1. Expression of CD10 by Human T Cells That Undergo Apoptosis Both In Vitro and In Vivo
by Giovanna Cutrona, Nicolò Leanza, Massimo Ulivi, Giovanni Melioli, Vito L. Burgio, Giovanni Mazzarello, Giovanni Gabutti, Silvio Roncella, and Manlio Ferrarini
Blood
Volume 94(9):
November 1, 1999
©1999 by American Society of Hematology

2. CD10 expression by H9 T cells after HIV infection (A), exposure to CD95 MoAb (C), and staurosporin or etoposide (D) for 24 hours.
CD10 expression by H9 T cells after HIV infection (A), exposure to CD95 MoAb (C), and staurosporin or etoposide (D) for 24 hours. The cells induced into apoptosis by one of these treatments were separated from nonapoptotic cells based on their FSC and SSC. Apoptotic and CD10+ cells were observed in the same gate (gate 1). Apoptosis was measured by PI staining of permeabilized cells (A through D) or Annexin-V staining (D) and flow cytometry. (D) Only the percentage of CD10+ and apoptotic cells gated in 1 is reported. Control cells (B), incubated with medium or with an irrelevant MoAb, were composed of a homogeneous cell population that did not express CD10 and was not apoptotic. The inset in A shows the flow cytometry profile for gp120 staining to document the ongoing HIV infection of the cells in vitro.
Giovanna Cutrona et al. Blood 1999;94:
©1999 by American Society of Hematology

3. Control tests to ascertain that CD10 is synthesized and expressed by apoptotic T cells.
Control tests to ascertain that CD10 is synthesized and expressed by apoptotic T cells. (A) Cells were exposed to CD95 MoAb for 18 hours, gated as indicated, and sorted. The sorted cells were analyzed for SSC and FSC or apoptosis by PI staining. (B) Cells were exposed to CD95 MoAb or medium for 48 hours and triple stained in isotonic medium with PI, Annexin-V-FITC, and CD10 MoAb-PerCP. The cells that excluded PI were gated and analyzed. (C) Cells were exposed to CD95 MoAb in the presence or absence of cycloheximide (50 μg/mL) and analyzed for CD10 expression and apoptosis by Annexin-V FITC staining.
Giovanna Cutrona et al. Blood 1999;94:
©1999 by American Society of Hematology

4. Inhibition of CD10 expression by treatment with VAD-FMK.
Inhibition of CD10 expression by treatment with VAD-FMK. H9 T cells were cultured with CD95 MoAb in the presence or absence of VAD-FMK (10 μmol/L, final concentration) for 24 or 48 hours. Control suspensions were cultured with medium alone. At the end of the culture period, the cells were obtained and stained with CD10 MoAb and Annexin-V-FITC or an irrelevant MoAb and Annexin-V-FITC.
Giovanna Cutrona et al. Blood 1999;94:
©1999 by American Society of Hematology

5. Expression of CD10 by CD4+ T cells induced into apoptosis in vitro.
Expression of CD10 by CD4+ T cells induced into apoptosis in vitro. (A) Purified CD4+ T cells were exposed to SEB and/or CD4 MoAb under cross-linking conditions in different combinations as indicated. The cells were obtained after 24 hours and CD10 expression and apoptosis (PI staining) were measured by flow cytometry. (B) RT-PCR analysis of CD10 mRNA expression in CD4+ T cells exposed to the indicated stimuli for 24 hours in vitro. Burkitt’s lymphoma B cells (LAM cell line) were used as positive control. (C) Alu I esonuclease digestion and analysis of the RT-PCR fragments extracted from the indicated cells. These results represent a typical experiment of the 3 performed.
Giovanna Cutrona et al. Blood 1999;94:
©1999 by American Society of Hematology

6. Expression of CD10 by apoptosing CD8-positive T-cell blasts.
Expression of CD10 by apoptosing CD8-positive T-cell blasts. T-cell blasts from IL-2–dependent continuous T-cell lines were cultured in the presence or absence of IL-2 (20 U/mL) for 48 hours. The cells were obtained, double stained as indicated, and analyzed by flow cytometry. The quadrants were drawn based on the analysis of negative controls stained with an irrelevant (isotype-matched) MoAb or analyzed in the absence of Annexin-V-FITC staining.
Giovanna Cutrona et al. Blood 1999;94:
©1999 by American Society of Hematology

7. Expression of CD10 by apoptosing T cells in vivo.
Expression of CD10 by apoptosing T cells in vivo. Lymph node cell suspensions from an HIV-seropositive individual were double or triple stained as indicated and analyzed by flow cytometry. CTR (control) indicates that the cells were exposed to an irrelevant MoAb.
Giovanna Cutrona et al. Blood 1999;94:
©1999 by American Society of Hematology

8. Expression of CD10 by T cells from the peripheral blood of HIV-seropositive individuals.
Expression of CD10 by T cells from the peripheral blood of HIV-seropositive individuals. Summary of the data obtained on the peripheral blood T cells from 10 HIV-seropositive patients and the average values obtained in 10 control individuals.
Giovanna Cutrona et al. Blood 1999;94:
©1999 by American Society of Hematology

9. Expression of CD10 by apoptosing peripheral blood T cells from HIV-seropositive patients.
Expression of CD10 by apoptosing peripheral blood T cells from HIV-seropositive patients. Peripheral blood MNC from 1 normal control or 2 HIV-seropositive patients were double or triple stained as indicated in Fig 6, and analyzed by flow cytometry. The quadrants were drawn based on the analysis of negative controls stained with an irrelevant (isotype-matched) MoAb or in the absence of Annexin-V-FITC.
Giovanna Cutrona et al. Blood 1999;94:
©1999 by American Society of Hematology