Non-inflammatory exudative lesions of Reinke’ pos space present the most common cause of phonation disorders in representatives of vocal professions. The leading role in pathogenesis of this disorder is given to the nearly complete absence of lymphatic drainage of Reinke space and local deposition of tissue fluid. The mechanisms of its progression are of particular importance when determining the condition of mucosaassociated laryngeal tissue. The aim of our work was to evaluate the parameters of immunohistochemical lymphocyte typing in exudative lesions from the Reinke’s space. Materials and methods: The main group consisted of 40 patients, at the mean age of 43.2±2.1 years, exhibiting tumor-like exudative lesions of Reinke’s space. The biopsy material was taken from the vocal folds, including polyps, vocal nodules and Reinke’s edema. Myxoid and angiomatous types of the polyps were separately assessed. Videofibrolaryngoscopy was performed using an Olympus TYPE 150 bronchofibroscope (Germany). Morphological studies were carried out using a DMRXA microscope (Leika, Germany) by means of the ImageScopeM computer program (Germany). The uniformly treated sections were stained with Hematoxylin & Eosin (Biovitrum, Russia). Immunohistochemical quantitative assessment of the main T cell populations (CD3+, CD4+, CD8+), B cells (CD20+), histiocytes (CD68+), and the cells expressing bcl-2 and p53 cell regulators was carried out automatically using the BenchMarkXT immunohistotainer (Ventana, USA). The results were expressed as U/mm2. Results and Discussion. Some special features of cellular immunophenotype were revealed in exudative lesions of Reinke’s space. Reinke’s edema was characterized by high content of CD3+ lymphocytes, CD4+, CD8+, CD20+, p53 positive cells of the basal epithelium, as well as low numbers of CD68+, bcl-2 positive lymphocytes and cells of the basal epithelium. Myxoid type of polyps was characterized by low content of CD3+, CD4+, CD8+, CD20+ lymphocytes, bcl-2 positive lymphocytes and basal epithelium cells, CD68+ monocyte-macrophage cells, and high amounts of p53 positive basal epithelial cells. In the angioma-type polyps, we have registered low contents of CD3+, CD4+, CD8+, CD20+ lymphocytes, high numbers of monocyte-macrophage CD68+ cells, MMP-9+, bcl-2 positive lymphocytes, and low content of p53 positive cells of basal epithelium. The samples from the vocal nodules were characterized by low content of CD3+, CD4+, CD8+, CD20+ and p53 positive basal epithelial cells; high numbers of CD68+ cells (monocyte-macrophage series), MMP-9+ and bcl-2 positive lymphocytes.
Exudative tumor-like neoplasms of the Reinke space in vocal folds are widespread in the population, more often among representatives of vocal professions, e.g., actors, singers, teachers, lecturers and represent a serious medical and social problem. In pathogenesis of such neoplasms, a key role is given to chronic phonotrauma, intoxication during smoking against the background of the nearly complete absence of lymphatic drainage in the Reinke space. Cellular factors of tissue and immune homeostasis are of great importance in morphogenesis of this disorder. Switching of immune responses aimed at maintaining tissue homeostasis is accompanied by increased production of pro-inflammatory cytokines, as well as chemotaxis regulators and growth factors. The aim of our work was to study the levels of chemokines (MIP-1, MIP-1, MIP-3, fractalkine, IL-8, I-TAC) in the tissues of exudative lesions from the Reinkes space (EPPR). Forty tissue samples of exudative lesions from Reinkes space, in particular, vocal fold polyps, vocal nodules and neoplasms presenting as Reinkes edema were taken as biological material for the study. The samples were taken intraoperatively when the neoplasm was removed, using an Olympus TYPE 150 fiber bronchoscope (Germany) using an integrated Lumenis Acu Pulse CO2 laser (Israel). The content of chemokines was determined in the supernatants of the tissues homogenates, using multiplex analysis with MAGPIX-100 immunoanalyzer (Bio-Rad, USA). Statistical processing was carried out using Statistica 10.0 for Windows software package. The results are presented as medians (Q0.25-Q0.75). Chronic phonotrauma and/or exposure to toxic factors leads to increased permeability of the vascular endothelium, tissue edema and activation of cells involved in tissue and immune homeostasis, e.g., fibroblasts, monocytes, endotheliocytes. The study of the chemokine expression in the tissues of various exudative lesions of Reinkes space enabled us to reveal the following features: (1) predominance of the CXC-chemokine content produced mainly by fibroblasts in dense neoplastic tissues, thus reflecting participation of the latter in the genesis of this pathology. (2) In the tissues of soft neoplasms with large proportion of liquid component, we have revealed increased concentrations of SS-chemokines and fractalkine. The latters are produced mainly by macrophages and endotheliocytes, thus, probably, reflecting a predominant role of these cells in development of myxoid polyps and Reinkes edema.
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