Antimicrobial peptide LL-37 and its pro-form, hCAP18, in desquamated epithelial cells of human whole saliva
(2020) In European Journal of Oral Sciences 128(1). p.1-6- Abstract
The antimicrobial peptide LL-37 is active against oral bacteria and has been demonstrated to be present in human saliva, but its distribution in different fractions of saliva is not known. LL-37 is formed from its intracellular pro-form, hCAP18, in an extracellular enzymatic reaction catalyzed by proteinase 3 and kallikrein 5. Here, we prepared cell-containing and cell-free fractions of unstimulated human whole saliva by centrifugation after depolymerization of mucins with dithiothreitol, and measured the levels of hCAP18/LL-37 in these fractions using ELISA. Cellular expression of hCAP18/LL-37 was determined by western blotting and immunocytochemistry. The ELISA analyses demonstrated that both cells and cell-free saliva contained... (More)
The antimicrobial peptide LL-37 is active against oral bacteria and has been demonstrated to be present in human saliva, but its distribution in different fractions of saliva is not known. LL-37 is formed from its intracellular pro-form, hCAP18, in an extracellular enzymatic reaction catalyzed by proteinase 3 and kallikrein 5. Here, we prepared cell-containing and cell-free fractions of unstimulated human whole saliva by centrifugation after depolymerization of mucins with dithiothreitol, and measured the levels of hCAP18/LL-37 in these fractions using ELISA. Cellular expression of hCAP18/LL-37 was determined by western blotting and immunocytochemistry. The ELISA analyses demonstrated that both cells and cell-free saliva contained hCAP18/LL-37. Western blot analysis of cell-pellet homogenates showed a strong band corresponding to hCAP18 at the correct molecular weight and a weak band corresponding to LL-37. Phase-contrast and light microscopy revealed that the cells consisted of desquamated epithelial cells. These cells expressed cytoplasmic immunoreactivity for hCAP18/LL-37. The peripheral part of the cytoplasm, corresponding to the plasma membrane, was particularly rich in hCAP18/LL-37 immunoreactivity. No immunoreactivity was observed after omission of the primary antibody. We conclude that desquamated epithelial cells of human whole saliva contain antimicrobial hCAP18/LL-37, suggesting that these cells may take part in the innate immune system by harboring and releasing these peptides.
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- author
- Aidoukovitch, Alexandra LU ; Dahl, Sara LU ; Fält, Felicia ; Nebel, Daniel LU ; Svensson, Daniel LU ; Tufvesson, Ellen LU and Nilsson, Bengt-Olof LU
- organization
- publishing date
- 2020-02
- type
- Contribution to journal
- publication status
- published
- subject
- in
- European Journal of Oral Sciences
- volume
- 128
- issue
- 1
- pages
- 6 pages
- publisher
- Wiley-Blackwell
- external identifiers
-
- pmid:31825534
- scopus:85076338607
- ISSN
- 0909-8836
- DOI
- 10.1111/eos.12664
- project
- Production and transport of the antimicrobial peptide LL-37 and other peptides in saliva: physiological and pathophysiological importance
- language
- English
- LU publication?
- yes
- additional info
- © 2019 Eur J Oral Sci.
- id
- 10c49954-09d9-4320-9c4b-9b634f78d058
- date added to LUP
- 2020-02-04 16:22:52
- date last changed
- 2024-08-21 15:04:00
@article{10c49954-09d9-4320-9c4b-9b634f78d058, abstract = {{<p>The antimicrobial peptide LL-37 is active against oral bacteria and has been demonstrated to be present in human saliva, but its distribution in different fractions of saliva is not known. LL-37 is formed from its intracellular pro-form, hCAP18, in an extracellular enzymatic reaction catalyzed by proteinase 3 and kallikrein 5. Here, we prepared cell-containing and cell-free fractions of unstimulated human whole saliva by centrifugation after depolymerization of mucins with dithiothreitol, and measured the levels of hCAP18/LL-37 in these fractions using ELISA. Cellular expression of hCAP18/LL-37 was determined by western blotting and immunocytochemistry. The ELISA analyses demonstrated that both cells and cell-free saliva contained hCAP18/LL-37. Western blot analysis of cell-pellet homogenates showed a strong band corresponding to hCAP18 at the correct molecular weight and a weak band corresponding to LL-37. Phase-contrast and light microscopy revealed that the cells consisted of desquamated epithelial cells. These cells expressed cytoplasmic immunoreactivity for hCAP18/LL-37. The peripheral part of the cytoplasm, corresponding to the plasma membrane, was particularly rich in hCAP18/LL-37 immunoreactivity. No immunoreactivity was observed after omission of the primary antibody. We conclude that desquamated epithelial cells of human whole saliva contain antimicrobial hCAP18/LL-37, suggesting that these cells may take part in the innate immune system by harboring and releasing these peptides.</p>}}, author = {{Aidoukovitch, Alexandra and Dahl, Sara and Fält, Felicia and Nebel, Daniel and Svensson, Daniel and Tufvesson, Ellen and Nilsson, Bengt-Olof}}, issn = {{0909-8836}}, language = {{eng}}, number = {{1}}, pages = {{1--6}}, publisher = {{Wiley-Blackwell}}, series = {{European Journal of Oral Sciences}}, title = {{Antimicrobial peptide LL-37 and its pro-form, hCAP18, in desquamated epithelial cells of human whole saliva}}, url = {{http://dx.doi.org/10.1111/eos.12664}}, doi = {{10.1111/eos.12664}}, volume = {{128}}, year = {{2020}}, }