Keratoplasty after mustard gas injury: clinical outcome and histology.

PURPOSE: Ocular injury by mustard gas can lead to severe eye damage with a delayed course. We report the corneal histology and follow-up after keratoplasty in a patient with mustard gas injury. METHODS: The patient presented with recurrent painful corneal inflammation in both eyes not improving under local therapy. Visual acuity impaired to handmovements. A penetrating keratoplasty was performed on the left eye and afterwards an autorotation keratoplasty on the right eye with a later corneal graft. RESULTS: After the operation of the left eye the patient was immediately painfree and the visual acuity improved to 0,4. So far there have been no signs for transplant rejection or inflammation. Histology of the cornea revealed massive stromal necrosis, and signs of chronic inflammation. Despite denervation of the cornea after autorotation keratoplasty the right eye was still painful and became only painfree after corneal transplantation. CONCLUSION: There has been not much experience with corneal transplantation after mustard gas injury and there is a high risk for transplant rejection due to inflammation and vascularisation of the cornea. Successful and painfree healing with keratoplasty seems only possible after complete removal of the necrotic material.

Low-dose anesthesia for corneal transplantation in mice.

BACKGROUND: Despite technical difficulties caused by the small dimensions, scientific and economic considerations stimulated activities in the murine keratoplasty model. Delicate surgery requires stable anesthesia with rapid postoperative waking. METHODS: For corneal transplantation, all animals were intraperitoneally injected with 0.08 ml of a 1:8 mixture of 2% xylazine and 2.5% esketamine hydrochloride. Thus they all received 0.18 mg of xylazine and 1.8 mg of ketamine. In other words, 4.5 mg/kg BW of xylazine and 45 mg/kg BW of ketamine were injected in a mouse weighing 25 g. The inhalation anesthetic isoflurane was used for short interventions like removing lid sutures or in the case of an animal awaking before the end of the operation. A small but effective set of apparatus comprising a glass bottle, a test tube and several interconnected syringes was developed to create a closed system and visualize the amount of isoflurane used in the animal. Air bubbles showed the amount of isoflurane escaping from the opened respiratory mask. RESULTS: The dose reduction of systemic anesthesia requires additional inhalation anesthesia with isoflurane in nearly all cases. The equipment developed is easy to handle and allows systemic anesthesia with a very low dose. CONCLUSION: This murine anesthesia model may be expected to help other surgeons performing corneal transplantations in mice.

Ipsilateral submandibular lymphadenectomy does not prolong orthotopic corneal graft survival in mice.

PURPOSE: To explore the impact of selective lymphadenectomy on the outflow from the outer eye and corneal graft survival in mice. METHODS: BALB/c mice underwent submandibular lymphadenectomy either on the left side 7 or 21 days before the experiments or bilaterally 7 days in advance. First, (99m)Tc colloidal albumin (Nanocoll) was given as a subcutaneous lower-lid, upper-lid or subconjunctival injection, and count rates were determined 24 h later in the eyes, submandibular lymph nodes, spleen, liver and blood. Second, corneal graft survival was assessed in lymphadenectomised and control mice, and IFN-gamma secretion was determined in draining lymph nodes at the time of transplant rejection. RESULTS: Following subconjunctival Nanocoll injection, count rates/min/mg tissue were significantly higher in the ipsilateral submandibular lymph node than in the other tissues or blood (<0.01). Ipsilateral lymphadenectomy prior to injection resulted in considerably increased count rates in contralateral nodes ( P<0.01), which were higher after injections into upper than into lower lids ( P=0.004). Bilateral submandibular lymphadenectomy led to enhanced count rates in the eyes, blood, spleen and liver (all P<0.01). Removal of the ipsilateral lymph node prior to corneal transplantation did not prolong allograft survival ( P>0.05) and considerably increased IFN-gamma secretion in contralateral nodes after prior removal of the ipsilateral ones paralleled transplant rejection. CONCLUSIONS: In mice, removed submandibular lymph nodes are functionally completely replaced by the contralateral nodes. These studies demonstrate for the first time lymphatic drainage crossing the midline of the body. Consequently, unilateral lymphadenectomy does not improve corneal graft survival.

Ballistic CTLA4 and IL-4 gene transfer into the lower lid prolongs orthotopic corneal graft survival in mice.

PURPOSE: To explore outflow from the eye and to determine and modulate the influence of lymphatic drainage on corneal graft survival in mice. METHODS: Tracer experiments were conducted in BALB/c mice using the (99m)Tc colloidal albumin Nanocoll. Count rates were determined in the eyes, submandibular lymph nodes, spleen, liver and blood 24 h after subconjunctival, intracorneal, intracameral (anterior chamber), intravenous and subcutaneous lower-lid or upper-lid injections ( n=6 each). Four groups of BALB/c mice ( n=8) received corneal transplants from C3H mice; two of them were treated ballistically with vector CTLA4+IL-4 onto the leg or the lower lid, one group was untreated and the other control group was treated with an empty minimalistic, immunologically defined, gene expression (MIDGE) vector. RESULTS: Radioactivity was detected in the liver, spleen and ipsilateral submandibular lymph node after intracameral injection as follows: 91.9%, 6.6% and 1.2% respectively. Radioactivity uptake of the ipsilateral submandibular lymph node was also low after intravenous injection (0.1%) but high after intracorneal (33.8%), lower-lid (62.0%) and subconjunctival (71.2%) injection. Vector CTLA4+IL-4 treatment of the lower lid but not of the leg prolonged graft survival ( P=0.004). CONCLUSION: These tracer studies confirmed for the first time identical lymphatic drainage from the cornea and the lower lid. Logically, lymphatic drainage could be manipulated and graft survival improved by gene transfer to the lower lid.

Vitrectomy and trabeculectomy combined with interferon alpha treatment in Adamantiades-Behçet's disease: a case report.

BACKGROUND: Adamantiades-Behçet's disease is a multisystem disorder with recurrent oral and/or genital ulcerations, skin lesions, and ocular involvement. Eye involvement is a common manifestation that affects the patients' quality of life more than any other. Left untreated, it leads to blindness and often to loss of the eye through secondary complications such as phthisis or painful glaucoma. METHODS: Case report of a 24-year-old patient of Turkish descent living in Germany since his birth was admitted to our Department of Ophthalmology in 1999 because of visual loss in his right eye. He could only perceive light in that eye and evidenced total retinal detachment. In the left eye he had 20/20 vision but showed signs of retinal vascular inflammation and secondary glaucoma. RESULTS: Treatment with interferon alpha (9x10(6) I.U./3x week s.c.) and prednisolone (100 mg/d p.o.) led to complete regression of the acute inflammation within 2 weeks prior to operation. Vitrectomy was then successfully performed in the right eye under the therapy of interferon alpha (9x10(6) I.U./3x week s.c.) and 10 mg prednisolone p.o. The prednisolone therapy was stopped 1 week following operation. The failure of conservative glaucoma treatment necessitated trabeculectomy in the left eye. The patient has had no further recurrence for 4 years under monotherapy with interferon alpha (3x10(6) I.U./3x week s.c.). CONCLUSION: Interferon alpha is a potent therapy for Adamantiades-Behçet's disease with ocular involvement. It also provides a basis for safe and reliable surgical interventions. There was no intra- or perioperative recurrence of inflammation, which is a common finding in these procedures.

Influence of ballistic gene transfer on antigen-presenting cells in murine corneas.

PURPOSE: The outcome of corneal transplantation may depend on passenger cells in corneal epithelium and stroma. Their presence in normal corneas is controversial. This study aimed at examining this question and elucidating the still unknown influence of ballistic gene transfer. METHODS: Central 2.5 mm discs of epithelial flatmounts and frozen stromal sections cut parallel to the outer corneal surface were stained for F4/80+ and MHC II+ cells. Corneas were immunohistologically examined in an untreated state and after gene gun treatment using minimalistic immunologically defined gene expression (MIDGE) vector DNA of IL-4 and CTLA4 ( n=6), or untreated/gene-gun-treated donor corneas (C3H mice) were orthotopically grafted into gene-gun-treated eyes (BALB/c mice), and their survival was investigated ( n=8). RESULTS: Untreated control corneas contained 115.7+/-33.7 F4/80+ and 106.8+/-46.2 MHC II+ cells in the epithelium, and 48.9+/-13.2 versus 7.3+/-5.5 in the stromal layer. Ballistic gene transfer caused migration of F4/80+ cells into the corneal stroma ( P<0.01). Graft survival (27.4+/-16.8 days) was not prolonged by gene gun transfection of donor and recipient corneas but increased significantly to 64+/-28 days ( P<0.01) after treating only the recipient. CONCLUSIONS: A multiplicity of F4/80+ and MHC II+ cells in normal murine corneas diminishes the immune privilege of the eye. Ballistic gene transfer impedes graft survival by triplicating these passenger cells in the stromal layer. However, ballistic transfer of MIDGE vector DNA of IL-4 and CTLA4 markedly improves graft survival when treating only the recipient eye.