IDENTIFICATION OF SIGNALING PATHWAYS BY WHICH CD40 STIMULATES AUTOPHAGY AND ANTI-MICROBIAL ACTIVITY AGAINST TOXOPLASMA GONDII IN MACROPHAGES

Infect Immun. 2016 Jun 27. pii: IAI.00101-16. [Epub ahead of print]

Liu E¹, Lopez Corcino Y², Portillo JC², Miao Y², Subauste CS³.

CD40 is an important stimulator of autophagy and autophagic killing anti-Toxoplasma gondii in host cells. In contrast to autophagy induced by nutrient deprivation or pattern recognition receptors, less is known about the effects of cell-mediated immunity on Beclin 1 and ULK1, key regulators of autophagy. Here we studied molecular mechanisms by which CD40 stimulates autophagy in macrophages. CD40 ligation caused biphasic JNK phosphorylation. The second phase of JNK phosphorylation was dependent on autocrine production of TNF-α. TNF-α and JNK signaling were required for CD40-induced increase in autophagy. JNK signaling downstream of CD40 caused Ser-87 phosphorylation of Bcl-2 and dissociation between Bcl-2 and Beclin 1, an event known to stimulate the autophagic function of Beclin 1. However, TNF-α alone was unable to stimulate autophagy. CD40 also stimulated autophagy via a pathway that included CaMKKβ, AMPK and ULK1. CD40 caused AMPK phosphorylation at its activating site Thr-172. This effect was mediated by CaMKKβ and was not impaired by neutralization of TNF-α. CD40 triggered AMPK-mediated Ser-555 phosphorylation of ULK1. CaMKKβ, AMPK and ULK1 were required for CD40-induced increase in autophagy. CD40-mediated autophagic killing of Toxoplasma gondii is known to require TNF-α. Knockdown of JNK, CaMKKβ, AMPK, or ULK1 prevented T. gondii killing in CD40-activated macrophages. The second phase of JNK phosphorylation - Bcl-2 phosphorylation - Bcl-2-Beclin 1 dissociation, and AMPK phosphorylation - ULK1 phosphorylation occurred simultaneously at approximately 4 h post-CD40 stimulation. Thus, CaMKKβ and TNF-α are upstream molecules by which CD40 acts on ULK1 and Beclin 1 to stimulate autophagy and killing of T. gondii.
Copyright © 2016, American Society for Microbiology. All Rights Reserved.

PMID:

27354443

DOI:

10.1128/IAI.00101-16

[PubMed - as supplied by publisher]