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Defective B-cell proliferation and maintenance of long-term memory in patients with chronic granulomatous disease  Nicola Cotugno, MD, Andrea Finocchi,

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Presentation on theme: "Defective B-cell proliferation and maintenance of long-term memory in patients with chronic granulomatous disease  Nicola Cotugno, MD, Andrea Finocchi,"— Presentation transcript:

1 Defective B-cell proliferation and maintenance of long-term memory in patients with chronic granulomatous disease  Nicola Cotugno, MD, Andrea Finocchi, MD, PhD, Alberto Cagigi, PhD, Gigliola Di Matteo, PhD, Maria Chiriaco, PhD, Silvia Di Cesare, PhD, Paolo Rossi, MD, PhD, Alessandro Aiuti, MD, PhD, Paolo Palma, MD, PhD, Iyadh Douagi, PhD  Journal of Allergy and Clinical Immunology  Volume 135, Issue 3, Pages e2 (March 2015) DOI: /j.jaci Copyright © Terms and Conditions

2 Fig 1 The B-cell phenotype of patients with CGD is characterized by lower memory and higher naive cell counts. Representative gates (A) and comparisons of B-cell percentages (B and C) and absolute counts (D) are shown. Horizontal bars represent medians. P values were determined with the nonparametric Mann-Whitney test. CD19+ cells established the B-cell population, and expression of IgD, CD27, CD21, CD24, CD38, and CD10 was used to define total naive (CD27−IgD+), total memory (CD27+IgD−), double-negative (DN; CD27−IgD−), resting memory (Rem), tissue-like (TL), activated memory (AM), naive (Na), and immature transitional (Tr) cells. FSC, Forward scatter; SSC, side scatter. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © Terms and Conditions

3 Fig 2 B-cell proliferation of patients with CGD is reduced if a single stimulus directed to BCR or TLR9 is given. Representative gate of carboxyfluorescein succinimidyl ester (CFSE)–diluted B cells (A) and comparisons of proliferating B-cell percentages between patients with CGD (n = 10) and HCs (n = 13; B) are shown in the scatter dot plots. Horizontal bars, Medians. Statistics have been defined with the nonparametric Mann-Whitney test, and only significant P values (P < .05) have been reported in the figure. SSC, Side scatter. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © Terms and Conditions

4 Fig 3 ROS activity is hampered in B cells affected by CGD. Representative gate (A) and comparisons (B) between patients with CGD and HCs in terms of modulation of ROS with and without PMA stimulation on gated CD19+ cells are shown. Modulation of ROS is defined as the ratio of the geometric mean of M2–2′,7′-dichlorodihydrofluorescein diacetate (DCFDA) before and after PMA stimulation. Means and SEMs are shown in the graph. Statistics have been defined with the nonparametric Mann-Whitney test. SSC, Side scatter. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © Terms and Conditions

5 Fig 4 In vitro and in vivo responses to measles in patients with CGD are lower compared with those seen in HCs. Representative ELISpot assay analysis (A) and comparisons (B) between patients with CGD and HCs are shown in the graph. Open triangles, Average of the IgG spots; horizontal bars, medians. Specific antibodies for measles and total IgG (C) and correlation (Spearman test) with age in patients with CGD (D) and HCs (E) are shown. KLH, Keyhole limpet hemocyanin. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © Terms and Conditions

6 Fig 5 Patients with CGD not producing measles-specific ASCs have lower memory and higher naive B cells. Among patients with CGD, 2 groups were created according to numbers of ASCs for measles. Responders and nonresponders have been selected according to the number of ASCs/106 cells greater than or less than the mean plus SD. The nonparametric Mann-Whitney test was used to analyze data. Box plots show means and SEMs. Na, Naive cells; Rem, resting memory cells; Tr, immature transitional cells. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © Terms and Conditions

7 Fig 6 Selective advantage of CGD carriers relates to B-cell defects. Representative gate (A) and contingency graph (B) of gp91phox-positive B cells in mothers with CGD are shown. Comparisons of B-cell phenotype (C and D) and proliferation (E) between HCs, patients with CGD, and CGD carriers are also shown. Different dot colors refer to the different CGD carriers according to selective advantage shown in Fig 6, B. The Mann-Whitney test was used to analyze data. Means and SDs are shown in the graphs. AM, Activated memory cells; Na, naive cells; Rem, resting memory cells; TL, tissue-like cells. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © Terms and Conditions

8 Fig E1 Proliferation of memory and naive B cells. Representative gates (A) and graphs of gated memory (B) and naive (C) B cells are shown. The Mann-Whitney test was used to analyze data. Means and SDs are shown in the graphs. CFSE, Carboxyfluorescein isothiocyanate; SSC, side scatter. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © Terms and Conditions

9 Fig E2 ROS production in gated and sorted B cells and myelocytic cells. A, Median fluorescence intensity of gated and sorted B cells are shown in unstimulated cells and after PMA stimuli. B, Modulation of ROS in myelocytic cells and B cells of patients with CGD and HCs. DCFDA, 2′,7′-Dichlorodihydrofluorescein diacetate. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © Terms and Conditions

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