Real-Time Polymerase Chain Reaction (RQ-PCR) for the Monitoring of Epstein-Barr Virus (EBV) Load in Peripheral Blood Mononuclear Cells

2000 ◽  
Vol 212 (4) ◽  
pp. 206-210 ◽  
Author(s):  
H. J. Wagner ◽  
W. Jabs ◽  
F. Smets ◽  
M. Wessel ◽  
L. Fischer ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Peripheral Blood Mononuclear Cells ◽  
Epstein Barr Virus ◽  
Mononuclear Cells ◽  
Chain Reaction ◽  
Peripheral Blood Mononuclear ◽  
Barr Virus ◽  
Blood Mononuclear Cells ◽  
Polymerase Chain ◽  
Epstein Barr

scholarly journals In vitro effects of Epstein-Barr virus on peripheral blood mononuclear cells from patients with rheumatoid arthritis and normal subjects.

1978 ◽  
Vol 148 (5) ◽  
pp. 1429-1434 ◽  
Author(s):  
L Slaughter ◽  
D A Carson ◽  
F C Jensen ◽  
T L Holbrook ◽  
J H Vaughan
Keyword(s):  
Rheumatoid Arthritis ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Epstein Barr Virus ◽  
Mononuclear Cells ◽  
Peripheral Blood Mononuclear ◽  
Barr Virus ◽  
Blood Mononuclear Cells ◽  
Epstein Barr

Peripheral blood mononuclear cells from 10 patients with rheumatoid arthritis and 9 control subjects were cultured in vitro for 30 days with and without infection by Epstein-Barr virus. All cultures showed polyclonal stimulation of B cells as indicated by rising levels of IgM in the culture supernates, reaching maximal at 18-24 days, and with no quantitative or kinetic difference between the RA and control cells. IgM anti-IgG was also produced in both groups and maximally at 18-24 days, but in greater quantity by the RA lymphocytes. The anti-IgG made by the RA lymphocytes was more easily absorbed by solid phase IgG than was the anti-IgG made by the normal lymphocytes and thus was judged to be of higher affinity. RA lymphocytes uninfected with EBV had higher transformation scores than did the normal controls and developed spontaneously into permanent cell lines in six instances.

Detection of tumor cells in purged bone marrow and peripheral-blood mononuclear cells by polymerase chain reaction amplification of bcl-2 translocations.

1994 ◽  
Vol 12 (5) ◽  
pp. 1021-1027 ◽  
Author(s):  
R S Negrin ◽  
J Pesando
Keyword(s):  
Bone Marrow ◽  
Polymerase Chain Reaction ◽  
Tumor Cells ◽  
Peripheral Blood Mononuclear Cells ◽  
Mononuclear Cells ◽  
Chain Reaction ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Before And After ◽  
Polymerase Chain

PURPOSE To compare bone marrow (BM) before and after purging with monoclonal antibodies (MAbs) and complement with peripheral-blood mononuclear cells (PBMNCs) for tumor-cell contamination by amplification of t(14;18) sequences using the polymerase chain reaction (PCR). PATIENTS AND METHODS Sixty patients with non-Hodgkin's lymphoma (NHL) undergoing autologous BM transplantation were evaluated. Six BM biopsies were performed at the time of harvesting and evaluated morphologically for tumor involvement. The harvested BM was treated with a panel of anti-B-cell MAbs directed against CD9, CD10, CD19, and CD20, followed by rabbit complement. Clonogenic assays were performed before and after purging. DNA was extracted and t(14;18) sequences amplified by PCR. PBMNCs collected by apheresis for back-up purposes were similarly evaluated. RESULTS Fifteen patients (25%) were PCR-positive before BM purging. Following MAb- and complement-mediated purging, there was a reduction in the PCR-amplified signal in 10 patients (67%). There was no reduction in colony-forming unit granulocyte-macrophage (CFU-GM) colony growth following purging. Eight of these 15 patients (53%) had morphologic evidence of BM involvement at the time of harvesting. In these eight patients, only three had a reduction in the PCR-amplified products, as compared with all seven who were morphologically negative at the time of BM harvesting (P = .026). Fourteen of these 15 patients had PBMNCs collected near the time of BM harvesting and 12 (86%) were PCR-positive. CONCLUSION BM purging with MAbs and complement results in reduction in the number of t(14;18)-positive tumor cells, especially in those patients who have no morphologic evidence of BM disease at the time of harvesting. Purged BM was less contaminated with t(14;18)-positive cells than unpurged PBMNCs, which were frequently contaminated with tumor cells.

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