Identification of DNMT3B2 as the Predominant Isoform of DNMT3B in Porcine Alveolar Macrophages and Its Involvement in LPS-Stimulated TNF-α Expression

Zhang, Yanbing; Li, Hui; Xiang, Xiaojun; Lu, Yan; Sharma, Mona; Li, Zongjie; Liu, Ke; Wei, Jianchao; Shao, Donghua; Li, Beibei; Ma, Zhiyong; Qiu, Yafeng

doi:10.3390/genes11091065

Cited by 4 publications

(3 citation statements)

References 34 publications

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“…We are particularly interested in the role of epigenetic regulation by DNA methylation in the immune response of porcine macrophages. In a previous study, we identified DNMT3B2 as the predominant isoform of DNMT3B in porcine alveolar macrophages and demonstrated its potential role in the regulation of lipopolysaccharide (LPS)-stimulated tumor necrosis factor α (TNF-α) expression [26]. In this study, we characterized DNMT1 in porcine alveolar macrophages.…”

Section: Introductionmentioning

confidence: 88%

A Porcine DNMT1 Variant: Molecular Cloning and Generation of Specific Polyclonal Antibody

Zhu

Wang

Zhang

et al. 2023

Genes

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DNA methyltransferase 1 (DNMT1), the first-identified DNA methyltransferase in mammals, has been well studied in the control of embryo development and somatic homeostasis in mice and humans. Accumulating reports have demonstrated that DNMT1 plays an important role in the regulation of differentiation and the activation of immune cells. However, little is known about the effects of porcine DNMT1 on such functional regulation, especially the regulation of the biological functions of immune cells. In this study, we report the cloning of DNMT1 (4833 bp in length) from porcine alveolar macrophages (PAMs). According to the sequence of the cloned DNMT1 gene, the deduced protein sequence contains a total of 1611 amino acids with a 2 amino acid insertion, a 1 amino acid deletion, and 12 single amino acid mutations in comparison to the reported DNMT1 protein. A polyclonal antibody based on a synthetic peptide was generated to study the expression of the porcine DNMT1. The polyclonal antibody only recognized the cloned porcine DNMT1 and not the previously reported protein due to a single amino acid difference in the antigenic peptide region. However, the polyclonal antibody recognized the endogenous DNMT1 in several porcine cells (PAM, PK15, ST, and PIEC) and the cells of other species (HEK-293T, Marc-145, MDBK, and MDCK cells). Moreover, our results demonstrated that all the detected tissues of piglet express DNMT1, which is the same as that in porcine alveolar macrophages. In summary, we have identified a porcine DNMT1 variant with sequence and expression analyses.

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Section: Introductionmentioning

confidence: 88%

A Porcine DNMT1 Variant: Molecular Cloning and Generation of Specific Polyclonal Antibody

Zhu

Wang

Zhang

et al. 2023

Genes

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“…RAW264.7 cells (ATCC) were maintained in Dulbecco’s modified Eagle medium (DMEM) with high glucose (Gibco, Invitrogen Corp., Carlsbad, CA, USA) supplemented by 10% (vol/vol) heat-inactivated fetal bovine serum (FBS) (Invitrogen). Porcine alveolar macrophages (PAMs) were isolated from piglets as previously described and cultured in RPMI 1640 containing 10% FBS, penicillin, streptomycin, and GlutaMAX (all purchased from Thermo Fisher Scientific, Shanghai, China) at 37 °C with 5% CO 2 [ 27 ]. For cell stimulation, RAW264.7 cells and PAMs were grown in 12-well tissue culture plates at a concentration of 4 × 10 5 cells/well.…”

Section: Methodsmentioning

confidence: 99%

TLR4 Agonist Combined with Trivalent Protein JointS of Streptococcus suis Provides Immunological Protection in Animals

et al. 2021

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Streptococcus suis (S. suis) serotype 2 (SS2) is the causative agent of swine streptococcosis and can cause severe diseases in both pigs and humans. Although the traditional inactive vaccine can protect pigs from SS2 infection, novel vaccine candidates are needed to overcome its shortcomings. Three infection-associated proteins in S. suis—muramidase-released protein (MRP), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and DLD, a novel putative dihydrolipoamide dehydrogenase—have been previously identified by immunoproteomic assays. In this study, the effective immune protection of the recombinant trivalent protein GAPDH-MRP-DLD (JointS) against SS2, SS7, and SS9 was determined in zebrafish. To improve the immune efficacy of JointS, monophosphoryl lipid A (MPLA) as a TLR4 agonist adjuvant, which induces a strong innate immune response in the immune cells of mice and pigs, was combined with JointS to immunize the mice. The results showed that immunized mice could induce the production of a high titer of anti-S. suis antibodies; as a result, 100% of mice survived after SS2 infection. Furthermore, JointS provides good protection against virulent SS2 strain infections in piglets. Given the above, there is potential to develop JointS as a novel subunit vaccine for piglets to prevent infection by SS2 and other S. suis serotypes.

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“…Compared to wild-type mice, Tet2-deficient mice are more susceptible to endotoxic shock and dextran sulfate sodium-induced colitis, leading to aggravated inflammation and IL-6 storm [227] (Figure 6C). In addition, during the polarization process of porcine macrophages stimulated by LPS, the expression of DNTM3b is reduced, leading to a downregulation of the methylation level of the Tnf-α gene promoter, thereby promoting its transcription [228] . Similarly, porcine reproductive and respiratory syndrome virus (PRRSV) infection in porcine macrophages inhibits the expression of FTO, resulting in an increase in m 6 A methylation levels.…”

Section: Pro-inflammatory Cytokinesmentioning

confidence: 99%

Impacts of epigenetic reprogramming on innate immunity

Fu,

Wang

2024

Anim. nutriomics

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The innate immune response is the host’s first line of defense, promptly activated upon pathogen invasion. Its precise and rapid activation relies on innate immune cells (IICs). Upon recognizing danger signals postinfection or injury, they release various innate immune effectors to eliminate invading pathogens or damaged cells, thus supporting the host’s immune homeostasis. Epigenetic modifications, by shaping chromatin structures, orchestrate specific gene transcription patterns to regulate the lineage development, differentiation, and activation of IICs. This intricate process ultimately contributes to effective pathogen clearance and IICs’ healthy development and differentiation. To thoroughly elucidate the epigenetic mechanisms underlying the development and differentiation of IICs, this review first introduces the fundamental concepts and latest advancements in this field. We then delve into how the immune microenvironment or other signaling molecules shape the epigenetic landscapes of distinct IIC subsets during their lineage development and differentiation. Furthermore, we summarize how different epigenetic modification profiles mediate specific transcriptional patterns, thereby influencing the lineage development, differentiation, and activation of IICs in response to infections or injuries. Finally, we discuss several unresolved critical issues from the perspective of targeting epigenetic modifications to modulate the innate immune response. In summary, this review aims to uncover the molecular mechanisms underlying the development, differentiation, and activation of IICs from an epigenetic perspective, providing theoretical foundations for scientific and medical researchers pursuing disease treatments.

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Identification of DNMT3B2 as the Predominant Isoform of DNMT3B in Porcine Alveolar Macrophages and Its Involvement in LPS-Stimulated TNF-α Expression

Cited by 4 publications

References 34 publications

A Porcine DNMT1 Variant: Molecular Cloning and Generation of Specific Polyclonal Antibody

A Porcine DNMT1 Variant: Molecular Cloning and Generation of Specific Polyclonal Antibody

TLR4 Agonist Combined with Trivalent Protein JointS of Streptococcus suis Provides Immunological Protection in Animals

Impacts of epigenetic reprogramming on innate immunity

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