Immunoglobulin E antibodies of atopic individuals exhibit a broad usage of V_H‐gene families

Abstract

SUMMARYThe term`atopy' describes the genetically determined tendency to mount immunoglobulin E (IgE) antibody responses against per se harmless antigens (allergens). In this study we investigated the usage of V H families in the formation of IgE antibodies in 10 patients suffering from mucosal and/or skin manifestations of atopy. IgE antibody reactivities to exogenous allergen sources as well as to autoallergens were determined and, by immunoabsorption, it was demonstrated that allergenspeci®c IgE accounted for most of the total serum IgE levels in these patients. Using primers with speci®city for the V H 1±6 gene families and a primer speci®c for the ®rst constant region of human IgE, cDNAs coding for IgE heavy chain fragments were ampli®ed using the reverse transcription± polymerase chain reaction (RT±PCR) from peripheral blood lymphocytes of the 10 atopic individuals. Hybridization of the heavy chain-encoding cDNAs with an IgE-speci®c internal oligonucleotide probe revealed a broad usage of all V H -gene families in the atopic individuals. The spectrum of V H families used in a given atopic individual was neither associated with the type or severity of clinical symptoms nor with the number of allergens recognized. The fact that allergenspeci®c IgE antibodies in atopic individuals originate from a broad variety of B cells thus re¯ects the activation of multiple B-cell clones during allergen sensitization. This ®nding should be borne in mind if therapeutic strategies for Type I allergy are considered that aim at a clonal elimination of allergen-speci®c B cells.

The Human IgE-encoding Transcriptome to Assess Antibody Repertoires and Repertoire Evolution

“…52 In conclusion, H-EPSILON repertoires in general represent a normal part of the total immunoglobulin repertoire. Thus, the evidence from our study supports the notion of extensive diversity in H-EPSILON repertoires [27][28][29] over the idea suggesting that H-EPSILON repertoires are highly restricted with respect to V gene usage. [23][24][25][26] In fact, there is nothing particular about the composition of H-EPSILON repertoires as identified during active disease and B cells are in this phase not in general preferentially selected into the H-EPSILON-producing population based on e.g.…”

“…Furthermore, the development of combinatorial library approaches allows us to link given members of a repertoire with antigen specificity. Many studies [23][24][25][26][27][28][29][30][31][32][33][34][35] have addressed aspects of one or the other of these approaches in relation to IgE repertoires but not tried to combine them into a unit that investigates both aspects. We have now validated an approach comprised of methods to carry out random sequencing of the immunoglobulin heavy epsilon chain (H-EPSILON)-encoding transcriptome and isolation of allergen-specific sequences by combinatorial antibody library technology.…”

Characterization of VHε gene expressed in PBL from children with atopic diseases: detection of homologous VH1–69 derived transcripts from three unrelated patients

“…The frequently found germline segments were V H 1, 1-46, 1-69; V H 3, 3-11, 3-21; V H 4, 4-39, and 4-59 and all of these segments were also detected in previous studies [14,15]. Interestingly, V H 1-69, and V H 3-11 derived transcripts (V H 1-69; NK-2, SS-3, KY-21, V H 3-11; NK-1, 3, 4, 5, 6, SS-1, 2, KY-14, 20) were identified in all the patients, which had similar DJ sequences but carried different point mutations at CDR (complementarity-determining region) 1, 2 and FR (framework region).…”

“…With a new approach using V H family specific primers, the IgE V H repertoire of patients with allergic disease have been analyzed [12][13][14][15]. The dominance of V H 5 genes from asthma patients was observed [12,13], on the other hand, no bias for the usage of IgE V H was detected in AD patients [14].…”

Analysis of IgE Antibodies from a Patient with Atopic Dermatitis: Biased V Gene Usage and Evidence for Polyreactive IgE Heavy Chain Complementarity-Determining Region 3

“…This is not the first report of the use of the V H 6 gene family in the IgE repertoire. IGHV6-1 has been identified in IgE rearrangements among 9 of 10 patients suffering from a variety of allergic diseases, including atopic dermatitis (54). The other small H chain V gene family (V H 5) has also been observed encoding IgE Abs (22)(23)(24)27).…”