The Staphylococcus aureus and Staphylococcus epidermidis transferrin-binding proteins are expressed in vivo during infection.
Staphylococci express a 42 kDa cell-wall-associated protein which functions as a receptor for the mammalian iron-binding glycoprotein transferrin. To determine whether this transferrin-binding protein (TBP) is expressed during infection, Staphylococcus aureus and Staphylococcus epidermidis were grown in vivo in chambers implanted intraperitoneally in rats. SDS-PAGE and Western blotting of cell wall proteins prepared from staphylococci recovered directly from the chambers revealed the presence of both the TBP and bacterial-surface-associated rat transferrin. To obtain evidence for the in vivo expression of the staphylococcal TBPs in humans, sera and human peritoneal dialysate (HPD) from non-infected patients undergoing continuous ambulatory peritoneal dialysis (CAPD) and sera from healthy human volunteers were screened for anti-TBP antibodies. Western immunoblots revealed that three out of ten samples from the latter group, seven out of ten HPD samples and ten of ten CAPD patient serum samples contained antibodies to the TBP of both S. aureus and S. epidermidis. To gain further insights into the appearance of TBP antibodies, HPD samples were collected over time from CAPD patients whose HPD samples taken immediately after catheter insertion lacked anti-TBP antibodies. In two of these patients, each of whom experienced an episode of peritonitis due to S. epidermidis or Staphylococcus hominis, antibodies to the TBP appeared in the HPD collected immediately post-infection. To determine whether such TBP antibodies were capable of blocking interactions between transferrin and its staphylococcal receptor, HPD immunoglobulin fractions were purified using protein A-Sepharose beads. In competition assays, these immunoglobulins blocked the binding of 125I-labelled transferrin both to whole bacteria and to the isolated 42 kDa TBPs of S. aureus and S. epidermidis. These provide evidence to show that staphylococcal TBPs are expressed in vivo during infection.
Modun
Belinda J
BJ
2 Institute of Infections and Immunity University of Nottingham, University Hospital, Nottingham NG7 2UH, UK.|1 Department of Pharmaceutical Sciences, University of Nottingham, University Park, Nottingham NG7 2RD, UK.
Cockayne
Alan
A
3 Division of Microbiology, Clinical Laboratory Sciences University of Nottingham, University Hospital, Nottingham NG7 2UH, UK.|2 Institute of Infections and Immunity University of Nottingham, University Hospital, Nottingham NG7 2UH, UK.
Finch
Roger
R
3 Division of Microbiology, Clinical Laboratory Sciences University of Nottingham, University Hospital, Nottingham NG7 2UH, UK.
Williams
Paul
P
2 Institute of Infections and Immunity University of Nottingham, University Hospital, Nottingham NG7 2UH, UK.|1 Department of Pharmaceutical Sciences, University of Nottingham, University Park, Nottingham NG7 2RD, UK.
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology / Q000302:isolation & purification; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D015153:Blotting, Western; D002352:Carrier Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D033862:Iron-Binding Proteins; D010531:Peritoneal Dialysis, Continuous Ambulatory; D051381:Rats; D017208:Rats, Wistar; D011990:Receptors, Transferrin / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000737:chemistry* / Q000276:immunology; D013212:Staphylococcus epidermidis / Q000737:chemistry* / Q000276:immunology; D033863:Transferrin-Binding Proteins
D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; D033862:Iron-Binding Proteins; D011990:Receptors, Transferrin; D033863:Transferrin-Binding Proteins