Mucin adsorbed by E. coli can affect neutrophil activation in vitro
Bacteria colonizing human intestine adhere to the gut mucosa and avoid the innate immune system. We previously demonstrated that Escherichia coli isolates can adsorb mucin from a diluted solution in vitro. Here, we evaluated the effect of mucin adsorption by E. coli cells on neutrophil activation in...
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| Format: | Article |
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Wiley
2020-02-01
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| Series: | FEBS Open Bio |
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| Online Access: | https://doi.org/10.1002/2211-5463.12770 |
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| author | Elena Mikhalchik Nadezhda Balabushevich Tatiana Vakhrusheva Alexey Sokolov Julia Baykova Daria Rakitina Petr Scherbakov Sergey Gusev Alexander Gusev Zaira Kharaeva Olga Bukato Olga Pobeguts |
| author_facet | Elena Mikhalchik Nadezhda Balabushevich Tatiana Vakhrusheva Alexey Sokolov Julia Baykova Daria Rakitina Petr Scherbakov Sergey Gusev Alexander Gusev Zaira Kharaeva Olga Bukato Olga Pobeguts |
| author_sort | Elena Mikhalchik |
| collection | DOAJ |
| description | Bacteria colonizing human intestine adhere to the gut mucosa and avoid the innate immune system. We previously demonstrated that Escherichia coli isolates can adsorb mucin from a diluted solution in vitro. Here, we evaluated the effect of mucin adsorption by E. coli cells on neutrophil activation in vitro. Activation was evaluated based on the detection of reactive oxygen species production by a chemiluminescent reaction (ChL), observation of morphological alterations in neutrophils and detection of exocytosis of myeloperoxidase and lactoferrin. We report that mucin adsorbed by cells of SharL1 isolate from Crohn's disease patient's inflamed ileum suppressed the potential for the activation of neutrophils in whole blood. Also, the binding of plasma complement proteins and immunoglobulins to the bacteria was reduced. Desialylated mucin, despite having the same adsorption efficiency to bacteria, had no effect on the blood ChL response. The effect of mucin suggests that it shields epitopes that interact with neutrophils and plasma proteins on the bacterial outer membrane. Potential candidates for these epitopes were identified among the proteins within the bacterial outer membrane fraction by 2D‐PAGE, fluorescent mucin binding on a blot and HPLC‐MS/MS. In vitro, the following proteins demonstrated mucin adsorption: outer membrane porins (OmpA, OmpC, OmpD and OmpF), adhesin OmpX, the membrane assembly factor OmpW, cobalamine transporter, ferrum uptake protein and the elongation factor Ef Tu‐1. In addition to their other functions, these proteins are known to be bacterial surface antigens. Therefore, the shielding of epitopes by mucin may affect the dynamics and intensity of an immune response. |
| format | Article |
| id | doaj-art-c4be7db4be674829b28dba71a8d57182 |
| institution | OA Journals |
| issn | 2211-5463 |
| language | English |
| publishDate | 2020-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj-art-c4be7db4be674829b28dba71a8d571822025-08-20T02:31:29ZengWileyFEBS Open Bio2211-54632020-02-0110218019610.1002/2211-5463.12770Mucin adsorbed by E. coli can affect neutrophil activation in vitroElena Mikhalchik0Nadezhda Balabushevich1Tatiana Vakhrusheva2Alexey Sokolov3Julia Baykova4Daria Rakitina5Petr Scherbakov6Sergey Gusev7Alexander Gusev8Zaira Kharaeva9Olga Bukato10Olga Pobeguts11Federal Research and Clinical Center of Physical‐Chemical Medicine Moscow RussiaDepartment of Chemistry Lomonosov Moscow State University RussiaFederal Research and Clinical Center of Physical‐Chemical Medicine Moscow RussiaFederal Research and Clinical Center of Physical‐Chemical Medicine Moscow RussiaFederal Research and Clinical Center of Physical‐Chemical Medicine Moscow RussiaFederal Research and Clinical Center of Physical‐Chemical Medicine Moscow RussiaFederal Research and Clinical Center of Physical‐Chemical Medicine Moscow RussiaFederal Research and Clinical Center of Physical‐Chemical Medicine Moscow RussiaFederal Research and Clinical Center of Physical‐Chemical Medicine Moscow RussiaKabardino‐Balkarian State University Nalchik RussiaFederal Research and Clinical Center of Physical‐Chemical Medicine Moscow RussiaFederal Research and Clinical Center of Physical‐Chemical Medicine Moscow RussiaBacteria colonizing human intestine adhere to the gut mucosa and avoid the innate immune system. We previously demonstrated that Escherichia coli isolates can adsorb mucin from a diluted solution in vitro. Here, we evaluated the effect of mucin adsorption by E. coli cells on neutrophil activation in vitro. Activation was evaluated based on the detection of reactive oxygen species production by a chemiluminescent reaction (ChL), observation of morphological alterations in neutrophils and detection of exocytosis of myeloperoxidase and lactoferrin. We report that mucin adsorbed by cells of SharL1 isolate from Crohn's disease patient's inflamed ileum suppressed the potential for the activation of neutrophils in whole blood. Also, the binding of plasma complement proteins and immunoglobulins to the bacteria was reduced. Desialylated mucin, despite having the same adsorption efficiency to bacteria, had no effect on the blood ChL response. The effect of mucin suggests that it shields epitopes that interact with neutrophils and plasma proteins on the bacterial outer membrane. Potential candidates for these epitopes were identified among the proteins within the bacterial outer membrane fraction by 2D‐PAGE, fluorescent mucin binding on a blot and HPLC‐MS/MS. In vitro, the following proteins demonstrated mucin adsorption: outer membrane porins (OmpA, OmpC, OmpD and OmpF), adhesin OmpX, the membrane assembly factor OmpW, cobalamine transporter, ferrum uptake protein and the elongation factor Ef Tu‐1. In addition to their other functions, these proteins are known to be bacterial surface antigens. Therefore, the shielding of epitopes by mucin may affect the dynamics and intensity of an immune response.https://doi.org/10.1002/2211-5463.12770E. colimucinneutrophilsouter membrane proteinsplasma proteinsreactive oxygen species |
| spellingShingle | Elena Mikhalchik Nadezhda Balabushevich Tatiana Vakhrusheva Alexey Sokolov Julia Baykova Daria Rakitina Petr Scherbakov Sergey Gusev Alexander Gusev Zaira Kharaeva Olga Bukato Olga Pobeguts Mucin adsorbed by E. coli can affect neutrophil activation in vitro FEBS Open Bio E. coli mucin neutrophils outer membrane proteins plasma proteins reactive oxygen species |
| title | Mucin adsorbed by E. coli can affect neutrophil activation in vitro |
| title_full | Mucin adsorbed by E. coli can affect neutrophil activation in vitro |
| title_fullStr | Mucin adsorbed by E. coli can affect neutrophil activation in vitro |
| title_full_unstemmed | Mucin adsorbed by E. coli can affect neutrophil activation in vitro |
| title_short | Mucin adsorbed by E. coli can affect neutrophil activation in vitro |
| title_sort | mucin adsorbed by e coli can affect neutrophil activation in vitro |
| topic | E. coli mucin neutrophils outer membrane proteins plasma proteins reactive oxygen species |
| url | https://doi.org/10.1002/2211-5463.12770 |
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