绵羊痒螨钙网蛋白对兔外周血单个核细胞Th1/Th2和Th17/Treg免疫平衡的影响

doi:10.11843/j.issn.0366-6964.2025.04.038

Abstract

Abstract:

The aim of this study was to investigate the effects of Psoroptes ovis calreticulin (PsoCRT) on Th1/Th2 and Th17/Treg immune balance in the peripheral blood mononuclear cells (PBMC) of healthy New Zealand rabbits. In this study, PsoCRT gene without signal peptide was amplified from mixed-stage P. ovis total RNA by reverse transcription PCR, and its recombinant protein (rPsoCRT) was further expressed in prokaryotic expression and purified. Moreover, healthy rabbit PBMC were isolated and cultured, then added the different concentrations of rPsoCRT (5, 10, 20, 40, and 80 μg·mL^-1). Finally, the transcription levels of the primary cytokines (IFN-γ, IL-4, IL-17A, IL-10) and transcription factors (T-bet, GATA-3, RORc, Foxp3) of Th1, Th2, Th17 and Treg cells of rabbit PBMC were measured by qRT-PCR. Our results showed that the full-length of PsoCRT was 1 254 bp, and PsoCRT belonged to the type calreticulin family. The purified rPsoCRT was of the expected molecular size of ~64 ku. Rabbit PBMC treated with rPsoCRT upregulated the IFN-γ and T-bet mRNA expressions of Th1 cells, and decreased IL-4 mRNA expression while no different of GATA-3 mRNA expression in Th2 cells, which leaded to shifting T-cell differentiation towards Th1 bias with a drop in the IFN-γ/IL-4 and T-bet/GATA-3 ratios. Additionally, the mRNA expression of IL-17A in Th17 cells and Foxp3 in Treg cells were decreased, while the expressions of IL-10 and RORc remained unchanged, which resulted in shifting T-cell differentiation towards Treg bias with a drop in the IL-17A/IL-10 and RORc/Foxp3 ratios. These results indicated that rPsoCRT could cause imbalance in the transcription levels of Th1/Th2 and Th17/Treg primary cytokines and transcription factors in rabbit PBMC, resulting in host immunity shift to Th1 and Treg responses, respectively.

Key words: Psoroptes ovis, calreticulin, Th1/Th2, Th17/Treg, immune balance

CLC Number:

S582.746

LI Yan'e, LIANG Youping, FAN Jie, WU Fangyan, YAO Xiangyue, LIMAO Quehu, CIREN Cangjue, HAO Guiying, GU Xiaobin. Effects of Recombinant Calreticulin of Psoroptes ovis on Th1/Th2 and Th17/Treg Immune Balance in Rabbit Peripheral Blood Mononuclear Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(4): 1910-1918.

Figures/Tables 6

Table 1

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分类 Sort	基因名称及GenBank No. Gene name and GenBank No.	引物序列(5′→3′) Primer sequence	扩增长度/bp Length
内参 Internal reference	β-actin NM001101683	F: GGCATGGAGTCGTGTGGCATC R: CGTGTTGGCGTACAGGTCCTTG	90
细胞因子 Cytokine	IFN-γ AB010386	F: TTTCCCAAGGATAGCAGTGGT R: TGAAGCCAGAAGTCCTCAAAA	146
	IL-17A XM002714498	F: ACTCTCCACCGCAACGAGGATC R: TTCCCTTCGGCATTGACACAGC	92
	IL-10 NM001082045	F: ATGCCGCAAGCTGAGAACCAC R: CTTCACCTGCTCCACTGCCTTG	147
	IL-4 DQ852343	F: AGAGCTCGGTGACCTCAGAC R: CTTGCATGGCGGTCTTTAG	150
转录因子 Transcription factor	T-bet XM008271373	F: GATGCGCCAGGAAGTTTCATTT R: TCTCTTGGAAGGTGAAGACGTG	194
	GATA-3 XM008268070	F: CAGGGTCTCTAGTGCTGTGAAA R: CCGTGTCCTTCATGCTTTTGTT	139
	Foxp3 XM002719910	F: ACCACCACCTCCTCCTTGAACC R: CCCAGCCGCACCCCAGAG	95
	RORc XM008264444	F: ACAACGCCGACAACCACACAG R: GAGCGAGCGGGAGAAGTCAAAG	116

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Effects of Recombinant Calreticulin of Psoroptes ovis on Th1/Th2 and Th17/Treg Immune Balance in Rabbit Peripheral Blood Mononuclear Cells

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