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Prognosis of sepsis induced by cecal ligation and puncture in mice improved by anti-Clonorchis Sinensis cyclopholin a antibodies.

Song T, Yang M, Chen J, Huang L, Yin H, He T, Huang H, Hu X - Parasit Vectors (2015)

Bottom Line: Lung and mesentery tissues were stained with hematoxylin-eosin.Anti-CsCyPA antibodies could combine with MuCyPA and inhibit its peptidyl prolyl isomerase (PPIase) activity.Furthermore, the survival rate was elevated, ranging from 10.0 % to 45.0 % compared to the control group.

View Article: PubMed Central - PubMed

Affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong, China. songtianzhang@163.com.

ABSTRACT

Background: Cyclophilin A (CyPA), a ubiquitously distributed intracellular protein, is thought to be one of the important inflammatory factors and plays a significant role in the development process of sepsis. In the form of cytokine, CyPA deteriorates sepsis by promoting intercellular communication, apoptosis of endothelial cells and chemotactic effect on inflammatory cells. In our previous study, cyclophilin A of Clonorchis sinensis (CsCyPA), a type of excretory-secretory antigen, could induce the patients infected with Clonorchis sinensis to produce specific anti-CsCyPA antibodies. In this study, we investigated whether anti-CsCyPA antibodies could cross-react with CyPA and then play a protective role against sepsis, just like other anti-cytokine antagonists.

Methods: The mice model with sepsis was established with cecal ligation and puncture (CLP). Fifty mg/kg purified anti-CsCyPA antibodies were injected via the caudal vein 6 h after the CLP operation, and persistent observation was performed for 72 h. Blood samples and tissues were collected at 6 h, 12 h, 24 h, 48 h and 72 h after CLP. Cytokines in serum were measured by ELISA. Lung and mesentery tissues were stained with hematoxylin-eosin. Endothelial cells (ECs) isolated from murine aorta were co-cultured with CyPA of mice (MuCyPA) and anti-CsCyPAs for 24 h, then, viability was measured by Cell Counting Kit-8.

Results: Anti-CsCyPA antibodies could combine with MuCyPA and inhibit its peptidyl prolyl isomerase (PPIase) activity. In the antibodies treatment group, blood coagulation indicators including PT, aPTT, D-dimer and platelet count were obviously more ameliorative, the proinflammary factors like IL-6, TNF-α, IL-1β were significantly lower at 12 h and 24 h after surgery and the viability of ECs was significantly improved compared to those in the control group. Furthermore, the survival rate was elevated, ranging from 10.0 % to 45.0 % compared to the control group.

Conclusions: These antibodies may have a favorable effect on sepsis via inhibition of enzymic activity or protection of endothelial cells.

No MeSH data available.


Related in: MedlinePlus

Detection of the Viability of EC with rMuCyPA and Anti-CsCyPAs. X axis final supernatant content of rMuCyPA and anti-CsCyPAs in each speed; Y axis presented the absorbance at OD450nm in each well after incubated with 10 μl of CCK-8 solution at 37 °C for 2 h. Pro is the abbreviation of rMuCyPA; anti is the abbreviation of anti-CsCyPAs; *P < 0.05
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Fig9: Detection of the Viability of EC with rMuCyPA and Anti-CsCyPAs. X axis final supernatant content of rMuCyPA and anti-CsCyPAs in each speed; Y axis presented the absorbance at OD450nm in each well after incubated with 10 μl of CCK-8 solution at 37 °C for 2 h. Pro is the abbreviation of rMuCyPA; anti is the abbreviation of anti-CsCyPAs; *P < 0.05

Mentions: To evaluate the viability of ECs after being co-cultured with rMuCyPA, ECs were incubated in 0 μg (control), 0.1 μg, 1 μg and 10 μg MuCyPA for 24 h, and cell viability was measured using the CCK-8 assays in Fig 9. Compared with the control group, the viability of ECs exposed to rMuCyPA showed a dose-dependent decrease. In the inhibition test, 0.1 μg, 1 μg and 10 μg antibodies were co-cultured with 10 μg MuCyPA for 1 h before reaction. Compared with 10 μg MuCyPA group, the result showed significant increase on the percentage of cells in the 1 μg and 10 μg antibody groups in a dose-dependent manner (p < 0.05). There was no significant difference between 10 μg MuCyPA group and 0.1 μg antibodies group (p > 0.05).Fig 9


Prognosis of sepsis induced by cecal ligation and puncture in mice improved by anti-Clonorchis Sinensis cyclopholin a antibodies.

Song T, Yang M, Chen J, Huang L, Yin H, He T, Huang H, Hu X - Parasit Vectors (2015)

Detection of the Viability of EC with rMuCyPA and Anti-CsCyPAs. X axis final supernatant content of rMuCyPA and anti-CsCyPAs in each speed; Y axis presented the absorbance at OD450nm in each well after incubated with 10 μl of CCK-8 solution at 37 °C for 2 h. Pro is the abbreviation of rMuCyPA; anti is the abbreviation of anti-CsCyPAs; *P < 0.05
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4591565&req=5

Fig9: Detection of the Viability of EC with rMuCyPA and Anti-CsCyPAs. X axis final supernatant content of rMuCyPA and anti-CsCyPAs in each speed; Y axis presented the absorbance at OD450nm in each well after incubated with 10 μl of CCK-8 solution at 37 °C for 2 h. Pro is the abbreviation of rMuCyPA; anti is the abbreviation of anti-CsCyPAs; *P < 0.05
Mentions: To evaluate the viability of ECs after being co-cultured with rMuCyPA, ECs were incubated in 0 μg (control), 0.1 μg, 1 μg and 10 μg MuCyPA for 24 h, and cell viability was measured using the CCK-8 assays in Fig 9. Compared with the control group, the viability of ECs exposed to rMuCyPA showed a dose-dependent decrease. In the inhibition test, 0.1 μg, 1 μg and 10 μg antibodies were co-cultured with 10 μg MuCyPA for 1 h before reaction. Compared with 10 μg MuCyPA group, the result showed significant increase on the percentage of cells in the 1 μg and 10 μg antibody groups in a dose-dependent manner (p < 0.05). There was no significant difference between 10 μg MuCyPA group and 0.1 μg antibodies group (p > 0.05).Fig 9

Bottom Line: Lung and mesentery tissues were stained with hematoxylin-eosin.Anti-CsCyPA antibodies could combine with MuCyPA and inhibit its peptidyl prolyl isomerase (PPIase) activity.Furthermore, the survival rate was elevated, ranging from 10.0 % to 45.0 % compared to the control group.

View Article: PubMed Central - PubMed

Affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong, China. songtianzhang@163.com.

ABSTRACT

Background: Cyclophilin A (CyPA), a ubiquitously distributed intracellular protein, is thought to be one of the important inflammatory factors and plays a significant role in the development process of sepsis. In the form of cytokine, CyPA deteriorates sepsis by promoting intercellular communication, apoptosis of endothelial cells and chemotactic effect on inflammatory cells. In our previous study, cyclophilin A of Clonorchis sinensis (CsCyPA), a type of excretory-secretory antigen, could induce the patients infected with Clonorchis sinensis to produce specific anti-CsCyPA antibodies. In this study, we investigated whether anti-CsCyPA antibodies could cross-react with CyPA and then play a protective role against sepsis, just like other anti-cytokine antagonists.

Methods: The mice model with sepsis was established with cecal ligation and puncture (CLP). Fifty mg/kg purified anti-CsCyPA antibodies were injected via the caudal vein 6 h after the CLP operation, and persistent observation was performed for 72 h. Blood samples and tissues were collected at 6 h, 12 h, 24 h, 48 h and 72 h after CLP. Cytokines in serum were measured by ELISA. Lung and mesentery tissues were stained with hematoxylin-eosin. Endothelial cells (ECs) isolated from murine aorta were co-cultured with CyPA of mice (MuCyPA) and anti-CsCyPAs for 24 h, then, viability was measured by Cell Counting Kit-8.

Results: Anti-CsCyPA antibodies could combine with MuCyPA and inhibit its peptidyl prolyl isomerase (PPIase) activity. In the antibodies treatment group, blood coagulation indicators including PT, aPTT, D-dimer and platelet count were obviously more ameliorative, the proinflammary factors like IL-6, TNF-α, IL-1β were significantly lower at 12 h and 24 h after surgery and the viability of ECs was significantly improved compared to those in the control group. Furthermore, the survival rate was elevated, ranging from 10.0 % to 45.0 % compared to the control group.

Conclusions: These antibodies may have a favorable effect on sepsis via inhibition of enzymic activity or protection of endothelial cells.

No MeSH data available.


Related in: MedlinePlus