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Tissue Transglutaminase-Regulated Transformed Growth Factor-β1 in the Parasite Links Schistosoma japonicum Infection with Liver Fibrosis.

Tang J, Zhu X, Zhao J, Fung M, Li Y, Gao Z, Yan S, Li X, Ji X, Su F, Li Z - Mediators Inflamm. (2015)

Bottom Line: During tTG inhibition, TGF-β1 level decreased in sera and liver of infected mice.The TGF-β1 mature peptide cDNA sequence and its extended sequence were amplified through RT-PCR and RACE-PCR using adult worms as template, and sequence is analyzed and loaded to NCBI GenBank (number GQ338152.1).TGF-β1 transcript in Sj eggs was higher than in adult worms.

View Article: PubMed Central - PubMed

Affiliation: Guangzhou Hoffmann Institute of Immunology, School of Basic Sciences, Guangzhou Medical University, Guangzhou 510182, China.

ABSTRACT
Transforming growth factor (TGF-β1) is among the strongest factors of liver fibrogenesis, but its association with Schistosoma-caused liver fibrosis is controversial. Tissue transglutaminase (tTG) is the principal enzyme controlling TGF-β1 maturation and contributes to Sj-infected liver fibrosis. Here we aim to explore the consistency between tTG and TGF-β1 and TGF-β1 source and its correlation with liver fibrosis after Sj-infection. TGF-β1 was upregulated at weeks 6 and 8 upon liver fibrosis induction. During tTG inhibition, TGF-β1 level decreased in sera and liver of infected mice. TGF-β1 showed positive staining in liver containing Sj adult worms and eggs. TGF-β1 was also detected in Sj adult worm sections, soluble egg antigen and Sj adult worm antigen, and adult worms' culture medium. The TGF-β1 mature peptide cDNA sequence and its extended sequence were amplified through RT-PCR and RACE-PCR using adult worms as template, and sequence is analyzed and loaded to NCBI GenBank (number GQ338152.1). TGF-β1 transcript in Sj eggs was higher than in adult worms. In Sj-infected liver, transcriptional level of TGF-β1 from Sj, but not mouse liver, correlated with liver fibrosis extent. This study provides evidence that tTG regulates TGF-β1 and illustrates the importance of targeting tTG in treating Sj infection-induced fibrosis.

No MeSH data available.


Related in: MedlinePlus

TGF-β1 protein was expressed in Sj and was secreted. (a) Sj adult female and male worms were fixed in paraformaldehyde, embedded in paraffin, and then sliced and stained for TGF-β1. Representative staining (brown) is shown at 40x magnification (large panel) and ×200 (inset). Dotted red arrow: gut epithelial cells; solid red arrow: subtegumental cells. (b) Equal amounts of Sj soluble adult worm antigen and soluble egg antigen were tested for TGF-β1 by ELISA. Data were shown as means ± SD. Experiment was performed four times (∗P < 0.05; and ∗∗P < 0.01 compared with negative PBS control). (c) Twenty pairs of adult worms were freshly collected, washed thrice using PBS, transferred into 2 mL sterile RPMI 1640 medium supplemented with 1 mM glutamine, 1000 units/mL penicillin, and 1000 μg/mL streptomycin for 2 h, and finally cultured in 2 mL sterile RPMI 1640 medium supplemented with 20% FBS, 1 mM glutamine, 100 units/mL penicillin, and 100 μg/mL streptomycin for 16 h. Sj worm culture medium was collected for TGF-β1 detection by ELISA, and condition medium (sterile RPMI 1640 medium supplemented with 20% FBS, 1 mM glutamine, 100 units/mL penicillin, and 100 μg/mL streptomycin) was used as control. Experiment was performed four times (∗P < 0.05; and ∗∗P < 0.01 compared with control medium without Sj adult worms).
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fig3: TGF-β1 protein was expressed in Sj and was secreted. (a) Sj adult female and male worms were fixed in paraformaldehyde, embedded in paraffin, and then sliced and stained for TGF-β1. Representative staining (brown) is shown at 40x magnification (large panel) and ×200 (inset). Dotted red arrow: gut epithelial cells; solid red arrow: subtegumental cells. (b) Equal amounts of Sj soluble adult worm antigen and soluble egg antigen were tested for TGF-β1 by ELISA. Data were shown as means ± SD. Experiment was performed four times (∗P < 0.05; and ∗∗P < 0.01 compared with negative PBS control). (c) Twenty pairs of adult worms were freshly collected, washed thrice using PBS, transferred into 2 mL sterile RPMI 1640 medium supplemented with 1 mM glutamine, 1000 units/mL penicillin, and 1000 μg/mL streptomycin for 2 h, and finally cultured in 2 mL sterile RPMI 1640 medium supplemented with 20% FBS, 1 mM glutamine, 100 units/mL penicillin, and 100 μg/mL streptomycin for 16 h. Sj worm culture medium was collected for TGF-β1 detection by ELISA, and condition medium (sterile RPMI 1640 medium supplemented with 20% FBS, 1 mM glutamine, 100 units/mL penicillin, and 100 μg/mL streptomycin) was used as control. Experiment was performed four times (∗P < 0.05; and ∗∗P < 0.01 compared with control medium without Sj adult worms).

Mentions: Similar to the findings of Hirata et al. [24], our results showed that TGF-β subfamily immunoreactive molecules are probably expressed in adult worms and eggs of Sj (Figure 2(c)). We validated these results through an IHC assay using sections of male and female adult worms (Figure 3(a)). Moreover, we detected TGF-β1 protein in the SEA, SWA of Sj, and in the culture medium of Sj adult worm using ELISA (Figures 3(b) and 3(c)). Figure 3(a) shows that TGF-β1 immunoreactivity was apparent in subtegumental cells and the lining gut epithelial cells of male and female worms, especially in female worms. TGF-β1 concentrations in SEA and SWA were 17.9 and 20.7 pg/mL, respectively (Figure 3(b)). Furthermore, higher concentration of TGF-β1 was secreted in culture medium of adult worms than in the control medium (Figure 3(c)).


Tissue Transglutaminase-Regulated Transformed Growth Factor-β1 in the Parasite Links Schistosoma japonicum Infection with Liver Fibrosis.

Tang J, Zhu X, Zhao J, Fung M, Li Y, Gao Z, Yan S, Li X, Ji X, Su F, Li Z - Mediators Inflamm. (2015)

TGF-β1 protein was expressed in Sj and was secreted. (a) Sj adult female and male worms were fixed in paraformaldehyde, embedded in paraffin, and then sliced and stained for TGF-β1. Representative staining (brown) is shown at 40x magnification (large panel) and ×200 (inset). Dotted red arrow: gut epithelial cells; solid red arrow: subtegumental cells. (b) Equal amounts of Sj soluble adult worm antigen and soluble egg antigen were tested for TGF-β1 by ELISA. Data were shown as means ± SD. Experiment was performed four times (∗P < 0.05; and ∗∗P < 0.01 compared with negative PBS control). (c) Twenty pairs of adult worms were freshly collected, washed thrice using PBS, transferred into 2 mL sterile RPMI 1640 medium supplemented with 1 mM glutamine, 1000 units/mL penicillin, and 1000 μg/mL streptomycin for 2 h, and finally cultured in 2 mL sterile RPMI 1640 medium supplemented with 20% FBS, 1 mM glutamine, 100 units/mL penicillin, and 100 μg/mL streptomycin for 16 h. Sj worm culture medium was collected for TGF-β1 detection by ELISA, and condition medium (sterile RPMI 1640 medium supplemented with 20% FBS, 1 mM glutamine, 100 units/mL penicillin, and 100 μg/mL streptomycin) was used as control. Experiment was performed four times (∗P < 0.05; and ∗∗P < 0.01 compared with control medium without Sj adult worms).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: TGF-β1 protein was expressed in Sj and was secreted. (a) Sj adult female and male worms were fixed in paraformaldehyde, embedded in paraffin, and then sliced and stained for TGF-β1. Representative staining (brown) is shown at 40x magnification (large panel) and ×200 (inset). Dotted red arrow: gut epithelial cells; solid red arrow: subtegumental cells. (b) Equal amounts of Sj soluble adult worm antigen and soluble egg antigen were tested for TGF-β1 by ELISA. Data were shown as means ± SD. Experiment was performed four times (∗P < 0.05; and ∗∗P < 0.01 compared with negative PBS control). (c) Twenty pairs of adult worms were freshly collected, washed thrice using PBS, transferred into 2 mL sterile RPMI 1640 medium supplemented with 1 mM glutamine, 1000 units/mL penicillin, and 1000 μg/mL streptomycin for 2 h, and finally cultured in 2 mL sterile RPMI 1640 medium supplemented with 20% FBS, 1 mM glutamine, 100 units/mL penicillin, and 100 μg/mL streptomycin for 16 h. Sj worm culture medium was collected for TGF-β1 detection by ELISA, and condition medium (sterile RPMI 1640 medium supplemented with 20% FBS, 1 mM glutamine, 100 units/mL penicillin, and 100 μg/mL streptomycin) was used as control. Experiment was performed four times (∗P < 0.05; and ∗∗P < 0.01 compared with control medium without Sj adult worms).
Mentions: Similar to the findings of Hirata et al. [24], our results showed that TGF-β subfamily immunoreactive molecules are probably expressed in adult worms and eggs of Sj (Figure 2(c)). We validated these results through an IHC assay using sections of male and female adult worms (Figure 3(a)). Moreover, we detected TGF-β1 protein in the SEA, SWA of Sj, and in the culture medium of Sj adult worm using ELISA (Figures 3(b) and 3(c)). Figure 3(a) shows that TGF-β1 immunoreactivity was apparent in subtegumental cells and the lining gut epithelial cells of male and female worms, especially in female worms. TGF-β1 concentrations in SEA and SWA were 17.9 and 20.7 pg/mL, respectively (Figure 3(b)). Furthermore, higher concentration of TGF-β1 was secreted in culture medium of adult worms than in the control medium (Figure 3(c)).

Bottom Line: During tTG inhibition, TGF-β1 level decreased in sera and liver of infected mice.The TGF-β1 mature peptide cDNA sequence and its extended sequence were amplified through RT-PCR and RACE-PCR using adult worms as template, and sequence is analyzed and loaded to NCBI GenBank (number GQ338152.1).TGF-β1 transcript in Sj eggs was higher than in adult worms.

View Article: PubMed Central - PubMed

Affiliation: Guangzhou Hoffmann Institute of Immunology, School of Basic Sciences, Guangzhou Medical University, Guangzhou 510182, China.

ABSTRACT
Transforming growth factor (TGF-β1) is among the strongest factors of liver fibrogenesis, but its association with Schistosoma-caused liver fibrosis is controversial. Tissue transglutaminase (tTG) is the principal enzyme controlling TGF-β1 maturation and contributes to Sj-infected liver fibrosis. Here we aim to explore the consistency between tTG and TGF-β1 and TGF-β1 source and its correlation with liver fibrosis after Sj-infection. TGF-β1 was upregulated at weeks 6 and 8 upon liver fibrosis induction. During tTG inhibition, TGF-β1 level decreased in sera and liver of infected mice. TGF-β1 showed positive staining in liver containing Sj adult worms and eggs. TGF-β1 was also detected in Sj adult worm sections, soluble egg antigen and Sj adult worm antigen, and adult worms' culture medium. The TGF-β1 mature peptide cDNA sequence and its extended sequence were amplified through RT-PCR and RACE-PCR using adult worms as template, and sequence is analyzed and loaded to NCBI GenBank (number GQ338152.1). TGF-β1 transcript in Sj eggs was higher than in adult worms. In Sj-infected liver, transcriptional level of TGF-β1 from Sj, but not mouse liver, correlated with liver fibrosis extent. This study provides evidence that tTG regulates TGF-β1 and illustrates the importance of targeting tTG in treating Sj infection-induced fibrosis.

No MeSH data available.


Related in: MedlinePlus