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Mast-cell activation augments the late phase reaction in experimental immune-mediated blepharoconjunctivitis

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Abstract

Background

How the early phase allergic reaction affects the late phase reaction remains unclear. We examined this issue with an experimental model of allergic conjunctivitis that permits the two reactions to be disconnected from each other.

Methods

Experimental immune-mediated blepharoconjunctivitis (EC) was initiated in Brown Norway rats by transferring ovalbumin (OVA)-specific T cells and then challenging with OVA-containing eye drops. To induce early phase reaction, a mast-cell activator, C48/80, was challenged together with or without OVA. Rats were evaluated clinically and eyes were harvested for histologic examination and for evaluation of chemokine expression by reverse-transcriptase PCR.

Results

The rats challenged with OVA alone developed the T-cell-mediated late phase reaction histologically, but not clinically, in the absence of early phase reaction. While rats challenged with C48/80 with or without OVA exhibited clinical signs of the early phase reaction, the clinical late phase reaction was observed only in the OVA+C48/80 group. Eosinophilic infiltration into the conjunctiva during the late phase reaction of the OVA+C48/80 group markedly exceeded that of rats challenged with either OVA or C48/80 alone. RANTES (regulated on activation, normal T-cell expressed and secreted), an eosinophil attractant, was expressed both in the OVA+C48/80 and OVA groups, while eotaxin was expressed at equivalent levels in all three groups.

Conclusion

The mast-cell-mediated early phase reaction potentiates the T-cell-mediated late phase reaction, and RANTES is involved in eosinophilic infiltration induced by antigen-specific T cells. Other molecules induced by allergen-specific T cells activated in an as yet unknown manner by the mast cells may be responsible for the infiltration of eosinophils.

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Fig. 1A–C.
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Fig. 3A–C.
Fig. 4A–C.

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Abbreviations

Ab:

antibody

AC:

allergic conjunctivitis

BN:

Brown Norway

C48/80:

compound 48/80

CFA:

complete Freund's adjuvant

cpm:

counts per minute

dNTPs:

deoxyribonucleoside triphosphates

EC:

experimental immune-mediated blepharoconjunctivitis

ELISA:

enzyme-linked immunosorbent assay

FCS:

fetal calf serum

Ig:

immunoglobulin

IL:

interleukin

JRU:

Japan reference units

2-ME:

2-mercaptoethanol

OVA:

ovalbumin

RANTES:

regulated on activation, normal T-cell expressed and secreted

RT-PCR:

reverse-transcriptase polymerase chain reaction

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Acknowledgements

This work was partially supported by a Grant-in-Aid for Encouragement of Young Scientists from the Ministry of Education, Science and Culture, Japan (to A.F.), a Kochi Shinyokinko Anshintomonokai Award and a President Fund of Kochi Medical School Hospital.

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Correspondence to Atsuki Fukushima.

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Ozaki, A., Fukushima, A., Fukata, K. et al. Mast-cell activation augments the late phase reaction in experimental immune-mediated blepharoconjunctivitis. Graefe's Arch Clin Exp Ophthalmol 241, 394–402 (2003). https://doi.org/10.1007/s00417-003-0641-9

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  • DOI: https://doi.org/10.1007/s00417-003-0641-9

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