Time-lapse monitoring of TLR2 ligand internalization with newly developed fluorescent probes

Arai, Yohei; Yokoyama, Kazuhiko; Kawahara, Yukie; Qi, Feng; Ohta, Ippei; Shimoyama, Atsushi; Inuki, Shinsuke; Fukase, Koichi; Kabayama, Kazuya; Fujimoto, Yukari

doi:10.1039/c7ob03205f

Cited by 5 publications

(3 citation statements)

References 32 publications

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“…Probably, promotion of antigen uptake through the recognition by TLR is required for the initiation of immune responses. 67 It should also be mentioned that V1 produced antibodies that selectively recognize tri-STn over STn, as expected.…”

Section: Resultssupporting

confidence: 71%

Precise immunological evaluation rationalizes the design of a self-adjuvanting vaccine composed of glycan antigen, TLR1/2 ligand, and T-helper cell epitope

et al. 2022

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Section: Resultssupporting

confidence: 71%

Precise immunological evaluation rationalizes the design of a self-adjuvanting vaccine composed of glycan antigen, TLR1/2 ligand, and T-helper cell epitope

et al. 2022

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“…Indeed, TLR2 has been shown to be of importance in the signalling of the antimicrobial human beta-defensin (hBD)-3 34 . Since TLR2, an anionic membrane receptor expressed by leukocytes 35 that binds exogenous as well as endogenous agents 36 that may internalized leading to cell activation 37 and since CXCL2 expression upon Mtb infection involves TLR2 signaling 38 , we were wondering if IL-26 uses TLR2 to mediate chemokines and cytokine expression in these cells.…”

Section: Resultsmentioning

confidence: 99%

Interleukin-26 activates macrophages and facilitates killing of Mycobacterium tuberculosis

Hawerkamp

Geelen

Korte

et al. 2020

Sci Rep

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Tuberculosis-causing Mycobacterium tuberculosis (Mtb) is transmitted via airborne droplets followed by a primary infection of macrophages and dendritic cells. During the activation of host defence mechanisms also neutrophils and T helper 1 (TH1) and TH17 cells are recruited to the site of infection. The TH17 cell-derived interleukin (IL)-17 in turn induces the cathelicidin LL37 which shows direct antimycobacterial effects. Here, we investigated the role of IL-26, a TH1- and TH17-associated cytokine that exhibits antimicrobial activity. We found that both IL-26 mRNA and protein are strongly increased in tuberculous lymph nodes. Furthermore, IL-26 is able to directly kill Mtb and decrease the infection rate in macrophages. Binding of IL-26 to lipoarabinomannan might be one important mechanism in extracellular killing of Mtb. Macrophages and dendritic cells respond to IL-26 with secretion of tumor necrosis factor (TNF)-α and chemokines such as CCL20, CXCL2 and CXCL8. In dendritic cells but not in macrophages cytokine induction by IL-26 is partly mediated via Toll like receptor (TLR) 2. Taken together, IL-26 strengthens the defense against Mtb in two ways: firstly, directly due to its antimycobacterial properties and secondly indirectly by activating innate immune mechanisms.

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“…This improvement could help detect a relatively small amount of the probe for a long time without significant photobleaching, and this clarified the recycling pathway. Using a similar concept, we previously imaged fluorescently labeled ligands of Toll-like receptor 2 diluted to physiologically active concentrations and successfully visualized receptor expression-dependent endocytosis (23).…”

Section: Discussionmentioning

confidence: 99%

Temporal analysis of localization and trafficking of glycolipids

Arai

Kabayama

Kanie

et al. 2020

Preprint

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Glycolipid metabolism occurs in the Golgi apparatus, but the detailed mechanisms have not yet been elucidated. We used fluorescently labeled glycolipids to analyze glycolipid composition and localization changes and shed light on glycolipid metabolism. In a previous study, the fatty chain of lactosyl ceramide was fluorescently labeled with BODIPY (LacCer-BODIPY) before being introduced into cultured cells to analyze the cell membrane glycolipid recycling process. However, imaging analysis of glycolipid recycling is complicated because of limited spatial resolution. Therefore, we examined the microscopic conditions that allow the temporal analysis of LacCer-BODIPY trafficking and localization. We observed that the glycolipid fluorescent probe migrated from the cell membrane to intracellular organelles before returning to the cell membrane. We used confocal microscopy to observe co-localization of the glycolipid probe with endosomes and Golgi markers, demonstrating that it recycles mainly through the trans-Golgi network (TGN). Here, a glycolipid recycling pathway was observed that did not require the lipids to pass through the lysosome.

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Time-lapse monitoring of TLR2 ligand internalization with newly developed fluorescent probes

Cited by 5 publications

References 32 publications

Precise immunological evaluation rationalizes the design of a self-adjuvanting vaccine composed of glycan antigen, TLR1/2 ligand, and T-helper cell epitope

Precise immunological evaluation rationalizes the design of a self-adjuvanting vaccine composed of glycan antigen, TLR1/2 ligand, and T-helper cell epitope

Interleukin-26 activates macrophages and facilitates killing of Mycobacterium tuberculosis

Temporal analysis of localization and trafficking of glycolipids

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