title: Evidence for in vivo expression of transferrin-binding proteins in Haemophilus influenzae type b.
abstract: Haemophilus influenzae type b acquires transferrin-bound iron via a siderophore-independent mechanism involving direct contact between the human iron-binding glycoprotein and the bacterial cell surface. Evidence has accumulated to show that the transferrin receptor consists of at least two iron-regulated outer membrane transferrin-binding proteins (TBPs), of which one has a molecular mass of around 100 kDa (TBP1) and the other has a molecular mass of 60 to 90 kDa (TBP2). In H. influenzae type b strain Eagan, proteins of 76, 90, and 107 kDa appear to be involved in transferrin binding. To determine whether these TBPs are expressed during growth in vivo, strain Eagan was recovered without subculture from the intraperitoneal cavities of infected infant rats. By using a dot blot assay, outer membranes prepared from these in vivo-grown bacteria, unlike those grown in iron-sufficient broth, bound human transferrin and produced the 76-, 90-, and 107-kDa TBPs. Immunoblotting experiments using convalescent sera from infected rats also revealed the presence of antibodies to the 76- and 90-kDa strain Eagan TBPs. In addition, convalescent sera from three of four patients recovering from H. influenzae type b meningitis contained antibodies to the 90- and 105-kDa TBPs of the corresponding infecting strain. Furthermore, fresh clinical isolates of H. influenzae type b recovered from blood and cerebrospinal fluid showed constitutive expression of the TBPs, which became iron regulated only after prolonged in vitro subculture on iron-sufficient medium. This contrasted with the laboratory-adapted Eagan strain, in which the TBPs remained iron regulated even after animal passage. These findings indicate that the H. influenzae type b transferrin receptor is expressed during experimental animal and human infections.
authors.affiliation: Department of Pharmaceutical Sciences, University of Nottingham, United Kingdom.
authors.forename: J
authors.initials: J
authors.lastname: Holland
authors.forename: P R
authors.initials: PR
authors.lastname: Langford
authors.forename: K J
authors.initials: KJ
authors.lastname: Towner
authors.forename: P
authors.initials: P
authors.lastname: Williams
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D000907:Antibodies, Bacterial / Q000097:blood; D000917:Antibody Formation; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D002462:Cell Membrane / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006193:Haemophilus influenzae / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D033862:Iron-Binding Proteins; D051381:Rats; D011919:Rats, Inbred Strains; D033863:Transferrin-Binding Proteins
chemical_list: D000907:Antibodies, Bacterial; D002352:Carrier Proteins; D033862:Iron-Binding Proteins; D033863:Transferrin-Binding Proteins