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Change of PDL1 expression of bone marrow neutrophils
during sepsis in mice with different severity and possible mechanism |
SUN Hui1, YU Yao1, LIU Yishu1, LIU Lu1, QI Xinxin1, HUANG JiaSUN Hui1, YU Yao1, LIU Yi-shu1, LIU Lu1, QI Xin-xin1, HUANG Jia-min1, SUN Ran1, SUN Bing-wei2 |
(1. School of Medicine, Jiangsu University, Zhenjiang Jiangsu 212013; 2. Department of Burns and Plastic Surgery, Suzhou Hospital Affiliated to Nanjing Medical University, Suzhou Jiangsu 215008, China) |
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Abstract Objective: To evaluate programmed death ligand 1(PDL1) expression of bone marrow neutrophils during sepsis in mice with different severity and to explore possible mechanism. Methods: ① In vivo:Thirty C57/B6 mice(male) were randomly divided into sham operation group(Sham, n=10),mild cecal ligation perforation group(mild CLP,n=10) and severe cecal ligation perforation group(severe CLP,n=10).Liver and lung tissues were harvested 24 hours post operation to detect pathological changes. Meanwhile,the changes of PDL1 on mouse bone marrow neutrophils were detected in this model. With repeating above steps about groups and CLP model construction at 72 hours post operation,the survival rate analysis was performed.② In vitro: Mouse bone marrow neutrophils were isolated and then divided into control group,lipopolysaccharide(LPS,1μg/mL) stimulation group.The function of neutrophil chemotaxis and phagocytosis were detected.In addition,PDL1 on neutrophils was detected in control group, LPS stimulation group, SB 203580 (10 μmol/L) group and SB 203580 (10 μmol/L)+LPS group,respectively. Results: ① In vivo: The survival rate of the mild CLP model group(40%, P<0.05)and the severe survival rate (0%,P<0.01) were significantly lower than that in sham operation group(100%).At the same time pathological score was 1.250±0.677 0 in liver of mild CLP group(P<0.01), was 1.600±0.843 3 in lung of mild CLP group(P<0.01),was 2.900±0.614 6 in the liver of severe CLP group (P<0.01) and was 3.000±0.666 7 in the lung of severe CLP group(P<0.01), compared with the sham operation group. With detecting the ratio of bone marrow neutrophils, the proportion of mild CLP group was (37.57±1.89)%,while that of severe CLP group was (10.4±0.877 5)% ,which were significantly lower than that of sham operation group \[(59.27±1.168)%, P<0.01\].With detecting the ratio of PDL1 on neutrophils, the proportion of mild CLP group was(2.130±0.37)% while that of severe CLP group was (4.587±0.501 2)%,which were significantly lower than that of sham operation group \[(0.733 3± 0.1097)% ,P<0.01\]. ② In vitro: In terms of chemotaxis function, compared with the control group\[(1 101.00±99.78)μm\], there was a significant decrease of the chemotaxis distance when neutrophils stimulated with LPS\[(660.6±120.8)μm,P<0.01\].In terms of phagocytic function, compared with the control group(656.7±60.54),there was a significant increase of the phagocytic function in mean fluorescence intensity when neutrophils stimulated with LPS(998.7±46.01,P<0.01).The positive expression rate of PDL1 on bone marrow neutrophils was (72.27±1.656)% with the stimulation of LPS for 12 hours,which was significantly higher than that in the control group\[(1.713±0.519 6)%,P<0.01\]. However,with using the inhibitor of P38 MAPK SB 203580,PDL1 positive neutrophils stimulated with LPS were declined\[(37.17±1.38)%, P<0.01\]. Conclusion: The infiltration of neutrophils into tissues and organs were enhanced with the severity of sepsis and PDL1 of neutrophils was highly expressed with the severity of sepsis in the same way.PDL1 on the surface of neutrophils during sepsis was regulated by the P38 MAPK pathway.
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Received: 19 October 2018
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