Investigation of corneal autoantibodies in horses with immune mediated keratitis (IMMK).

Immune mediated keratitis (IMMK) is primarily a non-ulcerative keratitis in horses causing intermittent ocular pain, eventually resulting in visual impairment. Affected horses typically respond to immunomodulatory treatment. However, the underlying cause of the disease remains enigmatic. The current study was undertaken to investigate the presence of autoantibodies in horses with immune mediated keratitis. Using 28 horses with IMMK and 27 healthy controls screening for serum autoantibodies against the corneal proteome using indirect immunofluorescence, one-dimensional (1DE) and two-dimensional electrophoresis (2DE) with subsequent western blot analysis was performed followed by mass spectrometric identification of bands or spots of interest. Indirect immunofluorescence did not reveal a difference in immune response towards corneal proteins between healthy horses and those with IMMK. Using western blot analysis some horses affected by IMMK (4/28) showed a single band (1D) or a single spot (2DE) (5/28) not detected in healthy controls. The corresponding spot was identified as maspin (SERPINB5), a protein responsible for the inhibition of corneal vascularisation, cell migration and cell adhesion to the extracellular matrix. Tests with a recombinant human protein commercially available did not verify blot findings, but the human protein may not be fully cross-reactive. Still, maspin might play a role in some cases of equine IMMK. Further research is needed to clarify the etiology of this disease.

Impression cytology as diagnostic tool in horses with and without ocular surface disease.

BACKGROUND: Cytology plays a major role in the diagnosis of ocular surface diseases. OBJECTIVE: To compare 2 cytological sampling methods for obtaining corneal and conjunctival cell samples regarding irritation for the patient, invasiveness, sample quality and diagnostic equivalence. STUDY DESIGN: Observational prospective study. METHODS: In 5 healthy horses, conjunctival and corneal samples were taken bilaterally by impression cytology sampling (ICS) and cytobrush sampling (CBS). Irritation and invasiveness were assessed with an eye irritation and an epithelial damaging score system, respectively. Sample quality was evaluated via morphometric analysis and graded by a board certified clinical pathologist. For the assessment of diagnostic equivalence, 15 eyes of 14 client owned horses with ocular surface anomalies were sampled by ICS and CBS. The methods were compared regarding the types of inflammatory cells and/or infectious agents detected and if the correct diagnosis could be achieved. Histopathology served as gold standard when available. RESULTS: ICS was significantly less invasive and less irritating for the horses. Both methods retrieved cells of overall high quality; the cell quantity was significantly higher in IC samples. ICS preserved the natural cellular layout. There was a fair, but no statistically significant agreement between the diagnostic outcomes between sampling methods although CBS resulted in a slightly greater variability of inflammatory cell types compared to ICS. MAIN LIMITATIONS: None. CONCLUSIONS: Because of its low irritability and invasiveness ICS can be recommended for cell sampling in fragile corneas and for experimental studies. ICS is especially useful in cases where preservation of cellular layout is advantageous. CBS samples are easier to obtain because of the small equine palpebral fissure. Additionally, the identification of inflammatory cells within conjunctival cell samples is easier. Overall, CBS is still an appropriate method in clinical settings.

[Ophthalmic examination of small animals]. / Die Augenuntersuchung bei Kleintieren.

This article comprises a practical approach to the small animal ophthalmic examination. Initially, the signalment is evaluated followed by the general and specific clinical history and a general physical examination. The ocular examination should be performed in a detailed systematic manner and always comprises both eyes. A thorough examination technique is rewarding as a diagnosis can be reached in most cases. The subsequent search for the underlying aetiology is mandatory.