Biomarker for judging efficacy of immune checkpoint inhibitor
A technology of immune checkpoints and substances, applied in the field of treatment of malignant tumors
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Embodiment 1
[0430] Example 1: Collection of Lymphocytes from Tumor Tissue and Identification of T Cells by Flow Cytometry
[0431] Tumor tissue was collected from cancer patients prior to administration of nivolumab, and tumor infiltrating lymphocytes were isolated by disrupting the tissue with a gentleMACSDissociator (MiltenyiBiotec). Peripheral blood mononuclear cells were prepared by density gradient centrifugation using Ficoll from the peripheral blood of the same patients prior to administration of substances that inhibit immune checkpoints. The isolated tumor-infiltrating lymphocytes and peripheral blood mononuclear cells were suspended in PBS (hereinafter referred to as "FACS buffer") added so that the final concentration of fetal calf serum was 2%, and human Fc Receptor binding inhibitor (ThermoFisher), and the mixture was stored at 4°C for 10 minutes. Thereafter, fluorescently-labeled antibodies against various cell surface markers (CD3, CD8, CD4, CD45RA, and PD-1) were added ...
Embodiment 2
[0433] Example 2: Based on Treg cells and CD8 + The respective PD-1 expression intensity in T cells, assessed for the determination of NiV Biomarkers of mAb effectiveness (non-small cell lung cancer)
[0434] Treg cells, Treg cells (Fr. + The expression intensity of PD-1 in T cells, and calculate its mean fluorescence intensity (MFI).
[0435] Among patients with non-small cell lung cancer, 7 patients showed the effect of CR or PR and 7 patients showed SD or PD by nivolumab administration. One remaining patient could not be evaluated and was therefore excluded from the following evaluations.
[0436] figure 1 and 2 The calculated values for each of the patient group showing CR or PR (hereinafter referred to as "CR / PR group") and the patient group showing SD or PD (hereinafter referred to as "SD / PD group") are shown. + The respective results of the ratio of PD-1 expression MFI in T cells to Treg cells and PD-1 expression MFI in Treg cells (Fr.II). In addition, th...
Embodiment 3
[0441] Example 3: Based on Treg cells and CD8 + The respective PD-1 expression intensity in T cells, assessed for the determination of NiV Biomarkers of mAb effectiveness (gastric cancer)
[0442] Treg cells, Treg cells (Fr.II) and CD8 were measured by flow cytometry in tumor tissues derived from 17 patients with gastric cancer + The expression intensity of PD-1 in T cells and calculate its MFI.
[0443] Among patients with gastric cancer, 3 patients showed an effect of PR and 14 patients showed SD or PD by the administration of nivolumab.
[0444] image 3 and 4 Shows the relative CD8 calculated in the group of patients showing PR and the "SD / PD group + The respective results of the ratio of PD-1 expression MFI in T cells to Treg cells and PD-1 expression MFI in Treg cells (Fr.II). In addition, the upper and lower bounds of the 95% confidence intervals were calculated based on the means and standard deviations of the same ratios in each group. Table 2 shows the r...
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