title: Human neutrophils express the alpha 1-antitrypsin gene and produce alpha 1-antitrypsin.
abstract: The potent serine protease, neutrophil elastase (NE), is stored in neutrophil azurophilic granules, where it is available to degrade phagocytosed material and can be released by the cell to assist in tissue migration and help clear tissue debris. While neutrophils carry NE, they cannot produce it; the NE gene is expressed only in bone marrow granulocyte precursor cells. Protection of normal tissues from the destructive capacity of NE is provided by alpha 1-antitrypsin (alpha 1 AT), a 52-Kd serine antiprotease produced by hepatocytes and mononuclear phagocytes. In the context of the broad destructive capacity of NE, we evaluated the concept that human neutrophils may be able to modulate the extracellular activity of NE by synthesizing and secreting alpha 1AT. Immunocytochemical analysis demonstrated that the neutrophil contains alpha 1AT. Northern analysis and in situ hybridization with alpha 1AT-specific probes demonstrated the presence of alpha 1AT messenger RNA transcripts within neutrophils. [35S]methionine-labeling of neutrophils followed by immunoprecipitation of the supernatant with an anti-alpha 1AT antibody and sodium dodecyl sulfate-acrylamide gel analysis demonstrated that neutrophils can synthesize alpha 1AT de novo and secrete the synthesized molecule. In the presence of major neutrophil degranulation, the antiprotease effect of neutrophil alpha 1AT is overwhelmed, allowing the NE to act unopposed in the extracellular microenvironment. However, in conditions where small amounts of NE are released by neutrophils, at least some of the secreted newly synthesized alpha 1AT was capable of complexing with NE. Thus, despite the fact that the neutrophil cannot synthesize NE, it can synthesize and secrete alpha 1AT, the inhibitor of NE, ie, the neutrophil is capable, to some extent, of modulating NE activity in the local milieu without the help of antiproteases produced by other cells.
authors.affiliation: Pulmonary Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892.
authors.forename: R M
authors.initials: RM
authors.lastname: du Bois
authors.forename: J F
authors.initials: JF
authors.lastname: Bernaudin
authors.forename: P
authors.initials: P
authors.lastname: Paakko
authors.forename: R
authors.initials: R
authors.lastname: Hubbard
authors.forename: H
authors.initials: H
authors.lastname: Takahashi
authors.forename: V
authors.initials: V
authors.lastname: Ferrans
authors.forename: R G
authors.initials: RG
authors.lastname: Crystal
mesh_terms: D015870:Gene Expression; D006801:Humans; D007124:Immunoenzyme Techniques; D007163:Immunosorbent Techniques; D019272:Leukocyte Elastase; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000737:chemistry / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D010196:Pancreatic Elastase / Q000378:metabolism; D015347:RNA Probes; D012333:RNA, Messenger / Q000032:analysis; D000515:alpha 1-Antitrypsin / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D015347:RNA Probes; D012333:RNA, Messenger; D000515:alpha 1-Antitrypsin; D009240:N-Formylmethionine Leucyl-Phenylalanine; D010196:Pancreatic Elastase; D019272:Leukocyte Elastase