Expression of key glycosphingolipid biosynthesis-globo series pathway genes inEscherichia coliF18-resistant andEscherichia coliF18-sensitive piglets

Dong, Wende; Dai, Chaohui; Sun, Li; Wang, J.; Sun, Shengyuan; Zhu, Guoqiang; Wu, Shenglong; Bao, Wenbin

doi:10.1111/age.12428

Cited by 9 publications

(6 citation statements)

References 22 publications

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“…The P1 antigen and blood group H type 2 antigens were 13.2 (1.0) and 10.9 (0.7) to 12.7 (1.3) in IgM, respectively, but the signal intensity toward high-mannose carbohydrates were negative. The comparison of ganglio- (Table 3) 32,[36][37][38][40][41][42] and globo-series (Table 4) 34,35,[43][44][45] glycolipid signal intensity in naïve monkeys suggested that certain carbohydrates in upstream of the cascades (GA1, GA2, GM2, GD3, or Gb3) showed higher signals than others (Figure 3). In the hierarchical clustering ( Figure 2), GM2 was clustered with GD3 in IgM, and with GD3 and GD2 in IgG.…”

Section: Antibody Signals Toward Non-α-gal Pig Antigens and Its Anamentioning

confidence: 99%

Profiling natural serum antibodies of non‐human primates with a carbohydrate antigen microarray

Nanno

Sterner

Gildersleeve

et al. 2019

Xenotransplantation

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Background: Engineering of α-Galactosyltransferase gene-knockout pigs circumvented hyperacute rejection of pig organs after xenotransplantation in non-human primates. Overcoming this hurdle revealed the importance of non-α-Gal carbohydrate antigens in the immunobiology of acute humoral xenograft rejection. Methods: This study analyzed serum from seven naïve cynomolgus monkeys (blood type O/B/AB = 3/2/2) for the intensity of natural IgM and IgG signals using carbohydrate antigen microarray, which included historically reported α-Gal and non-α-Gal carbohydrate antigens with various modifications.Results: The median (range) of IgM and IgG signals were 12.71 (7.23-16.38) and 9.05 (7.23-15.90), respectively. The highest IgM and IgG signals with narrowest distribution were from mono-and disaccharides, followed by modified structures. Natural anti-α-Gal antibody signals were medium to high in IgM (11.2-15.9) and medium in IgG (8.5-11.6) spectra, and was highest with Lac core structure (Galα1-3Galβ1-4Glc, iGb3) and lowest with LacNAc core structure (Galα1-3Galβ1-4GlcNAc). Similar signal intensities (up to 15.8 in IgM and up to 11.8 in IgG) were observed for historically detected natural non-α-Gal antigens, which included Tn antigen, T antigen, GM2 glycolipid, and Sd a antigen. The hierarchical clustering analysis revealed the presence of clusters of anti-A antibodies and was capable of distinguishing between the blood group B and AB non-human primates. Conclusions:The results presented here provide the most comprehensive evaluation of natural antibodies present in cynomolgus monkeys. K E Y W O R D S carbohydrate antigen microarray, natural serum antibodies, non-human primates, xenotransplantation GM3(Gc)−Sp − 05 CT/Sda−Sp − 05

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Section: Antibody Signals Toward Non-α-gal Pig Antigens and Its Anamentioning

confidence: 99%

Profiling natural serum antibodies of non‐human primates with a carbohydrate antigen microarray

Nanno

Sterner

Gildersleeve

et al. 2019

Xenotransplantation

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“…The GAPDH and β-actin genes were used as internal references. The primers for GAPDH were: forward 5'-ACATCATCCCTGCTTC-TACTGG-3', reverse 5'-CTCGGACGCCTGCTTCAC-3'; those for β-actin were: forward 5'-TGGCGCCCAGCAC-GATGAAG-3', reverse 5'-GATGGAGGGGCCGGACTCG-3' (Dong et al, 2016). Each sample was analyzed in triplicate, and relative gene expression was calculated using the 2 -ΔΔCt method (Livak and Schmittgen, 2001).…”

Section: Animals and Sample Collectionmentioning

confidence: 99%

Age-associated changes in DNA methylation and expression of the TNFα gene in pigs

et al. 2018

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DNA methylation is an important mediator of gene expression regulation and has been shown to be closely linked to aging. Immune-related genes tend to be influenced by DNA methylation at different ages. To explore DNA methylation changes in the porcine TNFα gene and analyze their potential effects on gene expression, we measured the methylation level of the TNFα promoter and TNFα mRNA expression in the spleen of Meishan piglets at six developmental stages (1, 7, 14, 21, 28 and 35 days old) by bisulfite sequencing PCR and quantitative PCR. The results revealed a trend for TNFα promoter methylation level to increase and mRNA expression to decrease with age. Correlation analysis showed a significant negative association between methylation level and mRNA expression (Pearson's r = − 0.775, P = 4.87E-07). In addition, the transcription factor Sp1 was revealed to bind with the TNFα promoter and regulate TNFα expression. DNA methylation in the TNFα promoter was found to decrease the promoter's activity, and methylation inhibition could enhance the expression level of TNFα, providing functional evidence that promoter methylation controls TNFα expression. Together, our data provide insights into age-associated changes in promoter methylation of the TNFα gene in the spleen and contribute to our understanding of regulatory mechanisms for TNFα expression in the immune system of pigs.

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“…Furthermore, these DEGs were enriched in 20 pathways, including 41 significantly enriched Toll-like receptor signaling pathways (i.e., p < 0.05) (see Supplementary Table S3 ). Among them, the pathway of the “Glycosphingolipid biosynthesis-lacto and neolacto series” (ko00940) was enriched in the shFUT8 and shNC samples ( Figure 4 D), which probably played an important role in the regulation of E. coli F18 receptor formation [ 20 , 21 ].…”

Section: Resultsmentioning

confidence: 99%

“…Additionally, we found that FUT8 also significantly affected the glycosphingolipid biosynthesis pathways. Studies showed that glycosphingolipid biosynthesis correlated with the generation of the receptor for E. coli F18 [ 19 , 20 ]. Thus, we speculated that FUT8 most likely regulated E. coli F18 susceptibility via the activation or suppression of TLR signaling (which is related to immune response) and glycosphingolipid biosynthesis (which is related to the formation of the E. coli F18 receptor).…”

Section: Discussionmentioning

confidence: 99%

Effect and Mechanism Analysis of Pig FUT8 Gene on Resistance to Escherichia coli F18 Infection

Wang

et al. 2022

IJMS

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Post-weaning diarrhea caused by enterotoxigenic Escherichia coli F18 (E. coli F18) causes significant economic losses for pig producers. Fucosyltransferase 8 (FUT8) is a glycosyltransferase that catalyzes core fucosylation; however, its role in mediating the resistance to E. coli F18 infection in pigs remains unknown. In this study, we systematically verified the relationship between FUT8 expression and E. coli resistance. The results showed that FUT8 was expressed in all detected tissues of Meishan piglets and that its expression was significantly increased in the duodenum and jejunum of E. coli F18-sensitive individuals when compared to E. coli F18-resistant individuals. FUT8 expression increased after exposure to E. coli F18 (p < 0.05) and decreased significantly after LPS induction for 6 h (p < 0.01). Then, the IPEC-J2 stable cell line with FUT8 interference was constructed, and FUT8 knockdown decreased the adhesion of E. coli F18ac to IPEC-J2 cells (p < 0.05). Moreover, we performed a comparative transcriptome study of IPEC-J2 cells after FUT8 knockdown via RNA-seq. In addition, further expression verification demonstrated the significant effect of FUT8 on the glycosphingolipid biosynthesis and Toll-like signaling pathways. Moreover, the core promoter of FUT8, which was located at −1213 bp to −673 bp, was identified via luciferase assay. Interestingly, we found a 1 bp C base insertion mutation at the −774 bp region, which could clearly inhibit the transcriptional binding activity of C/EBPα to an FUT8 promoter. Therefore, it is speculated that FUT8 acts in a critical role in the process of E. coli infection; furthermore, the low expression of FUT8 is conducive to the enhancement of E. coli resistance in piglets. Our findings revealed the mechanism of pig FUT8 in regulating E. coli resistance, which provided a theoretical basis for the screening of E. coli resistance in Chinese local pig breeds.

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Expression of key glycosphingolipid biosynthesis-globo series pathway genes inEscherichia coliF18-resistant andEscherichia coliF18-sensitive piglets

Cited by 9 publications

References 22 publications

Profiling natural serum antibodies of non‐human primates with a carbohydrate antigen microarray

Profiling natural serum antibodies of non‐human primates with a carbohydrate antigen microarray

Age-associated changes in DNA methylation and expression of the <i>TNFα</i> gene in pigs

Effect and Mechanism Analysis of Pig FUT8 Gene on Resistance to Escherichia coli F18 Infection

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