Selective increase in blood dendritic cell antigen-3-positive dendritic cells in bronchoalveolar lavage fluid in allergic patients.

Dendritic cells (DCs) are specific antigen-presenting cells that play critical roles in the initiation and polarization of immune responses. DCs residing in the lungs might be detected in the bronchoalveolar lavage fluid (BALF). We analysed DC compartment in the peripheral blood and BALF of patients with allergy and in controls. Plasmacytoid and four distinct subsets of myeloid DCs [characterized by the expression of blood dendritic cell antigen (BDCA)-1+ and -3+ and CD16 positivity or negativity] were detected in both tested compartments. We further evaluated the expression of C-type lectins [mannose receptor (MR), dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) and dendritic and epithelial cells (DEC)-205] relevant to the pathogenesis of asthma. Interestingly, we found a selective increase in the frequency of myeloid DC-expressing BDCA-3 and MR particularly in BALF from allergic patients. Specific and highly statistically significant increase in BDCA-3+ and/or MR+ DCs brings a novel characteristic to BAL analysis in allergic patients.

Assessment of lymphocyte proliferation: CFSE kills dividing cells and modulates expression of activation markers.

The measurement of cell proliferation after mitogenic stimulation is an important parameter used in diagnosis of immunodeficiencies in clinical laboratory as well as in various fields of lymphocyte research. Recent methods try to overcome the radioactive assay using tritiated thymidine ((3)H) by flow-cytometric methods using different fluorochromes such as CFSE or even to substitute these direct methods by tracing the expression of cell-membrane activation markers associated with various steps of proliferation cycle. In our study we compared the (3)H assay with CFSE-staining method and expression of activation markers (CD69, HLA-DR, CD25, CD27, CD71, CD152, CD134 and CD195) on a sample of 128 consecutive patients and healthy controls evaluated in clinical laboratory. We also tested various concentrations of CFSE and its impact on proliferation activity and expression of activation markers. We found that CFSE in concentration from 37nM to 10microM decreases the proliferative capacity (expressed in cpm (3)H assay) due to the decreased viability of proliferating cells (measured as 7-AAD+) in concentration-dependent manner. Moreover, CFSE substantially modulates the expression of activation molecules (decreasing CD69, HLA-DR, CD25 the majority of examinated molecules). We found a good correlation between CFSE-staining method with (3)H assay, if CFSE low population is gated on CD3+ population (correlation coefficient 0.801), but only in samples with stimulation index (SI) higher then 25. In poorly proliferating samples (SI25) no correlation was found due to several false positive results in CFSE test. Statistically significant correlation between proliferation assessed as (3)H-thymidine incorporation and expression of activation markers was found in the case of CD25, CD27, CD38, CD152, CD71, still only in samples with higher proliferation activity (SI>25). No correlation was found with CD134, CD195, HLA-DR and CD69. We conclude that standard assay with (3)H-thymidine incorporation is unreplaceable assay in diagnosis of severe cellular immunodeficiencies as CFSE assay have high proportion of false positive results. Researchers tracing cell-membrane bound molecules on dividing cells stained by CFSE must take into account that CFSE may substantially modulate the expression of these markers and decrease the viability of stained cells.

[Experience with implantation of posterior chamber lenses at the Ophthalmology Clinic of the Martin Medical School Hospital]. / Skúsenosti s implantáciou zadnokomorových sosoviek na Ocnej klinike Martinskej fakultnej nemocnice.

The authors refer to their experience with implantation of 23 IOL following the extraction of cataract, using the Baikoff's method (envelope technique). The ratio men:women in the treated group of patient was 13:10. In 19 patients the resultant vision acuity achieved the value of 6/18, or even better. Postoperative complications were represented by a temporary increase of intraocular pressure (occurred in 3 patients), fibrin reaction (occurred in 5 patients), deformation of the pupil (occurred in 3 patients) and vitreous body hemorrhage (occurred in 1 patient). The observation period lasted 3-6 months. The reason of deteriorated vision acuity which occurred in 4 cases was the presence of glaucoma chronicum simplex and degeneratio maculae luteae. (Tab. 3, Ref. 5.)