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J. Gao, K. F. Siemasko, J. Y. Niederkorn, V. L. Calder, M. Calonge, S. C. Pflugfelder, L. A. Wheeler, M. E. Stern; Corneas Exposed to Desiccating Stress Are Immunogenic and Induce T Cell Proliferation in Mice With Experimental Lacrimal Keratoconjunctivitis (LKC). Invest. Ophthalmol. Vis. Sci. 2008;49(13):192. doi: https://doi.org/.
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Environmental desiccating stress (DS) has been found to initiate CD4+ T cell-mediated lacrimal keratoconjunctivitis (LKC). Infiltrating T cells were restricted to the Lacrimal Functional Unit (LFU, cornea, conjunctiva and lacrimal gland) after adoptively transferred from mice exposed to DS to T cell deficient nude mice. We hypothesize that this ocular surface (OS) inflammation is engendered by DS-induced epithelial immunogenicity in the components of LFU.
From naive or DS mice (BALB/c), CD4+ T cells were purified from spleen/cervical lymph nodes and co-cultured with full-thickness cornea wedges for 4 days. T cell proliferation was quantified by WST assay. Co-culture was carried out in the presence or absence of anti-mouse I-A/I-E mAb (1 µg/106 CD4+ T cells) to determine if T cell response to DS cornea stimulation was due to MHC class II-bearing antigen presenting cells present in the DS cornea. This was further studied in vivo by i.p. injection to mice with the same anti-MHC II mAb on days-3, +1 and +3 of DS induction (100 µg Ab/mouse/dose). Corneas were replaced by dendritic cells (DC, differentiated from mouse bone marrow and pulsed with total corneal proteins) in the CD4+ T cells co-culture (DC:CD4+=1:10) to investigate the potential antigenic role of ocular surface in LKC. Cytokines in the culture supernatant were determined by Luminex.
CD4+ T cell proliferation was induced by corneas of DS mice in comparison to corneas of naive mice (52% increase, p=2.18E-05). DS cornea-induced CD4+ T cell proliferation was inhibited when co-culture was carried out in the presence of anti MHC class II mAbs (80% decrease, p=0.0044) as compared to the isotype control. In vivo administration of anti-MHC class II Ab in naïve BALB/c mice prior to expose to DS also inhibited CD4+ T cell proliferation (41% decrease, p=0.012). Cytokines (IL-6, TNF-a, and GM-CSF) were significantly increased (p<0.02) in the supernatants from CD4+ T cells and DS cornea co-culture, but decreased in cultures treated with anti-MHC II mAbs. When DS cornea was replaced by DCs-paused with DS corneal proteins, a similar proliferative response from CD4+ T cells was detected.
The enhanced lymphoproliferation induced by DS corneas suggests an immunogenic basis for the desiccated ocular surface in promoting T cell activation in the pathogenesis of LKC.
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