Interleukin-27 promotes autophagy in human serum-induced primary macrophages via an mTOR- and LC3-independent pathway

Abstract

Interleukin-27 (IL-27) is a cytokine that suppresses human immunodeficiency virus (HIV)-1 infection in macrophages and is considered as an immunotherapeutic reagent for infectious diseases. It is reported that IL-27 suppresses autophagy in Mycobacterium tuberculosis-infected macrophages; however, a role for IL-27 on autophagy induction has been less studied. In this study, we investigated the impact of IL-27 in both autophagy induction and HIV-1 infection in macrophages. Primary human monocytes were differentiated into macrophages using human AB serum (huAB) alone, macrophage-colony stimulating factor (M-CSF) alone, or a combination of IL-27 with huAB or M-CSF. Electron microscopy and immunofluorescence staining demonstrated that a 20-fold increase in autophagosome formation was only detected in IL-27 + huAB-induced macrophages. Western blot analysis indicated that the autophagosome induction was not linked to either dephosphorylation of the mammalian target of rapamycin (mTOR) or lipidation of microtubule-associated protein 1A/1B-light chain 3 (LC3), an autophagosomal marker, implying that IL-27 can induce autophagy through a novel non-canonical pathway. Here we show for the first time that IL-27 induces autophagy during monocyte-to-macrophage differentiation in a subtype-dependent manner.

Escherichia coli grown in cost-effective conical tubes produces more plasmid DNA

“…To determine whether the increased output effect was E. coli strain- or plasmid size-dependent, we compared extraction of a 15.5 kbp plasmid from the STABL3 strain (Invitrogen) [9]. STABLE3 growth was similar levels in F-Round-PP, O-Conical-PP, and F-Conical-PS and the amounts of plasmid DNA from those three tubes were comparable (Figure 1C).…”

“…Glycerol stock of Escherichia coli ( E. coli ) (TOP10 strain, Invitrogen, Thermo Fisher Science, Waltham, MA, USA) transformed with plasmid pCR2.1 (Invitrogen) containing 3.6 kbp DNA fragment (a total DNA size is 7.5 kbp) [8], or Stbl3 (Invitrogen) with plasmid pUC19 (Invitrogen) containing 12.8 kbp DNA (total size is 15.5 kbp) [9] was streaked on Luria-Bertani (LB) media (Quality Biological, Germantown, MD, USA)-Bacto Agar (Becton Dickinson, Franklin Lakes, NJ, USA) plate containing 100µg/ml ampicillin (MilliporeSigma, Burlington, MA, USA) (LA-plate), Five single colonies were individually inoculated in 5 mL of LA medium in 50 mL Falcon polypropylene (PP) tube (Cat# 352098) and then cultured for overnight at 37°C at 225 rpm. The next day, one % of the overnight culture was reinoculated into tubes containing warm LA (1/5 vol of each tube capacity; detail of the volume is described in the results and discussion) and cultured for 16 hr at 37°C at 225 rpm, and then, 1 mL of each bacteria suspension was subjected to isolate plasmid DNA using the S.N.A.P.…”

PM2.5 induces autophagy‐dependent ferroptosis by endoplasmic reticulum stress in SH‐SY5Y cells

“…During autophagy, LC3I is decomposed to produce LC3II with a lower molecular weight, and the content of LC3II is proportional to the degree of autophagy. P62 is a specific substrate marker for autophagy, which can interact with LC3 to infiltrate into autophagosomes and effectively degrade by autophagolysosomes (Laverdure et al, 2021; Shi et al, 2019; Wu et al, 2018). The result reflected that beclin‐1 expression was significantly increased in the 80 and 160 μg/mL PM 2.5 exposure groups, indicating that autophagy was massively activated.…”

Molecular Regulation Mechanism of Microglial Autophagy in the Pathology of Alzheimer's Disease

“…For example, IL-27 promoted the survival of Mycobacterium tuberculosis-infected macrophages by inhibiting autophagy [23]. Meanwhile, Laverdure et al reported that IL-27 promotes autophagy in stimulated primary human macrophages [24]. These contrasting effects of IL-27 reported in previous studies warrant further investigation into the matter.…”