PMID- 11349030
OWN - NLM
STAT- MEDLINE
DCOM- 20010628
LR  - 20231105
IS  - 0019-9567 (Print)
IS  - 1098-5522 (Electronic)
IS  - 0019-9567 (Linking)
VI  - 69
IP  - 6
DP  - 2001 Jun
TI  - Expression of cytokine and chemokine genes by human middle ear epithelial cells 
      induced by formalin-killed Haemophilus influenzae or its lipooligosaccharide htrB 
      and rfaD mutants.
PG  - 3678-84
AB  - To define the role of nontypeable Haemophilus influenzae (NTHI) 
      lipooligosaccharide (LOS) in the induction of proinflammatory cytokine gene 
      expression during otitis media, we compared the abilities of formalin-killed NTHI 
      strain 2019 and its LOS htrB and rfaD mutants to stimulate human middle ear 
      epithelial (HMEE) cell cytokine and chemokine gene expression and production in 
      vitro. Strain DK-1, an rfaD gene mutant, expresses a truncated LOS consisting of 
      only three deoxy-D-manno-octulosonic acid residues, a single heptose, and lipid 
      A. Strain B29, an isogenic htrB mutant, possesses an altered oligosaccharide core 
      and an altered lipid A. HMEE cells were incubated with formalin-killed NTHI 2019, 
      B29, or DK-1. The supernatants and the cells were collected at 2, 4, 8, and 24 h 
      after stimulation. Expression of genes for the cytokines tumor necrosis factor 
      alpha (TNF-alpha), interleukin lbeta (IL-1beta), and IL-6 and for the chemokines 
      macrophage inflammatory protein 1beta (MIP-1beta), monocyte chemotactic peptide 1 
      (MCP-1), and IL-8 was quantitated by real-time PCR. NTHI B29 did not 
      significantly stimulate any cytokine or chemokine mRNA expression in HMEE cells. 
      In striking contrast, NTHI 2019 induced up to 105-, 139-, and 187-fold increases 
      in HMEE cell expression of IL-1beta, TNF-alpha, and MIP-1beta, respectively (P < 
      0.01 [2019 versus B29]). NTHI 2019 also induced upregulation of IL-8, IL-6, and 
      MCP-1 mRNA expression (by 26-, 44-, and 14-fold, respectively [P < 0.05 (2019 
      versus B29)]). The significant induction of cytokine genes was confirmed by 
      quantitating the secretion of cytokines in culture supernatants with an 
      enzyme-linked immunosorbent assay. There were no significant differences in mRNA 
      expression of IL-8, IL-6, and MCP-1 between the 2019- and DK-1-treated groups. 
      The low levels of gene transcripts observed after incubation of HMEE cells with 
      B29 indicate that products of the disrupted NTHI htrB LOS gene may play a major 
      role in induction of these particular inflammatory mediators.
FAU - Tong, H H
AU  - Tong HH
AD  - Department of Otolaryngology, College of Medicine and Public Health, The Ohio 
      State University, Columbus 43210, USA.
FAU - Chen, Y
AU  - Chen Y
FAU - James, M
AU  - James M
FAU - Van Deusen, J
AU  - Van Deusen J
FAU - Welling, D B
AU  - Welling DB
FAU - DeMaria, T F
AU  - DeMaria TF
LA  - eng
GR  - 5 R01 DC00090-28/DC/NIDCD NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Infect Immun
JT  - Infection and immunity
JID - 0246127
RN  - 0 (Chemokines)
RN  - 0 (Cytokines)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (lipid-linked oligosaccharides)
RN  - 1HG84L3525 (Formaldehyde)
RN  - EC 5.1.3.- (ADP-L-glycero-D-mannoheptose-6-epimerase)
RN  - EC 5.1.3.- (Carbohydrate Epimerases)
SB  - IM
MH  - Adult
MH  - Carbohydrate Epimerases/genetics/*immunology
MH  - Cells, Cultured
MH  - Chemokines/genetics/metabolism
MH  - Cytokines/genetics/*metabolism
MH  - Ear, Middle/*cytology/immunology/microbiology
MH  - Epithelial Cells/*immunology/microbiology
MH  - Formaldehyde/pharmacology
MH  - Haemophilus influenzae/drug effects/*immunology
MH  - Humans
MH  - Kinetics
MH  - Lipopolysaccharides/*immunology
MH  - Mutation
PMC - PMC98367
EDAT- 2001/05/12 10:00
MHDA- 2001/06/29 10:01
PMCR- 2001/06/01
CRDT- 2001/05/12 10:00
PHST- 2001/05/12 10:00 [pubmed]
PHST- 2001/06/29 10:01 [medline]
PHST- 2001/05/12 10:00 [entrez]
PHST- 2001/06/01 00:00 [pmc-release]
AID - 0009 [pii]
AID - 10.1128/IAI.69.6.3678-3684.2001 [doi]
PST - ppublish
SO  - Infect Immun. 2001 Jun;69(6):3678-84. doi: 10.1128/IAI.69.6.3678-3684.2001.