title: Tissue-based immune monitoring I: tumor core needle biopsies allow in-depth interrogation of the tumor microenvironment.
abstract: We sought to assess the feasibility and reproducibility of performing tissue-based immune characterization of the tumor microenvironment using CT-compatible needle biopsy material. Three independent biopsies were obtained intraoperatively from one metastatic epithelial ovarian cancer lesion of 7 consecutive patients undergoing surgical cytoreduction using a 16-gauge core biopsy needle. Core specimens were snap-frozen and subjected to immunohistochemistry (IHC) against human CD3, CD4, CD8, and FoxP3. A portion of the cores was used to isolate RNA for 1) real-time quantitative (q)PCR for CD3, CD4, CD8, FoxP3, IL-10 and TGF-beta, 2) multiplexed PCR-based T cell receptor (TCR) CDR3 Vβ region spectratyping, and 3) gene expression profiling. Pearson's correlations were examined for immunohistochemistry and PCR gene expression, as well as for gene expression array data obtained from different tumor biopsies. Needle biopsy yielded sufficient tissue for all assays in all patients. IHC was highly reproducible and informative. Significant correlations were seen between the frequency of CD3+, CD8+ and FoxP3+ T cells by IHC with CD3ε, CD8A, and FoxP3 gene expression, respectively, by qPCR (r=0.61, 0.86, and 0.89; all p< 0.05). CDR3 spectratyping was feasible and highly reproducible in each tumor, and indicated a restricted repertoire for specific TCR Vβ chains in tumor-infiltrating T cells. Microarray gene expression revealed strong correlation between different biopsies collected from the same tumor. Our results demonstrate a feasible and reproducible method of immune monitoring using CT-compatible needle biopsies from tumor tissue, thereby paving the way for sophisticated translational studies during tumor biological therapy.
authors.affiliation: Ovarian Cancer Research Center, Division of Gynecologic Oncology, University of Pennsylvania, Philadelphia, USA.
authors.forename: Andrea R
authors.initials: AR
authors.lastname: Hagemann
authors.forename: Mark
authors.initials: M
authors.lastname: Cadungog
authors.forename: Ian S
authors.initials: IS
authors.lastname: Hagemann
authors.forename: Rachel
authors.initials: R
authors.lastname: Hammond
authors.forename: Sarah F
authors.initials: SF
authors.lastname: Adams
authors.forename: Christina S
authors.initials: CS
authors.lastname: Chu
authors.forename: Steven C
authors.initials: SC
authors.lastname: Rubin
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.forename: Kathakali
authors.initials: K
authors.lastname: Addya
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Birrer
authors.forename: Phyllis A
authors.initials: PA
authors.lastname: Gimotty
authors.forename: George
authors.initials: G
authors.lastname: Coukos
mesh_terms: D000328:Adult; D000368:Aged; D001707:Biopsy, Needle / Q000379:methods; D017252:CD3 Complex / Q000235:genetics / Q000276:immunology; D015704:CD4 Antigens / Q000235:genetics / Q000276:immunology; D016827:CD8 Antigens / Q000235:genetics / Q000276:immunology; D000077216:Carcinoma, Ovarian Epithelial; D005240:Feasibility Studies; D005260:Female; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000276:immunology; D005629:Frozen Sections; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D008875:Middle Aged; D015166:Monitoring, Immunologic / Q000379:methods*; D009375:Neoplasms, Glandular and Epithelial / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis; D010051:Ovarian Neoplasms / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D010053:Ovary / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D046888:Tissue Array Analysis / Q000379:methods*; D059016:Tumor Microenvironment / Q000235:genetics / Q000276:immunology*
chemical_list: D017252:CD3 Complex; D015704:CD4 Antigens; D016827:CD8 Antigens; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D011948:Receptors, Antigen, T-Cell