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Am. J. Respir. Cell Mol. Biol., Volume 19, Number 3, September, 1998 507-512

Combined Nasal Challenge with Diesel Exhaust Particles and Allergen Induces In Vivo IgE Isotype Switching

Shigeharu Fujieda, David Diaz-Sanchez, and Andrew Saxon

The Hart and Louise Lyon Laboratory, Division of Clinical Immunology/Allergy, Department of Medicine, The Jonsson Comprehensive Cancer Center Institute, and The Molecular Biology Institute, University of California, Los Angeles School of Medicine, Los Angeles, California; and Department of Otorhinolaryngology, Fukui Medical University, Japan

In this study we undertook to provide evidence for local in vivo isotype switching to IgE following nasal challenges. Detection of deleted switch circular DNA (switch circles) by a novel nested polymerase chain reaction-based approach was employed as definitive molecular evidence of Ig isotype switching. Nasal challenge in humans with diesel exhaust particles (DEP) plus ragweed antigen has been shown to enhance local IgE production, stimulate local cytokine production, and markedly increase mucosal IgE antibody to ragweed. Four days after combined intranasal DEP plus ragweed challenge, we detected and characterized clones of deleted switch circular DNA (Svarepsilon /Sµ) representing switching from µ to varepsilon  from nasal lavage cells. No switch circular DNA was detected in nasal lavage cells following challenge with DEP alone nor with ragweed allergen alone. These results indicate that the combination of mucosal stimulation with DEP and ragweed allergen is capable of driving in vivo isotype switching to IgE in humans with ragweed allergy. These results are the first direct demonstration of in vivo IgE isotype switching in humans.




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