Anti tim 3 pe clone 344823

A single flow cytometry panel was tested on all HIV-infected individuals and healthy control subjects as seen in Buggert et al. The antibodies used in the assay were: anti-CD3 APC-H7 (Clone SK7), anti-CD14 V500 (Clone M5E2), anti-CD19 V500 (Clone B43) (BD Bioscience); anti-CD45RO ECD (clone UCHL1) (Beckman Coulter); anti-CD4 BV650 (Clone OKT4), anti-PD-1 BV421 (clone EH12.2H7), anti-CD27 BV785 (clone O323), anti-CD28 PerCP-Cy5.5 (clone CD28.2), anti-CD38 APC (clone HIT2), anti-CD57 FITC (clone HCD57) (Biolegend); anti-HLA-DR PE-Cy7 (clone LN3) (eBioscience); anti-CD8 Qd565 (Clone 3B5) (Life Technologies) and anti-Tim-3 PE (clone 344823) (R&D Systems). LIVE/DEAD Aqua amine dye (Life Technologies) was used to discriminate dead cells and debris.

Prior to co-culture with transduced DCs, 10⁶ T cells were harvested, washed and stained with anti-CD8-APC.H7 (clone SK1), anti-CD3-Percp-Cy5.5 (clone SK7), anti-PD-1-APC (clone MIH4), anti-CD38-FITC (cloneHIT2) (all from BD Bioscience, UK), anti-CD160-PE (clone 688327), anti-TIM3-PE (clone 344823)(both from R&D, UK) and anti-LAG3-FITC (clone 17B4, Enzo Life Sciences, UK) for 20 mins at room temperature. Following this incubation, cells were washed and fixed with BD stabilising fixative (BD Biosciences). One hundred thousand cells were acquired within a live gate using a 3-laser configuration LSRII flow cytometer (BD Biosciences, USA) and analysed by FlowJo (Tree Star Inc, USA).
To assess gag protein production, transduced DCs were harvested, washed and stained with anti-gag-PE (clone KC57, Beckman Coulter UK) for 20 mins at room temperature. Following this incubation cells were washed and fixed with BD stabilising fixative (BD Biosciences) and cells were acquired on a forward scatter versus PE plot using a 3-laser configuration LSRII flow cytometer (BD Biosciences, USA) and analysed by FlowJo (Tree Star Inc, USA).