Application of paraffin embedding slice fluorescent staining method in the experimental xenogenic corneal transplantation immune diagnosis

Abstract

AIM: Using monoclonal antibodies indirectly immunofluorescence to detect the changes of CD4 and CD8 T lymphocytes in the different time points in corneal tissue after corneal transplantation. METHODS: Normal rabbit cornea, axillary lymph node tissue were taken. Get corneal tissue at one week, two weeks after ostrich-rabbit heterogeneous corneal transplantation, fixed by formaldehyde, conventional paraffin embedding sectioning dewaxing after water, gastric enzyme digestion method for antigen repair, use mouse anti rabbit CD4, CD8 monoclonal antibody and fluorescent tagging sheep anti mouse IgG for immunohistochemical assay, and observe the expression of CD4+, CD8+T lymphocytes in the cornea tissue in fluorescence microscope.RESULTS: The expression of CD4+, CD8+T cells in the cornea tissue was negative one week after the ostrich-rabbit heterologous cornea transplantation, CD4 T lymphocytes showed strong positive expression after the second week, CD8 was negative. CONCLUSION: Indirect immunofluorescence method can be used for determination of related antigen expression in local corneal tissue after heterologous corneal transplantation, and it can lay the foundation for the studies of heterogeneous cornea transplantation rejection mechanism and postoperative drug screening.

Keywords

• heterologous cornea transplantation
• rejection
• paraffin embedding organizations
• indirect immunofluorescence staining
• CD4 and CD8 T lymphocytes