TY - JOUR
T1 - Impact of Escherichia coli K12 and O18:K1 on human platelets
T2 - Differential effects on platelet activation, RNAs and proteins
AU - Fejes, A V
AU - Best, M G
AU - van der Heijden, W A
AU - Vancura, A
AU - Verschueren, H
AU - de Mast, Q
AU - Wurdinger, T
AU - Mannhalter, C
N1 - Funding Information: We want to thank M. Schuster, F. Schischlik (CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna), A. Assinger and her team, particularly J. B. Kral, W. C. Schrottmaier and M. Salzmann (Medical University of Vienna) and G. Leitner (University Clinic for Blood Group Serology and Transfusion Medicine, Vienna) for their scientific and technical support and helpful discussions. The project was funded by the Austrian Science Fund (FWF) PhD program ‘Cell Communication in Health and Disease’, Project APW01205FW_07. Publisher Copyright: © 2018, The Author(s).
PY - 2018/11/1
Y1 - 2018/11/1
N2 - Blood platelets can interact with bacteria, possibly leading to platelet activation, cytokine and microparticle release and immune signalling. Besides, bacteria can also affect the platelet RNA content. We investigated the impact of non-pathogenic K12 and pathogenic O18:K1 Escherichia (E.) coli strains on platelet activation, RNA expression patterns, and selected proteins. Depending on bacteria concentration, contact of platelets with E. coli K12 lead to an increase of P-selectin (24-51.3%), CD63 (15.9-24.3%), PAC-1 (3.8-14.9%) and bound fibrinogen (22.4-39%) on the surface. E. coli O18:K1 did not affect these markers. Sequencing analysis of total RNA showed that E. coli K12 caused a significant concentration change of 103 spliced mRNAs, of which 74 decreased. For the RNAs of HMBS (logFC = +5.73), ATP2C1 (logFC = -3.13) and LRCH4 (logFC = -4.07) changes were detectable by thromboSeq and Tuxedo pipelines. By Western blot we observed the conversion of HMBS protein from a 47 kDA to 40 kDa product by E. coli K12, O18:K1 and by purified lipopolysaccharide. While ATP2C1 protein was released from platelets, E. coli either reduced the secretion or broke down the released protein making it undetectable by antibodies. Our results demonstrate that different E. coli strains influence activation, RNA and protein levels differently which may affect platelet-bacteria crosstalk.
AB - Blood platelets can interact with bacteria, possibly leading to platelet activation, cytokine and microparticle release and immune signalling. Besides, bacteria can also affect the platelet RNA content. We investigated the impact of non-pathogenic K12 and pathogenic O18:K1 Escherichia (E.) coli strains on platelet activation, RNA expression patterns, and selected proteins. Depending on bacteria concentration, contact of platelets with E. coli K12 lead to an increase of P-selectin (24-51.3%), CD63 (15.9-24.3%), PAC-1 (3.8-14.9%) and bound fibrinogen (22.4-39%) on the surface. E. coli O18:K1 did not affect these markers. Sequencing analysis of total RNA showed that E. coli K12 caused a significant concentration change of 103 spliced mRNAs, of which 74 decreased. For the RNAs of HMBS (logFC = +5.73), ATP2C1 (logFC = -3.13) and LRCH4 (logFC = -4.07) changes were detectable by thromboSeq and Tuxedo pipelines. By Western blot we observed the conversion of HMBS protein from a 47 kDA to 40 kDa product by E. coli K12, O18:K1 and by purified lipopolysaccharide. While ATP2C1 protein was released from platelets, E. coli either reduced the secretion or broke down the released protein making it undetectable by antibodies. Our results demonstrate that different E. coli strains influence activation, RNA and protein levels differently which may affect platelet-bacteria crosstalk.
KW - Antigens, Bacterial/genetics
KW - Blood Platelets/metabolism
KW - Calcium-Transporting ATPases/blood
KW - Escherichia coli Infections/blood
KW - Escherichia coli K12/genetics
KW - Gene Expression Regulation, Bacterial/genetics
KW - Humans
KW - Lipopolysaccharides/genetics
KW - Nerve Tissue Proteins/genetics
KW - P-Selectin/genetics
KW - Platelet Activation/genetics
KW - RNA/blood
KW - Sequence Analysis, RNA
KW - Tetraspanin 30/genetics
KW - Uroporphyrinogen III Synthetase/genetics
UR - http://www.scopus.com/inward/record.url?scp=85055900523&partnerID=8YFLogxK
U2 - https://doi.org/10.1038/s41598-018-34473-w
DO - https://doi.org/10.1038/s41598-018-34473-w
M3 - Article
C2 - 30385858
SN - 2045-2322
VL - 8
SP - 16145
JO - Scientific reports
JF - Scientific reports
IS - 1
M1 - 16145
ER -