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Published on: February 25, 2016
Quantitative nitric oxide production by rat, bovine and porcine macrophages
Petra Zelnickova1, Jan Matiasovic, Barbora Pavlova
1Department of Immunology, Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech Republic.
Abstract:
The aim of this work was to compare in vitro nitric oxide (NO) production by rat, bovine and porcine macrophages. NO production was induced by lipopolysaccharide (LPS) or by phorbol 12-myristate 13-acetate (PMA) with ionomycin or recombinant interferon gamma (rIFN-gamma) and was assessed by Griess reaction. NO synthase type II (NOS II) expression was quantified by immunocytochemistry, Western blot and real-time polymerase chain reaction (RT-PCR). There were differences in NO production by pulmonary alveolar macrophages (PAM) in all species tested. The largest amounts of NO were produced by rat PAM. Less NO was produced by bovine PAM. Moreover, PAM in rats and cows differed in their abilities to respond to various stimulators. Neither porcine PAM nor Kupffer cells produced NO. Stimulation of porcine PAM with alternative concentrations of LPS did not lead to inducing NO production. Stimulation of porcine PAM with rIFN-gamma together with LPS led to a significant increase in the expression of NOS II mRNA, albeit without detectable NO production or NOS II expression on the protein level.
Insights
Rat and bovine macrophages produce nitric oxide (NO), unlike porcine macrophages. Macrophage responses to stimulators like lipopolysaccharide (LPS) vary significantly across species, impacting NO production and inducible NO synthase (NOS II) expression.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Macrophages play a crucial role in immune responses, including the production of nitric oxide (NO).
- Species-specific differences in macrophage function can impact disease susceptibility and therapeutic strategies.
- Inducible nitric oxide synthase (NOS II) is a key enzyme in NO production.
Purpose of the Study:
- To compare in vitro nitric oxide (NO) production by rat, bovine, and porcine macrophages.
- To investigate the expression of inducible NO synthase type II (NOS II) in response to various stimuli.
- To elucidate species-specific differences in macrophage activation and NO generation.
Main Methods:
- Pulmonary alveolar macrophages (PAM) and Kupffer cells were isolated from rats, bovines, and pigs.
- NO production was induced using lipopolysaccharide (LPS), phorbol 12-myristate 13-acetate (PMA) with ionomycin, or recombinant interferon gamma (rIFN-gamma).
- NO levels were assessed using the Griess reaction, and NOS II expression was quantified via immunocytochemistry, Western blot, and real-time polymerase chain reaction (RT-PCR).
Main Results:
- Rat PAM produced the highest levels of NO, followed by bovine PAM, while porcine PAM and Kupffer cells produced no detectable NO.
- Rat and bovine PAM exhibited differential responses to various stimulators.
- Porcine PAM showed increased NOS II mRNA expression upon stimulation with rIFN-gamma and LPS, but no detectable NO or protein-level NOS II.
Conclusions:
- Significant species-specific differences exist in NO production by macrophages from rats, bovines, and pigs.
- Porcine macrophages appear to lack the capacity for NO production despite inducible NOS II mRNA expression.
- These findings highlight the importance of considering species of origin when studying macrophage function and NO signaling.
