Abstract
Background
One of the biggest caveats in diffuse glioma therapy is anti-tumor immune response suppression. By using cell-cell interactions like secreting cytokines and grow factors that modulate recruited macrophages and resident microglia, neoplastic cells do manage to promote favorable immediate milieus within healthy brain tissue. The aim of this study is to estimate proportions of immune- related cells in diffuse gliomas.
Material and Methods
Starting with fresh tumor biopsies, tissue integrity is mechanically disrupted followed by an enzymatic digestion. After initial output filtering, cell suspension is cleared from debris by gradient centrifugation. Immune cell fraction is enriched by magnetic cell sorting using anti-CD45 beads (leucocyte common antigen). CD45+ (positive) or column retained fraction, contains majority of the immune cells present in the tumor biopsy whereas CD45- (negative) or column eluted fraction, is comprised by tumor cells and other non-immune system related cells. Flow cytometry based cell-phenotyping approach has been used aiming for T lymphocyte, B lymphocyte, macrophage and granulocyte cell types estimation.
Results
Accordingly, we can point out proportion differences between different diffuse gliomas entities like an anaplastic astrocytoma (AA) grade III World Health Organization and glioblastoma (GBM) grade IV. Our results show a strong macrophage-related component and reveal a substantial granulocyte component in a GBM patient compared to the AA patient
Conclusion
Altogether, this novel approach allows portraying preliminary immune proportions differences from AA grade III and a GBM grade IV patients, helping into the grounding step towards further understanding on how immune response is conformed in diffuse gliomas.