Analysis of Immunoglobulin (Ig) Isotype Diversity and Igm/D Memory in the Response to Phenyl-Oxazolone

doi:10.1084/jem.191.12.2209

Published online 19 June 2000.

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© The Rockefeller University Press, 0022-1007/2000/6/2209/ $5.00
The Journal of Experimental Medicine, Volume 191, Number 12, June 19, 2000 2209-2220

Original Article

Analysis of Immunoglobulin (Ig) Isotype Diversity and Igm/D Memory in the Response to Phenyl-Oxazolone

Harry White^a and David Gray^b

^a Department of Molecular Immunology, Institute of Child Health, London WC1N 1EH, United Kingdom
^b Institute of Cell, Animal and Population Biology, University of Edinburgh, Edinburgh EH9 3JT, United Kingdom
Dept. of Molecular Immunology, Institute of Child Health, 30 Guilford St., London WC1N 1EH, UK.44-171-831-436644-171-905-2210

h.white{at}ich.ucl.ac.uk

The distribution of immunoglobulin (Ig) isotypes within specificB cell clones in vivo after immunization is not well defined.Using an IgV_H/CDR3- and isotype-specific reverse transcriptionpolymerase chain reaction method, we have carried out a surveyof the diversification of the isotype in a splenic responseto phenyl-oxazolone (phOx) on a chicken serum albumin carrier.The phOx-specific V_H (V_HOx-1 with specific CDR3 motif) is associatedwith all of the heavy chains (µ, ${delta}$ , ${alpha}$ , ${gamma}$ , and ${varepsilon}$ ) after simpleimmunization with antigen in alum. The kinetics of expressionof each isotype are distinct and reproducible. Focusing mainlyon the expression of secretory Ig transcripts, IgM, IgG1, andIgE are found after priming, whereas IgD and IgA appear afterboosting. Secretory IgD transcripts are found reproducibly atmoderate levels and may, therefore, contribute significantlyto the secreted Ig response in mice. Most crucially, we findenhanced levels of secretory IgM/V_HOx-1 transcripts (with ‘phOx-specific’CDR3) after boosting, strongly indicating the existence of IgMmemory cells that give rise to an enhanced specific IgM secretionin the secondary response.

Key Words: B cell memory • immunoglobulin isotype • repertoire • IgD • oxazolone

Abbreviations used in this paper: CSA, chicken serum albumin;mem, membrane; phOx, phenyl-oxazolone; RT, reverse transcription;sec, secretory.

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