Marek Fol; Magdalena Kowalewicz-Kulbat; El ż bieta Ograczyk; Marcin W ł odarczyk; Krzysztof Krawczyk
Volume 13, Issue 2 , June 2016, , Pages 132-140
Abstract
Background: The immunomagnetic separation technique is the basis of monocyte isolation and further generation of monocyte-derived dendritic cells. Objective: To compare the efficiency of monocyte positive and negative separation, concentration of beads, and their impact on generated dendritic cells. ...
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Background: The immunomagnetic separation technique is the basis of monocyte isolation and further generation of monocyte-derived dendritic cells. Objective: To compare the efficiency of monocyte positive and negative separation, concentration of beads, and their impact on generated dendritic cells. Methods: Monocytes were obtained using monoclonal antibody-coated magnetic beads followed the Ficoll-Paque gradient separation of mononuclear cell fraction from the peripheral blood of 6 healthy volunteers. CD14 expression was analyzed by flow cytometry. Results: The percentage of MDDCs generated from monocytes obtained by positive and negative selection was comparable (51.8 ± 15.0 and 46.7 ± 3.4, respectively; p=0.885). The median values for the number of MDDCs obtained from monocytes after positive selection (3.9 × 106) and for MDDCs obtained from monocytes after negative selection (3.1 × 106) were comparable (p=0.194). The use of the recommended or half of the amount of beads for both types of separation had no significant influence on the percentage of isolated cells. Conclusions: Both types of magnetic separation including recommended and reduced concentrations of beads did not affect the yield and the purity of monocytes and their surface CD14 expression. However, DCs originated from the “positively” separated monocytes had noticeable higher expression of CD80.
Ali Shams Shahemabadi; Ahmad Zavaran Hosseini; Shapour Shaghasempour; Mohammad Reza Masjedi; Majid Rayani; Majid Shams; Nasrin Esphandyari; Majid Pouramiri
Volume 7, Issue 1 , March 2010, , Pages 57-63
Abstract
Background: Mycobacterium tuberculosis lipid antigens take part in pathogenicity of the bacterium but the response of monocytes/macrophages to these antigens in tubercu-losis is not well known. Objective: The aim of current investigation was to study the M. tuberculosis lipid antigens in tuberculosis ...
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Background: Mycobacterium tuberculosis lipid antigens take part in pathogenicity of the bacterium but the response of monocytes/macrophages to these antigens in tubercu-losis is not well known. Objective: The aim of current investigation was to study the M. tuberculosis lipid antigens in tuberculosis pathogenesis. Methods: In the present study M. tuberculosis lipid antigens were extracted. Monocytes and macrophages from mul-tidrug-resistant tuberculosis (MDR-TB), TB patients, asymptomatic healthy individuals with positive tuberculin skin test positive and healthy individuals with negative tubercu-lin skin test were collected using magnetic cell sorting. The cells were stimulated by M. tuberculosis total lipid antigens and IL-12 and IL-10 in their supernatants were meas-ured by enzyme-linked immunosorbent assay. Results: The IL-12 production by mono-cytes in response to M. tuberculosis total sonicate antigens in the MDR-TB patients did not show a considerable difference with the PPD positive healthy subjects, whereas in the active TB patients, IL-12 levels significantly decreased (p<0.05). IL-10 production by monocytes in TB patients in response to total lipid antigens showed a significant in-crease in comparison to MDR-TB patients and healthy individuals. Conclusion: In the MDR-TB patients, IL-10 and IL-12 production by monocytes in response to M. tubercu-losis lipid antigens are similar to the healthy subjects.
