How far can we simplify in vitro diagnostics for Fagales tree pollen allergy? A study with three whole pollen extracts and purified natural and recombinant allergens

Clin Exp Allergy. 1999 Jun;29(6):848-55. doi: 10.1046/j.1365-2222.1999.00521.x.

Abstract

Background: Current diagnostic tests for Fagales tree pollen allergy are often composed of mixtures of pollen of birch, alder and hazel. Their complex composition hampers accurate standardization.

Objective: The aim of this study was to investigate whether mixtures of tree pollen extracts can be replaced by a single pollen species, and whether a single pollen species can be replaced by a limited number of purified natural or recombinant major allergens.

Methods: Sera (n = 1725) were selected on ground of a general suspicion for inhalant allergy, and tested in a RAST for birch, alder and hazel pollen. Sera with > 0.5 RU/mL for any of the three species were tested in a RAST for natural Bet v 1 and Bet v 2 as well as for recombinant versions of both allergens.

Results: Specific IgE antibodies (> 0.3 RU/mL) against birch, alder and hazel were found in 242, 298 and 292 sera, respectively. All sera with a positive RAST for alder and/or hazel and a negative RAST for birch were low-responder sera on alder and hazel, only five sera having a RAST value > 1.0 (all < 2.0). For all sera with a RAST > 0.5 RU/mL (n = 250), the mean of individual ratio's alder/birch and hazel/birch was 1.02 and 0.54, respectively. Of 223 of these sera, 63.2% had specific IgE against natural Bet v 1 and 63.7% against natural Bet v 2. When responses to both allergens were combined 93.7% were positive. The mean ratios Bet v 1 + 2/extract were 1.00, 1.04 and 2. 11 in case of birch, alder and hazel, respectively. For 211 sera the same analysis was performed with recombinant Bet v 1 and Bet v 2. Only six sera with Bet v 1-specific IgE (all < 0.5 RU/mL) were negative (< 0.3 RU/mL) on recombinant Bet v 1. For Bet v 2, 77/132 sera with specific IgE to the natural allergen did not react to the recombinant version. Twelve false-negatives had RAST values > 1.0 RU/mL. The mean of the individual recombinant/natural ratios was 0. 98 for Bet v 1 and 0.38 for Bet v 2 (P < 0.001). The mean ratio rBet v 1 + 2/birch was 0.75 with 17.5% false-negatives on the combination of recombinant allergens.

Conclusion: Reliable in vitro diagnosis is possible with a single tree pollen extract (birch or alder). The same is true for purified natural Bet v 1 and Bet v 2. A combination of recombinant molecules is slightly less efficient.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allergens / immunology
  • Antigens, Plant
  • Carbohydrates / immunology
  • Contractile Proteins*
  • Epitopes / immunology
  • Humans
  • Hypersensitivity / diagnosis*
  • Immunoglobulin E / immunology
  • Microfilament Proteins / immunology
  • Plant Extracts / immunology
  • Plant Proteins / immunology
  • Pollen / chemistry
  • Pollen / immunology*
  • Profilins
  • Recombinant Proteins
  • Trees / immunology*

Substances

  • Allergens
  • Antigens, Plant
  • Carbohydrates
  • Contractile Proteins
  • Epitopes
  • Microfilament Proteins
  • Plant Extracts
  • Plant Proteins
  • Profilins
  • Recombinant Proteins
  • Bet v 1 allergen, Betula
  • Immunoglobulin E