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. 2002 Jan 7;195(1):113–123. doi: 10.1084/jem.20010956

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Copyright © 2002, The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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Figure 2. — Cytotoxicity of PD5-induced CTL lines. PD5-specific CTL lines, induced by culturing PBMCs with PD5-loaded APCs for 12–14 d, were tested for their cytotoxicity against PD5 or control peptide-loaded autologous BCL targets at different effector/target (E/T) ratios. HBc18–27, an HLA-A2-restricted irrelevant epitope, was used as control. Displayed are mean specific lysis of triplicate cultures. (A) PBC patients PBC9 and PBC11. (B) Control donors CL2 (granulomatous liver disease) and H2 (healthy donor). (C) Cytotoxic activity of a PD5-induced CTL line against target cells presenting endogenously processed PDC-E2 Ag. The CTL line from patient PBC1 was tested for cytotoxicity against autologous BCL targets infected with a PDC-E2–expressing vaccinia vector (VVrPDC-E2), wild-type vaccinia virus (VVwild) alone, loaded with PD5 or a control peptide HBc18–27. Displayed are mean specific lysis of triplicate testing at an E/T ratio of 40:1. (D) Phenotype analysis of PD5-induced CTLs. CD4⁺ cell or CD8⁺ cells were depleted respectively from a CTL line derived from patient PBC2 using anti-CD4 or anti-CD8–coated magnetic beads before testing for cytotoxic activity with PD5 or control peptide-loaded autologous BCL targets. Unfractionated cells were also tested in parallel. Specific cytotoxicity was calculated by subtracting the cytotoxicity of the effector cells against control peptide-pulsed BCLs from that against PD5 peptide-pulsed BCLs. Displayed are mean specific lysis of triplicate testing at an E/T ratio of 40:1 according to the cell counts before depletion.

Figure 2. — Cytotoxicity of PD5-induced CTL lines. PD5-specific CTL lines, induced by culturing PBMCs with PD5-loaded APCs for 12–14 d, were tested for their cytotoxicity against PD5 or control peptide-loaded autologous BCL targets at different effector/target (E/T) ratios. HBc18–27, an HLA-A2-restricted irrelevant epitope, was used as control. Displayed are mean specific lysis of triplicate cultures. (A) PBC patients PBC9 and PBC11. (B) Control donors CL2 (granulomatous liver disease) and H2 (healthy donor). (C) Cytotoxic activity of a PD5-induced CTL line against target cells presenting endogenously processed PDC-E2 Ag. The CTL line from patient PBC1 was tested for cytotoxicity against autologous BCL targets infected with a PDC-E2–expressing vaccinia vector (VVrPDC-E2), wild-type vaccinia virus (VVwild) alone, loaded with PD5 or a control peptide HBc18–27. Displayed are mean specific lysis of triplicate testing at an E/T ratio of 40:1. (D) Phenotype analysis of PD5-induced CTLs. CD4⁺ cell or CD8⁺ cells were depleted respectively from a CTL line derived from patient PBC2 using anti-CD4 or anti-CD8–coated magnetic beads before testing for cytotoxic activity with PD5 or control peptide-loaded autologous BCL targets. Unfractionated cells were also tested in parallel. Specific cytotoxicity was calculated by subtracting the cytotoxicity of the effector cells against control peptide-pulsed BCLs from that against PD5 peptide-pulsed BCLs. Displayed are mean specific lysis of triplicate testing at an E/T ratio of 40:1 according to the cell counts before depletion.