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Updated: Jun 4, 2026
A Method for Generating Pulmonary Neutrophilia Using Aerosolized Lipopolysaccharide
Published on: December 15, 2014
[The changes in secretory function of pulmonary intravascular macrophages after challenge of lipopolysaccharide]
Sheng-liang Li1, Zhi-hong Wu, Shu-qin Zhang
1Department of Respiration, the 517th Hospital of PLA, Kelan 036301, Shanxi, China. LSL5516@163.com
Objective:
To investigate the role of pulmonary intravascular macrophages (PIMs) in the pathogenesis of acute lung injury (ALI) due to infection.
Methods:
Porcine pulmonary blood vessels were flushed by modified Morton method, and PIMs were isolated and cultured. The adhered PIMs were collected with adhesion method and incubated in RPMI 1640 medium. They were challenged with lipopolysaccharide (LPS, 10 mg/L). The activity of interleukin-1β (IL-1β), and contents of IL 6 and IL 8 in the culture supernatant were measured by method of thymocyte proliferation and enzyme linked immunoadsorbent assay (ELISA).
Results:
The released contents of IL-1β, IL-6 and IL-8 from PIMs were increased significantly compared with those before LPS challenge , and they peaked at 2 hours [IL-1β activity: (10 400 ± 2 389) scintillant count/min], 4 hours [IL-6 content: (0.80 ± 0.36) μg/L], and 6 hours [IL-8 content: (4.94 ± 1.19) μg/L ] after LPS challenge , and the differences were significant compared with hose before LPS challenge [IL-1β activity: (213 ± 85) scintillant count/min, IL-6 content: (0.27 ± 0.12) μg/L, IL-8 content: (1.84 ± 0.53) μg/L, all P <0.01].
Conclusion:
Among the cytokines released from PIMs after LPS challenge , the increase in IL-1β occurred earlier in comparison with that of IL-6 and IL-8, suggesting that the former might play an important role at the early stage of ALI; on the other hand, though the increase in IL-6 and IL-8 contents occurred later than that of IL- 1β but it lasted for a longer duration, suggesting that they might be associated with the advancement of ALI. The Results also suggested that interaction of these cytokines played a more important role in the pathogenesis of ALI.
Insights
Pulmonary intravascular macrophages (PIMs) release inflammatory cytokines like IL-1β, IL-6, and IL-8 upon challenge, contributing to acute lung injury (ALI). IL-1β appears early, while IL-6 and IL-8 indicate later stages and progression of ALI.
Area of Science:
- Immunology
- Pathology
- Respiratory Medicine
Background:
- Acute lung injury (ALI) is a severe condition with significant morbidity and mortality.
- Pulmonary intravascular macrophages (PIMs) are key immune cells residing in lung vasculature.
- The specific role of PIMs in ALI pathogenesis remains incompletely understood.
Purpose of the Study:
- To investigate the role of pulmonary intravascular macrophages (PIMs) in the pathogenesis of acute lung injury (ALI).
- To analyze the cytokine release profile of PIMs following lipopolysaccharide (LPS) challenge.
Main Methods:
- Isolation and culture of porcine PIMs.
- Stimulation of PIMs with lipopolysaccharide (LPS).
- Quantification of interleukin-1β (IL-1β), IL-6, and IL-8 using thymocyte proliferation and ELISA.
Main Results:
- LPS challenge significantly increased IL-1β, IL-6, and IL-8 release from PIMs.
- IL-1β levels peaked at 2 hours post-LPS.
- IL-6 and IL-8 levels peaked at 4 and 6 hours, respectively, with prolonged elevation.
Conclusions:
- Early IL-1β release suggests a role in the initial phase of ALI.
- Later, sustained IL-6 and IL-8 release may contribute to ALI progression.
- The interplay of these cytokines is crucial in ALI pathogenesis.
