Expression of TNF inhibitor gene in the lacrimal gland promotes recovery of tear production and tear stability and reduced immunopathology in rabbits with induced autoimmune dacryoadenitis

BACKGROUND: The most common cause of ocular morbidity in developed countries is dry eye, many cases of which are due to lacrimal insufficiency. Dry eye affects approximately 10 million in the United States., most of whom are women. In the U.S. alone, an estimated 2 million Sjögren's syndrome patients have dysfunctional lacrimal glands and severe dry eye, and there is no satisfactory treatment. These patients would benefit if their lacrimal tissue function could be restored. METHODS: The effect of adenovirus-mediated transfer of tumor necrosis factor (TNF)-α inhibitor gene on induced autoimmune dacryoadenitis was evaluated in a rabbit model. Soluble transgene protein was detected in tears by ELISA for 7 days following transduction. RESULTS: Two weeks after induction of disease with activated lymphocytes, tear production, as determined by Schirmer testing, was reduced by about 40%, while tear film stability, as measured by tear breakup time (BUT), declined by 43%. Adenovirus-mediated gene therapy using AdTNFRp55-Ig given 2 weeks after disease induction, resulted in the return of tear production to normal levels by week 4. In the treated disease group, tear BUT improved significantly by week 4. Rose bengal scores, an indicator of corneal surface defects, increased after disease induction and declined after gene therapy. In the lacrimal gland, the CD4 to CD8 T cell ratio was 4:1 in the disease group compared to 1:2 in the treated group. Infiltration of T cells and CD18+ cells was reduced approximately 50% after gene therapy. CONCLUSION: We concluded that therapeutic levels of soluble TNF inhibitor were achieved in the lacrimal gland and on the corneal surface. Anti-inflammatory cytokine gene expression might offer a potential therapeutic modality for the treatment of autoimmune dacryoadenitis, once suitable vectors become available

Interacting Influences of Pregnancy and Corneal Injury on Rabbit Lacrimal Gland Immunoarchitecture and Function

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Corneal topographic changes in premenopausal and postmenopausal women

<p>Abstract</p> <p>Background</p> <p>To asses the effect of menopause on the corneal curvature changes using corneal computerized videokeratography (CVK) in premenopausal and postmenopausal healthy women.</p> <p>Methods</p> <p>Thirty-six postmenopausal women with mean ages of 49.2 (range 39 to 57) were enrolled in this randomized, prospective study, comparing with 26 healthy controls with mean ages of 38.5 +/- 4.9 (range 32 to 49). Subjects were determined to be postmenopausal, by the Gynecology and Obstetrics Department, based on blood Follicular Stimulating Hormone (FSH), Luteinizing Hormone (LH), Estradiol, Progesterone levels and clinical complaints. Complete ophthalmic examination and CVK using Haag-Streit System was performed in both premenopausal and postmenopausal women.</p> <p>Results</p> <p>Mean horizontal curvature and vertical curvature of central corneal power in premenopausal women were 43.5 +/- 1.25 Diopter (D), and 44.1 +/- 1.53 D. Mean horizontal curvature and vertical curvature of central corneal power in postmenopausal women were 43.9 +/- 1.4 D, and 44.6 +/- 1.3 D. The mean keratometric astigmatisms of premenopausal and postmenopausal women were 0.81 +/- 0.57 D (4–179 degrees), 0.74 degrees +/- 0.5 D (1–180 degrees) respectively. No significant corneal curvature changes were detected between premenopausal and postmenopausal groups (P > 0.05). On the other hand, we only found negative but significant correlation between horizontal corneal curvature and estrogen level of postmenopausal women (r = -0.346, p = 0.038).</p> <p>Conclusion</p> <p>Menopause is physiologic process and may also affect corneal topographic changes. In postmenopausal women, corneal steeping was observed minimally compared to premenopausal women. The results suggest that changes in estrogen level of women with menopause are associated with slightly alteration of horizontal curvature of cornea.</p

Nitric oxide differentially regulates renal ATP-binding cassette transporters during endotoxemia

Nitric oxide (NO) is an important regulator of renal transport processes. In the present study, we investigated the role of NO, produced by inducible NO synthase (iNOS), in the regulation of renal ATP-binding cassette (ABC) transporters in vivo during endotoxemia. Wistar–Hannover rats were injected with lipopolysaccharide (LPS+) alone or in combination with the iNOS inhibitor, aminoguanidine. Controls received detoxified LPS (LPS−). After LPS+, proximal tubular damage and a reduction in renal function were observed. Furthermore, iNOS mRNA and protein, and the amount of NO metabolites in plasma and urine, increased compared to the LPS− group. Coadministration with aminoguanidine resulted in an attenuation of iNOS induction and reduction of renal damage. Gene expression of 20 ABC transporters was determined. After LPS+, a clear up-regulation in Abca1, Abcb1/P-glycoprotein (P-gp), Abcb11/bile salt export pump (Bsep), and Abcc2/multidrug resistance protein (Mrp2) was found, whereas Abcc8 was down-regulated. Up-regulation of Abcc2/Mrp2 was accompanied by enhanced calcein excretion. Aminoguanidine attenuated the effects on transporter expression. Our data indicate that NO, produced locally by renal iNOS, regulates the expression of ABC transporters in vivo. Furthermore, we showed, for the first time, expression and subcellular localization of Abcb11/Bsep in rat kidney

Traffic of Major Histocompatibility Complex Class II Molecules in Rabbit Lacrimal Gland Acinar Cells

Purpose. It has been suggested that lacrimal gland acinar cells, which have been induced to express major histocompatibility complex class II (MHC II) molecules, might initiate local autoimmunity by using mechanisms similar to those operating in the specialized antigenpresenting cells to process and present autoantigens. Surface-labeling experiments indicate that constituents of the acinar cell plasma membrane participate in a rapid recycling traffic. The authors have surveyed the subcellular distribution of MHC II molecules and have evaluated their participation in the traffic between plasma membranes and intracellular compartments. Methods. Acinar cells were isolated from rabbit lacrimal glands and maintained for two nights in a serum-free, hormone-supplemented culture medium containing 10 fxM carbachol. MHC II molecules were detected with a monoclonal antibody (MAB 2C4), biotinylated goat-antimouse IgG (BGAM), and avidin-ferritin (AvFe) or streptavidin-gold (SAvAu) conjugates. Results. Postembedding labeling with MAB 2C4, BGAM, and AvFe revealed MHC II molecules at the surface membranes, in cytoplasmic vesicles, and in secretory vesicles. When cells were chilled to 4°C and subjected to preembedding labeling with MAB 2C4, BGAM, and AvFe, surface MHC II molecules were specifically labeled. Labeled complexes were rapidly internalized upon warming to 37°C. Postembedding labeling with MAB 2C4, BGAM, and SAvAu revealed additional intracellular MHC II molecules, with a distribution overlapping that of the MHC II molecules labeled during the preembedding procedure. When cells were cultured overnight in the presence of MAB 2C4 and subjected to postembedding labeling widi BGAM and AvFe, label was detectable in small vesicles and in secretory vesicles. However, the extent of labeling appeared less than obtained with postembedding labeling with MAB 2C4, BGAM, and AvFe. Preembedding labeling of cells that had been incubated overnight with MAB 2C4 indicated that the cells continued to express MHC II molecules at their surface membranes, and rapid internalization of label upon warming to 37°C confirmed that MHC II molecule traffic continued in the presence of MAB 2C4. Postembedding labeling with MAB 2C4, BGAM, and SAvAu indicated die continued presence of a large intracellular pool of MHC II molecules. Conclusions. MHC II molecules in lacrimal acinar cells are present in large intracellular and small surface pools. They move rapidly between these two pools, but further work will be required to determine whether the MHC II molecule traffic represents recycling or turnover and whether recycling pools and sequestered pools coexist. The presently available data make it reasonable to propose that the traffic of MHC II molecules to plasma membranes provides a mechanism by which acinar cells display intracellularly generated autoantigens to potentially reactive helper T lymphocytes. Invest Ophthalmol Vis Sci. 1994;35:3943-3951

Data for: Pregnancy Probabilistically Augments Potential Precursors to Lacrimal Gland Chronic, Immune-Mediated Infiltrates or Autoimmune Infiltrates

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