A favorable follicular helper CD4+ T cell programming characterizes neutralization activity in chronic HIV infection
Eirini Moysi, Ashish A. Sharma, Sijy O’Dell, Spiros Georgakis, Perla Mariana Del Rio Estrada, Ghneim Khader, Alonso Arana, Fernanda Torres-Ruiz, Mauricio González Navarro, Yara Andrea Luna Villalobos, Santiago Avila Rios, Gustavo Reyes-Teran, Margaret H. Beddall, Sung Hee Ko, Frida Belinky, Michail Orfanakis, Laurence de Leval, Ana B. Enriquez, Clarisa M. Buckner, Susan Moir, Helen Lindsay, Raphael Gottardo, Nicole Doria-Rose, Eli A. Boritz, John R. Mascola, Rafick-Pierre Sekaly, Richard A. Koup, Constantinos Petrovas
Eirini Moysi, Ashish A. Sharma, Sijy O’Dell, Spiros Georgakis, Perla Mariana Del Rio Estrada, Ghneim Khader, Alonso Arana, Fernanda Torres-Ruiz, Mauricio González Navarro, Yara Andrea Luna Villalobos, Santiago Avila Rios, Gustavo Reyes-Teran, Margaret H. Beddall, Sung Hee Ko, Frida Belinky, Michail Orfanakis, Laurence de Leval, Ana B. Enriquez, Clarisa M. Buckner, Susan Moir, Helen Lindsay, Raphael Gottardo, Nicole Doria-Rose, Eli A. Boritz, John R. Mascola, Rafick-Pierre Sekaly, Richard A. Koup, Constantinos Petrovas
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Research Article AIDS/HIV Immunology

A favorable follicular helper CD4+ T cell programming characterizes neutralization activity in chronic HIV infection

Abstract

A subset of people living with HIV (PLWH) can produce broadly neutralizing antibodies (bNAbs) against HIV, but the lymph node (LN) dynamics that promote the generation of these Abs are poorly understood. Here, we explored LN-associated histological, immunological, and virological determinants of bNAb generation in a cohort of antiretroviral therapy–naive PLWH. We found that participants who produce bNAbs, termed “neutralizers” (Ns), have a better-preserved LN-associated B cell follicle architecture than do PLWH who do not. The former was associated with a substantially higher in situ prevalence of B-cell lymphoma 6 (Bcl-6hi) follicular helper CD4+ T cells (Tfh), expressing a molecular program that favors their differentiation and stemness, and substantially reduced IL-10 follicular suppressor CD4+ T cells. Furthermore, our data reveal possible molecular targets mediating Tfh–B cell interactions in Ns. Together, we identify germinal center cellular and molecular signatures that could contribute to the development of bNAbs in PLWH.

Authors

Eirini Moysi, Ashish A. Sharma, Sijy O’Dell, Spiros Georgakis, Perla Mariana Del Rio Estrada, Ghneim Khader, Alonso Arana, Fernanda Torres-Ruiz, Mauricio González Navarro, Yara Andrea Luna Villalobos, Santiago Avila Rios, Gustavo Reyes-Teran, Margaret H. Beddall, Sung Hee Ko, Frida Belinky, Michail Orfanakis, Laurence de Leval, Ana B. Enriquez, Clarisa M. Buckner, Susan Moir, Helen Lindsay, Raphael Gottardo, Nicole Doria-Rose, Eli A. Boritz, John R. Mascola, Rafick-Pierre Sekaly, Richard A. Koup, Constantinos Petrovas

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Figure 3

Tfh cells are characterized by a molecular profile favoring their differentiation and longevity in cross-neutralization participants.

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Tfh cells are characterized by a molecular profile favoring their differ...
(A) scRNA analysis workflow. DEG, differentially expressed gene. (B) UMAP projections of total CD4+ T cells with color-coded cell populations and CD4+ T cell subpopulations identified and their percentages in NNs versus Ns. (C and D) Volcano plots and dot plots are shown to illustrate transcriptomic differences and pathway enrichment between Ns (Neut) and NNs (NonNeut). DEGs were identified using the MAST (hurdle model–based) test implemented in the FindMarkers function of the Seurat package in R. P values were adjusted by the Benjamini-Hochberg method, and genes meeting an adjusted P < 0.05 and |log2fold change [FC]| > 0.5 are highlighted in the volcano plots. Blue and red dots in the volcano plots represent genes that are significantly downregulated or upregulated in Ns, respectively. The accompanying dot plots depict average scaled expression (z scores) and the percentage of cells expressing each gene within Tfh or Treg subsets. Color intensity reflects relative expression, while dot size indicates the fraction of expressing cells. NESs from the GSEA are shown separately for selected pathways and transcription factor target gene sets. (E) Representative confocal images showing FOXP3hi CD4+ T cells in extrafollicular and follicular areas in NNs versus Ns. FoxP3hi CD4+ T cells (white arrows) were defined by means of concurrent CD4 (green) and FoxP3 (red) expression within CD20 (cyan) and Ki67 (yellow) double-positive areas. Original magnification, ×40; scale bars: 100 μm and 20 μm (enlarged images corresponding to the white dotted enclosures). Superplots showing the normalized per area imaged FoxP3hi CD4+ T cell numbers of both individual (faint background symbols) and mean (bold foreground symbols) follicular measurements (± SEM) in NNs (n = 6, blue) compared with Ns (n = 6, red), as measured by quantitative imaging analysis and histocytometry. The different symbols represent different donors. For statistical comparisons using the mean measurements, a Mann-Whitney U test was used. P = 0.1797. Superplots of CD25hiFoxP3hi (F, right) and IL-10hiCD25hiFoxP3hi (G, right) CD4+ T cells of both individual (faint background symbols) and mean (bold foreground symbols) follicular measurements in NNs (n = 6, blue) versus Ns (n = 5, red), as measured by quantitative imaging analysis and histocytometry. The different symbols represent different donors. For statistical comparisons using the mean measurements, a Mann-Whitney U test was applied (P = 0.426, P = 0.0823). Dot plots for extrafollicular (F, left) and whole tissue (G, left) data, where each participant tissue is represented by a circle of a different color. Mean values are shown in dot plots (E–G) and quartiles in box plots (F and G).