Detecting EBV-Infected Cell Populations

Infected cells were detected and isolated as described previously [37 (link)]. Briefly, the PBMCs obtained from patients were isolated by density gradient centrifugation using Separate-L (Muto Pure Chemical, Tokyo, Japan) and were sorted into CD19-, CD4-, CD8-, or CD56-positive fractions using antibody-conjugated magnetic beads (Miltenyi Biotec, Bergisch Gladbach, Germany; 130-050-301, 130-045-101, 130-045-201, 130-090-875). After that, the EBV DNA levels in each fraction were evaluated by real-time PCR using the TaqMan System (Applied Biosystems, Foster City, CA) [38 (link)].

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Onozawa E., Shibayama H., Takada H., Imadome K.I., Aoki S., Yoshimori M., Shimizu N., Fujiwara S., Koyama T., Miura O, & Arai A. (2018). STAT3 is constitutively activated in chronic active Epstein-Barr virus infection and can be a therapeutic target. Oncotarget, 9(57), 31077-31089.

Publication 2018

antibody-conjugated magnetic beads TaqMan System

Antibody Cells Density gradient centrifugation Patients Real time pcr

Corresponding Organization :

Other organizations : Tokyo Medical and Dental University, National Center For Child Health and Development, Nihon University

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Variable analysis

independent variables

Cell fractions (CD19-, CD4-, CD8-, or CD56-positive)

dependent variables

EBV DNA levels in each cell fraction

control variables

PBMCs obtained from patients
Density gradient centrifugation using Separate-L
Antibody-conjugated magnetic beads for cell sorting

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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