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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 gene in Sj was cloned and identified. (a) TGF-β1 mature peptide cDNA in Sj was amplified using Sj adult worm cDNA collected from infected mice as template and primers designed according to mouse TGF-β1 mature peptide sequence, and the 5′ and 3′ ends of TGF-β1 in Sj were extended via RACE. The alignment of TGF-β1 cDNA sequences of Sj and mouse is shown (Query is Sj TGF-β1 and Sbjct. is mouse TGF-β1 cDNA sequence). Solid lines: specific mouse TGF-β1 primer pairs for PCR amplification; dotted lines: specific Sj TGF-β1 primer pairs for PCR amplification. Gray: TGF-β1 mature peptide cDNA sequence. (b) The alignment of TGF-β1 amino acid sequences of Sj and mouse is shown (Query is Sj TGF-β1 and Sbjct. is mouse TGF-β1 amino acid sequence). Gray: TGF-β1 mature peptide amino acid sequence. (c) TGF-β1 primers of mouse- or Sj-specific source were identified through PCR using Sj adult worms or normal mice liver cDNA as template, respectively. M: DL2000 DNA Marker; 1: mouse cDNA as template and specific Sj TGF-β1 primer; 2: mouse cDNA as template and specific mouse TGF-β1 primer; 3: Sj cDNA as template and specific Sj TGF-β1 primer; 4: Sj cDNA as template and specific mouse TGF-β1 primer.
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fig4: TGF-β1 gene in Sj was cloned and identified. (a) TGF-β1 mature peptide cDNA in Sj was amplified using Sj adult worm cDNA collected from infected mice as template and primers designed according to mouse TGF-β1 mature peptide sequence, and the 5′ and 3′ ends of TGF-β1 in Sj were extended via RACE. The alignment of TGF-β1 cDNA sequences of Sj and mouse is shown (Query is Sj TGF-β1 and Sbjct. is mouse TGF-β1 cDNA sequence). Solid lines: specific mouse TGF-β1 primer pairs for PCR amplification; dotted lines: specific Sj TGF-β1 primer pairs for PCR amplification. Gray: TGF-β1 mature peptide cDNA sequence. (b) The alignment of TGF-β1 amino acid sequences of Sj and mouse is shown (Query is Sj TGF-β1 and Sbjct. is mouse TGF-β1 amino acid sequence). Gray: TGF-β1 mature peptide amino acid sequence. (c) TGF-β1 primers of mouse- or Sj-specific source were identified through PCR using Sj adult worms or normal mice liver cDNA as template, respectively. M: DL2000 DNA Marker; 1: mouse cDNA as template and specific Sj TGF-β1 primer; 2: mouse cDNA as template and specific mouse TGF-β1 primer; 3: Sj cDNA as template and specific Sj TGF-β1 primer; 4: Sj cDNA as template and specific mouse TGF-β1 primer.

Mentions: To clarify whether or not TGF-β1 gene exists in Sj, we designed a pair of primers according to the cDNA sequence of mouse TGF-β1 mature peptide because the amino acid sequence of TGF-β1 mature peptide is highly conserved. This primer pair and Sj adult worm cDNA isolated from mice as template were used for PCR amplification of Sj TGF-β1 mature peptide cDNA, and a 339 bp product was obtained, sequenced, and analyzed using bioinformatics. The 5′ and 3′ ends of the fragment were extended via RACE-PCR using primers designed within the known sequence. The extended TGF-β1 cDNA sequence (792 bp long) was loaded into National Center for Biotechnology Information (NCBI) GenBank with number GQ338152.1. The deduced amino acid sequence comprised 263 amino acid residues and contained partial sequence of TGF-β1 propeptide and complete TGF-β1-like domain. The nucleotide sequence of extended TGF-β1 gene from Sj was 85% identical to that from mouse (Figure 4(a)), whereas the amino acid sequence of TGF-β1 was 88% identical (Figure 4(b)), as revealed by BLAST search results. The nucleotide sequence of Sj TGF-β1 mature peptide was different from that of mouse by multiple nucleotides, but the amino acid sequence of Sj TGF-β1 mature peptide had merely two amino acid differences compared with that of mouse (Figure 4(b)). To identify species-specific TGF-β1, we designed and identified primers of Sj and mouse-specific primers (the primer sequences have been underlined in solid and dashed lines, resp., as shown in Figure 4(a)). Figure 4(c) shows that positive bands appeared in the agarose gel only when we used specific primer pairs and the corresponding templates for PCR amplification. Moreover, the transcription level of TGF-β1 was higher in eggs than in adult Sj worms. These results suggested the existence of TGF-β1 gene in Sj adult worms and eggs.


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 gene in Sj was cloned and identified. (a) TGF-β1 mature peptide cDNA in Sj was amplified using Sj adult worm cDNA collected from infected mice as template and primers designed according to mouse TGF-β1 mature peptide sequence, and the 5′ and 3′ ends of TGF-β1 in Sj were extended via RACE. The alignment of TGF-β1 cDNA sequences of Sj and mouse is shown (Query is Sj TGF-β1 and Sbjct. is mouse TGF-β1 cDNA sequence). Solid lines: specific mouse TGF-β1 primer pairs for PCR amplification; dotted lines: specific Sj TGF-β1 primer pairs for PCR amplification. Gray: TGF-β1 mature peptide cDNA sequence. (b) The alignment of TGF-β1 amino acid sequences of Sj and mouse is shown (Query is Sj TGF-β1 and Sbjct. is mouse TGF-β1 amino acid sequence). Gray: TGF-β1 mature peptide amino acid sequence. (c) TGF-β1 primers of mouse- or Sj-specific source were identified through PCR using Sj adult worms or normal mice liver cDNA as template, respectively. M: DL2000 DNA Marker; 1: mouse cDNA as template and specific Sj TGF-β1 primer; 2: mouse cDNA as template and specific mouse TGF-β1 primer; 3: Sj cDNA as template and specific Sj TGF-β1 primer; 4: Sj cDNA as template and specific mouse TGF-β1 primer.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4493306&req=5

fig4: TGF-β1 gene in Sj was cloned and identified. (a) TGF-β1 mature peptide cDNA in Sj was amplified using Sj adult worm cDNA collected from infected mice as template and primers designed according to mouse TGF-β1 mature peptide sequence, and the 5′ and 3′ ends of TGF-β1 in Sj were extended via RACE. The alignment of TGF-β1 cDNA sequences of Sj and mouse is shown (Query is Sj TGF-β1 and Sbjct. is mouse TGF-β1 cDNA sequence). Solid lines: specific mouse TGF-β1 primer pairs for PCR amplification; dotted lines: specific Sj TGF-β1 primer pairs for PCR amplification. Gray: TGF-β1 mature peptide cDNA sequence. (b) The alignment of TGF-β1 amino acid sequences of Sj and mouse is shown (Query is Sj TGF-β1 and Sbjct. is mouse TGF-β1 amino acid sequence). Gray: TGF-β1 mature peptide amino acid sequence. (c) TGF-β1 primers of mouse- or Sj-specific source were identified through PCR using Sj adult worms or normal mice liver cDNA as template, respectively. M: DL2000 DNA Marker; 1: mouse cDNA as template and specific Sj TGF-β1 primer; 2: mouse cDNA as template and specific mouse TGF-β1 primer; 3: Sj cDNA as template and specific Sj TGF-β1 primer; 4: Sj cDNA as template and specific mouse TGF-β1 primer.
Mentions: To clarify whether or not TGF-β1 gene exists in Sj, we designed a pair of primers according to the cDNA sequence of mouse TGF-β1 mature peptide because the amino acid sequence of TGF-β1 mature peptide is highly conserved. This primer pair and Sj adult worm cDNA isolated from mice as template were used for PCR amplification of Sj TGF-β1 mature peptide cDNA, and a 339 bp product was obtained, sequenced, and analyzed using bioinformatics. The 5′ and 3′ ends of the fragment were extended via RACE-PCR using primers designed within the known sequence. The extended TGF-β1 cDNA sequence (792 bp long) was loaded into National Center for Biotechnology Information (NCBI) GenBank with number GQ338152.1. The deduced amino acid sequence comprised 263 amino acid residues and contained partial sequence of TGF-β1 propeptide and complete TGF-β1-like domain. The nucleotide sequence of extended TGF-β1 gene from Sj was 85% identical to that from mouse (Figure 4(a)), whereas the amino acid sequence of TGF-β1 was 88% identical (Figure 4(b)), as revealed by BLAST search results. The nucleotide sequence of Sj TGF-β1 mature peptide was different from that of mouse by multiple nucleotides, but the amino acid sequence of Sj TGF-β1 mature peptide had merely two amino acid differences compared with that of mouse (Figure 4(b)). To identify species-specific TGF-β1, we designed and identified primers of Sj and mouse-specific primers (the primer sequences have been underlined in solid and dashed lines, resp., as shown in Figure 4(a)). Figure 4(c) shows that positive bands appeared in the agarose gel only when we used specific primer pairs and the corresponding templates for PCR amplification. Moreover, the transcription level of TGF-β1 was higher in eggs than in adult Sj worms. These results suggested the existence of TGF-β1 gene in Sj adult worms and eggs.

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