Detection of rhinovirus infection of the nasal mucosa by oligonucleotide in situ hybridization.
Human rhinoviruses (HRVs) cause the common cold and often induce lower airway symptoms such as cough and wheezing. Although HRV infection is presumed to involve primarily ciliated epithelial cells, this has not been confirmed in vivo, and the cellular distribution and spread of infection as well as the pathogenesis of cold related nasal and chest symptoms remain speculative. We have developed in situ hybridization (ISH) to explore localization of the virus to airway tissues, employing HRV 16-derived oligonucleotide probes after sequencing part of the genome of this serotype. A reverse transcription-polymerase chain reaction was used to generate DNA from HRV 16 for sequencing; this yielded 305 nucleotide bases that showed considerable homology to other HRVs. The HRV 16 sequence was used to design oligonucleotides functioning as antisense and sense probes. These probes as well as random sequence and pathogen control oligonucleotides were applied to HRV-infected cell-clot complexes and finally to sections from six paired nasal biopsies obtained before, during, or after HRV-proven colds. Specificity of hybrids was established by the absence of signal in uninfected tissue, in cells infected with other viruses, after RNase pretreatment, and with application of control probes. Hybridization signals were observed in epithelial cells in three of six biopsies obtained during a cold, using probes to viral (+) strand; intermediate (-) strand, implying viral replication, was present in one biopsy. Evidence for infection of nonepithelial cells was inconclusive. HRVs cause productive infection of nasal epithelium during a cold and their intracellular localization may produce perturbation of inflammatory mediators and cytokine profiles.(ABSTRACT TRUNCATED AT 250 WORDS)
Bardin
P G
PG
Immunopharmacology Group, Southampton General Hospital, United Kingdom.
Johnston
S L
SL
Sanderson
G
G
Robinson
B S
BS
Pickett
M A
MA
Fraenkel
D J
DJ
Holgate
S T
ST
D001483:Base Sequence; D003139:Common Cold / Q000175:diagnosis*; D015342:DNA Probes; D004279:DNA, Viral; D006367:HeLa Cells; D006801:Humans; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D009297:Nasal Mucosa / Q000382:microbiology* / Q000473:pathology; D012229:Rhinovirus / Q000235:genetics / Q000302:isolation & purification*
D015342:DNA Probes; D004279:DNA, Viral