共载丹参多糖和Mn2+的PLGA纳米粒作为灭活PCV2的佐剂效果评价

doi:10.11843/j.issn.0366-6964.2025.08.040

Abstract

Abstract:

In this study, PLGA was used to encapsulate Salvia miltiorrhiza polysaccharide and Mn²⁺ with potent immune stimulation effect and good biosafety, and the immune enhancement effect of the system as an inactivated PCV2 adjuvant was evaluated. Firstly, we prepared a PLGA nano-adjuvant delivery system (SM-PLGA) co-loaded with Salvia miltiorrhiza polysaccharide and Mn²⁺ by double emulsion solvent evaporation method. The SM-PLGA was mixed with PCV2 antigen, and 100 μL was injected subcutaneously respectively into the left and right thighs of mice for the first immunization. At the same time, PCV2 group, Alhydrogel (Algel)-PCV2 group and the Control group were established, and on the 14th day after the first immunization, mice were immunized twice. Then, the activation of DCs in popliteal lymph nodes and inguinal lymph nodes were evaluated by flow cytometry on the 5th day after the first immunization. The serum of mice was collected on the 21st, 28th, 35th and 42nd day after the first immunization, and the PCV2 specific antibodies IgG, IgG1 and IgG2a were dynamically monitored to evaluate the enhancement effect of the adjuvant on PCV2 specific humoral immune response. The spleen lymphocytes of mice were collected on the 21st and 28th day after the first immunization, and the activation of CD4⁺T cells, CD8⁺T cells, CTL and memory T cells were measured, and the IFN-β, IFN-γ and IL-6 in the supernatant of the cells on the 28th day were measured. Finally, the lymph nodes of mice were collected on the 35th day after the first immunization to determine the activation of germinal center B cells. Through the results of the above three periods, the enhancement effect of SM-PLGA on cellular immune response was evaluated. SM-PLGA could significantly promote the activation of lymph node DCs on the 5th day after the first immunization, and continuously stimulate the secretion of PCV2-specific antibody IgG and its subtypes on the 21st to 42nd day after the first immunization. SM-PLGA could significantly promote the spleen CD4⁺T cells, CD8⁺T cells and CTL response on the 21st day after the first immunization, and still had a strong stimulating effect on memory T cells and Germinal center B cells on the 28th and 35th day after the first immunization. At the same time, it significantly promoted the secretion of related cytokines, IFN-β, IFN-γ and IL-6. The results showed that SM-PLGA, as an adjuvant for inactivating PCV2, could induce a potent Th1/Th2 immune response and provide a reference for the prevention of PCV2.

Key words: Salvia miltiorrhiza polysaccharide, Mn²⁺, PLGA, adjuvant, cellular immunity, humoral immunity

CLC Number:

S852.65

ZHU Yixuan, GU Pengfei, ZHAO Qi, XU Panpan, FAN Yingsai, BAO Yongzhan, WANG Xiao, SHI Wanyu. Evaluation of PLGA Nanoparticles Co-loaded with Salvia miltiorrhiza Polysaccharide and Mn²⁺ as an Adjuvant for Inactivating PCV2[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(8): 4031-4041.

Figures/Tables 6

Table 1

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时间 Time	目标细胞 Target cells	染色抗体^[29] Staining antibody	细胞因子 Cytokine
第21天 Day 21	CD3⁺CD4⁺; CD3⁺CD8a⁺	CD3-FITC; CD4-APC; CD8a-PE	颗粒酶B
	细胞毒性T细胞	CD8a-PE; CD107a(lamp-1)-FITC; CD178 FASL-APC
第28天 Day 28	记忆T细胞	CD4-APC；CD8a-PE；CD44-PE-Cyanine7；CD62L(L-Selectin)-FITC；	IFN-γ IFN-β IL-6

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Evaluation of PLGA Nanoparticles Co-loaded with Salvia miltiorrhiza Polysaccharide and Mn²⁺ as an Adjuvant for Inactivating PCV2

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Evaluation of PLGA Nanoparticles Co-loaded with Salvia miltiorrhiza Polysaccharide and Mn2+ as an Adjuvant for Inactivating PCV2

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Evaluation of PLGA Nanoparticles Co-loaded with Salvia miltiorrhiza Polysaccharide and Mn²⁺ as an Adjuvant for Inactivating PCV2