RESUMEN
Adhesion is critical for the maintenance of cellular structures as well as intercellular communication, and its dysfunction occurs prevalently during cancer progression. Recently, a growing number of studies indicated the ability of oxygen to regulate adhesion molecules expression, however, the influence of physiological hypoxia (physioxia) on cell adhesion remains elusive. Thus, here we aimed: (i) to develop an optical tweezers based assay to precisely evaluate single diffuse large B-cell lymphoma (DLBCL) cell adhesion to neighbor cells (mesenchymal stromal cells) and extracellular matrix (Matrigel) under normoxia and physioxia; and, (ii) to explore the role of integrins in adhesion of single lymphoma cell. We identified the pronouncedly reduced adhesive properties of lymphoma cell lines and primary lymphocytes B under physioxia to both stromal cells and Matrigel. Corresponding effects were shown in bulk adhesion assays. Then we emphasized that impaired ß1, ß2 integrins, and cadherin-2 expression, studied by confocal microscopy, account for reduction in lymphocyte adhesion in physioxia. Additionally, the blockade studies conducted with anti-integrin antibodies have revealed the critical role of integrins in lymphoma adhesion. To summarize, the presented approach allows for precise confirmation of the changes in single cell adhesion properties provoked by physiological hypoxia. Thus, our findings reveal an unprecedented role of using physiologically relevant oxygen conditioning and single cell adhesion approaches when investigating tumor adhesion in vitro.
Asunto(s)
Médula Ósea/patología , Matriz Extracelular/metabolismo , Hipoxia/patología , Linfoma de Células B Grandes Difuso/patología , Pinzas Ópticas , Antígenos CD/metabolismo , Cadherinas/metabolismo , Adhesión Celular , Moléculas de Adhesión Celular/metabolismo , Línea Celular Tumoral , Proliferación Celular , Supervivencia Celular , Colágeno/metabolismo , Combinación de Medicamentos , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Integrina beta1/metabolismo , Laminina/metabolismo , Rayos Láser , Linfoma de Células B Grandes Difuso/metabolismo , Células Madre Mesenquimatosas/metabolismo , Proteoglicanos/metabolismo , Análisis de la Célula Individual , Células del Estroma/patología , Factores de TiempoRESUMEN
AIM: The aim was to study the association between the phage neutralization of patients' sera and the clinical outcome of phage therapy (PT). PATIENTS: About 62 patients with various bacterial infections receiving PT as well as 30 healthy volunteers were studied. MATERIALS & METHODS: Antiphage activity of sera (AAS) was examined using the phage neutralization test of different types of phages before and during PT in relation to the route of phage administration and correlated with the results of PT. RESULTS & CONCLUSION: The analysis of the association between AAS level and clinical results indicated that the level of AAS is not correlated with the outcome of PT.