Cd3 clone ucht1

Cells were surface stained with anti-human antibodies (Abs) to CD11b (clone ICRF44), CD14 (clone TuK4), CD31 (clone M89D3), CD45 (clone 2D1), CD163 (clone GHI/61), IL-6 (clone MQ2-39C3), TNF-α (clone MAb11), vimentin (clone RV202) (BD Pharmingen, San Diego, CA), IL-8 (clone E8N1), CD326 (EpCAM, clone 9C4) (Biolegend, San Diego, CA), and CD3 (clone UCHT1, Beckman-Coulter, Miami, FL). Antibodies conjugated to the following fluorochromes were used in these studies: Fluorescein isothiocyanate (FITC), Phycoerythrin (PE), Peridinin chlorophyll protein (PerCP)-Cy5.5, PE-Cy7, Energy Coupled Dye or PE-Texas-Red conjugate (ECD), Pacific Blue, Brilliant Violet (BV) 570, BV605, BV650, Quantum dot (QD) 800, Alexa 647, allophycocyanin (APC)-Alexa 700 and APC-H7.

Cells were stained with mAbs to CD3 (clone UCHT1), CD69 (clone TPI-55-3) (Beckman-Coulter, Miami, FL), CD8 (clone HIT8a), CD161 (clone DX12), IL-10 (clone JES3-9D7), interferon (IFN)-γ (clone B27), tumor necrosis factor (TNF)-α (clone MAb11) (BD Pharmingen, San Diego, CA, USA), CD14 (clone TuK4), CD45 (clone H130) (Invitrogen, Carlsbad, CA), and HLA-G (clone 87G), CD85j (ILT2) (clone GHI/75), and TCRα7.2 (clone 3C10)(Biolegend, San Diego, CA). Polyclonal antibody to the Salmonella Common Structural Antigens (CSA) (KPL, Gaithersburg, MD, USA) was used to measure S. Typhi infection. Antibodies conjugated to the following fluorochromes were used in these studies: Fluorescein isothiocyanate (FITC), Phycoerythrin (PE), PE-Cy5, Peridinin chlorophyll protein (PerCP)-Cy5.5, PE-Cy7, Pacific Blue, Brilliant Violet (BV) 570, BV605, BV650, Quantum dot (QD) 800, Alexa 647, allophycocyanin (APC)-Alexa 700, or APC-H7.

T-cell clonal expansion was detected using TCR Vβ repertoire analysis by means of an IOTest Beta Mark TCR V Kit (Beckman Coulter, Brea, CA, USA). Relative frequencies of 24 Vβ T cell receptor families were analyzed in CD4+ and CD8+ T cells by flow cytometry in the peripheral blood of 17 recipients of ASCT from the LTR cohort. TCR Vβ panels included antibodies against CD3 (clone UCHT1), CD4 (clone 13B8.2), and CD8 (clone T8) (Beckman Coulter). Cells were acquired on a BD FACSCanto II cytometer and data were analyzed with FlowJo Software v.10 (BD Biosciences, San Jose, CA, USA).