Induction of iNOS in human monocytes infected with different Legionella species.
The contribution of nitric oxide (NO) radicals to the suppression of intracellular replication of Legionella has been well established in rodents but remained questionable in humans. Considering the fact that human monocytes do not exhibit a high-output NO production, we used sensitive methods such as detection of inducible NO synthase (iNOS) mRNA by reverse transcription-PCR and demonstration of iNOS protein expression by means of flow cytometry and Western blot to compare the levels of iNOS induced by Legionella species which, in accordance to their human prevalence, show different multiplication rates within human monocytic cells. The expression of iNOS in Mono Mac 6 (MM6) cells showed an only moderate inverse correlation to the intracellular replication rate of a given Legionella species in the protein expression assays. However, stimulation of host cells with 1,25-dihydroxyvitamin D(3) to enhance NO production and inhibition of NO production by treatment of host cells with N(G)-methyl-L-arginine were not able to modify the intracellular multiplication of legionellae within MM6 cells. Therefore, NO production does not seem to play a crucial role for the restriction of intracellular replication of Legionella bacteria within human monocytic cells. Rodent models in investigations which are supposed to clarify the involvement of NO radicals in defense mechanisms against Legionella infections in humans are of doubtful significance.
Neumeister
B
B
Abteilung Transfusionsmedizin, Otfried-Müller-Strasse 4/1, Universitätsklinikum Tübingen, 72076, Tübingen, Germany. birgid.neumeister@med.uni-tuebingen.de
Bach
V
V
Faigle
M
M
Northoff
H
H
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D004790:Enzyme Induction; D005434:Flow Cytometry; D006801:Humans; D007875:Legionella / Q000254:growth & development / Q000502:physiology*; D009000:Monocytes / Q000201:enzymology* / Q000276:immunology / Q000378:metabolism / Q000382:microbiology*; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012333:RNA, Messenger; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase