基于转录组学探究日粮添加酵母β-葡聚糖对新城疫疫苗免疫鸡肠道免疫的影响

doi:10.11843/j.issn.0366-6964.2025.03.042

Abstract

Abstract:

The aim of this study is to investigate the effect of adding yeast β-glucan (G70) to the diet on intestinal immune function in chickens immunized Newcastle disease (ND) vaccine. Sixteen 1-day-old healthy black-bone chickens were randomly divided into 2 groups, the Vaccine group and the G70+Vaccine group, and in the G70+Vaccine group, yeast β-glucan was added at 1 g·kg^-1 in the basal diet. At 14 and 28 days of age, ND vaccine was administered for the 1^st immunization and booster immunization. At 35 days of age, jejunum tissue was collected to determine the number of IgA⁺cells, the proportions of CD4⁺CD8⁺double-positive T cells, and the mRNA expression levels of immune-related genes in jejunum, then transcriptome sequencing was performed, and the differentially expressed genes were analyzed by GO function and KEGG pathway. The results showed that, compared with the Vaccine group, dietary G70 significantly increased the number of IgA⁺cells (P < 0.05) and the proportions of CD4⁺CD8⁺double-positive T cells in the jejunum (P < 0.05), and the mRNA expression levels of GATA-3, MHC-Ⅰ, MHC-Ⅱ, CCR7 and IFN-γ were significantly up-regulated (P < 0.05). RNA-seq showed that there were 559 differentially expressed genes in G70+Vaccine group, of which 54 were up-regulated and 505 were down-regulated, the GO analysis revealed that most of the differentially expressed genes were annotated to the GO entries related to amine metabolism and catabolism, protein synthesis, stimulation of TGF-β, and regulation of enzyme activities. KEGG signaling pathway analysis revealed that the differentially expressed genes were mainly enriched in signaling pathways related to cell synthesis and metabolism, such as ribosomes, MAPK, cell adhesion and local adhesion. In summary, the addition of yeast β-glucan (G70) to the diet increased the number of IgA⁺cells, promoted the proportions of CD4⁺CD8⁺double-positive T cells, and up-regulated the expression of immune-related genes in jejunum. The possible mechanism of improving chicken's intestinal immune function might be affecting the IL-17 receptor, TGF-β and MAPK signaling pathways by regulating the expression of immune-related genes such as IL-17RD, TGF-β and TMEM158.

Key words: yeast β-glucan, Newcastle disease vaccine, intestinal immune function, transcriptomics

CLC Number:

S831.7

LI Changying, LI Jun, LI Xifeng, BI Shicheng, CAO Liting. Effect of Dietary Yeast β-glucan Supplementation on Intestinal Immune Function in Chickens Immunized against Newcastle Disease Vaccine based on Transcriptomic[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(3): 1441-1452.

Figures/Tables 12

Table 1

Table 2

Primer sequences for RT-qPCR"

基因 Genes	引物序列(5′→3′) Primer sequences(5′→3′)	扩增长度/bp Length
β-actin	5′-GAGAAATTGTGCGTGACATCA-3′ 5′-CCTGAACCTCTCATTGCCA-3′	134
TGF-β	F: 5′-TTCCAACACCAGGTCCTACTCCAG-3′ R: 5′-AAGCAGACAGGTCCAGCAATAACAG-3′	88
IL-6	F: 5′-GAAATCCCTCCTCGCCAATCTGAAG-3′ R: 5′-GCCCTCACGGTCTTCTCCATAAAC-3′	108
IFN-γ	F: 5′-ACGACACCATCCTGGACACC-3′ R: 5′-TTTGGCGTTGGCTGTCGTTC-3′	129
NF-κB	F: 5′-CTTGCCAAGCGTCACTGCAA-3′ R: 5′-GCAGTGAGATGGCGCTGAAC-3′	89
CD40	F: 5′-GGGCTCGTGGTGAAGGTGAAAG-3′ R: 5′-GGATCAGCACTGACAGCGATGAG-3′	85
CD80	F: 5′-TCGTTCAGAGTCTCCAGTCTTCACC-3′ R: 5′-CAGCGGTAACAAAGAGGGTCACAG-3′	84
CCR7	F: 5′-CCGACGACTATGACGCCAACAC-3′ R: 5′-GGAGGAAGGCAGCACGGAAATC-3′	96
CCR-9	F: 5′-AGTGCTCCTCCTTGGGTGACAG-3′ R: 5′-GGCATGAGGCAGGAACAACAGAG-3′	132
TLR-3	F: 5′-GCAAGCTATTGAGCAAAGTCGAGAC-3′ R: 5′-GCCAGTTCAAGATGCAGCAAGATC-3′	128
TLR-4	F: 5′-CATCCCAACCCAACCACAGTAGC-3′ R: 5′-CCACTGAGCAGCACCAATGAGTAG-3′	119
TLR-5	F: 5′-ACTCCCTTCCTTCCCACATCTGAC-3′ R: 5′-TGTGTTGCTACTATTGCCGTGTGAG-3′	87
GATA-3	F: 5′-CTACTTGTGTAACGCCTGTGGACTC-3′ R: 5′-GTGGTGGTGGTCTGACAGTTAGC-3′	132
J-CHAIN	F: 5′-GGTTCGTCCTTGTGGCAGGTTATC-3′ R: 5′-GAGGTCACCGTTACGCACTTACAC-3′	88
pIgR	F: 5′-AAGGGCTCTCCAACAGGGTCAG-3′ R: 5′-AGAACTTCCTCGTGCTGGCATAATC-3′	144
MHC-Ⅰ	F: 5′-GCACAGCCCCATCCTCT-3′ R: 5′-TGGCCCATCATTTTATTTCA-3′	101
MHC-Ⅱ	F: 5′-CAGCGTTCTTCTTCTGCGGT-3′ R: 5′-GGTTGTAGATTTCCCGTTCCAG-3′	90
AN04	5′-CAGCCCTTCCAAAGATGATGACTCC-3′ 5′-TTCCTCTCCCACCACTTCCAACC-3′	94
ASPA	5′-AATCTTGGACAGACAGTGGTGGAAG-3′ 5′-TGTTAGCAGTGGTGTTGTGAAGGTC-3′	136
IL-17RD	5′-GCCTTCCTCCTCGCTCTCCTC-3′ 5′-ATTGCTCTTGGTGACAGATGACAGG-3′	135
MYH	5′-AGCCGAAGCACAAGCCAATCTG-3′ 5′-GCCAATATCCCTTGACTTGCTCCTC-3′	133
CLDN3	5′-CTTCATCGGCAACAACATCGTGAC-3′ 5′-CCAGCATGGAGTCGTACACCTTG-3′	113
TMEM158	5′-GCTCAACTTCTGCTGCTTGGATTTC-3′ 5′-CCACGCTCCACACGATGATGAC-3′	134
RRS1	5′-GGCAAGAAACGGCGCTTCCAG-3′ 5′-CTCGCGGAGCTGCTTGTTGAC-3′	138
MT3	5′-ACCTGCACGTGTGGAGACAA-3′ 5′-GCACACTTGGCACATCCTGC-3′	101

Table 2

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Table 3

Down- and up-regulated differentially expressed genes"

基因ID Gene ID	基因名称 Gene name	差异倍数 log₂FC	基因描述 Gene characterization
下调基因 Down-regulated genes
ENSGALG00000053991	SERF2	-3.070914732	small EDRK-rich factor 2
ENSGALG00000039489	5-8S rRNA	-3.027282516	5_8S ribosomal RNA
ENSGALG00000030890	RRS1	-2.198902274	ribosome biogenesis regulator homolog
ENSGALG00000034326	TMEM158	-2.198603339	transmembrane protein 158
ENSGALG00000041752	CD164L2	-1.965298427	CD164 molecule like 2
ENSGALG00000049538	RPS28	-1.921395818	ribosomal protein S28
ENSGALG00000002517	TMEM35B	-1.741653236	transmembrane protein 35B
ENSGALG00000043641	JUND	-1.618482633	JunD proto-oncogene
ENSGALG00000037713	FNDC10	-1.458563877	fibronectin type Ⅲ
ENSGALG00000049153	PDF	-1.400669956	peptide deformylase
ENSGALG00000029768	PTGDR	-1.375135892	prostaglandin D2 receptor-like
ENSGALG00000037716	UBA52	-1.330577817	ubiquitin A-52
ENSGALG00000031067	TMEM132A	-1.305910743	transmembrane protein 132A
ENSGALG00000030407	IL-6R	-1.268521422	interleukin 6 receptor
ENSGALG00000041232	TMEM223	-1.142077014	transmembrane protein 223
上调基因 Up-regulated genes
ENSGALG00000011614	ANO4	2.392598397	anoctamin 4
ENSGALG00000015358	MYH15	1.956091863	myosin, heavy chain 15
ENSGALG00000051773	JHY	1.822528177	chromosome 11 open reading frame 63
ENSGALG00000005499	IL17-RD	1.615012462	interleukin 17 receptor
ENSGALG00000003876	TIMD4	1.524616906	T-cell immunoglobulin and mucin domain containing 4
ENSGALG00000043027	HEPHL1	1.417362402	hephaestin like 1
ENSGALG00000005360	CA4	1.377146878	carbonic anhydrase 4
ENSGALG00000049288	CD86	1.278387892	CD86 molecule
ENSGALG00000028790	DNASE2B	1.267707784	deoxyribonuclease 2 beta
ENSGALG00000019651	AGR3	1.241116654	anterior gradient 3
ENSGALG00000030703	CYBRD1	1.197753577	cytochrome b reductase 1
ENSGALG00000002390	CA6	1.123467369	carbonic anhydrase 6
ENSGALG00000037077	CCR5	1.038338392	C-C motif chemokine receptor 5
ENSGALG00000013748	CD226	1.02970962	CD226 molecule

Table 3

Fig. 6

Fig. 7

Table 4

Fig. 8

References 31

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日粮组成 Feed ingredient	1~21日龄 1~21 day-old	22-35日龄 22~35 day-old
玉米Corn	55.34	60.53
麸皮Wheat bran	2.70	2.70
植物油Vegetable oil	3.17	3.83
鱼粉Fish meal	5.01	4.90
豆粕Soybean meal	19.00	12.00
胡麻饼Hemp cake	4.50	5.20
棉籽粕Cottonseed meal	3.20	4.10
菜籽粕Rapeseed meal	2.70	3.06
DL-蛋氨酸DL-Methionine	0.12	0.02
磷酸氢钙CaHPO₄	3.13	2.46
石粉Limestone	0.15	0.33
食盐NaCl	0.27	0.26
氯化胆碱Choline chloride	0.12	0.02
预混料^a Premix^a	0.50	0.50
合计Total	100	100
营养水平^b Nutrient content^b
粗蛋白质Crude protein	20.00	18.90
代谢能/(MJ·kg^－1)ME	12.11	12.54
蛋氨酸Methionine	0.48	0.38
赖氨酸Lysine	1.09	0.94
钙Calcium	1.00	0.90
有效磷Effective phosphorus	0.45	0.41

通路 Pathway	数量 Count	q值 q-value	基因 Gene
核糖体 Ribosome	11	0.001	RPS28/MRPL9、MRPS14/MRPL17、MRPL4/UBA52、MRPL20/RPS19、MRPS12/RPL35A、MRPS21
MAPK信号通路 MAPK signaling pathway	9	0.23	JUND/ECSIT NFKB2/TGFA/GADD45G/PGF
细胞黏附分子 Cell adhesion molecules	7	0.032	CLDN3/BLB1、CD40/ITGB7、CD86/CD226
黏着斑 Focal adhesion	7	0.26	PPP1R12C/ITGB7/PGF

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Effect of Dietary Yeast β-glucan Supplementation on Intestinal Immune Function in Chickens Immunized against Newcastle Disease Vaccine based on Transcriptomic

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