Nuclear factor-kappa B activation in mouse lung lavage cells in response to Streptococcus pneumoniae pulmonary infection

Objectives: To assess the state and activation kinetics of the nuclear tran scription regulatory protein nuclear factor-kappa B (NF-kappa B) in lung la vage cells in a murine pneumococcal pneumonia model and to determine how th e virulence of the infecting organisms altered the activation state of NF-k appa B. Design: Experimental, comparative study of three Streptococcus pneumoniae s trains that induced three distinct pulmonary diseases. Setting: Experimental laboratory in a university-based medical center. Subjects: Female BALB/cby mice, 8-10 wks of age. Interventions: We randomly divided the mice into the following five groups: a) the control group; b) animals infected by virulent encapsulated S. pneu moniae P4241 strain; c) animals infected by avirulent encapsulated S. pneum oniae P15986 strain; d) animals infected by avirulent unencapsulated S. pne umoniae R6 strain; e) animals infected by virulent lysed S. pneumoniae P424 1 strain. Animals were anesthetized and infected by intratracheal delivery of 4 x 10(5) colony-forming units (CN) of S. pneumoniae per mouse or bacter ial components equivalent to 4 x 10(5) CN for lysed S. pneumoniae challenge . After intratracheal challenge with virulent encapsulated strain P4241, mi ce developed acute pneumonia, became bacteremic, and died within 3 to 5 day s. None of the mice infected with the avirulent encapsulated strain P15986 or the avirulent unencapsulated strain R6 died. After collection of lung la vage cells and nuclear extraction, NF-kappa B activation was determined 1 h r, 4 hrs, 6 hrs and 24 hrs after pneumococcal infection. At the same time, pulmonary and blood clearance, bronchoalveolar lavage cells population, and tumor necrosis factor-or production were assessed (six mice per time point ). Measurements and Main Results:NF-KB was constitutively expressed within nuc lear extracts of lung lavage cells from uninfected control mice. A signific ant increase in NF-kappa B activation was detected within 1 hr after inject ion of virulent lysed S. pneumoniae P4241 strain (bacterial components equi valent to 4 x 105 CFU), and was still present 24 hrs after the injection. A fter live pneumococcal challenge, significant NF-kappa B activation was det ected within 4 hrs with a peak at 24 hrs. Responses to all three strains (P 4241, P15986 and R6) were time-dependent (p < .0001), as NF-kappa B activat ion gradually increased during the first 24 hrs. Moreover, compared with th e control uninfected mice, the intensity of the retarded kappa B oligonucle otide, as determined by densitometry, was increased approximately four- to five-fold and seven-fold in reactions containing nuclear extracts isolated 24 hrs after infection with the avirulent strains P15986 or R6 and the viru lent strain P4241, respectively. With the virulent strain P4241, responses were significantly stronger than with the avirulent strains P15986 and (p < .01). Responses were of similar order with avirulent strains P15986 and R6 (p > .05). Conclusion: Pulmonary infection by S. pneumoniae induced delayed and time-d ependent activation of NF-kappa B in mouse lung lavage cells. The degree of NF-kappa B activation in lung lavage cells correlated with the virulence o f the infecting organisms. Our results suggest that the more severe the inf ection, the higher the rise in NF-kappa B.