Analysis of Human T Cell Clones Reactive with Group V Grass Pollen Allergens

Müller, Wolf‐Dieter; Karamfilov, Theodor; Fahlbusch, B; Vogelsang, Harald; Jäger, Lothar

doi:10.1159/000236789

Cited by 24 publications

(24 citation statements)

References 11 publications

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“…The demonstration of cross-reactivity of Pas n 1 peptide-specific TCC suggests that the concurrent clinical reactivity to P. notatum, L. perenne and C. dactylon in BaGP-allergic subjects that we observed could be induced by T cell cross-reactivity. However, the different cross-reactivity patterns observed for Pas n 1 peptide-specific TCC in this study, and as reported previously for other grass pollen allergens [33,34,35,36], demonstrates that unique species-specific TCC also exist and that multiple T cell epitopes, unequally represented in related allergens, may be recognized by allergen-specific T cells of grass pollen-allergic patients.…”

Section: Discussionsupporting

confidence: 71%

Unique and Cross-Reactive T Cell Epitope Peptides of the Major Bahia Grass Pollen Allergen, Pas n 1

Etto

Boer

Prickett

et al. 2012

Int Arch Allergy Immunol

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Background: Bahia grass pollen (BaGP) is a major cause of allergic rhinitis. Subcutaneous allergen-specific immunotherapy is effective for grass pollen allergy, but is unsuitable for patients with moderate to severe asthma due to the risk of anaphylaxis. T cell-reactive but IgE nonreactive peptides provide a safer treatment option. This study aimed to identify and characterize dominant CD4⁺ T cell epitope peptides of the major BaGP allergen, Pas n 1. Methods: Pas n 1-specific T cell lines generated from the peripheral blood of BaGP-allergic subjects were tested for proliferative and cytokine response to overlapping 20-mer Pas n 1 peptides. Cross-reactivity to homologous peptides from Lol p 1 and Cyn d 1 of Ryegrass and Bermuda grass pollen, respectively, was assessed using Pas n 1 peptide-specific T cell clones. MHC class II restriction of Pas n 1 peptide T cell recognition was determined by HLA blocking assays and peptide IgE reactivity tested by dot blotting. Results: Three Pas n 1 peptides showed dominant T cell reactivity; 15 of 18 (83%) patients responded to one or more of these peptides. T cell clones specific for dominant Pas n 1 peptides showed evidence of species-specific T cell reactivity as well as cross-reactivity with other group 1 grass pollen allergens. The dominant Pas n 1 T cell epitope peptides showed HLA binding diversity and were non-IgE reactive. Conclusions: The immunodominant T cell-reactive Pas n 1 peptides are candidates for safe immunotherapy for individuals, including those with asthma, who are allergic to Bahia and possibly other grass pollens.

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Section: Discussionsupporting

confidence: 71%

Unique and Cross-Reactive T Cell Epitope Peptides of the Major Bahia Grass Pollen Allergen, Pas n 1

Etto

Boer

Prickett

et al. 2012

Int Arch Allergy Immunol

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“…Phl p 5–specific TCL were obtained as described by Müller et al [31]with several modifications: 1×10 5 PBMC and 10 µg Phl p 5/ml Ultraculture were seeded in a 96–well round–bottom culture plate in a total volume of 100 µl/well. After 6 days the cultures were fed by adding 100 µl Ultraculture supplemented with 25 U IL–2/ml (PBH, Hannover, Germany).…”

Section: Methodsmentioning

confidence: 99%

Reactivity of T Cells with Grass Pollen Allergen Extract and Allergoid

Kähler¹,

Stüwe²,

Cromwell³

et al. 1999

Int Arch Allergy Immunol

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Background: Successful allergen–specific immunotherapy is achieved with progressively increasing doses of allergen or allergoid. In order to gain further insight into the mechanism of action of allergoids several in vitro investigations were conducted. Methods: Peripheral blood mononuclear cells (PBMC) from grass pollen allergic and nonallergic subjects were stimulated with either grass pollen extract or allergoid and the proliferation and cytokine production (IL–5, IFN–γ) were measured. Similar investigations were performed with Phl p 5–specific T cell lines (TCL) and clones (TCC). Dendritic cells and PBMC were compared in terms of their relative efficacies as antigen–presenting cells. Results: Both allergen and allergoid induced proliferation and Th2 and Th1 cytokine synthesis by PBMC of allergic subjects, whereas PBMC of nonallergic subjects did not produce IL–5. The maximum level of IL–5 was obtained with a lower concentration than was necessary for maximal IFN–γ production. Higher stimulation doses of allergen and allergoid shifted the cytokine profiles towards a Th1 phenotype. TCL and TCC clearly showed reactivity with both allergen and allergoid when using autologous PBMC for antigen presentation, but compared with the native allergen the reactivity of the allergoid was reduced with most of the TCC. Using dendritic cells for antigen presentation a pronounced increase of stimulation of the TCC especially for the allergoids becomes obvious. Conclusion: In common with grass pollen allergen the corresponding allergoids possess a strong allergen–specific T cell–stimulating capacity. However, the degree of T cell stimulation by the allergoid seems to be dependent on the type of the antigen–presenting cell. Both, allergen and allergoid, can modulate T cell responses in a dose–dependent manner.

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“…Phi p 5-rcactive TCC were obtained by a technique described re cently [6], Briefly, peripheral blood mononuclear cells (PBMC) were stimulated in 24-well (final volume I ml) flat-bottom plates (NUNC, Roskilde, Denmark) using an optimal concentration (5 10 pg/ml) of purified [9] Phi p 5 for 7 days in RPMI-1640 medium (Gibco, Paisley, UK) supplemented with 2 mmol/l ¿-glutamine (Sigma, St. Louis, Mo., USA), 1% v/v antibiotic-antimycotic solution (Sigma), and 10% inac tivated human AB scrum (Sigma). After a further stimulation period of 7 days in the presence of interleukin 2 (IL-2; 25 lU/ml; Pharma Bio technology, Hannover, Germany), lymphoblasts were enriched on density gradient (Lymphoprep; Nycomed, Oslo, Norway).…”

Section: Isolation O F Phi P 5-reactive Tcc/proliferation Assaysmentioning

confidence: 99%

“…Initially group 5 reactivities of TCCs were assessed as published recently [6]. After collection of a sufficient panel of Pill p 5-rcactivc TCCs proliferative responses to recombinant Phi p 5 allergen and frag ments as well as to natural Lol p 1 allergen were tested as follows.…”

Section: Proliferation Assaysmentioning

confidence: 99%