Tumor cell and immune cell profiles in primary human glioblastoma: Impact on patient outcome

© 2020 The Authors.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 (GFAPCD45) 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. CD3T-cells and their TCD4, TCD8, TCD4CD8, and (CD25CD127) regulatory (T-regs) subsets, (CD19CD20) 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 TCD4CD8 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.Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, Madrid, Spain and fondos FEDER, Grant/Award Number: CB16/12/00400 and ISCIII PI16/0476; Consejería de Sanidad Junta de Castilla y León, Gerencia Regional de Salud, Spain, Grant/Award Number: GRS2049/A/1

Sensitivity of the early life stages of a mayfly to fine sediment and orthophosphate levels

The ecological effects of interacting stressors within lotic ecosystems have been widely acknowledged. In particular, the ecological effects of elevated fine sediment inputs and phosphate have been identified as key factors influencing faunal community structure and composition. However, while knowledge regarding adult and larval life stage responses to environmental stressors has grown, there has been very limited research on their eggs. In this study, the eggs of the mayfly Serratella ignita (Ephemerellidae: Ephemeroptera) were collected and incubated in laboratory aquaria to hatching under differing concentrations of inert suspended sediment (SS) and orthophosphate (OP), individually and in combination. Results indicate that SS and OP have greater effects on egg hatching in combination than when either were considered in isolation. SS displayed a greater effect on egg survival than OP in isolation or when OP was added to elevated SS treatments. Egg mortality in control treatments was around 6% compared to 45% in treatments with 25 mg 1⁻¹ SS and 52% in 0.3 mg 1⁻¹ OP treatments. Even relatively modest levels of each stressor (10 mg 1⁻¹ SS; 0.1 mg 1⁻¹ OP), below national legal thresholds, had significant effects on egg survival to hatching. The results support calls for legal levels of SS to be reassessed and suggest that more research is required to assess the impacts of pollution on invertebrate egg development given their different sensitivity and exposure pathways compared to other life stages