title: Detailed characterization of tumor infiltrating lymphocytes in two distinct human solid malignancies show phenotypic similarities.
abstract: BACKGROUND
  We examined the phenotype and function of lymphocytes collected from the peripheral blood (PBL) and tumor (TIL) of patients with two different solid malignancies: colorectal cancer liver metastases (CRLM) and ovarian cancer (OVC).
  
  
  METHODS
  Tumor and corresponding peripheral blood were collected from 16 CRLM and 22 OVC patients; immediately following resection they were processed and analyzed using a multi-color flow cytometry panel. Cytokine mRNA from purified PBL and TIL CD4(+) T cells were also analyzed by qPCR.
  
  
  RESULTS
  Overall, we found similar changes in the phenotypic and cytokine profiles when the TIL were compared to PBL from patients with two different malignancies. The percentage of Treg (CD4(+)/CD25(+)/FoxP3(+)) in PBL and TIL was similar: 8.1% versus 10.2%, respectively in CRLM patients. However, the frequency of Treg in primary OVC TIL was higher than PBL: 19.2% versus 4.5% (p <0.0001). A subpopulation of Treg expressing HLA-DR was markedly increased in TIL compared to PBL in both tumor types, CRLM: 69.0% versus 31.7% (p = 0.0002) and OVC 74.6% versus 37.0% (p <0.0001), which suggested preferential Treg activation within the tumor. The cytokine mRNA profile showed that IL-6, a cytokine known for its immunosuppressive properties through STAT3 upregulation, was increased in TIL samples in patients with OVC and CRLM. Both TIL populations also contained a significantly higher proportion of activated CD8(+) T cells (HLA-DR(+)/CD38(+)) compared to PBL (CRLM: 30.2% vs 7.7%, (p = 0.0012), OVC: 57.1% vs 12.0%, (p <0.0001)).
  
  
  CONCLUSIONS
  This study demonstrates that multi-color flow cytometry of freshly digested tumor samples reveals phenotypic differences in TIL vs PBL T cell sub-populations. The TIL composition in primary and metastatic tumors from two distinct histologies were remarkably similar, showing a greater proportion of activated/suppressive Treg (HLA-DR(+), CD39(+), CTLA-4(+) and Helios(+)) and activated cytotoxic T cells (CD8(+)/HLA-DR(+)/CD38(+)) when compared to PBL and an increase in IL-6 mRNA from CD4 TIL.
authors.affiliation: Earle A. Chiles Research Institute, Providence Cancer Center, Portland Providence Medical Center, 4805 NE Glisan St, Portland, Oregon 97213 USA.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Kovacsovics-Bankowski
authors.affiliation: Earle A. Chiles Research Institute, Providence Cancer Center, Portland Providence Medical Center, 4805 NE Glisan St, Portland, Oregon 97213 USA.
authors.forename: Lana
authors.initials: L
authors.lastname: Chisholm
authors.affiliation: Earle A. Chiles Research Institute, Providence Cancer Center, Portland Providence Medical Center, 4805 NE Glisan St, Portland, Oregon 97213 USA.
authors.forename: Jonna
authors.initials: J
authors.lastname: Vercellini
authors.affiliation: Earle A. Chiles Research Institute, Providence Cancer Center, Portland Providence Medical Center, 4805 NE Glisan St, Portland, Oregon 97213 USA ; Agonox Inc, 4805 NE Glisan St, Portland, Oregon 97213 USA.
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Tucker
authors.affiliation: Agonox Inc, 4805 NE Glisan St, Portland, Oregon 97213 USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Montler
authors.affiliation: Earle A. Chiles Research Institute, Providence Cancer Center, Portland Providence Medical Center, 4805 NE Glisan St, Portland, Oregon 97213 USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Haley
authors.affiliation: Earle A. Chiles Research Institute, Providence Cancer Center, Portland Providence Medical Center, 4805 NE Glisan St, Portland, Oregon 97213 USA.
authors.forename: Philippa
authors.initials: P
authors.lastname: Newell
authors.affiliation: Providence Gynecologic Oncology, Providence Cancer Center, Portland Providence Medical Center, 4805 NE Glisan St, Portland, Oregon 97213 USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Ma
authors.affiliation: Providence Gynecologic Oncology, Providence Cancer Center, Portland Providence Medical Center, 4805 NE Glisan St, Portland, Oregon 97213 USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Tseng
authors.affiliation: Providence Surgical Oncology, Providence Cancer Center, Portland Providence Medical Center, 4805 NE Glisan St, Portland, Oregon 97213 USA.
authors.forename: Ronald
authors.initials: R
authors.lastname: Wolf
authors.affiliation: OHSU, division of oncological surgery and OHSU Knight Cancer Center, 3303 SW Bond Ave, Portland, OR 97239 USA.
authors.forename: John T
authors.initials: JT
authors.lastname: Vetto
authors.affiliation: Providence Surgical Oncology, Providence Cancer Center, Portland Providence Medical Center, 4805 NE Glisan St, Portland, Oregon 97213 USA.
authors.forename: Chet
authors.initials: C
authors.lastname: Hammill
authors.affiliation: Providence Surgical Oncology, Providence Cancer Center, Portland Providence Medical Center, 4805 NE Glisan St, Portland, Oregon 97213 USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Hansen
authors.affiliation: Earle A. Chiles Research Institute, Providence Cancer Center, Portland Providence Medical Center, 4805 NE Glisan St, Portland, Oregon 97213 USA ; Agonox Inc, 4805 NE Glisan St, Portland, Oregon 97213 USA.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Weinberg
keywords: Regulatory T cells; Tumor infiltrating lymphocytes