免疫调节剂对球虫感染鸡的免疫学指标的影响

为探讨免疫调节剂对鸡球虫病的免疫反应调节作用 ,设计并进行了免疫调节剂对球虫感染鸡的免疫学指标影响的研究。测定指标包括 :淋巴细胞玫瑰花环细胞形成率 ,淋巴细胞转化率 ,红细胞免疫复合物花环形成率 ,红细胞 C3b受体花环形成率 ,脾脏重 /体重相对重 ;血清中 Ig G,Ig M,Ig A的 EL ISA P/ N值。结果各项免疫学指标的差异显著。表明胸腺素、黄芪多糖和卡介苗等免疫调节剂可提高球虫感染鸡的淋巴细胞玫瑰花形成率、淋巴细胞转化率。     

中药免疫调节剂体外细胞毒性测定

采用形态学观察和MTT两种方法,测试了中药免疫调节剂(TJJ)在鸡胚成纤维细胞(CEF)和小鼠纤维母细胞(L929)体外培养中的最大安全浓度。结果表明:⑴TJJ的细胞毒性很低,对CEF和L929的最大无毒浓度分别达3.12g/L和1.56g/L;⑵两种细胞比较,L929对药物毒性的反应性较CEF更敏感;⑶两种方法比较,MTT法更灵敏、更客观;⑷在同一种细胞的不同生长状态下(形成单层前后)加入TJJ,药物对细胞的毒性结果无变化,说明药物对细胞的毒性与细胞种类有关,而与细胞的生长状态关系不明显。     

Synthesis and characterization of Azadirachta indica constructed silver nanoparticles and their immunomodulatory activity in fish

Green synthesis of nanoparticles by using different biological agents has emerged as an alternative to overcome the toxic effect of chemically synthesized nanoparticles. Among various biological agents, plants are mostly preferred. This study describes an eco‐friendly and green synthesis of silver nanoparticles (G‐AgNPs) using Azadirachta indica (neem) as a reducing agent. UV–Vis spectral analysis proved the wavelength of sample to be 420 nm, approaching the surface resonance peak specific for G‐AgNPs. Dynamic light scattering (DLS) analysis showed the mean diameter of particles as 35.4 nm with zeta potential +34.6 mV. TEM results revealed the compact and spherical shape of the particles. Fourier transform infrared spectroscopies (FT‐IR) demonstrate the presence of possible functional groups involved in synthesis of the silver nanoparticles. The functional activity of immunological parameters, such as nitroblue tetrazolium assay, myeloperoxidase activity, phagocytic activity, anti‐protease and lysozyme activity, increased significantly (P < 0.05) in fish treated with G‐AgNPs. Relative percentage survival (74%) and enhanced disease resistance were observed in G‐AgNP‐treated Cirrhinus mrigala fingerlings challenged with Aeromonas hydrophila. In summary, present results demonstrate biosynthesized silver nanoparticles have immunomodulatory and antibacterial activity.     

Pharmacokinetics of low-dose methotrexate in horses

This study aimed to investigate both the pharmacokinetic behavior and tolerance of methotrexate (MTX) in horses to design a specific dosing regimen as a new immunomodulatory drug for long-term treatment. To determine the primary plasma pharmacokinetic variables after single intravenous, subcutaneous or oral administration, six horses were administered 0.3 mg/kg MTX in a crossover design study. After a 10-week washout, MTX was administered subcutaneously to three of the six previously treated horses at a dose of 0.3 mg/kg once per week for 3 months. In both studies, MTX and metabolite concentrations were measured using LC-MS/MS. The absolute bioavailability of MTX was 73% following subcutaneous administration but less than 1% following oral administration. The plasma clearance was 1.54 ml min⁻¹ kg⁻¹ (extraction ratio = 2%). After 24 hr, plasma concentrations were below the LOQ. No adverse effects were noted except for a moderate reversible elevation in liver enzymes (GLDH). With regards to the main metabolites of MTX, very low concentrations of 7-hydroxy-MTX were found, whereas polyglutamated forms (mainly short chains) were found in red blood cells. A subcutaneous dose of 0.2 mg kg⁻¹ week⁻¹ may be safe and relevant in horses, although this has yet to be clinically confirmed.     

In vitro and in vivo modulation of the equine immune response by parapoxvirus ovis

REASON FOR PERFORMING STUDY: While immune modulators are used routinely in equine medicine, their mechanism of action is not always known. OBJECTIVES: To determine the effect of a commercial preparation of inactivated parapoxvirus ovis (Orf virus; PPVO) on cytokine gene expression by equine peripheral blood mononuclear cells (PBMC) both in vitro and in vivo. METHODS: PBMC were prepared from 6 mixed-breed yearlings and cultured in vitro with PPVO with or without Concanavalin A (Con A) for 24 h. Effects on the expression of IFNalpha, IFNbeta IFNgamma, TNFalpha and IL-18 were analysed by real time quantitative PCR (RT-PCR). In addition, 12 yearling horses were treated with PPVO and whole blood RNA samples were prepared at regular intervals to assess effects on in vivo cytokine gene expression. Six of those yearlings were later treated with saline and served as treatment controls. Nine additional yearlings were injected intradermally with a single dose and their injection sites biopsied at 24 and 48 h for cytokine expression. RESULTS: In vitro culture of PBMC with PPVO led to a significant increase in IFNalpha and IFNbeta gene expression compared to mock-stimulated cultures. In addition, expression of IFNgamma and TNFalpha was significantly higher in PBMC stimulated with PPVO and Con A, than those stimulated with Con A alone. No changes were observed in IL-18 gene expression in vitro. Treatment of horses with a 3-dose regimen of PPVO resulted in elevation of IFNgamma gene expression, which was detected 24 h after the first dose and declined thereafter. Intradermal inoculation led to increased expression of IFNgamma along with IFNbeta, IL-15 and IL-18. CONCLUSIONS: Together these results indicate that PPVO stimulated IFNgamma production both in vitro and in vivo. Increased cytokine expression could account for its immunomodulatory activity. POTENTIAL RELEVANCE: The absence of adverse reactions and clear indications of increased expression of cytokine gene expression supports previous clinical uses for this immune modulator in those situations when increased expression of IFNgamma is warranted.     

免疫调节剂与抗球虫药联合应用的抗球虫效果

为探讨免疫调节剂对鸡球虫病的免疫反应调节作用,设计并进行了免疫调节剂与抗球虫药合用的效果的研究。试验结果以抗球虫指数的测定来判定抗球虫药与免疫调节剂合用的效果。结果表明,免疫调节剂与抗球虫药合用状态下,抗球虫指数检测显示免疫调节剂卡介苗和黄芪多糖可明显提高抗球虫指数、改善球虫感染鸡的临床症、提高增重速度等,为控制球虫病提供了新的途径。     

脂质体作为免疫增强剂在动物疾病防治方面的应用现状

脂质体作为药物递送系统,能够定向地将药物送至靶位,并能保持药物的活性避免其被降解;作为免疫调节剂,可以同时增强体液和细胞介导的免疫应答反应,能显著增强疫苗效果。论文通过总结近几年的有关文献,对脂质体作为药物递送系统和免疫调节剂在疾病治疗中的关键作用进行分析,对其作用机制及应用现状加以归纳总结,并探讨了现存问题以及未来展望。     

缓苗增蘖剂和免疫剂在盐碱地水稻上的应用

为提高盐碱地水稻的植株成活率,采用对照试验的方法,探讨盐碱地水稻缓苗增蘖剂和水稻盐碱免疫剂在盐碱地水稻上的应用效果,为其大面积推广应用提供科学依据。     

Development of Highly Selective Kv1.3-Blocking Peptides Based on the Sea Anemone Peptide ShK

ShK, from the sea anemone Stichodactyla helianthus, is a 35-residue disulfide-rich peptide that blocks the voltage-gated potassium channel Kv1.3 at ca. 10 pM and the related channel Kv1.1 at ca. 16 pM. We developed an analog of this peptide, ShK-186, which is currently in Phase 1b-2a clinical trials for the treatment of autoimmune diseases such as multiple sclerosis and rheumatoid arthritis. While ShK-186 displays a >100-fold improvement in selectivity for Kv1.3 over Kv1.1 compared with ShK, there is considerable interest in developing peptides with an even greater selectivity ratio. In this report, we describe several variants of ShK that incorporate p-phophono-phenylalanine at the N-terminus coupled with internal substitutions at Gln16 and Met21. In addition, we also explored the combinatorial effects of these internal substitutions with an alanine extension at the C-terminus. Their selectivity was determined by patch-clamp electrophysiology on Kv1.3 and Kv1.1 channels stably expressed in mouse fibroblasts. The peptides with an alanine extension blocked Kv1.3 at low pM concentrations and exhibited up to 2250-fold selectivity for Kv1.3 over Kv1.1. Analogs that incorporates p-phosphono-phenylalanine at the N-terminus blocked Kv1.3 with IC₅₀s in the low pM range and did not affect Kv1.1 at concentrations up to 100 nM, displaying a selectivity enhancement of >10,000-fold for Kv1.3 over Kv1.1. Other potentially important Kv channels such as Kv1.4 and Kv1.6 were only partially blocked at 100 nM concentrations of each of the ShK analogs.     

中药免疫调节剂对免疫抑制模型小鼠的保护作用

【目的】研究中药免疫调节剂对免疫抑制小鼠的组织保护作用。【方法】用环磷酰胺(Cyclophospha-mide,Cy)构建小鼠免疫抑制病理模型。分别给正常小鼠和环磷酰胺所致免疫抑制小鼠饮服自制中药免疫调节剂玉屏风散加减(ZY)和经典方剂玉屏风散(YPF),检测ZY、YPF对不同免疫状态下小鼠血液学指标(白细胞计数(WBC)、红细胞计数(RBC)、血红蛋白(HGB)、红细胞压积(HCT)、平均血红蛋白含量(MCH)、平均红细胞体积(MCV))、体质量、脾脏指数以及肝脾组织病理学的影响。【结果】(1)ZY和YPF对健康小鼠血液学指标影响不显著(P0.05);ZY对Cy所致的免疫抑制小鼠WBC、RBC、HGB、HCT、MCH、MCV值有显著影响(P0.05),YPF对Cy所致的免疫抑制小鼠RBC、HGB和MCH值有显著影响(P0.05),且ZY的作用效果优于YPF(P0.05)。(2)ZY和YPF对正常小鼠体质量及脾脏指数的影响不明显(P0.05),而对Cy所致的免疫抑制小鼠体质量影响显著(P0.05),小鼠的脾脏指数显著低于Cy所致的免疫抑制小鼠(P0.05)。(3)ZY+Cy组和YPF+Cy组小鼠脾脏白髓大而明显,肝细胞轻度变性坏死;而模型对照组(Cy组)小鼠脾脏白髓不明显,肝细胞变性坏死严重。【结论】ZY对正常小鼠血细胞数、体质量、脾脏指数的影响均不显著,而对免疫抑制小鼠血细胞数、体质量、脾脏指数以及脾、肝组织病理学均表现出一定的正向调节作用。