The specific aims of the experiments proposed here are: 1. to identify the subsets of lymphocytes present in corneal immune reactions; 2. to determine the functions of the specific lymphocyte subpopulations present in corneal immune reactions; 3. to develop a reproducible and quantitative alternative model to the experimental cornea allograft procedure; and 4. to compare the effectiveness of topical cyclosporine with topical steroids in preventing or suppressing corneal immune reactions. Inbred rats of one strain will be given intracorneal injections of allogeneic keratocytes from a second inbred strain. The fate of the injected keratocytes and the host response to the allogeneic keratocytes will be determined. Enumeration and identification of specific lymphocyte subsets of host origin in the keratocyte-injected corneas will be accomplished by histological and immunohistochemical methods. The immunoperoxidase staining technique, coupled with antibody specific for lymphocyte subset markers, will permit accurate analysis of the recipient's response to the allogeneic keratocytes. Specific functions of intracorneal lymphocytes will be determined by a modified plaque assay technique developed to permit study of B and T lymphocytes. Immunologically competent lymphocytes synthesize and secrete biologically potent mediators that exert a regulatory action on effector cells. The modified plaque assay technique will be used to identify intracorneal lymphocytes that secrete specific mediators. A new experimental analog of the cornea transplant will be developed and the lymphocyte subset(s) responsible for this reaction will be identified by cell fractionation techniques. The time course of this model and the optimum nunber of lymphocytes necessary to establish the model will be determined. Cyclosporine, the promising new immunosuppressive agent, will be tested as a topical immunosuppressive in the alternative corneal allograft model, in an effort to establish the potential usefulness of this drug in clinical ophthalmology. Corneal immune reactions, such as the allograft reaction, are serious clinical problems. By developing and investigating a variety of experimental systems and by determining the cellular and humoral components of corneal immune reactions, it will be possible to better understand and medically treat such reactions in humans. The availability of reproducible and relevant experimental corneal immune reactions will facilitate the development and testing of new therapeutic strategies for controlling or preventing deleterious corneal immune reactions.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
3R01EY003150-09S1
Application #
3257412
Study Section
(SSS)
Project Start
1979-07-01
Project End
1990-09-29
Budget Start
1987-09-30
Budget End
1990-09-29
Support Year
9
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Louisiana State University Hsc New Orleans
Department
Type
Schools of Medicine
DUNS #
782627814
City
New Orleans
State
LA
Country
United States
Zip Code
70112
Clarke, B L; Gebhardt, B M; Blalock, J E (1993) Mitogen-stimulated lymphocytes release biologically active corticotropin. Endocrinology 132:983-8
Frantz, J M; Gebhardt, B M; Reidy, J J et al. (1991) Immunogenicity of epikeratophakia tissue lenses containing living donor keratocytes. Refract Corneal Surg 7:141-5
Gebhardt, B M; Hill, J M (1990) Cellular neuroimmunologic responses to ocular herpes simplex virus infection. J Neuroimmunol 28:227-36
Gebhardt, B M; Salmeron, B; McDonald, M B (1990) Effect of excimer laser energy on the growth potential of corneal keratocytes. Cornea 9:205-10
Reidy, J J; Gebhardt, B M; Kaufman, H E (1990) The collagen shield. A new vehicle for delivery of cyclosporin A to the eye. Cornea 9:196-9
Chen, Y F; Gebhardt, B M; Reidy, J J et al. (1990) Cyclosporine-containing collagen shields suppress corneal allograft rejection. Am J Ophthalmol 109:132-7
Gebhardt, B M (1990) The role of class II antigen-expressing cells in corneal allograft immunity. Invest Ophthalmol Vis Sci 31:2254-60
Xie, L; Gebhardt, B M (1989) A simplified technique for the short-term tissue culture of rabbit corneal cells. In Vitro Cell Dev Biol 25:20-2
Gebhardt, B M (1988) The effect of tissue dose and site of grafting on immunity to corneal allografts. Invest Ophthalmol Vis Sci 29:1663-70
Johnson, M K; Gebhardt, B M; Berman, M B (1988) Appearance of collagenase in pneumolysin-treated corneal fibroblast cultures. Curr Eye Res 7:951-3

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