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IgE+ cells in the peripheral blood of atopic, nonatopic, and bee venom–hypersensitive individuals exhibit the phenotype of highly differentiated B cells 

Published byΚλυμένη Μπότσαρης Modified over 5 years ago

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Presentation on theme: "IgE+ cells in the peripheral blood of atopic, nonatopic, and bee venom–hypersensitive individuals exhibit the phenotype of highly differentiated B cells "— Presentation transcript:

1 IgE+ cells in the peripheral blood of atopic, nonatopic, and bee venom–hypersensitive individuals exhibit the phenotype of highly differentiated B cells  Peter J. Donohoe, PhDa, Robert J. Heddle, MD, PhD a, Pamela J. Sykes, PhDb, Michael Fusco, BSca, Loretta R. Flego, BSca, Heddy Zola, PhDa  Journal of Allergy and Clinical Immunology  Volume 95, Issue 2, Pages (February 1995) DOI: /S (95) Copyright © 1995 Mosby, Inc. Terms and Conditions

2 FIG. 1 Reduction of adsorbtion of immunoglobulin to lymphocytes. Cells were incubated at room temperature for 30 minutes with neat normal sheep serum. Data are presented on PBLs from an atopic donor (A) without and (B) with preincubation. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

3 FIG. 1 Reduction of adsorbtion of immunoglobulin to lymphocytes. Cells were incubated at room temperature for 30 minutes with neat normal sheep serum. Data are presented on PBLs from an atopic donor (A) without and (B) with preincubation. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

4 FIG. 2 Gating parameters used to analyze lymphocytes (R1) from 10,000 PBLs (upper left). IgE fluorescence distribution in the lymphocyte population (R1) (upper right). Real-time gating histogram showing the range for data collection (R2) and the gating (R3) used for data analysis (center left). Dot plot of expression of kappa and lambda light chains on IgE real-time–gated cells with gate R1 and the brightly fluorescent population R3 (center right). At lower left and right are dot plots showing kappa and lambda expression on the less brightly stained populations in R4 and R5. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

5 FIG. 2 Gating parameters used to analyze lymphocytes (R1) from 10,000 PBLs (upper left). IgE fluorescence distribution in the lymphocyte population (R1) (upper right). Real-time gating histogram showing the range for data collection (R2) and the gating (R3) used for data analysis (center left). Dot plot of expression of kappa and lambda light chains on IgE real-time–gated cells with gate R1 and the brightly fluorescent population R3 (center right). At lower left and right are dot plots showing kappa and lambda expression on the less brightly stained populations in R4 and R5. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

6 FIG. 2 Gating parameters used to analyze lymphocytes (R1) from 10,000 PBLs (upper left). IgE fluorescence distribution in the lymphocyte population (R1) (upper right). Real-time gating histogram showing the range for data collection (R2) and the gating (R3) used for data analysis (center left). Dot plot of expression of kappa and lambda light chains on IgE real-time–gated cells with gate R1 and the brightly fluorescent population R3 (center right). At lower left and right are dot plots showing kappa and lambda expression on the less brightly stained populations in R4 and R5. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

7 FIG. 2 Gating parameters used to analyze lymphocytes (R1) from 10,000 PBLs (upper left). IgE fluorescence distribution in the lymphocyte population (R1) (upper right). Real-time gating histogram showing the range for data collection (R2) and the gating (R3) used for data analysis (center left). Dot plot of expression of kappa and lambda light chains on IgE real-time–gated cells with gate R1 and the brightly fluorescent population R3 (center right). At lower left and right are dot plots showing kappa and lambda expression on the less brightly stained populations in R4 and R5. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

8 FIG. 2 Gating parameters used to analyze lymphocytes (R1) from 10,000 PBLs (upper left). IgE fluorescence distribution in the lymphocyte population (R1) (upper right). Real-time gating histogram showing the range for data collection (R2) and the gating (R3) used for data analysis (center left). Dot plot of expression of kappa and lambda light chains on IgE real-time–gated cells with gate R1 and the brightly fluorescent population R3 (center right). At lower left and right are dot plots showing kappa and lambda expression on the less brightly stained populations in R4 and R5. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

9 FIG. 2 Gating parameters used to analyze lymphocytes (R1) from 10,000 PBLs (upper left). IgE fluorescence distribution in the lymphocyte population (R1) (upper right). Real-time gating histogram showing the range for data collection (R2) and the gating (R3) used for data analysis (center left). Dot plot of expression of kappa and lambda light chains on IgE real-time–gated cells with gate R1 and the brightly fluorescent population R3 (center right). At lower left and right are dot plots showing kappa and lambda expression on the less brightly stained populations in R4 and R5. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

10 FIG. 3 Histograms of IgE+ cells after gating on the lymphocyte population. Examples from each of the donor groups are shown with data from 100,000 cells. IgE population was selected with the criteria outlined in the legend to Fig. 2. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

11 FIG. 3 Histograms of IgE+ cells after gating on the lymphocyte population. Examples from each of the donor groups are shown with data from 100,000 cells. IgE population was selected with the criteria outlined in the legend to Fig. 2. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

12 FIG. 3 Histograms of IgE+ cells after gating on the lymphocyte population. Examples from each of the donor groups are shown with data from 100,000 cells. IgE population was selected with the criteria outlined in the legend to Fig. 2. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

13 FIG. 4 Direct enumeration of IgE+ cells in PBLs from each of the three donor groups. Filled circles, Cell count carried out on 100,000 cells, n = 3; open circles, cell count carried out on 10,000 cells, n = 20 (see text). Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

14 FIG. 5 Histograms showing the percent expression of CD19, CD38, CD32, CD45RO, and CD45RA on cells collected by real-time gating on the IgE+ lymphocyte population. The lower right histogram shows expression of IgE on PLA-2+ population. The negative control (×63) is superimposed on each graph (unshaded area). Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

15 FIG. 5 Histograms showing the percent expression of CD19, CD38, CD32, CD45RO, and CD45RA on cells collected by real-time gating on the IgE+ lymphocyte population. The lower right histogram shows expression of IgE on PLA-2+ population. The negative control (×63) is superimposed on each graph (unshaded area). Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

16 FIG. 5 Histograms showing the percent expression of CD19, CD38, CD32, CD45RO, and CD45RA on cells collected by real-time gating on the IgE+ lymphocyte population. The lower right histogram shows expression of IgE on PLA-2+ population. The negative control (×63) is superimposed on each graph (unshaded area). Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

17 FIG. 6 Expression of CD38 on IgE + cells with three-color staining. Top, Histogram of CD3 expression on lymphocytes triple-stained with anti-IgE, CD38, and CD3. Region R2 shows the gating for collection of cells, and R3 shows the gating used to generate the dot plot in the bottom panel. Bottom, Dot plot of cells stained with OKT10 and IgE after gating out the CD3-negative population. Quadrants set for IgE+ parameters were selected by the generation of a histogram as described in the legend to Fig. 2. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

18 FIG. 6 Expression of CD38 on IgE + cells with three-color staining. Top, Histogram of CD3 expression on lymphocytes triple-stained with anti-IgE, CD38, and CD3. Region R2 shows the gating for collection of cells, and R3 shows the gating used to generate the dot plot in the bottom panel. Bottom, Dot plot of cells stained with OKT10 and IgE after gating out the CD3-negative population. Quadrants set for IgE+ parameters were selected by the generation of a histogram as described in the legend to Fig. 2. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

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