Distinct patterns of Bcl-2 expression occur in R5- and X4-tropic HIV-1-producing lymphoid tissue cells infected ex vivo

Michelle K. Haas, David Levy, Joy M. Folkvord, Elizabeth Connick

Research output: Contribution to journalArticle

Abstract

Most HIV-1 replication occurs in secondary lymphoid tissues in T cells within B cell follicles. Mechanisms underlying the accumulation of HIV-1-producing cells at these sites are not understood. Antiapoptotic proteins such as Bcl-2 could promote follicular CD4+ T cell survival, contributing to sustained virus production. Tonsils obtained from subjects without known HIV infection were disaggregated and analyzed for Bcl-2 expression in follicular (CXCR5+) and extrafollicular (CXCR5-) CD3+CD4+ cells by flow cytometry. Additional tonsil cells were cultured with phytohemagglutinin (PHA) and interleukin-2 (IL-2) for 2 days, infected with either CCR5(R5) or CXCR4-tropic (X4) GFP reporter viruses, and analyzed for Bcl-2 expression. In freshly disaggregated CD3+CD4+ tonsil cells, mean florescence intensity (MFI) for Bcl-2 was higher in CXCR5+ (median, 292) compared to CXCR5- cells (median, 194; p=0.001). Following in vitro stimulation with PHA and IL-2, Bcl-2 MFI was higher in both CXCR5+ cells (median, 757; p=0.03) and CXCR5- cells (median, 884; p=0.002) in uninfected cultures compared to freshly isolated tonsil cells. Bcl-2 MFI was higher in GFP+CD3+CD8- R5-producing cells (median, 554) than in X4-producing cells (median, 393; p=0.02). Bcl-2 MFI was higher in R5-producing CXCR5+ cells (median, 840) compared to all other subsets including R5-producing CXCR5- cells (median, 524; p=0.04), X4-producing CXCR5+ cells (median, 401; p=0.02), and X4-producing CXCR5- cells (median, 332; p=0.008). Bcl-2 expression is elevated in R5 HIV-1-producing CXCR5+ T cells in vitro, which may contribute to propagation of R5 virus in B cell follicles in vivo.

Original languageEnglish (US)
Pages (from-to)298-304
Number of pages7
JournalAIDS Research and Human Retroviruses
Volume31
Issue number3
DOIs
StatePublished - Mar 1 2015

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Lymphoid Tissue
HIV-1
Lymphocytes
Palatine Tonsil
Phytohemagglutinins
Viruses
T-Lymphocytes
Interleukin-2
B-Lymphocytes
HIV Infections
Cultured Cells
Cell Survival
Flow Cytometry

ASJC Scopus subject areas

  • Immunology
  • Virology
  • Infectious Diseases

Cite this

Distinct patterns of Bcl-2 expression occur in R5- and X4-tropic HIV-1-producing lymphoid tissue cells infected ex vivo. / Haas, Michelle K.; Levy, David; Folkvord, Joy M.; Connick, Elizabeth.

In: AIDS Research and Human Retroviruses, Vol. 31, No. 3, 01.03.2015, p. 298-304.

Research output: Contribution to journalArticle

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abstract = "Most HIV-1 replication occurs in secondary lymphoid tissues in T cells within B cell follicles. Mechanisms underlying the accumulation of HIV-1-producing cells at these sites are not understood. Antiapoptotic proteins such as Bcl-2 could promote follicular CD4+ T cell survival, contributing to sustained virus production. Tonsils obtained from subjects without known HIV infection were disaggregated and analyzed for Bcl-2 expression in follicular (CXCR5+) and extrafollicular (CXCR5-) CD3+CD4+ cells by flow cytometry. Additional tonsil cells were cultured with phytohemagglutinin (PHA) and interleukin-2 (IL-2) for 2 days, infected with either CCR5(R5) or CXCR4-tropic (X4) GFP reporter viruses, and analyzed for Bcl-2 expression. In freshly disaggregated CD3+CD4+ tonsil cells, mean florescence intensity (MFI) for Bcl-2 was higher in CXCR5+ (median, 292) compared to CXCR5- cells (median, 194; p=0.001). Following in vitro stimulation with PHA and IL-2, Bcl-2 MFI was higher in both CXCR5+ cells (median, 757; p=0.03) and CXCR5- cells (median, 884; p=0.002) in uninfected cultures compared to freshly isolated tonsil cells. Bcl-2 MFI was higher in GFP+CD3+CD8- R5-producing cells (median, 554) than in X4-producing cells (median, 393; p=0.02). Bcl-2 MFI was higher in R5-producing CXCR5+ cells (median, 840) compared to all other subsets including R5-producing CXCR5- cells (median, 524; p=0.04), X4-producing CXCR5+ cells (median, 401; p=0.02), and X4-producing CXCR5- cells (median, 332; p=0.008). Bcl-2 expression is elevated in R5 HIV-1-producing CXCR5+ T cells in vitro, which may contribute to propagation of R5 virus in B cell follicles in vivo.",
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