TY - JOUR
T1 - Tumor cell and immune cell profiles in primary human glioblastoma
T2 - impact on patient outcome
AU - González-Tablas Pimenta, María
AU - Otero, Álvaro
AU - Arandia Guzman, Daniel Angel
AU - Pascual-Argente, Daniel
AU - Ruíz Martín, Laura
AU - Sousa-Casasnovas, Pablo
AU - García-Martin, Andoni
AU - Roa Montes de Oca, Juan Carlos
AU - Villaseñor-Ledezma, Javier
AU - Torres Carretero, Luis
AU - Almeida, Maria
AU - Ortiz, Javie
AU - Nieto, Adelaida
AU - Orfao, Alberto
AU - Tabernero, María Dolores
N1 - Funding Information:
This work was supported by CIBERONC (grant CB16/12/00400 CIBERONC, Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, Madrid, Spain) the ISCIII PI16/0476 grant (Instituto de Salud Carlos III), Fondos FEDER and GRS2049/A/19 grant (Consejería de Sanidad Junta de Castilla y León, Gerencia Regional de Salud, Spain).
Publisher Copyright:
© 2020 The Authors. Brain Pathology published by John Wiley & Sons Ltd on behalf of International Society of Neuropathology
PY - 2021/3
Y1 - 2021/3
N2 - The distribution and role of tumor-infiltrating leucocytes in glioblastoma (GBM) remain largely unknown. Here, we investigated the cellular composition of 55 primary (adult) GBM samples by flow cytometry and correlated the tumor immune profile with patient features at diagnosis and outcome. GBM single-cell suspensions were stained at diagnosis (n = 44) and recurrence following radiotherapy and chemotherapy (n = 11) with a panel of 8-color monoclonal antibody combinations for the identification and enumeration of (GFAP+CD45−) tumor and normal astrocytic cells, infiltrating myeloid cells —i.e. microglial and blood-derived tumor-associated macrophages (TAM), M1-like, and M2-like TAM, neutrophils. and myeloid-derived suppressor cells (MDSC)— and tumor-infiltrating lymphocytes (TIL) —i.e. CD3+T-cells and their TCD4+, TCD8+, TCD4−CD8−, and (CD25+CD127lo) regulatory (T-regs) subsets, (CD19+CD20+) B-cells, and (CD16+) NK-cells—. Overall, GBM samples consisted of a major population (mean ± 1SD) of tumor and normal astrocytic cells (73% ± 16%) together with a significant but variable fraction of immune cells (24% ± 18%). Within myeloid cells, TAM predominated (13% ± 12%) including both microglial cells (10% ± 11%) and blood-derived macrophages (3% ± 5%), in addition to a smaller proportion of neutrophils (5% ± 9%) and MDSC (4% ± 8%). Lymphocytes were less represented and mostly included TCD4+ (0.5% ± 0.7%) and TCD8+ cells (0.6% ± 0.7%), together with lower numbers of TCD4−CD8− T-cells (0.2% ± 0.4%), T-regs (0.1% ± 0.2%), B-lymphocytes (0.1% ± 0.2%) and NK-cells (0.05% ± 0.05%). Overall, three distinct immune profiles were identified: cases with a minor fraction of leucocytes, tumors with a predominance of TAM and neutrophils, and cases with mixed infiltration by TAM, neutrophils, and T-lymphocytes. Untreated GBM patients with mixed myeloid and lymphoid immune infiltrates showed a significantly shorter patient overall survival versus the other two groups, in the absence of gains of the EGFR gene (p = 0.02). Here we show that immune cell infiltrates are systematically present in GBM, with highly variable levels and immune profiles. Patients with mixed myeloid and T-lymphoid infiltrates showed a worse outcome.
AB - The distribution and role of tumor-infiltrating leucocytes in glioblastoma (GBM) remain largely unknown. Here, we investigated the cellular composition of 55 primary (adult) GBM samples by flow cytometry and correlated the tumor immune profile with patient features at diagnosis and outcome. GBM single-cell suspensions were stained at diagnosis (n = 44) and recurrence following radiotherapy and chemotherapy (n = 11) with a panel of 8-color monoclonal antibody combinations for the identification and enumeration of (GFAP+CD45−) tumor and normal astrocytic cells, infiltrating myeloid cells —i.e. microglial and blood-derived tumor-associated macrophages (TAM), M1-like, and M2-like TAM, neutrophils. and myeloid-derived suppressor cells (MDSC)— and tumor-infiltrating lymphocytes (TIL) —i.e. CD3+T-cells and their TCD4+, TCD8+, TCD4−CD8−, and (CD25+CD127lo) regulatory (T-regs) subsets, (CD19+CD20+) B-cells, and (CD16+) NK-cells—. Overall, GBM samples consisted of a major population (mean ± 1SD) of tumor and normal astrocytic cells (73% ± 16%) together with a significant but variable fraction of immune cells (24% ± 18%). Within myeloid cells, TAM predominated (13% ± 12%) including both microglial cells (10% ± 11%) and blood-derived macrophages (3% ± 5%), in addition to a smaller proportion of neutrophils (5% ± 9%) and MDSC (4% ± 8%). Lymphocytes were less represented and mostly included TCD4+ (0.5% ± 0.7%) and TCD8+ cells (0.6% ± 0.7%), together with lower numbers of TCD4−CD8− T-cells (0.2% ± 0.4%), T-regs (0.1% ± 0.2%), B-lymphocytes (0.1% ± 0.2%) and NK-cells (0.05% ± 0.05%). Overall, three distinct immune profiles were identified: cases with a minor fraction of leucocytes, tumors with a predominance of TAM and neutrophils, and cases with mixed infiltration by TAM, neutrophils, and T-lymphocytes. Untreated GBM patients with mixed myeloid and lymphoid immune infiltrates showed a significantly shorter patient overall survival versus the other two groups, in the absence of gains of the EGFR gene (p = 0.02). Here we show that immune cell infiltrates are systematically present in GBM, with highly variable levels and immune profiles. Patients with mixed myeloid and T-lymphoid infiltrates showed a worse outcome.
KW - Glioblastoma
KW - Immune cells
KW - Lymphocytes
KW - Microenvironment
KW - Microglia
KW - Myeloid cells
UR - http://www.scopus.com/inward/record.url?scp=85099162663&partnerID=8YFLogxK
U2 - 10.1111/bpa.12927
DO - 10.1111/bpa.12927
M3 - Article
C2 - 33314398
AN - SCOPUS:85099162663
SN - 1015-6305
VL - 31
SP - 365
EP - 380
JO - Brain Pathology
JF - Brain Pathology
IS - 2
ER -
