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Differential Internalization of Thrombin-Derived Host Defense Peptides into Monocytes and Macrophages

Hansen, Finja C. LU ; Nadeem, Aftab LU ; Browning, Kathryn L. ; Campana, Mario ; Schmidtchen, Artur LU and Van Der Plas, Mariena J.A. LU (2022) In Journal of Innate Immunity 14(5). p.418-432
Abstract

Proteolytic cleavage of thrombin generates C-terminal host defense peptides exerting multiple immunomodulatory effects in response to bacterial stimuli. Previously, we reported that thrombin-derived C-terminal peptides (TCPs) are internalized in monocytes and macrophages in a time- and temperature-dependent manner. In this study, we investigated which endocytosis pathways are responsible for the internalization of TCPs. Using confocal microscopy and flow cytometry, we show that both clathrin-dependent and clathrin-independent pathways are involved in the internalization of the prototypic TCP GKY25 in RAW264.7 and human monocyte-derived M1 macrophages, whereas the uptake of GKY25 in monocytic THP-1 cells is mainly dynamin-dependent.... (More)

Proteolytic cleavage of thrombin generates C-terminal host defense peptides exerting multiple immunomodulatory effects in response to bacterial stimuli. Previously, we reported that thrombin-derived C-terminal peptides (TCPs) are internalized in monocytes and macrophages in a time- and temperature-dependent manner. In this study, we investigated which endocytosis pathways are responsible for the internalization of TCPs. Using confocal microscopy and flow cytometry, we show that both clathrin-dependent and clathrin-independent pathways are involved in the internalization of the prototypic TCP GKY25 in RAW264.7 and human monocyte-derived M1 macrophages, whereas the uptake of GKY25 in monocytic THP-1 cells is mainly dynamin-dependent. Internalized GKY25 was transported to endosomes and finally lysosomes, where it remained detectable for up to 10 h. Comparison of GKY25 uptake with that of the natural occurring TCPs HVF18 and FYT21 indicates that the pathway of TCP endocytosis is not only cell type-dependent but also depends on the length and composition of the peptide as well as the presence of LPS and bacteria. Finally, using neutron reflectometry, we show that the observed differences between HVF18 and the other 2 TCPs may be explained partially by differences in membrane insertion. Taken together, we show that TCPs are differentially internalized into monocytes and macrophages.

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author
; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Bacterial infection, Caveolin-dependent endocytosis, Clathrin-dependent endocytosis, Host defense peptides, Lysosomes, Thrombin
in
Journal of Innate Immunity
volume
14
issue
5
pages
418 - 432
publisher
Karger
external identifiers
  • pmid:34937021
  • scopus:85121767245
ISSN
1662-811X
DOI
10.1159/000520831
language
English
LU publication?
yes
id
49dd4bc1-7b6e-414d-b0fc-b693c0f16776
date added to LUP
2022-03-23 12:13:29
date last changed
2025-01-06 17:26:44
@article{49dd4bc1-7b6e-414d-b0fc-b693c0f16776,
  abstract     = {{<p>Proteolytic cleavage of thrombin generates C-terminal host defense peptides exerting multiple immunomodulatory effects in response to bacterial stimuli. Previously, we reported that thrombin-derived C-terminal peptides (TCPs) are internalized in monocytes and macrophages in a time- and temperature-dependent manner. In this study, we investigated which endocytosis pathways are responsible for the internalization of TCPs. Using confocal microscopy and flow cytometry, we show that both clathrin-dependent and clathrin-independent pathways are involved in the internalization of the prototypic TCP GKY25 in RAW264.7 and human monocyte-derived M1 macrophages, whereas the uptake of GKY25 in monocytic THP-1 cells is mainly dynamin-dependent. Internalized GKY25 was transported to endosomes and finally lysosomes, where it remained detectable for up to 10 h. Comparison of GKY25 uptake with that of the natural occurring TCPs HVF18 and FYT21 indicates that the pathway of TCP endocytosis is not only cell type-dependent but also depends on the length and composition of the peptide as well as the presence of LPS and bacteria. Finally, using neutron reflectometry, we show that the observed differences between HVF18 and the other 2 TCPs may be explained partially by differences in membrane insertion. Taken together, we show that TCPs are differentially internalized into monocytes and macrophages. </p>}},
  author       = {{Hansen, Finja C. and Nadeem, Aftab and Browning, Kathryn L. and Campana, Mario and Schmidtchen, Artur and Van Der Plas, Mariena J.A.}},
  issn         = {{1662-811X}},
  keywords     = {{Bacterial infection; Caveolin-dependent endocytosis; Clathrin-dependent endocytosis; Host defense peptides; Lysosomes; Thrombin}},
  language     = {{eng}},
  number       = {{5}},
  pages        = {{418--432}},
  publisher    = {{Karger}},
  series       = {{Journal of Innate Immunity}},
  title        = {{Differential Internalization of Thrombin-Derived Host Defense Peptides into Monocytes and Macrophages}},
  url          = {{http://dx.doi.org/10.1159/000520831}},
  doi          = {{10.1159/000520831}},
  volume       = {{14}},
  year         = {{2022}},
}