Inducible nitric oxide synthase in pulmonary alveolar macrophages from patients with tuberculosis.
The high-output pathway of nitric oxide production helps protect mice from infection by several pathogens, including Mycobacterium tuberculosis. However, based on studies of cells cultured from blood, it is controversial whether human mononuclear phagocytes can express the corresponding inducible nitric oxide synthase (iNOS;NOS2). The present study examined alveolar macrophages fixed directly after bronchopulmonary lavage. An average of 65% of the macrophages from 11 of 11 patients with untreated, culture-positive pulmonary tuberculosis reacted with an antibody documented herein to be monospecific for human NOS2. In contrast, a mean of 10% of bronchoalveolar lavage cells were positive from each of five clinically normal subjects. Tuberculosis patients' macrophages displayed diaphorase activity in the same proportion that they stained for NOS2, under assay conditions wherein the diaphorase reaction was strictly dependent on NOS2 expression. Bronchoalveolar lavage specimens also contained NOS2 mRNA. Thus, macrophages in the lungs of people with clinically active Mycobacterium tuberculosis infection often express catalytically competent NOS2.
Nicholson
S
S
Beatrice and Samuel A. Seaver Laboratory, Division of Hematology-Oncology Cornell University Medical College, New York 10021, USA.
Bonecini-Almeida
M da G
Mda G
Lapa e Silva
J R
JR
Nathan
C
C
Xie
Q W
QW
Mumford
R
R
Weidner
J R
JR
Calaycay
J
J
Geng
J
J
Boechat
N
N
Linhares
C
C
Rom
W
W
Ho
J L
JL
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001483:Base Sequence; D002460:Cell Line; D002478:Cells, Cultured; D017931:DNA Primers; D008058:Dihydrolipoamide Dehydrogenase / Q000032:analysis / Q000378:metabolism; D004730:Endothelium, Vascular / Q000201:enzymology; D006801:Humans; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000032:analysis / Q000096:biosynthesis; D008168:Lung; D016676:Macrophages, Alveolar / Q000201:enzymology*; D017737:Macrophages, Peritoneal / Q000201:enzymology; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D019001:Nitric Oxide Synthase / Q000032:analysis* / Q000096:biosynthesis; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D012016:Reference Values; D014158:Transcription, Genetic; D014397:Tuberculosis, Pulmonary / Q000201:enzymology*
D000906:Antibodies; D017931:DNA Primers; D007527:Isoenzymes; D009842:Oligopeptides; D010446:Peptide Fragments; D012333:RNA, Messenger; D011994:Recombinant Proteins; D019001:Nitric Oxide Synthase; D008058:Dihydrolipoamide Dehydrogenase