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. 2000 Sep 5;192(5):637–646. doi: 10.1084/jem.192.5.637

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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graphic file with name JEM000695.f3a.jpg — Characterization of the replication-competent virus associated with PBMC-derived B cells of HIV-infected patients. (A) B cells and T cells isolated from three HIV-infected individuals were pretreated with pronase followed by coculture with indicator T cells. (B, top) Flow cytometric comparison of CD21 expression on representative HIV-negative B cells and presorted B cells of patient 17, followed by (B, bottom) post-sorted CD20⁺/CD21⁺ and CD20⁺/CD21⁻ fractions of patient 17. (C) B cells of three HIV-infected patients sorted into CD21⁺ and CD21⁻ fractions (and CD32⁺ versus CD32⁻ fractions in the case of patient 19) and cocultured with indicator T cells. Colors identify the sorted fraction and symbols identify the number of coculture wells plated for a given sorted fraction. (D) Flow cytometric comparison of CD21 expression on pronase-treated and -untreated B cells of patient 9. (E) B cells of three HIV-infected patients preincubated with mAbs FE8 and BL13 and cocultured with indicator T cells. (F) FE8-displaced HIV from 2 × 10⁶ B cells of three HIV-infected patients, treated with rabbit anti–human C3d (C3d Ab) or normal rabbit serum (Normal Ab), and captured with magnetized anti-rabbit antibodies. Fractions captured on magnet (+ bead) and fractions not captured (− bead) were tested for HIV-1 RNA by bDNA.

graphic file with name JEM000695.f3b.jpg — Characterization of the replication-competent virus associated with PBMC-derived B cells of HIV-infected patients. (A) B cells and T cells isolated from three HIV-infected individuals were pretreated with pronase followed by coculture with indicator T cells. (B, top) Flow cytometric comparison of CD21 expression on representative HIV-negative B cells and presorted B cells of patient 17, followed by (B, bottom) post-sorted CD20⁺/CD21⁺ and CD20⁺/CD21⁻ fractions of patient 17. (C) B cells of three HIV-infected patients sorted into CD21⁺ and CD21⁻ fractions (and CD32⁺ versus CD32⁻ fractions in the case of patient 19) and cocultured with indicator T cells. Colors identify the sorted fraction and symbols identify the number of coculture wells plated for a given sorted fraction. (D) Flow cytometric comparison of CD21 expression on pronase-treated and -untreated B cells of patient 9. (E) B cells of three HIV-infected patients preincubated with mAbs FE8 and BL13 and cocultured with indicator T cells. (F) FE8-displaced HIV from 2 × 10⁶ B cells of three HIV-infected patients, treated with rabbit anti–human C3d (C3d Ab) or normal rabbit serum (Normal Ab), and captured with magnetized anti-rabbit antibodies. Fractions captured on magnet (+ bead) and fractions not captured (− bead) were tested for HIV-1 RNA by bDNA.

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