Analysis of Tears in Patients with Atopic Keratoconjunctivitis, Using Flow Cytometry

Abstract

Atopic keratoconjunctivitis (AKC) is a chronic allergic eye disease. Although the pathogenesis is not fully understood, some impairment in cell-mediated immunity was suggested by histopathological findings in conjunctival specimens obtained from affected individuals. T-cell infiltration and an enhanced T-helper/T-suppressor cell ratio in conjunctival biopsy specimens were observed previously by immunofluorescence procedures. We analyzed the cells in tears of patients with AKC using flow cytometry (FCM) and compared the results to those of normal subjects to identify the role of T lymphocytes in the pathogenesis of the disease. The tear samples of the patients and normal subjects were collected with capillary tubes, and the surface receptors of cells were detected with FCM. Statistical analyses were performed with Student’s t test. The percentages of T cells, activated B cells, and T-helper/T-suppressor cell ratios were found to be higher in the tears of patients with AKC than in controls. We propose that a decreased T-suppressor cell concentration in tears may enhance immunoglobulin-E production of B cells, and the signs and symptoms are provoked by inflammatory mediators liberated from mast cell degranulation.

Diagnostic Tests Based on Human Basophils: More Potentials and Perspectives than Pitfalls

“…The utility of BAT in hymenoptera venom hypersensitivity [34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56]has been confirmed hitherto by many more than the two studies quoted by Kleine-Tebbe et al [1] (table 2). The main benefit of BAT in that indication is that it may confirm clinical history in the absence of positive skin tests or positive allergen-specific IgE.…”

“…Many of these studies have been reviewed recently [8,9,10]. Herewith, tables 1, 2, 3, 4, 5, 6, 7summarize the results of validated clinical studies performed with various allergens such as inhalant allergens [25,26,27,28,29,30,31,32,33] (table 1), hymenoptera venoms [34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56] (table 2), latex [57,58,59,60,61,62,63] (table 3), food [64,65,66,67,68,69,70,71,72,73,74,75,76,77,78,79,80] (table 4) and drugs [81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97,98,99,100,101,…”

Basophil Activation Test in Allergy: Time for an Update?

“…In these first attempts, correlation between expression of a membrane molecule and a degranulation event was of the utmost interest. A very good correlation between the histamine release test (HRT) and CD63-positive basophils was observed in latex [21], hymenoptera venom [22,23], grass pollen [6], food [7] and drug allergy [24], so that flow cytometry was proposed for routine diagnosis of allergic reactions [25,26]. In any case, basophil flow cytometry also presented an opportunity to study basophilic cells in general and to aid in separating them from peripheral blood [27,28].…”

T-cell characterization in chronic allergic eye disease

“…In AKC, the percentages of T cells, activated B cells, and Thelper/T-suppressor cell ratios were higher in the tears than in normal controls [11]. A decreased T-suppressor cell concentration in tears might enhance IgE production by B cells, and augment production of inflammatory mediators liberated by the increased mast-cell degranulation.…”