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Blood, Vol. 107, Issue 9, 3665-3668, May 1, 2006
A family with Papillon-Lefèvre syndrome reveals a requirement for cathepsin C in granzyme B activation and NK cell cytolytic activity
Blood Meade et al.
107: 3665
Supplemental Figures for: Meade et al
Files in this Data Supplement:
- Figure S1. Lymphocyte cell surface phenotype in Papillon-Lefèvre Syndrome as determined by flow cytometry (PDF, 31.4 KB) -
Cell surface markers were analyzed in the two PLS patients (Patients 1 and 2), the Heterozygote (Het) and a healthy control sample (Control). Lymphocytes were analysed using CD56 staining (x-axis) in conjunction with either CD3, CD94, CD16, CD158a or CD158b for NK cells or CD19 for B-cells (y-axis). T-cells can be visualized in the CD56 and CD3 staining panel as the CD3+ population.
- Figure S2. Intracellular expression of perforin in Papillon-Lefèvre Syndrome NK cells (PDF, 34.5 KB) -
Intracellular perforin expression within NK cells was analysed in a PLS patient (Patient 2) and a healthy control sample (control). Cells were surface stained for CD56 and CD3 expression and then fixed, permeabilized and stained with an anti-perforin antibody. The plots shown are gated on the CD3 negative population.
- Figure S3. A complex between granzyme B and proteinase inhibitor-9 (PI-9) in unstimulated and IL-2 activated NK cells (PDF, 71.8 KB) -
Upper panel: This western blot is also shown in cropped form in the right hand panel of Figure 2E. The blot was probed with the anti-granzyme B antibody 2C5 which has been shown to detect uncomplexed granzyme B and granzyme B:PI-9 complexes.23 The blot indicates the position of the uncomplexed granzyme B, the granzyme B:PI-9 complexes and major degradation products (in these complexes, both the inhibitor and the granzyme B are believed to undergo severe distortion, leading to their degradation).23
Lower panel: Enlargement of area of western blot showing granzyme B:PI-9 complexes and degradation products in the different samples. These are visible in control and patient IL-2 stimulated samples and control unstimulated samples whereas no complexes can be seen in the lysates from unstimulated patient NK cells consistent with a lack of active granzyme B in this sample. Stimulation with IL-2 increases the synthesis of granzyme B and this may lead to differences in the pattern and/or stability of complexes and degradation products compared to unstimulated samples.
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