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  • Brief Communications Arising
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Evidence that CD32a does not mark the HIV-1 latent reservoir

Nature volume 561, pages E20–E28 (2018)Cite this article

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A Publisher Correction to this article was published on 12 October 2018

This article has been updated

arising from B. Descours et al. Nature 543, 564–567 (2017); https://doi.org/10.1038/nature21710

A recent report by Descours et al.1 suggests that the cell surface expression of the low affinity Fc receptor CD32a (also known as FcγRIIa) marks the replication-competent HIV-1 reservoir in CD4+ T cells from 12 HIV-1-infected participants receiving suppressive anti-retroviral therapy (ART)1. We have undertaken considerable efforts to replicate these findings using peripheral blood mononuclear cells (PBMCs) from 20 HIV-1-infected, ART-suppressed participants (Extended Data Table 1). We found no evidence to suggest that CD32a marks a CD4+ T cell population enriched in either HIV-1 DNA or replication-competent HIV-1 in our study participants. There is a Reply to this Comment by Descours, B. et al. Nature 561, https://doi.org/10.1038/s41586-s41586-018-0496-1 (2018).

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Fig. 1: CD32-expressing CD4+ T cells are not enriched in HIV-1 DNA and express markers of B cell origin.
Fig. 2: Flow cytometry imaging of sorted CD32-expressing T cells.

Change history

  • 12 October 2018

    In this Brief Communications Arising Comment, the first three authors (Osuna, Lim and Kublin) should have been listed as equally contributing authors; this has been corrected online.

References

  1. Descours, B. et al. CD32a is a marker of a CD4 T-cell HIV reservoir harbouring replication-competent proviruses. Nature 543, 564–567 (2017).

    Article  ADS  CAS  Google Scholar 

  2. Chun, T. W. et al. Quantification of latent tissue reservoirs and total body viral load in HIV-1 infection. Nature 387, 183–188 (1997).

    Article  ADS  CAS  Google Scholar 

  3. Finzi, D. et al. Identification of a reservoir for HIV-1 in patients on highly active antiretroviral therapy. Science 278, 1295–1300 (1997).

    Article  ADS  CAS  Google Scholar 

  4. Veri, M. C. et al. Monoclonal antibodies capable of discriminating the human inhibitory Fcγ-receptor IIB (CD32B) from the activating Fcγ-receptor IIA (CD32A): biochemical, biological and functional characterization. Immunology 121, 392–404 (2007).

    Article  CAS  Google Scholar 

  5. Boruchov, A. M. et al. Activating and inhibitory IgG Fc receptors on human DCs mediate opposing functions. J. Clin. Invest. 115, 2914–2923 (2005).

    Article  CAS  Google Scholar 

  6. Engelhardt, W., Matzke, J. & Schmidt, R. E. Activation-dependent expression of low affinity IgG receptors FcγRII(CD32) and FcγRIII(CD16) in subpopulations of human T lymphocytes. Immunobiology 192, 297–320 (1995).

    Article  CAS  Google Scholar 

  7. Sandilands, G. P. et al. Differential expression of CD32 isoforms following alloactivation of human T cells. Immunology 91, 204–211 (1997).

    Article  CAS  Google Scholar 

  8. Cohn, L. B. et al. Clonal CD4+ T cells in the HIV-1 latent reservoir display a distinct gene profile upon reactivation. Nat. Med. 24, 604–609 (2018).

    Article  CAS  Google Scholar 

  9. Cone, R. E., Sprent, J. & Marchalonis, J. J. Antigen-binding specificity of isolated cell-surface immunoglobulin from thymus cells activated to histocompatibility antigens. Proc. Natl Acad. Sci. USA 69, 2556–2560 (1972).

    Article  ADS  CAS  Google Scholar 

  10. Hwang, I. et al. T cells can use either T cell receptor or CD28 receptors to absorb and internalize cell surface molecules derived from antigen-presenting cells. J. Exp. Med. 191, 1137–1148 (2000).

    Article  CAS  Google Scholar 

  11. Wetzel, S. A., McKeithan, T. W. & Parker, D. C. Peptide-specific intercellular transfer of MHC class II to CD4+ T cells directly from the immunological synapse upon cellular dissociation. J. Immunol. 174, 80–89 (2005).

    Article  CAS  Google Scholar 

  12. Rosenits, K., Keppler, S. J., Vucikuja, S. & Aichele, P. T cells acquire cell surface determinants of APC via in vivo trogocytosis during viral infections. Eur. J. Immunol. 40, 3450–3457 (2010).

    Article  CAS  Google Scholar 

  13. Daubeuf, S. et al. Preferential transfer of certain plasma membrane proteins onto T and B cells by trogocytosis. PLoS One 5, e8716 (2010).

    Article  ADS  Google Scholar 

  14. Garside, P. et al. Visualization of specific B and T lymphocyte interactions in the lymph node. Science 281, 96–99 (1998).

    Article  ADS  CAS  Google Scholar 

  15. Okada, T. et al. Antigen-engaged B cells undergo chemotaxis toward the T zone and form motile conjugates with helper T cells. PLoS Biol. 3, e150 (2005).

    Article  Google Scholar 

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Author information

Author notes

  1. These authors contributed equally: Christa E. Osuna, So-Yon Lim, Jessica L. Kublin.

Authors and Affiliations

  1. Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA

    Christa E. Osuna, So-Yon Lim, Jessica L. Kublin, Elsa Chen & James B. Whitney

  2. Center for Human Immunology, National Institute of Allergy and Infectious Diseases, Bethesda, MD, USA

    Richard Apps

  3. Division of Infectious Diseases, Weill Department of Medicine, Weill Cornell Medical College, New York, NY, USA

    Talia M. Mota, Szu-Han Huang, Yanqin Ren, Nathaniel D. Bachtel, R. Brad Jones & Douglas F. Nixon

  4. Brigham and Women’s Hospital, Boston, Massachusetts Harvard Medical School, Boston, MA, USA

    Athe M. Tsibris

  5. Thayer School of Engineering, Dartmouth College, Hanover, NH, USA

    Margaret E. Ackerman

  6. Ragon Institute of MGH, MIT, and Harvard, Cambridge, MA, USA

    James B. Whitney

Authors
  1. Christa E. Osuna
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Contributions

D.F.N. and J.B.W. designed the studies. R.B.J., R.A., E.C., Y.R., N.D.B., C.E.O., R.T. and S.Y.L. led the virology assays. S.H.H., D.C., J.L.K., M.A. and C.E.O. led the immunology assays. J.B.W. led the studies and wrote the paper with all co-authors.

Corresponding author

Correspondence to James B. Whitney.

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Declared none.

Extended data figures and tables

Extended Data Fig. 1 Frequency and activation status of CD32-expressing CD4+ T cells and their HIV-1 DNA content.

a, The frequency of CD32high CD4+ T cells was measured by flow cytometry in PBMCs from ART-suppressed, HIV-1+ (n = 20) and HIV-1− (n = 8) donors. Bars denote median values. P values were determined by a Mann–Whitney test. b, DNA copies per cell in sorted subsets of total CD4+, CD4+CD32neg, CD4+CD32int and CD4+CD32high T cells are shown with median values and the IQR. The results are shown as either the actual HIV-1 DNA copies per million cells (filled symbols) or as estimated values calculated using the LOD and applied to the number of cells when the DNA input did not reach the threshold (open symbols). P values were determined by a Kruskal–Wallis test. c, The percentage of CD69, CD25 and HLA-DR expression was measured by flow cytometry on CD32neg and CD32high (FUN-2) CD4+ T cells from PBMCs from HIV-1+ participants (n = 20). Error bars show the median and IQR. P values were determined by Wilcoxon matched-pairs signed rank tests.

Extended Data Fig. 2 Detection of B cell proteins and mRNA in CD32-expressing CD4+ T cells.

a, CD32neg and CD32high (FUN-2) CD4+ T cells from human PBMCs were assessed by flow cytometry for the expression of CD19, CD40 and HLA-DR, and compared to B cells (CD3− CD14−CD19+ lymphocytes). Representative flow cytometry results of per cell antigen levels on B cells (top, blue histograms) and CD32neg and CD32high CD4+ T cells (bottom, grey and red histograms, respectively) from an HIV-1− donor. b, Representative CD32b staining of PBMCs from an HIV-1+, ART-suppressed participant. PBMCs were stained with an optimized concentration of the 2B6 monoclonal anti-CD32b antibody, followed by an antibody cocktail that included the FUN-2 monoclonal pan-CD32 antibody, as described in the Methods. Shown are the 2B6 and FUN-2 fluorescence minus one (FMO) antibody cocktail-stained samples and a sample co-stained with 2B6 and FUN-2. c, d, CD32 mRNA expression levels in CD4+CD32+ subsets. c, The relative expression of CD32A and CD32B mRNA isoforms in sorted CD4+CD32int and CD4+CD32high subsets from HIV-1+, ART-suppressed participants (n = 4). d, mRNA expression of CD32A and CD32B from patient G07W1610. e, T and B cell lineage-specific mRNA transcripts in sorted CD4+CD32+ subsets from participant G07W1610. Relative mRNA expression of target genes was normalized to ATCB using the comparative Ct method. Results are mean ± s.d. of each value from each participant (n = 4; c), or from values generated from two separate experiments using samples from the same patient (d).

Extended Data Fig. 3 Doublet composition of the sorted CD4+CD32high T cells.

Sorted B cells and CD4+CD32neg, CD4+CD32int and CD4+CD32high T cells from an HIV-1+, ART-suppressed participant were analysed using an Amnis imaging cytometer. Singlets and doublets were quantified using the aspect ratio and nuclear staining. a, The proportion of total singlet and doublet events among total nucleated cells detected on the Amnis cytometer in each sorted population was determined, and is shown as individual composite bar graphs for two patients (G07W1610 and OM5365). b, A composite bar graph of the proportion of conjugates, doublets and trogocytotic events that comprised the sorted CD4+CD32high population (n = 2).

Extended Data Table 1 Viral suppression of 20 HIV-1-infected participants on ART
Full size table
Extended Data Table 2 CD4+CD32high subset proportions and HIV-1 DNA compared to total CD4+ and CD32neg CD4+ T cells
Full size table

Supplementary information

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This file contains Supplementary Figure 1, Supplementary Methods and Supplementary References.

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Osuna, C.E., Lim, SY., Kublin, J.L. et al. Evidence that CD32a does not mark the HIV-1 latent reservoir. Nature 561, E20–E28 (2018). https://doi.org/10.1038/s41586-018-0495-2

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