Basophils from patients with allergic asthma show a primed phenotype W.Johan Lie, MD, Edward F. Knol, PhD, Frederik P.J. Mul, Henk M. Jansen, MD, PhD, Dirk Roos, PhD, Jaring S. van der Zee, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 104, Issue 5, Pages 1000-1007 (November 1999) DOI: 10.1016/S0091-6749(99)70081-3 Copyright © 1999 Mosby, Inc. Terms and Conditions

Fig. 1 Histamine release after addition of thapsigargin (2 μmol/L). Purified basophils (purified) as well as leukocyte suspensions obtained by lysis of erythrocytes with NH 4Cl (lysis) were incubated for 90 minutes with various concentrations of IL-3 at 37°C. The cells were then extensively washed, thapsigargin was added, and the cells were incubated for 30 minutes at 37°C. Histamine was determined in the cell-free supernatants as described in the “Methods” section. Results are means ± SD of 3 independent experiments. For each donor, the purified basophils and the total leukocytes were obtained from the same blood donation. Spontaneous histamine release of purified basophils was 3.3% ± 1.9%, and that of leukocytes was 10.4% ± 1.3% (mean ± SD). These values have not been subtracted from the results shown in the Fig. Journal of Allergy and Clinical Immunology 1999 104, 1000-1007DOI: (10.1016/S0091-6749(99)70081-3) Copyright © 1999 Mosby, Inc. Terms and Conditions

Fig. 2 Histamine release after addition of thapsigargin (2 μmol/L). After purification of basophil suspensions, thrombocytes (B+T) , monocytes (B+M) , lymphocytes (B+L) , or all of these cells from the same donor together (all) were added to the basophil fraction (B) . Leukocyte suspensions were incubated for 30 minutes with thapsigargin at 37°C. Histamine was determined in the cell-free supernatants as described in the “Methods” section. Results are means ± SD of 3 independent experiments. Spontaneous histamine release (mean ± SD) was 2.9% ± 2.8% for B , 4.6% ± 4.1% for B+T, 3.7% ± 0.8% for B+M, 5.6% ± 3.1% for B+L , and 7.4% ± 2.7% for all . These values have been substracted from the total histamine release. The resulting net histamine release is indicated in the Fig. Journal of Allergy and Clinical Immunology 1999 104, 1000-1007DOI: (10.1016/S0091-6749(99)70081-3) Copyright © 1999 Mosby, Inc. Terms and Conditions

Fig. 3 Histamine release after addition of thapsigargin (2 μmol/L). Leukocyte suspensions were incubated for 30 minutes with thapsigargin at 37°C. Histamine was determined in the cell-free supernatants as described in the “Methods” section. Spontaneous histamine release is indicated in Table II; these values have not been substracted from the results shown in the Fig. Journal of Allergy and Clinical Immunology 1999 104, 1000-1007DOI: (10.1016/S0091-6749(99)70081-3) Copyright © 1999 Mosby, Inc. Terms and Conditions