Pollen from perennial rye grass (Lolium perenne), a major cause of type-I allergy worldwide, contains a complex mixture of allergenic proteins among which Lol p 1 is one of the most important. We describe the expression, purification and characterization of a recombinant Lol p 1 overproduced in Escherichia coli. The recombinant allergen, expressed in high yields and purified in milligram amounts, bound to specific IgE antibodies from human sera, induced histamine release from sensitized human basophils, and elicited rabbit antisera that recognize specifically recombinant Lol p 1 and natural Lol p 1 of pollen extract. Recombinant Lol p 1 was used to develop ImmunoCAP assays for analysis of 150 sera that were Radioallergosorbent test positive to L. perenne pollen. In 130 of them (87%) the assay detected a significant level of IgE antibodies to Lol p 1, reaching on average 37% of the level obtained with a test for IgE to the whole grass pollen extract. To map epitopes on Lol p 1, we produced three deletion mutants [des-(116-240)-Lol p 1, des-(1-88)-Lol p 1 and des-(133-189)-Lol p 1], which were efficiently expressed in bacteria. These all showed a strong reactivity with the specific rabbit IgG antibodies, but lacked most or all the allergenic properties of recombinant Lol p 1. A study of the antigenic structure of Lol p 1 was performed using the three deletion mutants and a set of 17-18-residue overlapping synthetic peptides covering the whole allergen sequence. The results indicate that human IgE and rabbit IgG antibodies bind to distinct regions of Lol p 1, and that at least some important IgE epitopes are mainly conformational. The findings suggest that recombinant allergens constitute useful reagents for further development of serological diagnosis of allergy, and that it should be possible to produce immunogenic fragments of allergenic proteins without allergenic properties.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1111/j.1432-1033.1997.00886.x | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
A comprehensive understanding of the influence and molecular mechanism of exeA on the pathogenicity in Aeromonas hydrophila.
Int J Biol Macromol
January 2025
Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Integrative Science Center of Germplasm Creation in Western China (CHONGQING) Science City & Aquaculture Engineering Technology Research Center, College of Fisheries, Southwest University, Chongqing 400715, China. Electronic address:
Aeromonas hydrophila is a serious human and animal co-pathogenic bacterium. The type II secretion system (T2SS), a key virulence factor, is vital for the secretion of exotoxins from the bacterium. exeA gene is important for the assembly of the T2SS.
View Article and Find Full Text PDFCytoBas: Precision component-resolved diagnostics for allergy using flow cytometric staining of basophils with recombinant allergen tetramers.
Allergy
October 2021
Department of Immunology and Pathology, Central Clinical School, Monash University, Melbourne, VIC, Australia.
Background: Diagnostic tests for allergy rely on detecting allergen-specific IgE. Component-resolved diagnostics incorporate multiple defined allergen components to improve the quality of diagnosis and patient care.
Objective: To develop a new approach for determining sensitization to specific allergen components that utilizes fluorescent protein tetramers for direct staining of IgE on blood basophils by flow cytometry.
Effective Platform for the Production of Recombinant Outer Membrane Vesicles in Gram-Negative Bacteria.
J Microbiol Biotechnol
May 2022
Chulabhorn International College of Medicine, Thammasat University, Pathumthani 12120, Thailand.
Bacterial outer membrane vesicles (OMVs) typically contain multiple immunogenic molecules that include antigenic proteins, making them good candidates for vaccine development. In animal models, vaccination with OMVs has been shown to confer protective immune responses against many bacterial diseases. It is possible to genetically introduce heterologous protein antigens to the bacterial host that can then be produced and relocated to reside within the OMVs by means of the host secretion mechanisms.
View Article and Find Full Text PDFInterspecific and intraspecific hybrid Epichloë species symbiotic with the North American native grass Poa alsodes.
Mycologia
May 2018
b Noble Research Institute, LLC ., 2510 Sam Noble Parkway, Ardmore , Oklahoma 73401.
The endophyte presence and diversity in natural populations of Poa alsodes were evaluated along a latitudinal transect from the southern distribution range in North Carolina to New York. Two distinct Epichloë hybrid taxa were identified from 23 populations. Each taxon could easily be distinguished by polymerase chain reaction (PCR) genotyping with primers designed to mating type genes and alkaloid biosynthesis genes that encode key pathway steps for ergot alkaloids, indole-diterpenes, lolines, and peramine.
View Article and Find Full Text PDFEvaluation of molecular basis of cross reactivity between rye and Bermuda grass pollen allergens.
Allergol Int
December 2009
ARC Centre of Excellence for Integrative Legume Research, Faculty of Land and Food Resources, University of Melbourne, Parkville, Australia.
Background: Allergenic cross reactivity between the members of the Pooids (Lolium perenne, Phleum pratense, and Poa pratensis) and Chloridoids (Cynodon dactylon and Paspalum notatum) is well established. Studies using crude extracts in the past have demonstrated limited cross reactivity between the Pooids and the Chloridoids suggesting separate diagnosis and therapy. However, little is known regarding the molecular basis for the limited cross reactivity observed between the 2 groups of grasses.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!