title: Phenotypic and functional analysis of tumor-infiltrating lymphocytes compared with tumor-associated lymphocytes from ascitic fluid and peripheral blood lymphocytes in patients with advanced ovarian cancer.
abstract: To investigate and compare the phenotype and function of lymphocytes collected from patients harboring advanced ovarian cancer, leukocytes from peripheral blood (n = 18), ascitic fluid (n = 13) and tumor tissues (n = 13) were evaluated for the relative proportions of lymphocyte subsets, including CD3+, CD4+, CD8+, CD19+, CD56 and the early (CD25) and late (HLA-DR) activation markers on CD3+ T cells. The ability to synthesize type 1 cytokines (IFN-gamma and IL-2) and a type 2 cytokine (IL-4) was assessed by flow cytometry. In all patients, T cells (CD3+) were the major leukocyte population detected in each tissue, with CD4+ T cells being dominant in peripheral blood lymphocytes (PBL) and tumor-associated lymphocytes (TAL) but not in tumor-infiltrating lymphocytes (TIL) (CD4:CD8 ratios: 3.0 vs. 2.0 vs. 1.0, respectively). CD19+ lymphocytes (B cells) and CD56+ lymphocytes (NK cells) were significantly higher in PBL compared to TAL and TIL (p < 0.05). TAL and TIL had a higher proportion of T cells expressing the late activation marker HLA-DR compared to PBL. In contrast, no significant differences were detected in PBL, TAL and TIL in the expression of the early activation marker CD25. Type 1 cytokines were the dominant type produced by in vitro stimulated T cells for each population, with a greater proportion of IFN-gamma+ T cells in TAL and TIL compared to PBL (p < 0.01), and a higher proportion of IL-2+ T cells in PBL compared with TAL and TIL (p < 0.05). Low percentages of IL-4+ T cells (i.e. Th2) were detected in each tissue. Taken together, these data demonstrate the recruitment and accumulation of high concentrations of antigen-experienced T lymphocytes in TAL and TIL compared to PBL. However, low surface expression of IL-2 receptor (i.e. CD25), as well as depressed intracellular IL-2 production in chronically stimulated TAL and TIL suggests that the impaired antitumor function commonly detected in these lymphocyte populations may be secondary to an acquired dysregulation of the IL-2 pathway.
authors.affiliation: Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, University of Arkansas for Medical Sciences, Little Rock, AR 72205-7199, USA.
authors.forename: A D
authors.initials: AD
authors.lastname: Santin
authors.forename: P L
authors.initials: PL
authors.lastname: Hermonat
authors.forename: A
authors.initials: A
authors.lastname: Ravaggi
authors.forename: S
authors.initials: S
authors.lastname: Bellone
authors.forename: J J
authors.initials: JJ
authors.lastname: Roman
authors.forename: C V
authors.initials: CV
authors.lastname: Smith
authors.forename: S
authors.initials: S
authors.lastname: Pecorelli
authors.forename: A
authors.initials: A
authors.lastname: Radominska-Pandya
authors.forename: M J
authors.initials: MJ
authors.lastname: Cannon
authors.forename: G P
authors.initials: GP
authors.lastname: Parham
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001202:Ascitic Fluid / Q000276:immunology*; D016516:CD4-CD8 Ratio; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000032:analysis / Q000096:biosynthesis; D007376:Interleukin-2 / Q000032:analysis / Q000096:biosynthesis; D015847:Interleukin-4 / Q000032:analysis / Q000096:biosynthesis; D018655:Lymphocyte Count; D016131:Lymphocyte Subsets; D008214:Lymphocytes / Q000276:immunology* / Q000502:physiology; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology* / Q000502:physiology; D008875:Middle Aged; D010051:Ovarian Neoplasms / Q000276:immunology*
chemical_list: D007376:Interleukin-2; D015847:Interleukin-4; D007371:Interferon-gamma