The inflammaging microenvironment induces dysfunctional rewiring of Tfh cell differentiation
Cody S. Nelson, Manuel A. Podestà, Maya G. Gempler, Jeong-Mi Lee, Cole J. Batty, Peterson G. Mathenge, Asra Sainju, Matthew R. Chang, Hanzhong Ke, Pragya Chandrakar, Elsa Bechu, Sierra Richardson, Cecilia B. Cavazzoni, Stefan G. Tullius, Reza Abdi, Musie Ghebremichael, Marcia C. Haigis, Wayne A. Marasco, Peter T. Sage
Cody S. Nelson, Manuel A. Podestà, Maya G. Gempler, Jeong-Mi Lee, Cole J. Batty, Peterson G. Mathenge, Asra Sainju, Matthew R. Chang, Hanzhong Ke, Pragya Chandrakar, Elsa Bechu, Sierra Richardson, Cecilia B. Cavazzoni, Stefan G. Tullius, Reza Abdi, Musie Ghebremichael, Marcia C. Haigis, Wayne A. Marasco, Peter T. Sage
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Research Article Aging Immunology

The inflammaging microenvironment induces dysfunctional rewiring of Tfh cell differentiation

Abstract

Humoral immunity is orchestrated by follicular helper T (Tfh) cells, which promote cognate B cells to produce high-affinity, protective antibodies. In aged individuals, humoral immunity after vaccination is diminished despite the presence of Tfh cells, suggesting defects after initial Tfh cell formation. In this study, we utilized both murine and human systems to investigate how aging alters Tfh cell differentiation after influenza vaccination. We found that young Tfh cells underwent progressive differentiation after influenza vaccination, culminating in clonal expansion of effector-like cells in both draining lymph nodes and blood. In aging, early stages of Tfh cell development occurred normally. However, aging rewired the later stages of development in Tfh cells, resulting in a transcriptional program reflective of cellular senescence, sustained pro-inflammatory cytokine production, and metabolic reprogramming. We investigated the extent to which this rewiring of aged Tfh cells is due to the age-associated inflammatory (“inflammaging”) microenvironment and found that this setting was sufficient to both block the transition of Tfh cells to a post-effector resting state and skew Tfh cells toward the age-rewired state. Together, these data suggest that aging dampens humoral immunity by cytokine-mediated rewiring of late effector Tfh cell differentiation into an activated, yet less functional, cellular state.

Authors

Cody S. Nelson, Manuel A. Podestà, Maya G. Gempler, Jeong-Mi Lee, Cole J. Batty, Peterson G. Mathenge, Asra Sainju, Matthew R. Chang, Hanzhong Ke, Pragya Chandrakar, Elsa Bechu, Sierra Richardson, Cecilia B. Cavazzoni, Stefan G. Tullius, Reza Abdi, Musie Ghebremichael, Marcia C. Haigis, Wayne A. Marasco, Peter T. Sage

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Figure 7

The inflammaging microenvironment rewires Tfh cell differentiation to a hyperinflammatory and senescent-like transcriptional state.

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The inflammaging microenvironment rewires Tfh cell differentiation to a ...
(A) Experimental schematic to assess effects of the aged microenvironment on Tfh differentiation in vivo. Total splenic CD4+ T cells from OT-II+IL21FM/Rep (OT-II+Tg.Il21CreRosa26LoxSTOPLoxTdTomatoIl21VFP) were adoptively transferred to 8-week-old (young) or 80-week-old (aged) mice, which were given an NP-OVA vaccine. (B) The frequency of dLN OT-II+ cells from the total CD4+ population (left) and of fully differentiated effector Tfh (middle) and TfhEx (right) from the total OT-II+ population. Each point represents an individual mouse. Results are reflective of 1 representative experiment (left, middle) or concatenated data from 2 experiments (right). (C) Experimental schematic to assess effects of the inflammaging microenvironment on human late effector Tfh differentiation in vitro. CD4+CXCR5+ Tfh cells sorted from healthy donors were incubated with an inflammaging cocktail containing 10 ng/mL each of IL-6, IL-1β, and TNF-α. (D) Tfh cell count (left) and viability (right) by spectral flow cytometry. (E) Tfh cell polarization to Tfh1 (left; CXCR3+CCR6–), Tfh2 (middle; CXCR3–CCR6–), or Tfh17 (right; CXCR3–CCR6+). (F) Expression of Tfh activation markers PD-1 (left), ICOS (middle), and CD40L (right). (G) Concentration of IL-21 (left), IL-4 (middle), and IFN-γ (right) in culture supernatants. (H) (Left) Frequency of class-switched B cells from cultures as in C. (Right) Concentration of IgG in culture supernatant in experiments. (I) Heatmap of genes in Tfh cells cultured in control (blue) or inflammaging (gray) conditions assessed 7 days after culture. (J) Volcano plot of P value and fold-change for genes in Tfh cells (red triangles P < 0.01; fold-change > 2). (K) GSEA of selected Hallmark gene modules and senescence gene module (29) in Tfh cells from I. All displayed data points represent mean value of technical replicates. *P < 0.05, uncorrected Mann-Whitney U test or Wilcoxon’s signed-rank test.