ex vivo expansion of tumour infiltrating lymphocytes in high grade serous ovarian cancer Authors Caitlin A Waddell 1 Marcus J Price 1 Eleanor J Cheadle 1 Richard J Edmondson ID: 1042847
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1. Short term inhibition of immune checkpoint proteins increases ex vivo expansion of tumour infiltrating lymphocytes in high grade serous ovarian cancer Authors: Caitlin A. Waddell1, Marcus J. Price1, Eleanor J. Cheadle1 , Richard J. Edmondson1, 2, Gemma L. Owens 1, 2 1 Division of Cancer Sciences, University of Manchester, Manchester, United Kingdom2 Department of Gynaecological Oncology, Manchester University NHS Foundation Trust, Manchester, United Kingdom.Figures, Tables and Supplementary data
2. Supplementary Data Table S1NameFluorophoreCloneCompanyAnti-CD8APC R700RPA-T8BD Biosciences, UKAnti-CD4BV785OKT4Biolegend, UKAnti-CD14Pe-Cy7HCD14Biolegend, UKAnti-CD20Pe-Cy72H7Biolegend, UKAnti-CD56Pe-Cy7HCD56Biolegend, UKAnti-PD1PerCPEH12.2H7Biolegend, UKAnti-TIM3BV510F38-2E2Biolegend, UKAnti-LAG3FITC11C3C65Biolegend, UKAnti-ICOSAPCC398.4ABiolegend, UKAnti-OX40BV711ACT35BD Biosciences, UKAnti-CD28PE-DazzleCD28.2Biolegend, UKAnti-IL2*BV4215344.111BD Biosciences, UKAnti-TNFa*BV650Mab11Biolegend, UKAnti-IFNy*APC Fire4S.B3Biolegend, UKGranzyme B*PEGB11BD Biosciences, UK *Denotes intracellular antigens Table 1. Antibodies utilised for flow cytometry
3. Supplementary Data Figure S1Supplementary Figure S1. Dose response of inhibitory antibodies against PD-1, TIM-3 and LAG-3 on TIL expansion in ovarian cancer tumour samples. The optimal concentration for expansion of TIL was determined for each inhibitory antibody against PD-1, TIM-3 and LAG-3. TIL isolated from two samples were incubated in the presence of a range of concentrations of each antibody (0µg/mL to 50µg/mL) on the first day of TIL expansion only. 10µg/mL of Nivolumab was utilised as a positive control for dose response. In further experiments, 10µg/mL of each antibody was used for TIL expansion experiments, having shown the greatest effect on TIL expansion.N/A, 1 replicate? N numberN/A P valuesN/A Stats test