scholarly journals A sensitive fluorescent assay for measuring the cysteine protease activity of Der p 1, a major allergen from the dust mite Dermatophagoides pteronyssinus

1998 ◽  
Vol 51 (4) ◽  
pp. 222-224 ◽  
Author(s):  
O. Schulz ◽  
H. F. Sewell ◽  
F. Shakib
Keyword(s):  
Cysteine Protease ◽  
Protease Activity ◽  
Major Allergen ◽  
Dermatophagoides Pteronyssinus ◽  
Fluorescent Assay ◽  
Cysteine Protease Activity ◽  
Dust Mite ◽  
Der P 1

scholarly journals The Cysteine Protease Activity of the Major Dust Mite Allergen Der P 1 Selectively Enhances the Immunoglobulin E Antibody Response

1999 ◽  
Vol 190 (12) ◽  
pp. 1897-1902 ◽  
Author(s):  
Lucy Gough ◽  
Oliver Schulz ◽  
Herb F. Sewell ◽  
Farouk Shakib
Keyword(s):  
Cysteine Protease ◽  
Protease Activity ◽  
Immunoglobulin E ◽  
Specific Ige ◽  
Cysteine Protease Inhibitor ◽  
Cysteine Protease Activity ◽  
Dust Mite ◽  
Ige Synthesis ◽  
Der P 1

The house dust mite Dermatophagoides pteronyssinus allergen Der p 1 is the most immunodominant allergen involved in the expression of dust mite–specific immunoglobulin (Ig)E–mediated hypersensitivity. The reason for this potent IgE-eliciting property of Der p 1 remains unknown, but there is mounting in vitro evidence linking the allergenicity of Der p 1 to its cysteine protease activity. Here we demonstrate for the first time that immunization of mice with proteolytically active Der p 1 results in a significant enhancement in total IgE and Der p 1–specific IgE synthesis compared with animals immunized with Der p 1 that was irreversibly blocked with the cysteine protease inhibitor E-64. We conclude that the proteolytic activity of Der p 1 is a major contributor to its allergenicity.

scholarly journals An Engineered Hybrid Protein from Dermatophagoides pteronyssinus Allergens Shows Hypoallergenicity

2019 ◽  
Vol 20 (12) ◽  
pp. 3025 ◽  
Author(s):  
Dalgys Martínez ◽  
Marlon Munera ◽  
Jose Fernando Cantillo ◽  
Judith Wortmann ◽  
Josefina Zakzuk ◽  
...  
Keyword(s):  
Cysteine Protease ◽  
Protease Activity ◽  
Dermatophagoides Pteronyssinus ◽  
Basophil Activation ◽  
Allergen Extract ◽  
Hybrid Protein ◽  
Dot Blot ◽  
Cysteine Protease Activity ◽  
Der P 1 ◽  
Der P 2

The house dust mite (HDM) Dermatophagoides pteronyssinus is an important risk factor for asthma and rhinitis. Allergen specific immunotherapy that is based on recombinant proteins has been proposed for the safer and more efficient treatment of allergic diseases. The aim of this study was to design and obtain a hybrid protein (DPx4) containing antigenic regions of allergens Der p 1, Der p 2, Der p 7, and Der p 10 from this mite. DPx4 was produced in Escherichia coli and its folding was determined by circular dichroism. Non-denaturing dot-blot, ELISA, basophil activation test, dot blot with monoclonal antibodies, ELISA inhibition, and cysteine protease activity assays were performed. Mice that were immunized with DPx4 were also analyzed. We found that DPx4 had no cysteine protease activity and it showed significantly lower IgE reactivity than Der p 1, Der p 2, and D. pteronyssinus extract. DPx4 induced lower basophil activation than Der p 2 and the allergen extract. Immunized mice produced IgG antibodies that inhibited the binding of allergic patient’s IgE to the allergen extract and induced comparatively higher levels of IL-10 than the extract in peripheral blood mononuclear cells (PBMC) culture. These results suggest that DPx4 has immunological properties that are useful for the development of a mite allergy vaccine.

scholarly journals Human glutathione-S-transferase pi potentiates the cysteine-protease activity of the Der p 1 allergen from house dust mite through a cysteine redox mechanism

Redox Biology ◽  
2019 ◽  
Vol 26 ◽  
pp. 101256 ◽  
Author(s):  
Juan Carlos López-Rodríguez ◽  
Juliana Manosalva ◽  
J. Daniel Cabrera-García ◽  
María M. Escribese ◽  
Mayte Villalba ◽  
...  
Keyword(s):  
House Dust ◽  
House Dust Mite ◽  
Cysteine Protease ◽  
Protease Activity ◽  
Glutathione S Transferase ◽  
Redox Mechanism ◽  
Cysteine Protease Activity ◽  
Dust Mite ◽  
Der P 1

Recombinant Der p 1 and Der f 1 exhibit cysteine protease activity but no serine protease activity

2005 ◽  
Vol 328 (4) ◽  
pp. 944-952 ◽  
Author(s):  
Toshiro Takai ◽  
Takeshi Kato ◽  
Yasuhisa Sakata ◽  
Hiroshi Yasueda ◽  
Kenji Izuhara ◽  
...  
Keyword(s):  
Serine Protease ◽  
Cysteine Protease ◽  
Protease Activity ◽  
Cysteine Protease Activity ◽  
Serine Protease Activity ◽  
Der P 1

scholarly journals Proteolytic Cleavage of CD25, the α Subunit of the Human T Cell Interleukin 2 Receptor, by Der p 1, a Major Mite Allergen with Cysteine Protease Activity

1998 ◽  
Vol 187 (2) ◽  
pp. 271-275 ◽  
Author(s):  
Oliver Schulz ◽  
Herb F. Sewell ◽  
Farouk Shakib
Keyword(s):  
Cysteine Protease ◽  
Protease Activity ◽  
Human Peripheral Blood ◽  
Interleukin 2 ◽  
Interferon Γ ◽  
Th2 Cells ◽  
Α Subunit ◽  
Cysteine Protease Activity ◽  
Human T Cell ◽  
Der P 1

Recent reports have indicated that the cysteine protease activity of Der p 1 may play a significant role in its ability to elicit IgE antibody responses, mainly through cleavage of membrane CD23 on B cells and interleukin (IL)-4 synthesis and secretion from mast cells and basophils. Here we demonstrate for the first time that Der p 1 also cleaves the α subunit of the IL-2 receptor (IL-2R or CD25) from the surface of human peripheral blood T cells and, as a result, these cells show markedly diminished proliferation and interferon γ secretion in response to potent stimulation by anti-CD3 antibody. Given that the IL-2R is pivotal for the propagation of Th1 cells, its cleavage by Der p 1 may consequently bias the immune response towards Th2 cells, thereby creating an allergic microenvironment.

scholarly journals MONOMERIC ALLERGOID FROM HOUSE DUST MITE DERMATOPHAGOIDES PTERONYSSINUS: IMMUNOLOGICAL PROPERTIES

2016 ◽  
Vol 13 (4-5) ◽  
pp. 29-36
Author(s):  
A A Babakhin ◽  
A A Laskin ◽  
V V Smirnov ◽  
S M Andreev ◽  
K K Babievsky ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
House Dust ◽  
House Dust Mite ◽  
Tertiary Structure ◽  
Major Allergen ◽  
Dermatophagoides Pteronyssinus ◽  
Ige Binding ◽  
Sds Page ◽  
Dust Mite ◽  
Der P 1

Background. The purpose of this study was to characterize the immunologic properties of the monomeric allergoid obtained from house dust mite Dermatophagoides pteronyssinus (D. pteronyssinus) extract. Methods. To obtain monomeric allergoid the extract from D. pteronyssinus (D1) was chemically modified by succinilation and then the degree of modification and total protein content were determined. Analysis of proteins in native D1 or modified sD1 was performed by SDS-PAGE. Determination of the major allergen Der p 1 in the native extract D1 was done by mass-spectrometry method. For measurement of tertiary structure alterations of protein molecules in D1 or recombinant Der p 1 (rDer p 1) after their modification by succinilation the circular dichroism (CD) method was applied. Allergenicity of sD1 was estimated by inhibition of specific IgE binding in ELISA and chemiluminescent analysis using Phadia ImmunoCAP as well as by detection of histamine released from leukocytes of whole blood of Der p sensitized patients after its incubation with D1 or sD1. To evaluate immunogenicity of D1 and sD1 BALB/c mice were i.p. immunized 4 times in 3 week interval with D1 or sD1 with or without adjuvant Al(OH),. The levels of anti-Der p IgE, IgG1 and IgG2a in pulled mice sera were determined by ELISA. Results. Succinilation of D1 extract led to «unfolding» of allergen molecules in modified sD1. The degree ofmodification was 98,9%. Results of SDS-PAGE demonstrated that native D1 and succinilated sD1 extract showed bands at the same molecular mass confirming that succinilation does not influence the size of modified molecule which thus maintains the monomeric nature of the sD1. Mass-spectrometry analysis revealed the presence of major allergen Der p 1 in the native D1 extract. CD method demonstrated the alteration of tertiary structure of succinilated recombinant rDer p 1 that may be extrapolated on the other allergenic proteins in the sD1 extract. Allergenicity of sD1 was significantly reduced in compare to D1 that was confirmed by inhibition of IgE-binding and histamine release from leukocytes of whole blood, Immunization of BALB/c mice with sD1 (with or without Al(OH)3) induced anti-Der p IgE-response which was substantially lower than that of D1 induced. At the same time anti-Der p IgG1 and IgG2a responses after immunization with sD1 (with or without Al(OH)3) were significantly higher than that ofD1 induced. Conclusion. Modification of the extract from house dust mite D. pteronyssinus by succinilation leads to formation of monomeric allergoid possesses low allergenicity and preserved (or even increased) immunogenicity that may a new approach for creation of allergy vaccines for safe and effective allergen-specific immunotherapy.

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