Rhinitis, sinusitis, and upper airway diseaseAllergen specificity of IgG4-expressing B cells in patients with grass pollen allergy undergoing immunotherapy
Section snippets
Patient data
Five patients (Table I) receiving subcutaneous immunotherapy with an alum-absorbed 6-grass mix (Alutard; ALK-Abelló, Hørsholm, Denmark) were recruited at the Royal Brompton Hospital (London, United Kingdom). The study was approved by the London and City Research Ethics Committee, and blood samples were obtained after obtaining written informed consent. Levels of serum IgE and IgG4 specific for the grass pollen allergens Phl p 1, Phl p 2, Phl p 5, and Phl p 7 were determined by using ELISA (see
Molecular characteristics of IgG4
Immunoglobulin heavy- and light-chain V-region sequences were amplified from 14 individual IgG4+ memory B cells isolated from 5 patients (Table I) receiving grass pollen immunotherapy. Variable (V), diversity (D) and joining (J) IgH gene and V and J IgL gene sequences were compared with known immunoglobulin germline sequences by using IMGT/V-quest (Table II). IgH V gene use in the 14 IgG4 antibodies consisted of VH1 (n = 4), VH3 (n = 4), VH4 (n = 3), and VH5 (n = 3) families. The ratio of κ to
Discussion
We have described the use of single-cell RT-PCR for the molecular analysis and expression of IgG4 antibodies derived from 5 patients treated with grass pollen immunotherapy. Although we did not formally assess the clinical response to treatment, all participants reported an improvement in their hay fever symptoms. IgG4 is the least abundant subclass in adult human serum, ranging from 0.2 to 1 g/L compared with 5 to 12 g/L for IgG1.21 Allergen immunotherapy results in the production of high
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2017, Journal of Allergy and Clinical ImmunologyCitation Excerpt :In addition, many groups have generated anti-SEB mAbs by using either mouse hybridomas or phage display and pursued their use as therapeutic agents.42,43 In earlier work we produced a recombinant allergen (Phl p 7)–specific antibody with the native (“matched”) heavy and light chain by using single-cell PCR.34 To the best of our knowledge, the present work is the first report of recombinant anti-SAE antibodies containing matched heavy and light chains cloned by using single-cell RT-PCR.
Supported by a grant from Asthma UK (S.R.D. and H.J.G.) and a fellowship from the London Law Trust (L.K.J.). The authors acknowledge financial support from the Department of Health through a National Institute for Health Research (NIHR) comprehensive Biomedical Research Centre award to Guy's & St Thomas' NHS Foundation Trust in partnership with King's College London and King's College Hospital NHS Foundation Trust.
Disclosure of potential conflict of interest: A. J. Beavil has received research support from Asthma UK and the Medical Research Council UK. S. R. Durham has received lecture fees from ALK-Abelló and Merck, has received consultancy fees from Circassia, has received research support from Novartis and ALK-Abelló, has provided legal consultation/expert witness testimony for Merck, and is on the Immune Tolerance Network National Institute of Allergy and Infectious Diseases Steering Committee. The rest of the authors declare that they have no relevant conflicts of interest.