by multiplexed bead immunoassays (Genecast, Wuxi, China) ( Figure 4C ). The serum levels of MCP-1 did not change significantly during neoadjuvant therapy and increased during adjuvant therapy. The serum level of IL-10 was consistently decreased during neoadjuvant and adjuvant immunochemotherapy ( Figures 4D-K ).