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John T. Schroeder, PhD, Lawrence M. Lichtenstein, MD, PhD, Ellen M

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1 IL-4 production by human basophils found in the lung following segmental allergen challenge 
John T. Schroeder, PhD, Lawrence M. Lichtenstein, MD, PhD, Ellen M. Roche, BA, HuiQing Xiao, MD, Mark C. Liu, MD  Journal of Allergy and Clinical Immunology  Volume 107, Issue 2, Pages (February 2001) DOI: /mai Copyright © 2001 Mosby, Inc. Terms and Conditions

2 Fig. 1 Percentage of Alcian blue–positive cells in bronchoalveolar lavage obtained following segmental allergen challenge before (Pre) and after (cell fractions) density centrifugation on Percoll (d = 1.066/1.079 g/mL). Values are the mean ± SEM, n = 8. Eos, Eosinophil. Journal of Allergy and Clinical Immunology  , DOI: ( /mai ) Copyright © 2001 Mosby, Inc. Terms and Conditions

3 Fig. 2 IL-4 secretion (panel A ) by bronchoalveolar lavage cell fractions following a 4-hour culture in medium (n = 5), anti-IgE (10 ng/mL) (n = 4), ragweed allergen (10 ng/mL) (n = 2), or IL-3 (10 ng/mL) (n = 3). The percentage of histamine released by the same basophil-enriched cultures is shown in panel B , which also shows the histamine released in response to 1 μmol/L N-formyl-methyonyl-leucyl-phenylalanine (n = 3). Values are the mean ± SEM, when n ≥ 3. Journal of Allergy and Clinical Immunology  , DOI: ( /mai ) Copyright © 2001 Mosby, Inc. Terms and Conditions

4 Fig. 3 IL-4 secretion (panel A ) by bronchoalveolar lavage cell fractions following 4-hour culture in medium containing ionomycin (500 ng/mL) (n = 5), phorbol myristate acetate (PMA) (10 ng/mL) (n = 4), or the combination of these stimuli (Iono/PMA) (n = 4). The percentage of histamine released by the same basophil-enriched cultures is shown in panel B . Values are the mean ± SEM. Journal of Allergy and Clinical Immunology  , DOI: ( /mai ) Copyright © 2001 Mosby, Inc. Terms and Conditions

5 Fig. 4 Detection of intracellular IL-4 in bronchoalveolar lavage (BAL) basophils. BAL basophils were enriched on double-Percoll gradients. Cells were cultured in A, medium alone, B, anti–high affinity IgE receptor (FcεRI) (22E7, 10 ng/mL), C, ionomycin (500 ng/mL), or D, N-formyl-methyonyl-leucyl-phenylalanine (fMLP) (1 μmol/L) in the presence of 2 μmol/L monensin. After a 4-hour incubation, the cultures were harvested and fixed. Two-color flow cytometry was performed to assess intracellular IL-4 in IgE-bearing cells (ie, basophils). The scatter graphs are from a representative experiment with the net percentages shown being those above the isotype/phycoerythrin (PE) control. The values shown in parentheses are the mean ± SEM from additional experiments. Journal of Allergy and Clinical Immunology  , DOI: ( /mai ) Copyright © 2001 Mosby, Inc. Terms and Conditions

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