Bronchopulmonary dysplasia (BPD) is a major morbidity in preterm infants. Production of the chemokine interleukin (IL)-8 by alveolar macrophages (AMs) is crucial to the hyperoxia-induced pulmonary injury that recapitulates some aspects of Bpd. This proposal's hypothesis is that IL-1Beta and tumor necrosis factor a (TNFalpha) are competence factors for the in vivo oxygen induction of IL-8 in neonatal rabbit /AMs.
Its specific aims are to: 1. Measure the contributions of IL-1Beta, TNFalpha and oxygen to the regulation of IL-8 expression in the oxygen-stressed alveolar macrophage. Treatment of isolated rabbit AMs with exogenous IL-1Beta and TNFalpha will investigate the sufficiency of these cytokines to induce IL-8 production in response to oxygen. Interference with IL-1Beta and TNFalpha activity by the addition of IL-1 receptor antagonist (IL-1ra) and pentoxifylline, respectively, will test their necessity to the response in oxygen-exposed cells from the human U937 histiocytic cell line. 2. Determine the specific cis-and trans-activating factors responsible for the expression of IL-8 in the isolated rabbit alveolar macrophage. Isolation of a genomic clone of rabbit IL-8 will allow generation of genomic probes and a reporter construct containing the IL-8 promoter. Deletion mutagenesis, co-transfection experiments, electromobility shift analysis (EMSA) and site-directed mutagenesis will define specific cis- and trans-activating factors using the rabbit promoter in the U937 cell line and in transiently transfected rabbit Ams. 3. Determine the proximate events and trans-activating factors responsible for IL-8 expression in the rabbit alveolar macrophage exposed in vivo to oxygen stress. Adult rabbits will be exposed to >95% oxygen for 64 hours and AMs isolated. EMSA will measure differences from previous in vitro stimulation. IL-8, IL-1Beta and TNFalpha expression will be compared to in vitro stimulation. 4. Measure the differences between adult and newborn rabbit alveolar macrophages' regulation of IL-8 following oxygen stress. Newborn rabbit AMs will be co-stimulated with oxygen and exogenous IL-1Beta and/or TNFalpha. Adult and newborn AMs will be exposed to pulmonary lavage fluid from hyperoxia-exposed adult and newborn animals to assess independently possible host and cellular immaturities in the response.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Clinical Investigator Award (CIA) (K08)
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Special Emphasis Panel (ZHL1-CSR-Y (F1))
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University of Rochester
Schools of Dentistry
United States
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D'Angio, Carl T; LoMonaco, Michael B; Johnston, Carl J et al. (2004) Differential roles for NF-kappa B in endotoxin and oxygen induction of interleukin-8 in the macrophage. Am J Physiol Lung Cell Mol Physiol 286:L30-6
Waheed, Saira; D'Angio, Carl T; Wagner, Carol L et al. (2002) Transforming growth factor alpha (TGF(alpha)) is increased during hyperoxia and fibrosis. Exp Lung Res 28:361-72
D'Angio, Carl T; Maniscalco, William M (2002) The role of vascular growth factors in hyperoxia-induced injury to the developing lung. Front Biosci 7:d1609-23
D'Angio, Carl T; Basavegowda, Kumar; Avissar, Nelly E et al. (2002) Comparison of tracheal aspirate and bronchoalveolar lavage specimens from premature infants. Biol Neonate 82:145-9
Hastings, Randolph H; Ryan, Rita M; D'Angio, Carl T et al. (2002) Parathyroid hormone-related protein response to hyperoxic lung injury. Am J Physiol Lung Cell Mol Physiol 282:L1198-208
D'Angio, C T; Johnston, C J; Wright, T W et al. (1998) Chemokine mRNA alterations in newborn and adult mouse lung during acute hyperoxia. Exp Lung Res 24:685-702