Lipid bodies are cytoplasmic inclusions that develop in leukocytes, including eosinophils and mast cells, associated with allergic inflammation. Our investigations of the formation and function of leukocyte lipid bodies have revealed that they are distinct, inducible endoplasmic reticulum (ER)-derived, membrane- and ribosome-containing organelles with diverse functional roles in inflammatory responses of leukocytes pertinent to allergic inflammation. Leukocyte lipid bodies contain all enzymes required for synthesizing cyclooxygenase (COX)- and lipoxygenase (LO)-derived eicosanoids. Lipid body formation, rapidly inducible in vitro and in vivo by specific intracellular signaling pathways, enhances leukocyte formation of COX- and LO-derived eicosanoids. Lipid bodies are discrete sites of new eicosanoid synthesis, as documented for immunolocalized LTC4, LTB4 and PGE2. Lipid body-derived eicosanoids function as both paracrine and intracrine mediators of allergic inflammation. Based on combined proteomic and ultrastructural studies, leukocyte lipid bodies are complex organelles with internal membranes and ribosomes whose proteome includes proteins involved in protein synthesis. In IgE-activated cells, lipid bodies are early sites of agonist- elicited 5-LO mRNA localization and de novo 5-LO, LTC4 synthase and cytosolic phospholipase A2 protein synthesis. In addition, lipid bodies are sites of localization of cytokines, including TNF- demonstrable in vivo in tissue and blood leukocytes, although mechanisms and consequences of lipid body cytokine local- ization are unknown. Our central hypothesis is that leukocyte lipid bodies associated with allergic inflammation are inducible organelles that have major roles in the regulated formation of eicosanoids and cytokines that function as intra- and extracellular mediators of allergic inflammation. Our investigations have three aims: 1) Investigate the formation and function of eicosanoids within lipid bodies by evaluating the regulation of eicosanoid- synthesizing enzymes at lipid bodies, including 5-LO regulatory interactions with other proteins in lipid bodies and lipid bodies as sites of differential eicosanoid formation. 2) Investigate the formation and function of cytokines within lipid bodies by studying mechanisms regulating the formation and accumulation of cytokines within leukocyte lipid bodies and whether lipid body localized cytokines function locally within lipid bodies by signaling via cytokine receptors present within lipid bodies. 3) Investigate the mechanisms regulating leukocyte lipid body-related mRNA expression and function. The planned studies aim to provide a better understanding of mechanisms by which leukocytes contribute to inflammation pertinent to allergic and other diseases.
Asthma and related disorders due to allergic inflammation are increasingly prevalent diseases. The research will help in understanding mechanisms of allergic inflammation that are responsible for making asthma and allergic diseases major public health problems.
|Melo, Rossana C N; Morgan, Ellen; Monahan-Earley, Rita et al. (2014) Pre-embedding immunogold labeling to optimize protein localization at subcellular compartments and membrane microdomains of leukocytes. Nat Protoc 9:2382-94|
|Melo, Rossana C N; Weller, Peter F (2014) Unraveling the complexity of lipid body organelles in human eosinophils. J Leukoc Biol 96:703-12|
|Akuthota, Praveen; Capron, Kelsey; Weller, Peter F (2014) Eosinophil purification from peripheral blood. Methods Mol Biol 1178:13-20|
|Dias, Felipe F; Amaral, Kátia B; Carmo, Lívia A S et al. (2014) Human Eosinophil Leukocytes Express Protein Disulfide Isomerase in Secretory Granules and Vesicles: Ultrastructural Studies. J Histochem Cytochem 62:450-459|
|Kovalszki, Anna; Weller, Peter F (2014) Eosinophilia in mast cell disease. Immunol Allergy Clin North Am 34:357-64|
|Shamri, R; Young, K M; Weller, P F (2013) PI3K, ERK, p38 MAPK and integrins regulate CCR3-mediated secretion of mouse and human eosinophil-associated RNases. Allergy 68:880-9|
|Melo, Rossana C N; Paganoti, Guillherme F; Dvorak, Ann M et al. (2013) The internal architecture of leukocyte lipid body organelles captured by three-dimensional electron microscopy tomography. PLoS One 8:e59578|
|Akuthota, Praveen; Melo, Rossana C N; Spencer, Lisa A et al. (2012) MHC Class II and CD9 in Human Eosinophils Localize to Detergent-Resistant Membrane Microdomains. Am J Respir Cell Mol Biol 46:188-95|
|Bandeira-Melo, Christianne; Weller, Peter F; Bozza, Patricia T (2011) Identifying intracellular sites of eicosanoid lipid mediator synthesis with EicosaCell assays. Methods Mol Biol 717:277-89|
|Melo, Rossana C N; D'Avila, Heloisa; Bozza, Patricia T et al. (2011) Imaging lipid bodies within leukocytes with different light microscopy techniques. Methods Mol Biol 689:149-61|
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