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Ecto-NTPDase CD39 is a negative checkpoint that inhibits follicular helper cell generation
Wenqiang Cao, … , Cornelia M. Weyand, Jörg J. Goronzy
Wenqiang Cao, … , Cornelia M. Weyand, Jörg J. Goronzy
Published May 26, 2020
Citation Information: J Clin Invest. 2020;130(7):3422-3436. https://doi.org/10.1172/JCI132417.
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Research Article Aging

Ecto-NTPDase CD39 is a negative checkpoint that inhibits follicular helper cell generation

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Abstract

Vaccination is a mainstay in preventive medicine, reducing morbidity and mortality from infection, largely by generating pathogen-specific neutralizing antibodies. However, standard immunization strategies are insufficient with increasing age due to immunological impediments, including defects in T follicular helper (Tfh) cells. Here, we found that Tfh generation is inversely linked to the expression of the ecto-NTPDase CD39 that modifies purinergic signaling. The lineage-determining transcription factor BCL6 inhibited CD39 expression, while increased Tfh frequencies were found in individuals with a germline polymorphism preventing transcription of ENTPD1, encoding CD39. In in vitro human and in vivo mouse studies, Tfh generation and germinal center responses were enhanced by reducing CD39 expression through the inhibition of the cAMP/PKA/p-CREB pathway, or by blocking adenosine signaling downstream of CD39 using the selective adenosine A2a receptor antagonist istradefylline. Thus, purinergic signaling in differentiating T cells can be targeted to improve vaccine responses, in particular in older individuals who have increased CD39 expression.

Authors

Wenqiang Cao, Fengqin Fang, Timothy Gould, Xuanying Li, Chulwoo Kim, Claire Gustafson, Simon Lambert, Cornelia M. Weyand, Jörg J. Goronzy

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Figure 2

Transcriptional regulation of ENTPD1.

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Transcriptional regulation of ENTPD1.
(A–C) Human peripheral T cells wer...
(A–C) Human peripheral T cells were transduced with RUNX3-overexpressing lentivirus (A), RUNX3 shRNA lentivirus (B), or BCL6-overexpressing lentivirus (C). Cells were cultured for 5 days after anti-CD3/anti-CD28 Dynabead activation. CD39 expression in CD4+GFP+ cells was determined by flow cytometry. Results are shown as representative contour plots (left) and summary plots from 6 to 8 experiments (right). (D) Purified human naive CD4+ T cells were activated by anti-CD3/anti-CD28 Dynabeads in the presence or absence of the BCL6 inhibitor 79-6 for 5 days; CD39 expression was determined by flow cytometry. Representative contour plots and summary data from 5 experiments. (E) Predicted BCL6, RUNX3, and p-CREB binding sites adjacent to the ENTPD1 transcription start site. (F and G) Human memory CD4+ cells were activated by anti-CD3/anti-CD28 Dynabeads for 3 days. BCL6 (F) or RUNX3 (G) binding was determined by ChIP-PCR for indicated sites. Results are shown as mean ± SEM of 3 experiments. (H and I) HEK-293T cells were cotransfected with pGL3-basic or pGL3 reporter construct containing the ENTPD1 promoter together with mock or BCL6-overexpressing (H) or RUNX3-overexpressing (I) plasmids. Luciferase activity was determined after 48 hours. Results are shown as mean ± SEM of 3 experiments. (J) Reporter activity in HEK-293T cells cotransfected with pNL3.3-basic or the ENTPD1 enhancer construct together with BCL6 or mock plasmid. Data were compared by 2-tailed paired t test (A–D), 1-way ANOVA with post hoc Tukey’s test (F and G), or unpaired t test (H–J). *P < 0.05; **P ≤ 0.01; ***P ≤ 0.001. NS, not significant.

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ISSN: 0021-9738 (print), 1558-8238 (online)

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