Isolating Cell Types from HBoV1-Positive Tonsils

To investigate the cell types of HBoV1 persistence, three populations were isolated sequentially from HBoV1 DNA-positive tonsils (Prep 3) using CD antibody-coupled magnetic beads (Dynabeads; Invitrogen), according to the manufacturer’s instructions: T cells with anti-CD3 beads, B cells with anti-CD19 beads for the CD3-depleted cells, and monocytes with anti-CD14 beads for the CD3-CD19-depleted cells. To test the purity of the three isolates, cells from the beads were released with DETACHaBEADS (Invitrogen), and labeled with anti-human CD19-FITC (CD19 labeled with fluorescein isothiocyanate) (clone SJ25-C1; Invitrogen), CD3-PE/Cy5.5 (CD3 labeled with phycoerythrin or Cy5.5) (clone UCHT1; Abcam), or CD14-PE (clone MφP9; BD Biosciences) and analyzed on a BD Accuri C6 flow cytometer as described previously (26 (link)). Assessment of the cell fraction proportions is described in Text S1.

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Xu M., Perdomo M.F., Mattola S., Pyöriä L., Toppinen M., Qiu J., Vihinen-Ranta M., Hedman K., Nokso-Koivisto J., Aaltonen L.M, & Söderlund-Venermo M. (2021). Persistence of Human Bocavirus 1 in Tonsillar Germinal Centers and Antibody-Dependent Enhancement of Infection. mBio, 12(1), e03132-20.

Publication 2021

Dynabeads DETACHaBEADS CD14-PE (clone MφP9; BD Accuri C6 flow cytometer

Anti cd3 Antibody B cells Cell Clone Cy5 5 Fitc Fluorescein Human Isothiocyanate Monocytes Phycoerythrin Populations T cells Tonsils

Corresponding Organization :

Other organizations : University of Helsinki, University of Jyväskylä, University of Kansas Medical Center, Helsinki University Hospital

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Variable analysis

independent variables

Cell type isolation using CD antibody-coupled magnetic beads (Dynabeads; Invitrogen)

dependent variables

Cell type proportions

control variables

Manufacturer's instructions for using CD antibody-coupled magnetic beads

controls

Positive control: Not specified
Negative control: Not specified

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