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The effect of Streptococcus pneumoniae pneumolysin on human respiratory epithelium in vitro

The effect of Streptococcus pneumoniae pneumolysin on human respiratory epithelium in vitro
The effect of Streptococcus pneumoniae pneumolysin on human respiratory epithelium in vitro

Streptococcus pneumoniae culture filtrates and pneumolysin both slow human ciliary beating and damage respiratory epithelium in vitro. A polyclonal pneumolysin antibody bound to sepharose beads removed pneumolysin from culture filtrates and showed that pneumolysin alone was responsible for the effects on epithelium. In a 48-h organ culture pneumolysin caused ciliary slowing and epithelial disruption in a dose-dependent manner down to 5 ng/ml. Comparison of the ciliary slowing activity and pneumolysin concentration in filtrates in a continuous broth culture showed a maximal effect at 16 h (pneumolysin 7.5 micrograms/ml). Later the activity decreased while the pneumolysin concentration increased (8.8 micrograms/ml). This loss of activity was prevented by neutralisation of the acid pH of the culture medium. Eight different culture filtrates produced significant (P less than 0.05) ciliary slowing which correlated (r = 0.95) with simultaneously measured haemolytic (pneumolysin) activity. Substitution of tryptophan (position 433) by phenylalanine reduced the haemolytic and ciliary slowing activity of pneumolysin, but did not affect its ability to activate complement. There was no correlation between the ciliary slowing produced by the culture filtrate and that produced by the autolysate of a particular strain, nor between ciliary slowing and the extent of autolysis or the serotype of the strain.

Antibodies, Bacterial, Bacterial Proteins, Cell Movement, Culture Media, Dose-Response Relationship, Drug, Epithelium, Hemolysin Proteins, Humans, Mutagenesis, Site-Directed, Nasal Mucosa, Organ Culture Techniques, Recombinant Proteins, Streptococcus pneumoniae, Streptolysins, Journal Article, Research Support, Non-U.S. Gov't
0882-4010
275-284
Feldman, C.
f466c1a8-5663-4837-972d-1eb566eff1e5
Mitchell, T.J.
8e203ae1-74f1-4e5a-a049-98a8c69d15f3
Andrew, P.W.
48bbd6aa-7477-44ab-8b68-001b1e5deb5c
Boulnois, G.J.
a06298af-0bf9-4a06-ae2f-9f946d97c8f2
Read, R.C.
b5caca7b-0063-438a-b703-7ecbb6fc2b51
Todd, H.C.
02707687-9f39-4a4c-ad8b-3c5476d8a71e
Cole, P.J.
8d93f747-e6ed-4557-89ea-e12e0925bbf3
Wilson, R.
94eba368-b33f-4071-beb6-dac98ef23651
Feldman, C.
f466c1a8-5663-4837-972d-1eb566eff1e5
Mitchell, T.J.
8e203ae1-74f1-4e5a-a049-98a8c69d15f3
Andrew, P.W.
48bbd6aa-7477-44ab-8b68-001b1e5deb5c
Boulnois, G.J.
a06298af-0bf9-4a06-ae2f-9f946d97c8f2
Read, R.C.
b5caca7b-0063-438a-b703-7ecbb6fc2b51
Todd, H.C.
02707687-9f39-4a4c-ad8b-3c5476d8a71e
Cole, P.J.
8d93f747-e6ed-4557-89ea-e12e0925bbf3
Wilson, R.
94eba368-b33f-4071-beb6-dac98ef23651

Feldman, C., Mitchell, T.J., Andrew, P.W., Boulnois, G.J., Read, R.C., Todd, H.C., Cole, P.J. and Wilson, R. (1990) The effect of Streptococcus pneumoniae pneumolysin on human respiratory epithelium in vitro. Microbial Pathogenesis, 9 (4), 275-284. (doi:10.1016/0882-4010(90)90016-J).

Record type: Article

Abstract

Streptococcus pneumoniae culture filtrates and pneumolysin both slow human ciliary beating and damage respiratory epithelium in vitro. A polyclonal pneumolysin antibody bound to sepharose beads removed pneumolysin from culture filtrates and showed that pneumolysin alone was responsible for the effects on epithelium. In a 48-h organ culture pneumolysin caused ciliary slowing and epithelial disruption in a dose-dependent manner down to 5 ng/ml. Comparison of the ciliary slowing activity and pneumolysin concentration in filtrates in a continuous broth culture showed a maximal effect at 16 h (pneumolysin 7.5 micrograms/ml). Later the activity decreased while the pneumolysin concentration increased (8.8 micrograms/ml). This loss of activity was prevented by neutralisation of the acid pH of the culture medium. Eight different culture filtrates produced significant (P less than 0.05) ciliary slowing which correlated (r = 0.95) with simultaneously measured haemolytic (pneumolysin) activity. Substitution of tryptophan (position 433) by phenylalanine reduced the haemolytic and ciliary slowing activity of pneumolysin, but did not affect its ability to activate complement. There was no correlation between the ciliary slowing produced by the culture filtrate and that produced by the autolysate of a particular strain, nor between ciliary slowing and the extent of autolysis or the serotype of the strain.

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More information

Accepted/In Press date: 7 August 1990
Published date: October 1990
Keywords: Antibodies, Bacterial, Bacterial Proteins, Cell Movement, Culture Media, Dose-Response Relationship, Drug, Epithelium, Hemolysin Proteins, Humans, Mutagenesis, Site-Directed, Nasal Mucosa, Organ Culture Techniques, Recombinant Proteins, Streptococcus pneumoniae, Streptolysins, Journal Article, Research Support, Non-U.S. Gov't

Identifiers

Local EPrints ID: 416999
URI: https://eprints.soton.ac.uk/id/eprint/416999
ISSN: 0882-4010
PURE UUID: 348ca517-1e8a-4db0-bf78-0248bd222e23
ORCID for R.C. Read: ORCID iD orcid.org/0000-0002-4297-6728

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Date deposited: 16 Jan 2018 17:30
Last modified: 14 Mar 2019 01:36

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Contributors

Author: C. Feldman
Author: T.J. Mitchell
Author: P.W. Andrew
Author: G.J. Boulnois
Author: R.C. Read ORCID iD
Author: H.C. Todd
Author: P.J. Cole
Author: R. Wilson

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