The Burkholderia cenocepacia OmpA-like protein BCAL2958: identification, characterization, and detection of anti-BCAL2958 antibodies in serum from B. cepacia complex-infected Cystic Fibrosis patients.
Respiratory infections by bacteria of the Burkholderia cepacia complex (Bcc) remain an important cause of morbidity and mortality among cystic fibrosis patients, highlighting the need for novel therapeutic strategies. In the present work we have studied the B. cenocepacia protein BCAL2958, a member of the OmpA-like family of proteins, demonstrated as highly immunogenic in other pathogens and capable of eliciting strong host immune responses. The encoding gene was cloned and the protein, produced as a 6× His-tagged derivative, was used to produce polyclonal antibodies. Bioinformatics analyses led to the identification of sequences encoding proteins with a similarity higher than 96 % to BCAL2958 in all the publicly available Bcc genomes. Furthermore, using the antibody it was experimentally demonstrated that this protein is produced by all the 12 analyzed strains from 7 Bcc species. In addition, results are also presented showing the presence of anti-BCAL2958 antibodies in sera from cystic fibrosis patients with a clinical record of respiratory infection by Bcc, and the ability of the purified protein to in vitro stimulate neutrophils. The widespread production of the protein by Bcc members, together with its ability to stimulate the immune system and the detection of circulating antibodies in patients with a documented record of Bcc infection strongly suggest that the protein is a potential candidate for usage in preventive therapies of infections by Bcc.
iBB-Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, Torre Sul, Piso 6, 1049-001, Lisbon, Portugal. sousasilvia@tecnico.ulisboa.pt.
Sílvia A
SA
Sousa
iBB-Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, Torre Sul, Piso 6, 1049-001, Lisbon, Portugal.|Zoology Department, Faculty of Science, Helwan University, Cairo, Egypt.
Mostafa
M
Morad
iBB-Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, Torre Sul, Piso 6, 1049-001, Lisbon, Portugal.
Joana R
JR
Feliciano
iBB-Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, Torre Sul, Piso 6, 1049-001, Lisbon, Portugal.
Tiago
T
Pita
Zoology Department, Faculty of Science, Helwan University, Cairo, Egypt.
Soad
S
Nady
Zoology Department, Faculty of Science, Helwan University, Cairo, Egypt.
Rehab E
RE
El-Hennamy
Zoology Department, Faculty of Science, Helwan University, Cairo, Egypt.
Mona
M
Abdel-Rahman
Cystic Fibrosis Center, Department of Paediatrics, Hospital de D. Estefânia, Centro Hospitalar de Lisboa Central, Rua Jacinta Marto, 1169-045, Lisbon, Portugal.
José
J
Cavaco
Cystic Fibrosis Center, Department of Paediatrics, Hospital de Santa Maria, Centro Hospitalar Lisboa Norte, Av. Prof. Egas Moniz, 1649-028, Lisbon, Portugal.
Luísa
L
Pereira
Cystic Fibrosis Center, Department of Paediatrics, Hospital de Santa Maria, Centro Hospitalar Lisboa Norte, Av. Prof. Egas Moniz, 1649-028, Lisbon, Portugal.
Celeste
C
Barreto
iBB-Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, Torre Sul, Piso 6, 1049-001, Lisbon, Portugal. jorgeleitao@tecnico.ulisboa.pt.
Jorge H
JH
Leitão
Bcc positive CF serum samples; Burkholderia cepacia complex (Bcc); Cystic fibrosis (CF); Neutrophil activation; OmpA-like proteins