A phase II trial of autologous dendritic cell vaccination and radiochemotherapy following fluorescence-guided surgery in newly diagnosed glioblastoma patients

Table 4 Analysis of the inflammatory infiltrate in tumor samples

	CD4+ cells		CD8+ cells		CD4+ cells		CD8+ cells
	Basal (%)	Relapse (%)	Basal (%)	Relapse (%)	Basal (%)	Relapse (%)	Basal (%)	Relapse (%)
CD69	90.6	97.4	77	83.4	87.7	84.2	94.72	92
HLA-DR	71.85	59	57	46.48	56.1	56.17	37.1	50.29
Central memory	14.3	32.5	0	8.05	6.97	6.89	2.09	1.59
Efector memory	78.6	62.5	13.08	38.6	55.78	58.67	34.74	20.76
Effector	7.4	0	56.5	40.3	23.22	10.32	54.15	36.5
PD1	53.1	75	10.77	52	43.9	63.2	52.5	38.3

	MFI in Tumor cells		MFI in Tumor cells
	Basal	Relapse	Basal	Relapse
HLA-expression	36,969	10,038	42,433	14,245

A single cell suspension were obtained from biopsy sample by a mechanical disaggregation process. After washing twice, the cells were freeze following standard protocols
For flow cytometry analysis, an aliquot of cells was thawed and after 2 h at 37°, a panel of monoclonal antibodies was used to identify different cell subpopulations. Results are expressed in percentage regarding total alive cells (CD3 and myeloid suppressor cells), and regarding CD3+ cells (CD4 and CD8). Activation markers (CD69 and HLA-DR) and a panel of markers to characterize different T cell population were evaluated in CD4 and CD8 positive cells. HLA-I expression (mean fluorescence intensity) were measure in tumor cells
In all cases, cells were incubated with monoclonal antibodies conjugated to fluorochromes during 15 min at room temperature and in the dark and then the cells were washed. Cells stained with isotype control antibody were used as negative control. Cells were acquired in FACSCALIBUR cytometer (Becton–Dickinson) and then analyzed using Flow Jo software

ISSN: 1479-5876