Cd3 fitc clone sk

The phenotypes of cells in the blood were analyzed by flow cytometry according to standard procedures at the Hematopathology Unit, Dept. of Pathology, Karolinska University Hospital, using 6 color flow cytometry to detect T, B, NK cells and CD3− CD4+ cells (monocytes and dendritic cells). Flow cytometry was performed on a CANTO 1 flow cytometer (BD, Becton-Dickinson, Europe).
For data acquisition and analysis, a CANTO 1 flow cytometer (BD, Becton Dickinson, Europe) was used with Cell Quest software (Becton Dickinson, Franklin Lakes, New Jersey, USA). All samples were analyzed by setting appropriate side and forward scatter gates to identify the mononuclear cell population, using CD45 and forward and side scatter for gate setting. Consistency of analysis parameters was ascertained by calibrating the flow cytometer with calibrating beads and FacsComp software, both from Becton Dickinson. The results are reported as percentage of gated cells positive for each antibody. The following fluorochrome conjugated antibodies, all from BD, were used: CD4 PE, CD3 PerCP-Cy5.5, CD19 PE-Cy7, CD8 APC and CD45 APC-H7. We also used BD Multitest 6-Color TBNK Reagent containing CD3 FITC clone SK7, CD16 PE clone B73, CD56 PE clone NCAM 16.2, CD45 PerCP-Cy5.5 clone 2D1, CD4 PE-Cy7 clone SK3, CD19 APC clone SJ25C1 and CD8 APC-Cy7, clone SK1. The gating strategy is shown in Fig. S1.

Plasma HIV-1 viral loads (PVL), as well as CD4 and CD8 counts were determined in a laboratory operating under the Clinical Laboratory Improvement Amendment (CLIA). The plasma viral load was measured using the ultrasensitive Quantiplex HIV-1 bDNA version 3.0 (Bayer). CD4⁺ and CD8⁺ T-cell counts were determined by four-color flow cytometry. The BD Multitest (BD Biosciences) that was used includes the following Abs: CD3^FITC (clone SK7); CD4^APC (clone SK3); CD8^PE (clone SK1); and CD45^PerCP (clone 2D1). Samples were acquired on either FACSCalibur or FACSCanto (both BD Biosciences). CD4⁺ cell counts were calculated as percent of CD4⁺ CD3⁺ cells within CD45⁺ lymphocytes divided by 1% of the white blood cell count. The corresponding calculation was performed for CD8⁺ cell counts.