Summary:Two cases of GVHD with severe dry eyes are reported where conventional therapy failed to control ocular signs and symptoms. Autologous serum tears, however, resulted in a beneficial clinical effect with marked attenuation of the symptoms. This therapy proved to be safe during 10 months of treatment. Bone Marrow Transplantation (2000) 25, 1101-1103. Keywords: GVHD; dry eye; autologous serum tears; allogeneic PBPC transplantation The treatment of ocular disorders with biological fluids has long been advocated. 1 More recently, autologous serum (AS) tears have been indicated for the treatment of dry eye in Sjögren's syndrome. The rationale for this is based upon the fact that vitamins and growth factors present in tears are also present in serum. 2,3 In this report, we describe the treatment of dry eyes with AS tears in two adult women with extensive chronic graft-versus-host disease (GVHD) following allogeneic peripheral blood progenitor cell transplantation (PBPCT). Case 1A 44-year-old woman underwent PBPCT for chronic myeloid leukemia (CML) in the first chronic phase and developed sicca syndrome along with extensive chronic GVHD. Her previous ophthalmic history included only spectacles for myopia. The GVHD was treated with cyclosporin A and prednisone systemically and with commercial artificial tears, including preservative-free tears for 6 months, with minimal improvement in the ocular symptoms which included the sensation of presence of a foreign body, and redness.Examination revealed a corrected visual acuity of 20/25 for both eyes, conjunctival hyperemia and severe punctate At this point, we introduced AS tears, prepared as previously described, 2 with instructions to keep the bottles in a refrigerator and apply the drops several times a day. Four weeks later, the patient returned and reported an improvement in her symptoms from the day she started using the preparation. Compared to her previous tear substitute, the effect of AS tears lasted longer, allowing her to apply fewer drops throughout the day and also provided considerably more comfort in the morning, which was the most critical part of the day. Examination of her eyes showed a dramatic improvement in punctate staining, a reduction in hyperemia

Alkali burns to the cornea are among the most devastating injuries to the eye. The purpose of this study was to evaluate the effects of dexamethasone (Dex) or doxycycline (Doxy) on protease activity and corneal complications in a combined model (CM) of alkali burn and dry eye. C57BL/6 mice were subjected to the CM for 2 or 5 days (D). Mice were topically treated either with Dex (0.1%), Dox (0.025%) or vehicle QID and observed daily for appearance of corneal perforation. Quantitative real time PCR was performed to measure expression of inflammation cytokines and matrix metalloproteinases (MMP) in whole cornea lysates. No perforations were observed in the Dex-treated corneas. All wounds in Doxy-treated corneas were closed 2D post-injury, and they had significantly lower corneal opacity scores at days 4 and 5 post-injury compared to BSS treatment. Dex-treated corneas had the lowest corneal opacity scores. Dex treatment significantly decreased expression of IL-1β, IL-6, MMPs -1, -9, -13, and TIMP-1 after 2 days but increased levels of MMP-8, while Doxy treatment significantly decreased IL-1β, IL-6, MMP-8, and -9, compared to vehicle. Decreased MMP -1, -9 and -13 immunoreactivity and gelatinolytic activity were seen in corneas treated with Doxy and Dex compared to vehicle. Increased neutrophil infiltration and myeloperoxidase activity was noted in the vehicle group compared to Dex 2 days post-injury. These findings demonstrate that early initiation of anti-inflammatory therapy is very efficacious in preserving corneal clarity and facilitating wound healing, while modulating MMP production and suppressing neutrophil infiltration.

IntroductionThe lacrimal gland (LG) of the CD25-/- model of Sjögren’s syndrome (SS) has high interleukin (IL)-17, IL-13 and interferon-gamma (IFN-γ) cytokines. The specific contribution of these cytokines to the onset and severity of dacryoadenitis in the CD25-/- mice has not been evaluated.MethodsCD25−/−IL-17A−/−, CD25−/−IL-17−/−IFN-γ−/− and CD25−/−IFN-γ−/− were used at 4, 8, 12, 16 weeks (W). Total lymphocytic infiltration was evaluated by histology and characterized by flow cytometry. Epidermal growth factor (EGF) concentration was measured in tears. Immunofluorescent staining evaluated expression of IFN-γ receptor (IFN-γR) and apoptosis. Real-time PCR evaluated inflammatory and T cell-related cytokines expression in LG. Caspase-3, -8, -9 activities was assayed in LG lysates. T helper cytokines were measured in serum by Luminex assay.ResultsThe greatest total LG infiltration at 8 W was seen in CD25−/−IL-17A−/− (95%), followed by CD25−/− (71%) and IL-17−/− (12%). Tear EGF concentration was in normal range in CD25−/− at 4 W and in very low levels in both CD25−/− and CD25−/−IL-17A−/−. CD25−/− had high levels of inflammatory cytokines transcripts in LG compared to IL-17−/− mice; however, CD25−/−IL-17A−/− had even higher IL-1β, IFN-γR, caspase-3, -8, -9 mRNA levels, greater immunoreactivity to IFN-γR in LG acini, greater number of apoptotic+ cells and greater caspases activities in the LG at 8 W. CD25−/−IL-17A−/− had lower IL-13 concentration and lower IL-13/IFN-γ ratio compared to CD25−/− in serum. CD25−/−IFN-γ−/− had lower number of apoptotic+ cells and decreased caspase-3 expression in LG. CD25−/−IL-17−/−IFN-γ−/− had lower total lymphocytic cell infiltration at 8 W (48%), CD4+T cell infiltration and expression of IFN-γR and apoptotic+ cells in the LG and increased tear EGF concentration in tears.ConclusionsIFN-γ is critical for LG destruction and secretory dysfunction in the CD25−/− model of SS. Altered balance between IFN-γ and IL-13 in the CD25−/−IL-17A−/− mice accelerates LG destruction by increasing glandular apoptosis and facilitating apoptosis through increased expression of IFN-γR by glandular epithelium and activation of caspases. Targeting both IFN-γ and IL-17 may be beneficial for treating the LG inflammation in SS.

IntroductionTo investigate the role of interferon-gamma (IFN-γ) in the onset and severity of dacryoadenitis in the CD25 knockout (KO) mouse model of Sjögren Syndrome.MethodsCD25/IFN-γ double KO (γDKO) mice were created by crossbreeding CD25KO and IFN-γKO mice. Mice were used at 8, 12, and 16 weeks. Lacrimal gland (LG) infiltrating lymphocytes were characterized with flow cytometry. Tear epidermal growth factor (EGF) concentration was measured with enzyme-linked immunosorbent assay (ELISA). Quantitative polymerase chain reaction (PCR) evaluated T-cell-related cytokines in LGs. Serum autoantibodies against M3R in LG lysates were detected with Western blot.ResultsγDKO LG showed lower lymphocytic infiltration at 8 weeks than in the CD25KO parental strain (˜20% versus ˜60%, respectively), which increased to CD25KO levels at 16 weeks. Flow-cytometry analysis showed an increase in CD4+ and CD8+ T cells with aging in γDKO LG, similar to that in CD25KO. γDKO had lower levels of interleukin (IL)-17A, transforming growth-factor (TGF)-β1, IL-21, and CCL20, and higher IL-1β and IL-13 mRNA transcripts in the LG than in the parental CD25KO strain. Autoantibodies to M3R were observed in both strains and significantly increased with aging in both strains. CD25KO mice had very low tear EGF concentrations at all ages, whereas the ear EGF concentration in γDKO mice significantly decreased with aging and inversely correlated with the presence of M3R autoantibodies and the degree of LG CD4 and CD8+ T-cell infiltration.ConclusionsThe deletion of IFN-γ in the CD25KO mice strain delays glandular destruction and preserves glandular function. M3R autoantibodies increased with aging in both the γDKO and the CD25KO strains. The decrease in LG function in γDKO correlated with the degree of T-cell infiltration and the presence of M3R autoantibodies.