HAEMOPHILUS-INFLUENZAE RESIDES AND MULTIPLIES INTRACELLULARLY IN HUMAN ADENOID TISSUE AS DEMONSTRATED BY IN-SITU HYBRIDIZATION AND BACTERIAL VIABILITY ASSAY
J. Forsgren et al., HAEMOPHILUS-INFLUENZAE RESIDES AND MULTIPLIES INTRACELLULARLY IN HUMAN ADENOID TISSUE AS DEMONSTRATED BY IN-SITU HYBRIDIZATION AND BACTERIAL VIABILITY ASSAY, Infection and immunity, 62(2), 1994, pp. 673-679
The DNA oligomer 5'-d(TGCGGCCTCTCAGTCCCGCACTTTCATCTTCC)-3 specifically
recognizes Haemophilus influenzae 16S rRNA. We report here the use of
this oligonucleotide, with a fluorescein label tagged on its 5' end,
as a probe for the in situ detection of nonencapsulated nontypeable H.
influenzae in sections of adenoid tissue from 10 children who were cl
inically infection free but were having their adenoids removed because
of nasal obstruction. In some cases, the reticular crypt epithelium w
as focally infiltrated by H. influenzae. The reservoir for these bacte
rial colonizations, in all likelihood long standing, seemed to be macr
ophage-like cells found in the subepithelial layers in all 10 cases. T
hese mononuclear cells contained up to 200 intracellular H. influenzae
cells. In the transmission electron microscope, macrophage-like cells
with intracellular bacteria with coccoid morphology, at least some of
which were dividing, were seen. Adenoid cell suspensions, enriched fo
r macrophages by use of paramagnetic beads coated with monoclonal anti
bodies against the CD14 marker, yielded up to 1,100 CFU of nontypeable
H. influenzae per 10(5) cells after killing of extracellular bacteria
with gentamicin followed by mechanical lysis of the cells.