Identification and Characterization of Anaplasma phagocytophilum Proteins Involved in Infection of the Tick Vector, Ixodes scapularis.
Anaplasma phagocytophilum is an emerging zoonotic pathogen transmitted by Ixodes scapularis that causes human granulocytic anaplasmosis. Here, a high throughput quantitative proteomics approach was used to characterize A. phagocytophilum proteome during rickettsial multiplication and identify proteins involved in infection of the tick vector, I. scapularis. The first step in this research was focused on tick cells infected with A. phagocytophilum and sampled at two time points containing 10-15% and 65-71% infected cells, respectively to identify key bacterial proteins over-represented in high percentage infected cells. The second step was focused on adult female tick guts and salivary glands infected with A. phagocytophilum to compare in vitro results with those occurring during bacterial infection in vivo. The results showed differences in the proteome of A. phagocytophilum in infected ticks with higher impact on protein synthesis and processing than on bacterial replication in tick salivary glands. These results correlated well with the developmental cycle of A. phagocytophilum, in which cells convert from an intracellular reticulated, replicative form to the nondividing infectious dense-core form. The analysis of A. phagocytophilum differentially represented proteins identified stress response (GroEL, HSP70) and surface (MSP4) proteins that were over-represented in high percentage infected tick cells and salivary glands when compared to low percentage infected cells and guts, respectively. The results demonstrated that MSP4, GroEL and HSP70 interact and bind to tick cells, thus playing a role in rickettsia-tick interactions. The most important finding of these studies is the increase in the level of certain bacterial stress response and surface proteins in A. phagocytophilum-infected tick cells and salivary glands with functional implication in tick-pathogen interactions. These results gave a new dimension to the role of these stress response and surface proteins during A. phagocytophilum infection in ticks. Characterization of Anaplasma proteome contributes information on host-pathogen interactions and provides targets for development of novel control strategies for pathogen infection and transmission.
SaBio, Instituto de Investigación en Recursos Cinegéticos IREC-CSIC-UCLM-JCCM, Ronda de Toledo s/n, 13005, Ciudad Real, Spain.
Margarita
M
Villar
SaBio, Instituto de Investigación en Recursos Cinegéticos IREC-CSIC-UCLM-JCCM, Ronda de Toledo s/n, 13005, Ciudad Real, Spain.
Nieves
N
Ayllón
Department of Veterinary Pathobiology, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, Oklahoma, 74078, United States of America.
Katherine M
KM
Kocan
Centro Nacional de Investigaciones Cardiovasculares, Melchor Fernández Almagro 3, 28029, Madrid, Spain.
Elena
E
Bonzón-Kulichenko
SaBio, Instituto de Investigación en Recursos Cinegéticos IREC-CSIC-UCLM-JCCM, Ronda de Toledo s/n, 13005, Ciudad Real, Spain.
Pilar
P
Alberdi
Department of Veterinary Pathobiology, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, Oklahoma, 74078, United States of America.
Edmour F
EF
Blouin
The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, United Kingdom; The Pirbright Institute, Ash Road, Pirbright, Woking, GU24 0NF, United Kingdom.
Sabine
S
Weisheit
SaBio, Instituto de Investigación en Recursos Cinegéticos IREC-CSIC-UCLM-JCCM, Ronda de Toledo s/n, 13005, Ciudad Real, Spain.
Lourdes
L
Mateos-Hernández
SaBio, Instituto de Investigación en Recursos Cinegéticos IREC-CSIC-UCLM-JCCM, Ronda de Toledo s/n, 13005, Ciudad Real, Spain; Center for Infection and Immunity of Lille (CIIL), INSERM U1019 - CNRS UMR 8204, Université Lille Nord de France, Institut Pasteur de Lille, Lille, France.
Alejandro
A
Cabezas-Cruz
The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, United Kingdom; The Pirbright Institute, Ash Road, Pirbright, Woking, GU24 0NF, United Kingdom.
Lesley
L
Bell-Sakyi
Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, Branišovská 31, České Budějovice, CZ-37005, Czech Republic; Faculty of Science, University of South Bohemia, Branišovská 31, České Budějovice, CZ-37005, Czech Republic.
Marie
M
Vancová
Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, Branišovská 31, České Budějovice, CZ-37005, Czech Republic; Faculty of Science, University of South Bohemia, Branišovská 31, České Budějovice, CZ-37005, Czech Republic.
Tomáš
T
Bílý
CIRAD, UMR CMAEE, Site de Duclos, Prise d'eau, F-97170, Petit-Bourg, Guadeloupe, France; INRA, UMR1309 CMAEE, F-34398, Montpellier, France.
Damien F
DF
Meyer
Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, Branišovská 31, České Budějovice, CZ-37005, Czech Republic.
Jan
J
Sterba
SaBio, Instituto de Investigación en Recursos Cinegéticos IREC-CSIC-UCLM-JCCM, Ronda de Toledo s/n, 13005, Ciudad Real, Spain.
Marinela
M
Contreras
Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, Branišovská 31, České Budějovice, CZ-37005, Czech Republic.
Nataliia
N
Rudenko
Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, Branišovská 31, České Budějovice, CZ-37005, Czech Republic; Faculty of Science, University of South Bohemia, Branišovská 31, České Budějovice, CZ-37005, Czech Republic.
Libor
L
Grubhoffer
Centro Nacional de Investigaciones Cardiovasculares, Melchor Fernández Almagro 3, 28029, Madrid, Spain.
Jesús
J
Vázquez
SaBio, Instituto de Investigación en Recursos Cinegéticos IREC-CSIC-UCLM-JCCM, Ronda de Toledo s/n, 13005, Ciudad Real, Spain; Department of Veterinary Pathobiology, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, Oklahoma, 74078, United States of America.
José
J
de la Fuente
D041081:Anaplasma phagocytophilum; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D018834:Chaperonin 60 / Q000235:genetics* / Q000378:metabolism; D005260:Female; D041981:Gastrointestinal Tract / Q000382:microbiology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D054884:Host-Pathogen Interactions; D018884:Ixodes / Q000382:microbiology*; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D058977:Molecular Sequence Annotation; D020543:Proteome / Q000235:genetics* / Q000378:metabolism; D012469:Salivary Glands / Q000382:microbiology; D015398:Signal Transduction; D013312:Stress, Physiological
D001426:Bacterial Proteins; D018834:Chaperonin 60; D018840:HSP70 Heat-Shock Proteins; D008565:Membrane Proteins; D020543:Proteome