Characterization of tumor-infiltrating immune cells in relation to microbiota in colorectal cancers
Several articles have recently reported that certain colon microbiota can improve the efficacy of cancer immunotherapy. To develop new treatment strategies, including immunotherapy for colorectal cancer (CRC), we evaluated the correlations between subpopulations of tumor-infiltrating immune cells (TIICs) and intestinal microbiota in CRC.
Fresh surgically resected specimens, formalin-fixed paraffin-embedded whole tissue samples, and stool samples were collected. TIICs including Tregs, Th17 cells and tumor-associated macrophages (TAMs) in the surgically resected specimens were analyzed using flow cytometry. FOXp3, CD8, CD163, and phosphorylated-STAT1-positive TIICs in the whole tissue samples were analyzed using IHC, and intestinal microbiota in the stool samples was analyzed using 16S metagenome sequencing. TIICs subpopulations in the normal mucosa and tumor samples were evaluated, and the correlations between the TIIC subpopulations and intestinal microbiota were analyzed.
FOXp3lowCD45RA+ Tregs were significantly reduced (p = 0.02), FOXp3lowCD45RA− Tregs were significantly increased (p = 0.006), and M1 TAMs were significantly reduced in the tumor samples (p = 0.03). Bacteroides (phylum Bacteroidetes) and Faecalibacterium (phylum Firmicutes) were increased in the patients with high numbers of Tregs and clearly high distribution of FOXp3highCD45RA− Tregs, which are the effector Tregs. Faecalibacterium, Ruminococcaceae, Eubacterium (phylum Firmicutes), and Bacteroides were increased in patients with a high distribution of M1 TAMs.
The findings of the present study indicate that immune responses to tumors are suppressed in the tumor microenvironment of CRC depending on the increment of Tregs and the reduction of M1 TAMs and that intestinal microbiota might be involved in immunosuppression.
KeywordsColorectal cancer Microbiota Tumor-associated macrophage Treg
Principal coordinates analysis
Tumor-infiltrating immune cells
TK, KM, TT, and KK contributed to the study conception and design. TK, MA, HO, EE, KS, WS, SF, HE, MS, TM, ZS, and SO contributed to the acquisition of patient samples. TK, KM, MA, HO, WS, SF, HE, MS, TM, ZS, and SO performed flow cytometry and analyzed the flow cytometry data. KS, KY, and TT performed the 16S metagenome sequencing and analyzed the microbiota data. EE and KS performed IHC and evaluated the IHC staining. TK, KM, KS, KY, TT, and KK drafted the manuscript.
No relevant funding.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval and standards
This study was conducted in accordance with the ethical principles that have their origin in the Declaration of Helsinki and was approved by the Fukushima Medical University Research Ethics Committee (Receipt No. 29020).
Written informed consent was obtained from all patients included in the study for the use of their specimens and clinical data for research and publication prior to collecting the specimen at Fukushima Medical University Hospital.
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