Ageing promotes early T follicular helper cell differentiation by modulating expression of RBPJ.

Ageing profoundly changes our immune system and is thought to be a driving factor in the morbidity and mortality associated with infectious disease in older people. We have previously shown that the impaired immunity to vaccination that occurs in aged individuals is partly attributed to the effect of age on T follicular helper (Tfh) cell formation. In this study, we examined how age intrinsically affects Tfh cell formation in both mice and humans. We show increased formation of Tfh precursors (pre-Tfh) but no associated increase in germinal centre (GC)-Tfh cells in aged mice, suggesting age-driven promotion of only early Tfh cell differentiation. Mechanistically, we show that ageing alters TCR signalling which drives expression of the Notch-associated transcription factor, RBPJ. Genetic or chemical modulation of RBPJ or Notch rescues this age-associated early Tfh cell differentiation, and increased intrinsic Notch activity recapitulates this phenomenon in younger mice. Our data offer mechanistic insight into the age-induced changes in T-cell activation that affects the differentiation and ultimately the function of effector T cells

Optimized immunofluorescence staining protocol for imaging germinal centers in secondary lymphoid tissues of vaccinated mice

Summary: Location of immune cells that form the germinal center reaction within secondary lymphoid tissues can be characterized using confocal microscopy. Here, we present an optimized immunofluorescence staining protocol to image germinal center structures in fixed/frozen spleen sections from ChAdOx1 nCoV-19 immunized mice. This protocol can be adapted to identify other cell types within secondary lymphoid tissues.For complete information on the generation and use of this protocol to examine immune responses to the COVID vaccine ChAdOx1 nCoV-19, please refer to Silva-Cayetano et al. (2020)

Spatial dysregulation of T follicular helper cells impairs vaccine responses in aging.

Acknowledgements: We thank C. Vinuesa and A. Liston for critical feedback on this paper. We thank the Babraham Institute Biological Support Unit staff, who performed in vivo treatments of our animals and took care of animal husbandry. We thank the staff of the Babraham Flow Cytometry and Imaging Facilities for their technical support. The National Institute for Health Research Cambridge Biomedical Research Center is a partnership between Cambridge University Hospitals NHS Foundation Trust and the University of Cambridge, funded by the National Institute for Health Research. We thank the National Institute for Health Research Cambridge Biomedical Research Center volunteers for their participation and thank staff for their contribution in coordinating the vaccinations and venipuncture. This study was supported by funding from the Biotechnology and Biological Sciences Research Council (grant nos. BB/W001578/1, BBS/E/B/000C0407, BBS/E/B/000C0427 to M.A.L.; grant no. BBSRC BB/N011740/1 to A.E.D; and the Campus Capability Core Grant to the Babraham Institute), the European Union’s Horizon 2020 research and innovation program ‘ENLIGHT-TEN’ under the Marie Sklodowska-Curie grant agreement no. 675395 to M.A.L., a grant from IdEx Université de Paris (grant no. ANR-18-IDEX-0001 to M.E.) and by an ANR PRC grant (grant no. ANR-17-CE14-0019 to K.B.). M.A.L. is an EMBO Young Investigator and a Lister Institute Prize Fellow. D.B. was supported by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Emmy Noether Programs BA 5132/1-1 and BA 5132/1-2 (grant no. 252623821 to D.B.), SFB 1054 Project B12 (grant no. 210592381 to D.B.) and Germany’s Excellence Strategy EXC2151 (grant no. 390873048 to D.B.). P.A.R. was supported by the Human Frontier Science Program (grant no. RGP0033/2015 to P.A.R.) and a PhD fellowship granted by École Normale Supérieure de Lyon. J.L.L. is supported by a National Science Scholarship (PhD) by the Agency for Science, Technology and Research, Singapore. D.L.H. received a National Health and Medical Research Council Australia Early-Career Fellowship (grant no. APP1139911). A.R.B. received a Sir Henry Wellcome Postdoctoral Fellowship (grant no. 222793/Z/21/Z). J.P.L. was a recipient of the People Program (Marie Curie Actions) of the European Union’s Seventh Framework Program (FP7/2007-2013) under REA grant agreement no. PCOFUND-GA-2013-609102.The magnitude and quality of the germinal center (GC) response decline with age, resulting in poor vaccine-induced immunity in older individuals. A functional GC requires the co-ordination of multiple cell types across time and space, in particular across its two functionally distinct compartments: the light and dark zones. In aged mice, there is CXCR4-mediated mislocalization of T follicular helper (TFH) cells to the dark zone and a compressed network of follicular dendritic cells (FDCs) in the light zone. Here we show that TFH cell localization is critical for the quality of the antibody response and for the expansion of the FDC network upon immunization. The smaller GC and compressed FDC network in aged mice were corrected by provision of TFH cells that colocalize with FDCs using CXCR5. This demonstrates that the age-dependent defects in the GC response are reversible and shows that TFH cells support stromal cell responses to vaccines