PMID- 12110480
OWN - NLM
STAT- MEDLINE
DCOM- 20020926
LR  - 20220311
IS  - 0928-8244 (Print)
IS  - 0928-8244 (Linking)
VI  - 33
IP  - 3
DP  - 2002 Jul 12
TI  - Identification of ABC transporters in vancomycin-resistant Enterococcus faecium 
      as potential targets for antibody therapy.
PG  - 179-89
AB  - The occurrence of an outbreak of septicaemias due to vancomycin-resistant 
      Enterococcus faecium (VRE), in Manchester, UK, provided an opportunity to examine 
      the antibody responses in patients infected by the same strain. Immunoblotting 
      sera from 24 cases, six of whom died, showed an immunodominant cluster of 
      antigens at 34, 54 and 97 kDa, with a statistically significant correlate between 
      survival and immunoglobulin G to the 34 and 97 kDa bands (P<0.05). Screening a 
      genomic expression library of VRE with seropositive serum and peritoneal 
      dialysate from a survivor gave a recombinant clone with two contiguous open 
      reading frames, the derived amino acid sequences of which both showed sequence 
      homologue with ABC transporters, with a Walker A and Walker B motif and the 
      signature sequence LSGGQ. The first open reading frame (putative VRE ABC1) showed 
      57% homologue with YbxA from Bacillus subtilis. A partial sequence (putative VRE 
      ABC2) was also obtained, in the same recombinant clone, of a second ABC 
      transporter with 72% homologue with ybaE from B. subtilis. Affinity selection 
      with the seropositive serum and peritoneal dialysate used to screen the library 
      showed that the eluted antibody bound to the 97, 54, 34 and 30 kDa bands. Direct 
      amino acid sequencing identified this as a possible ABC transporter. Rabbit 
      antiserum against peptides representing Walker A and an area adjacent to the 
      Walker B site cross-reacted with bands at 34, 54, 97, 110 kDa and at 30, 34 and 
      54 kDa respectively. This therefore appeared to be an immunodominant complex of 
      ABC transporters of which the smallest was the 30 kDa antigen. Epitope mapping of 
      this antigen with seropositive patients' sera delineated three linear epitopes 
      (KVGIV, FGPKNF and RVAI). The Walker A site represented by peptide 1 
      (GHNGSGKSTLAKTIN), epitope RVAI represented by peptides 2 (MRRVAIAGVLAMPRE) and 3 
      (ELSGGQMRRVAIAGV), epitope KVGIV represented by peptide 4 (LKPIRKKVGIVFQFP), and 
      recombinant VRE ABC1 and VRE ABC2 expressed in Escherichia coli pBAD were then 
      used to isolate human genetically recombinant antibodies from a phage antibody 
      display library. An assessment of the protective potential of these antibodies 
      was carried out in a mouse model of the infection. This study suggests that an 
      ABC transporter homologue could be a target for antibody therapy against VRE 
      infections.
FAU - Burnie, James
AU  - Burnie J
AD  - Infectious Diseases Research Group, University of Manchester, Oxford Road, 
      Manchester M13 9WL, UK. jburnie@labmed.cmht.nwest.nhs.uk
FAU - Carter, Tracey
AU  - Carter T
FAU - Rigg, Gordon
AU  - Rigg G
FAU - Hodgetts, Samantha
AU  - Hodgetts S
FAU - Donohoe, Michael
AU  - Donohoe M
FAU - Matthews, Ruth
AU  - Matthews R
LA  - eng
PT  - Journal Article
PL  - England
TA  - FEMS Immunol Med Microbiol
JT  - FEMS immunology and medical microbiology
JID - 9315554
RN  - 0 (ATP-Binding Cassette Transporters)
RN  - 0 (Antibodies, Bacterial)
RN  - 0 (Antigens, Bacterial)
RN  - 0 (Immunodominant Epitopes)
RN  - 0 (Peptides)
RN  - 0 (Recombinant Proteins)
SB  - IM
MH  - ATP-Binding Cassette Transporters/chemistry/genetics/*immunology
MH  - Amino Acid Sequence
MH  - Animals
MH  - Antibodies, Bacterial/genetics/immunology/*therapeutic use
MH  - Antigens, Bacterial/immunology
MH  - Bacteremia/immunology/microbiology/therapy
MH  - Base Sequence
MH  - Enterococcus faecium/drug effects/*immunology
MH  - Epitope Mapping
MH  - Feces/microbiology
MH  - Gram-Positive Bacterial Infections/immunology/microbiology/*therapy
MH  - Humans
MH  - Immunoblotting
MH  - Immunodominant Epitopes
MH  - Mice
MH  - Molecular Sequence Data
MH  - Peptides/chemistry/genetics/immunology
MH  - Recombinant Proteins/genetics/immunology/*therapeutic use
MH  - *Vancomycin Resistance
EDAT- 2002/07/12 10:00
MHDA- 2002/09/27 06:00
CRDT- 2002/07/12 10:00
PHST- 2002/07/12 10:00 [pubmed]
PHST- 2002/09/27 06:00 [medline]
PHST- 2002/07/12 10:00 [entrez]
AID - S0928824402003048 [pii]
AID - 10.1111/j.1574-695X.2002.tb00589.x [doi]
PST - ppublish
SO  - FEMS Immunol Med Microbiol. 2002 Jul 12;33(3):179-89. doi: 
      10.1111/j.1574-695X.2002.tb00589.x.