Free DNA induces modification on the protein synthesis profile of human peripheral blood mononuclear cells of healthy donors.

2000 ◽  
Author(s):  
G A Martins ◽  
C R Gattas ◽  
M G da Costa Carvalho
Keyword(s):  
Protein Synthesis ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Peripheral Blood Mononuclear ◽  
Free Dna ◽  
Healthy Donors ◽  
Blood Mononuclear Cells

scholarly journals Human antibodies isolated from plasma by affinity chromatography increase the coxsackievirus B4-induced synthesis of interferon-α by human peripheral blood mononuclear cells in vitro

2001 ◽  
Vol 82 (8) ◽  
pp. 1899-1907 ◽  
Author(s):  
Wassim Chehadeh ◽  
Ahmed Bouzidi ◽  
Gunar Alm ◽  
Pierre Wattré ◽  
Didier Hober
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Interferon Α ◽  
Peripheral Blood Mononuclear ◽  
Coxsackievirus B4 ◽  
Healthy Donors ◽  
Blood Mononuclear Cells

Coxsackievirus B4 (CVB4) can be found in circulating blood of patients; however, the interaction of CVB4 with peripheral blood mononuclear cells (PBMCs) is poorly understood. CVB4 induced low levels of IFN-α synthesis in PBMCs from healthy donors. In contrast, preincubation of infectious CVB4 with plasma from these donors containing anti-CVB4 antibodies strongly enhanced the synthesis of IFN-α. IgG obtained from plasma by chromatography formed immune complexes with CVB4 and increased significantly the CVB4-induced production of IFN-α by PBMCs. These antibodies did not have a neutralizing effect on CVB4 infection of Hep-2 cells. The role of CVB and adenovirus receptor (CAR), FcγRII and FcγRIII in the increased synthesis of IFN-α induced by CVB4 preincubated with IgG was shown by inhibition with specific antibodies. The major interferon-α-producing cells in response to CVB4–IgG complexes were CD14+ cells and monocyte-enriched PBMCs. With the latter, detection of IFN-α by immunostaining was positive whereas in monocyte-depleted PBMCs it was not. This study shows that CVB4-induced synthesis of IFN-α by PBMCs can be enhanced by an antibody-dependent mechanism through interactions between the virus, non-neutralizing antivirus antibodies, FcγRII and III and CAR.

scholarly journals Easy-to-Use HPLC Method to Measure Intracellular Ascorbic Acid Levels in Human Peripheral Blood Mononuclear Cells and in Plasma

Antioxidants ◽  
2022 ◽  
Vol 11 (1) ◽  
pp. 134
Author(s):  
Gwendolyn van Gorkom ◽  
Birgit Gijsbers ◽  
Erik-Jan Ververs ◽  
Ahmed El Molla ◽  
Cindy Sarodnik ◽  
...  
Keyword(s):  
Ascorbic Acid ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Hplc Method ◽  
Peripheral Blood Mononuclear ◽  
Healthy Donors ◽  
Blood Mononuclear Cells ◽  
Reproducible Method

Given the growing interest in ascorbic acid (AA), there is a need for a reliable and reproducible method to measure AA status in the human body. Serum AA concentrations do not correlate well with tissue levels, but AA levels in leukocytes do. However, a standard method for clinical application is lacking. This present study describes a method to measure AA in the peripheral blood mononuclear cells (PBMCs) with hydrophilic interaction liquid chromatography (HILIC). The method can also be used in plasma and other leukocyte subsets. The measurements of AA in PBMCs and plasma were performed with HPLC with HILIC separation and UV detection. The sample preparation involved the isolation of PBMCs and lysis and precipitation with acetonitrile. European Medicine Agency guidelines for bioanalytic method validation were followed for the evaluation. A highly precise execution of the method was found with intra- and inter-assay variations at a maximum of 7.8%. In 40 healthy donors, a mean intracellular AA concentration of 7.9 microgram/108 cells was found in PBMCs. A correlation between plasma and PBMC AA concentration was not present (r = 0.22). In conclusion, we developed a convenient, reliable, and reproducible method for the quantitative determination of AA within PBMCs and plasma from human blood.

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