Mechanism of Innate Immune Response Induced by <i>Albizia julibrissin</i> Saponin Active Fraction Using C2C12 Myoblasts

<i>Albizia julibrissin</i> saponin active fraction (AJSAF), is a prospective adjuvant with dual Th1/Th2 and Tc1/Tc2 potentiating activity. Its adjuvant activity has previously been proven to be strictly dependent on its spatial co-localization with antigens, highlighting the role of loca...

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Main Authors: Jing Du, Xiang Meng, Tiantian Ni, Beibei Xiong, Ziyi Han, Yongliang Zhu, Jue Tu, Hongxiang Sun
Format: Article
Language:English
Published: MDPI AG 2023-10-01
Series:Vaccines
Subjects:
Online Access:https://www.mdpi.com/2076-393X/11/10/1576
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author Jing Du
Xiang Meng
Tiantian Ni
Beibei Xiong
Ziyi Han
Yongliang Zhu
Jue Tu
Hongxiang Sun
author_facet Jing Du
Xiang Meng
Tiantian Ni
Beibei Xiong
Ziyi Han
Yongliang Zhu
Jue Tu
Hongxiang Sun
author_sort Jing Du
collection DOAJ
description <i>Albizia julibrissin</i> saponin active fraction (AJSAF), is a prospective adjuvant with dual Th1/Th2 and Tc1/Tc2 potentiating activity. Its adjuvant activity has previously been proven to be strictly dependent on its spatial co-localization with antigens, highlighting the role of local innate immunity in its mechanisms. However, its potential targets and pathways remain unclear. Here, its intracellular molecular mechanisms of innate immune response were explored using mouse C2C12 myoblast by integrative analysis of the in vivo and in vitro transcriptome in combination with experimental validations. AJSAF elicited a temporary cytotoxicity and inflammation towards C2C12 cells. Gene set enrichment analysis demonstrated that AJSAF regulated similar cell death- and inflammatory response-related genes in vitro and in vivo through activating second messenger–MAPK–CREB pathways. AJSAF markedly enhanced the Ca<sup>2+</sup>, cAMP, and reactive oxygen species levels and accelerated MAPK and CREB phosphorylation in C2C12 cells. Furthermore, Ca<sup>2+</sup> chelator, CREB inhibitor, and MAPK inhibitors dramatically blocked the up-regulation of IL-6, CXCL1, and COX2 in AJSAF-treated C2C12 cells. Collectively, these results demonstrated that AJSAF induced innate immunity via Ca<sup>2+</sup>–MAPK–CREB pathways. This study is beneficial for insights into the molecular mechanisms of saponin adjuvants.
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spelling doaj.art-4ec7f4f360cd4322ae64bbcddeef073c2023-11-19T18:24:48ZengMDPI AGVaccines2076-393X2023-10-011110157610.3390/vaccines11101576Mechanism of Innate Immune Response Induced by <i>Albizia julibrissin</i> Saponin Active Fraction Using C2C12 MyoblastsJing Du0Xiang Meng1Tiantian Ni2Beibei Xiong3Ziyi Han4Yongliang Zhu5Jue Tu6Hongxiang Sun7College of Animal Sciences, Zhejiang University, Hangzhou 310058, ChinaCollege of Animal Sciences, Zhejiang University, Hangzhou 310058, ChinaCollege of Animal Sciences, Zhejiang University, Hangzhou 310058, ChinaCollege of Animal Sciences, Zhejiang University, Hangzhou 310058, ChinaCollege of Animal Sciences, Zhejiang University, Hangzhou 310058, ChinaLaboratory of Gastroenterology Department, Second Affiliated Hospital of School of Medicine, Zhejiang University, Hangzhou 310009, ChinaCollege of Animal Sciences, Zhejiang University, Hangzhou 310058, ChinaCollege of Animal Sciences, Zhejiang University, Hangzhou 310058, China<i>Albizia julibrissin</i> saponin active fraction (AJSAF), is a prospective adjuvant with dual Th1/Th2 and Tc1/Tc2 potentiating activity. Its adjuvant activity has previously been proven to be strictly dependent on its spatial co-localization with antigens, highlighting the role of local innate immunity in its mechanisms. However, its potential targets and pathways remain unclear. Here, its intracellular molecular mechanisms of innate immune response were explored using mouse C2C12 myoblast by integrative analysis of the in vivo and in vitro transcriptome in combination with experimental validations. AJSAF elicited a temporary cytotoxicity and inflammation towards C2C12 cells. Gene set enrichment analysis demonstrated that AJSAF regulated similar cell death- and inflammatory response-related genes in vitro and in vivo through activating second messenger–MAPK–CREB pathways. AJSAF markedly enhanced the Ca<sup>2+</sup>, cAMP, and reactive oxygen species levels and accelerated MAPK and CREB phosphorylation in C2C12 cells. Furthermore, Ca<sup>2+</sup> chelator, CREB inhibitor, and MAPK inhibitors dramatically blocked the up-regulation of IL-6, CXCL1, and COX2 in AJSAF-treated C2C12 cells. Collectively, these results demonstrated that AJSAF induced innate immunity via Ca<sup>2+</sup>–MAPK–CREB pathways. This study is beneficial for insights into the molecular mechanisms of saponin adjuvants.https://www.mdpi.com/2076-393X/11/10/1576AJSAFC2C12 myoblastCa<sup>2+</sup>–MAPK–CREB pathwayinflammationinnate immunitymicroarray
spellingShingle Jing Du
Xiang Meng
Tiantian Ni
Beibei Xiong
Ziyi Han
Yongliang Zhu
Jue Tu
Hongxiang Sun
Mechanism of Innate Immune Response Induced by <i>Albizia julibrissin</i> Saponin Active Fraction Using C2C12 Myoblasts
Vaccines
AJSAF
C2C12 myoblast
Ca<sup>2+</sup>–MAPK–CREB pathway
inflammation
innate immunity
microarray
title Mechanism of Innate Immune Response Induced by <i>Albizia julibrissin</i> Saponin Active Fraction Using C2C12 Myoblasts
title_full Mechanism of Innate Immune Response Induced by <i>Albizia julibrissin</i> Saponin Active Fraction Using C2C12 Myoblasts
title_fullStr Mechanism of Innate Immune Response Induced by <i>Albizia julibrissin</i> Saponin Active Fraction Using C2C12 Myoblasts
title_full_unstemmed Mechanism of Innate Immune Response Induced by <i>Albizia julibrissin</i> Saponin Active Fraction Using C2C12 Myoblasts
title_short Mechanism of Innate Immune Response Induced by <i>Albizia julibrissin</i> Saponin Active Fraction Using C2C12 Myoblasts
title_sort mechanism of innate immune response induced by i albizia julibrissin i saponin active fraction using c2c12 myoblasts
topic AJSAF
C2C12 myoblast
Ca<sup>2+</sup>–MAPK–CREB pathway
inflammation
innate immunity
microarray
url https://www.mdpi.com/2076-393X/11/10/1576
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