Alteration of the Cell Wall of Haemophilus influenzae Type b by Transformation with Cloned DNA: Association with Attenuated Virulence

Publisher Website
Google Scholar
Abstract
A virulent strain of Haemophilusinfluenzaetype b was used to construct a λ library of chromosomal DNA in Charon 4, amplified in Escherichia coli. From this library a recombinant (I-69) phage was isolated that contained a 10.2-kilobase-pair fragment of DNA eliciting H. influenzae transformants whose colonies had a distinctive opaque phenotype. Compared with their H. influenzaeparent strains the opaque I-69 transformants had two defined cell wall alterations: one in the lipopolysaccharide (greater mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and one in the outer membrane proteins. The I-69 transformant of virulent type b strain Rd−/b+ had stable expression of type b capsule. In contrast to strain Rd−/b+, the Rd−/b+/I-69 transformant was serum sensitive in vitro and avirulent in vivo in rats. Thus the potential of H. influenzae type b organisms to cause invasive infection can be substantially attentuated by altering the expression of one or more genes that affect the cell wall composition.