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Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front. Med.    2017, Vol. 11 Issue (4) : 536-547    https://doi.org/10.1007/s11684-017-0592-x
RESEARCH ARTICLE |
Irreversible phenotypic perturbation and functional impairment of B cells during HIV-1 infection
Jingjing Yan, Shuye Zhang, Jun Sun, Jianqing Xu(), Xiaoyang Zhang()
Shanghai Public Health Clinical Center & Institutes of Biomedical Sciences, Fudan University, Shanghai 201508, China
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Abstract

Human immunodeficiency virus type 1 (HIV-1) infection can damage humoral immunity. The knowledge of B cell perturbations during chronic HIV-1 infection and their recovery after combined antiretroviral therapy (cART) is not complete yet, and thus attempts to further improve humoral immunity are impeded. In this study, an HIV-1 chronically infected cohort with similar demographics, infection history, genetic background, and HIV-1 genotype was established to probe B cell perturbations. Results showed that the B cells from this cohort were highly activated and prone to cell death, and B cell compartments were altered significantly. Notably, although cART partially reversed the hyperactivation and reduced tissue-like memory B cells, other B cell perturbations, including impaired expression of survival factor Bcl-2, costimulatory molecules, and shrunken resting memory B cells, were irreversible. Further functional characterization revealed that the influenza HA-specific antibody-secreting cells were significantly lower during HIV-1 infection, whereas the recalled antibody response to HIV-1-specific antigens was decreased after cART. Finally, CpG plus R848 treatment increased the survival of B cells and memory B cells in vitro from HIV-1-infected patients. In conclusion, this study identified irreversible B cell immune perturbations in chronic HIV-1 infections regardless of cART and proposed the potential strategy to enhance B cell functions through the improvement of cell survival.

Keywords B cell      HIV-1      phenotype      functionality      antiretroviral therapy     
Corresponding Authors: Jianqing Xu,Xiaoyang Zhang   
Just Accepted Date: 30 October 2017   Issue Date: 04 December 2017
 Cite this article:   
Jingjing Yan,Shuye Zhang,Jun Sun, et al. Irreversible phenotypic perturbation and functional impairment of B cells during HIV-1 infection[J]. Front. Med., 2017, 11(4): 536-547.
 URL:  
http://academic.hep.com.cn/fmd/EN/10.1007/s11684-017-0592-x
http://academic.hep.com.cn/fmd/EN/Y2017/V11/I4/536
HIV negativeHIV-1 infected
ART naive ART treated
Number of participants123234
Age, year a25 (21–32)47.5 (34–66)50.5 (33–70)
Male sex, n (%)6 (50)17 (53)23 (68)
CD4+ T cell count (cell/mL) a756 (508–1181)390 (128–950) c390 (73–1020) c
CD8+ T cell count (cell/mLa)624 (289–913)1112 (306–3154) c855 (377–2516) b
B cell count (cell/mL) a190 (98–508)125 (26–428) b180 (40–794)
Duration of infection (month) aN/A89 (60–178)89 (13–166)
Duration of treatment (month) aN/AN/A29.5 (6–94)
HIV RNA (copy/mL) aN/A5600 (60–110 000)<50
Tab.1  Baseline characteristics of study participants
Fig.1  Peripheral lymphocyte counts in HIV-1-infected ART-naive and ART-treated individuals and HIV-negative individuals. (A) Peripheral CD4+ T cell counts (cell/mL) from three groups: HIV-negative, HIV-1-infected ART-naive, and ART-treated individuals. (B) Peripheral CD19+ B cell counts (cell/mL) from the three groups. (C) Spearman rank correlation of CD4+ T cell counts and B cell counts in the HIV-1-infected ART-naive group. Symbols represent individuals samples, horizontal bars represent means, and error bars show SEM. Differences were evaluated using the Mann–Whitney U test for intergroup comparisons and were considered significant at P<0.05(*) or P<0.01(**).
Fig.2  Altered phenotype of B cells during HIV-1 infection. (A) Representative gating strategy for molecule expression on CD19+CD20+ B cell. The scatter plots showed the percentage of activated molecules CD38 (B), cell turnover marker Ki-67 (C), apoptosis-associated molecules CD95 (D), Bcl-2 (E), PD-1 (F), and costimulatory molecules CD40 (G) and CD70 (H) among the HIV-negative, HIV-1-infected ART-naive, and HIV-1-infected ART-treated individuals. Symbols represent individual samples, horizontal bars represent means, and error bars show SEM. Differences were evaluated using the Mann–Whitney U test for intergroup comparisons and were considered significant at P<0.05(*) or P<0.01(**).
Fig.3  Perturbations of peripheral blood B cell subpopulations in chronic HIV-1 infection. HIV-1-infected ART-naive and ART-treated and HIV-negative individuals were investigated. (A) CD19+ B cells were gated from forward scatters and side scatters. B cell subpopulations were determined using flow cytometry as CD10CD27+ immature B cells, CD10CD21+CD27 naive B cells, CD10CD21+CD27+ resting memory B cells, CD10CD21CD27+ activated memory B cells, CD10CD21CD27 tissue-like memory B cells, and CD10CD21CD27++CD20 plasmablasts. The distribution of B cell subpopulations was shown in the representative HIV-negative individual. (B) The percentage of B cell subpopulations were shown among the three groups. Boxes represent the 25th–75th percentiles with medians, and whiskers represent min to max. Differences were evaluated using the Mann–Whitney U test for intergroup comparisons and were considered significant at P<0.05(*) or P<0.01(**).
Fig.4  Expression of activated, apoptosis-associated, and costimulatory molecules on B cell subpopulations in chronic HIV-1 infection. The bar plots represent the expression of activated molecule CD38 (A); cell turnover marker Ki-67 (B); apoptosis-associated molecules CD95 (C), Bcl-2 (D), and PD-1 (E); and costimulatory molecules CD40 (F) and CD70 (G) among the HIV-negative, HIV-1-infected ART-naive, and HIV-1-infected ART-treated individuals. The data were shown as mean with SEM. Differences were evaluated using the Mann–Whitney U test for intergroup comparisons and were considered significant at P<0.05(*) or P<0.01(**).
Fig.5  Impaired responses of B cells to polyclonal stimulation in chronic HIV-1 infection. (A) Strategy for evaluating functional responses of B cells from the HIV-negative, HIV-1-infected ART-naive, and HIV-1-infected ART-treated individuals. (B) Stimulated B cells were used for ELISpot to measure the frequencies of ASCs for total IgG and against soluble forms of monomeric HIV-1 gp120 envelopes and H1N1 influenza HA proteins. Spearman rank correlation was employed to analyze the correlation of CD95 expression and HA-specific ASCs among the three groups. (C) Culture supernatants were collected and assayed by CBA. The columns in the graphical chart represented the mean concentrations of each cytokine from B cells. Responses were measured in 12 HIV-negative, 12 ART-naive, and 12 ART-treated individuals. Symbols represent individuals samples, horizontal bars represent means, and error bars show SEM. Differences were evaluated using the Mann–Whitney U test for intergroup comparisons and were considered significant at P<0.05(*) or P<0.01(**).
Fig.6  CpG plus R848 increased the survival of B cells. B cells from 6 HIV-1-infected ART-naive and 6 ART-treated individuals were cultured in the absence (unsti) or presence (sti) of CpG (5mg/mL) and R848 (1mg/mL) for 24 h, followed by Annexin V staining. Of the 12 individuals, CD27+ B cells from 8 individuals were enriched and cultured in the absence or presence of CpG and R848 for 24 h, followed by Annexin V staining. Representative flow cytometric dot plots of Annexin V staining and means and interquartile ranges of the percentages of Annexin V-expressing cells are shown.
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