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Serine Protease Activity of Cur l 1 from Curvularia lunata Augments Th2 Response in Mice

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Journal of Clinical Immunology Aims and scope Submit manuscript

Abstract

Rationale

Studies with mite allergens demonstrated that proteolytic activity augments allergic airway inflammation. This knowledge is limited to few enzyme allergens.

Objective

The objective of this study is to investigate the effect of serine protease Cur l 1 from Curvularia lunata in airway inflammation/hyper-responsiveness.

Methods

Cur l 1 was purified and inactivated using a serine protease inhibitor. Balb/c mice were sensitized with enzymatically active Cur l 1 or C. lunata extract. Sensitized mice were given booster dose on day 14 with active or inactivated Cur l 1. Intranasal challenge was given on day 28, 29, and 30. Airway hyper-responsiveness was measured by plethysmography. Blood, bronchoalveolar lavage fluid (BALF), spleen, and lungs from mice were analyzed for cellular infiltration, immunoglobulins, and cytokine levels.

Results

Mice challenged with enzymatically active Cur l 1 demonstrated significantly higher airway inflammation than inactive Cur l 1 group mice (p < 0.01). There was a significant difference in serum IgE and IgG1 levels among mice immunized with active Cur l 1 and inactive Cur l 1 (p < 0.01). IL-4 and IL-5 were higher in BALF and splenocyte culture supernatant of active Cur l 1 than inactive Cur l 1 mice. Lung histology revealed increased eosinophil infiltration, goblet cell hyperplasia and mucus secretion in active group.

Conclusion

Proteolytic activity of Cur l 1 plays an important role in airway inflammation and the inactivated Cur l 1 has potential to be explored for immunotherapy.

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Abbreviations

BAEE:

α-benzoyl-l-arginine-ethyl ester hydrochloride

BALF:

bronchoalveolar lavage fluid

BSA:

bovine serum albumin

CBB:

Coomassie brilliant blue

EDTA:

ethylenediamine tetraacetic acid

EPO:

eosinophil peroxidase

FCS:

fetal calf serum

HE:

hematoxylin–eosin

HTAB:

hexadecyl trimethyl ammonium bromide

MTT:

3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide

OD:

optical density

OPD:

o-phenylenediamine

OVA:

ovalbumin

PAR:

protease activated receptors

PAS:

periodic acid-Schiff

PHA:

phytohemagglutinin

PMSF:

phenyl methyl sulphonyl fluoride

RPMI:

Roswell Park Memorial Institute

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Acknowledgments

One of the authors (Prabhanshu Tripathi) is a ‘Senior Research Fellow’ from Council of Scientific and Industrial Research, Delhi. The authors are thankful to CSIR task force project for financial support to this study.

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Authors and Affiliations

  1. Allergy and Immunology Section, Institute of Genomics and Integrative Biology, D.U. North Campus, Mall Road, Delhi, 110 007, India

    Prabhanshu Tripathi, Neetu Kukreja, B. P. Singh & Naveen Arora

  2. Department of Biotechnology, University of Pune, Ganeshkhind, Pune, 411 007, India

    Prabhanshu Tripathi

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  1. Prabhanshu Tripathi
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Correspondence to Naveen Arora.

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Tripathi, P., Kukreja, N., Singh, B.P. et al. Serine Protease Activity of Cur l 1 from Curvularia lunata Augments Th2 Response in Mice. J Clin Immunol 29, 292–302 (2009). https://doi.org/10.1007/s10875-008-9261-9

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  • DOI: https://doi.org/10.1007/s10875-008-9261-9

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