家蚕茧壳水解物对小鼠免疫功能的影响

以小鼠为实验动物研究家蚕茧壳水解物的免疫调节功能。分别给正常雄性BALB/c小鼠灌胃低剂量(575 mg/kg)、中剂量(1 150 mg/kg)和高剂量(2 300 mg/kg)家蚕茧壳水解物,30 d后测定小鼠胸腺、脾脏质量计算脏器/体质量比值,并通过脾淋巴细胞增殖试验、腹腔巨噬细胞吞噬鸡红细胞试验、乳酸脱氢酶法和血清溶血素法等,分别检测细胞免疫功能、巨噬细胞吞噬能力、NK细胞活性和体液免疫功能。与对照组相比,中剂量家蚕茧壳水解物灌胃组小鼠的脾指数显著增加(P0.05),低剂量组和高剂量组小鼠胸腺指数显著增加(P0.05);中、高剂量家蚕茧壳水解物灌胃组小鼠的脾淋巴细胞增殖能力、NK细胞杀伤活性、腹腔巨噬细胞对鸡红细胞的吞噬指数等显著(P0.05)或极显著提高(P0.01)。此外,家蚕茧壳水解物对小鼠体质量和血清溶血素含量没有显著影响。研究结果表明,家蚕茧壳水解物能促进正常小鼠的免疫器官质量增加,对小鼠的细胞免疫和非特异性免疫功能也有明显的促进作用。     

中药增效抗菌复方对小鼠免疫功能的影响

为明确中药增效抗菌复方的抗感染作用机制,本实验进行其对小鼠免疫功能影响的研究.将受试药物按每千克体质量1 g、5 g和10 g的剂量对小鼠连续腹腔注射15 d,采用脏器系数测定法、腹腔巨噬细胞吞噬鸡红细胞法、乳酸脱氢酶释放法和MTT法分别对小鼠免疫器官指数、腹腔巨噬细胞吞噬功能、NK细胞活性、血清溶血素水平和淋巴细胞增殖能力进行测定.结果表明,该药物可以显著增加小鼠免疫器官指数,增强腹腔巨噬细胞吞噬功能和NK细胞活性,也可以明显提高血清溶血素水平和淋巴细胞增殖能力.因此,中药增效抗菌复方对小鼠非特异性免疫和特异性免疫功能均有增强作用.     

硒化大蒜多糖对小鼠腹腔巨噬细胞功能的影响

【目的】 研究硒化大蒜多糖（sGPS）对小鼠腹腔巨噬细胞功能的影响,以期为硒化大蒜多糖的作用挖掘和临床应用提供依据。【方法】 依次通过分离、纯化得到大蒜多糖（GPS）,并经硝酸-亚硒酸钠硒化修饰得到sGPS₃、GPS₅和sGPS₆。以小鼠腹腔巨噬细胞为研究对象,用6.25、12.5、25、50、100 μg/mL sGPS₃、GPS₅、sGPS₆及10 μg/mL脂多糖（LPS组）处理小鼠腹腔巨噬细胞48 h,同时设置不加药物的细胞为对照组,用中性红法测定其吞噬功能,CCK-8法测定其増殖能力,筛选出活性最好的硒化大蒜多糖;然后用ELISA法检测活性最好的硒化大蒜多糖对巨噬细胞上清液中一氧化氮（NO）、肿瘤坏死因子-α（TNF-α）、干扰素-γ（IFN-γ）、白介素-1β（IL-1β）、白介素-6（IL-6）、白介素-12（IL-12）含量的影响;再通过小鼠碳廓清实验、小鼠腹腔巨噬细胞吞噬鸡红细胞实验测定空白对照组（生理盐水）及高（2 mg/mL）、中（1 mg/mL）、低（0.5 mg/mL）剂量活性最好的硒化大蒜多糖的吞噬指数与吞噬百分率。【结果】 除6.25 μg/mL sGPS₃外,6.25、12.5、25、50和100 μg/mL sGPS₃、sGPS₅、sGPS₆组小鼠腹腔巨噬细胞的A_{490 nm}值均显著高于对照组（P<0.05）,其中,50和100 μg/mL sGPS₆组的A_{490 nm}值显著高于LPS组（P<0.05）,因此选用sGPS₆进行后续试验;12.5、25、50和100 μg/mL sGPS₆组小鼠腹腔巨噬细胞上清中NO、TNF-α、IL-1β、IL-6含量均显著高于对照组,25、50和100 μg/mL sGPS₆组小鼠腹腔巨噬细胞上清中IFN-γ、IL-12显著高于对照组（P<0.05）,其中100 μg/mL sGPS₆组小鼠腹腔巨噬细胞上清中NO、TNF-α、IL-12显著高于LPS组（P<0.05）。2和1 mg/mL sGPS₆组的吞噬指数和吞噬率均显著高于空白对照组（P<0.05）。【结论】 硒化大蒜多糖能增强小鼠腹腔巨噬细胞的吞噬功能和増殖能力,促进细胞因子TNF-α、IL-1β和IL-6的分泌。     

酵母多糖对免疫抑制小鼠的免疫调节作用

为研究酵母多糖(YPS)对免疫抑制小鼠的免疫调节作用,将180只小鼠随机分成5组,分别为空白对照组、CY模型组、试验Ⅰ组(低剂量组)、试验Ⅱ组(中剂量组)、试验Ⅲ组(高剂量组),每组3个重复,每个重复12只.除空白对照组外,其余各组小鼠均按40 mg/(kg·w)连续3d腹腔注射环磷酰胺制备免疫力低下小鼠模型,造模后,试验Ⅰ、Ⅱ、Ⅲ组给小鼠分别每天灌服0.2、0.5、1 mg/mL的YPS 0.1 mL/10g体重,空白对照组和CY模型组灌服等量生理盐水.30 d后,检测各组小鼠免疫学指标.结果表明,试验Ⅰ组可显著提高免疫抑制小鼠的脾淋巴细胞增殖能力、迟发型变态反应、巨噬细胞吞噬率、巨噬细胞吞噬指数、NK细胞活性、血清中IL-4和IFN-γ的水平.试验Ⅱ、Ⅲ组显著提高免疫器官指数、抗体生成细胞溶血空斑数和血清溶血素抗体积数,极显著地提高免疫抑制小鼠的脾淋巴细胞增殖能力、迟发性变态反应、巨噬细胞吞噬率、巨噬细胞吞噬指数、NK细胞活性、血清中IL-4和IFN-γ的水平.说明YPS能不同程度的提高免疫抑制小鼠的免特异性和非特异性免疫功能.     

杨树花提取物对小鼠免疫功能的影响

为研究杨树花提取物对小鼠免疫功能的调节作用,每天给小鼠分别经口灌服0.1、0.2、0.4mg/kg的杨树花提取物和灭菌水30d后,测定其免疫器官指数、脾淋巴细胞增殖能力、抗体生成细胞水平、腹腔巨噬细胞吞噬功能、自然杀伤细胞(NK细胞)活性和IL-2的变化。结果表明,杨树花提取物的低剂量组可明显提高IL-2水平;中、高剂量组均可显著提高(P<0.05)小鼠的免疫器官指数、血清溶血素水平,极显著提高(P<0.01)脾淋巴细胞增殖能力、巨噬细胞吞噬率、巨噬细胞吞噬指数、NK细胞活性和IL-2水平。说明不同剂量的杨树花提取物均能提高小鼠免疫功能,中高剂量组效果更为显著。     

刺五加多糖纳米乳对免疫抑制小鼠的免疫增强作用

为给兽医临床提供一种有效的免疫增强剂，本试验通过建立小鼠免疫抑制模型，检测其免疫器官指数、脾细胞增殖活性、单核巨噬细胞的吞噬活性、自然杀伤(NK)细胞活性等来研究刺五加多糖纳米乳(ASPS-NE)对免疫抑制小鼠的免疫增强作用。结果显示：与CTX模型组比较，ASPS-NE高、中、低剂量组[100、50 mg/(kg·bw)和25 mg/(kg·bw)]均可不同程度提高免疫抑制小鼠的脏器指数，极显著增强刀豆球蛋白A(ConA)和脂多糖(LPS)诱导的小鼠脾淋巴细胞增殖活性，增强单核巨噬细胞的吞噬指数，提高NK细胞的杀伤率(P<0.01),尤其以ASPS-NE中剂量效果最佳(P<0.01)。结果表明ASPS-NE可增强免疫抑制小鼠的免疫力，发挥免疫增强作用。     

华蟾毒精对小鼠免疫细胞活性的影响

为观察华蟾毒精（CBG）对小鼠免疫细胞活性的影响,本研究采用MTT法检测小鼠脾淋巴细胞的增殖情况,检测小鼠腹腔巨噬细胞的吞噬功能以及小鼠NK细胞对靶细胞的杀伤作用。结果显示,CBG在一定剂量范围内单独或者协同非特异性丝裂原（Con A或LPS）作用能够显著增强小鼠脾淋巴细胞的增殖,CBG单独作用可以显著提高小鼠腹腔巨噬细胞的吞噬功能,并能够显著提高小鼠NK细胞对靶细胞的杀伤作用,表明CBG能够提高小鼠免疫细胞的活性。     

抗菌肽饲料添加剂对小鼠巨噬细胞吞噬功能的影响

对试验小鼠饲喂1‰和3‰抗菌肽饲料添加剂,鸡红细胞吞噬试验检测小鼠腹腔巨噬细胞吞噬功能,研究抗菌肽饲料添加剂对小鼠免疫功能的影响.结果表明,抗菌肽饲料添加剂可以显著提高小鼠腹腔巨噬细胞吞噬鸡红细胞的吞噬率和吞噬指数,增强小鼠非特异性免疫功能.     

甘草多糖对小鼠腹腔巨噬细胞吞噬功能的影响

为了测定甘草多糖对小鼠腹腔巨噬细胞吞噬能力的影响,试验从甘草中提取甘草多糖(gly-cyrrhizia polyaccharide,GPS),以不同浓度腹腔注射给小鼠,通过巨噬细胞吞噬试验观察小鼠腹腔巨噬细胞吞噬鸡红细胞情况。结果表明:小鼠腹腔注射不同浓度的GPS后,其吞噬指数、吞噬百分率均显著高于对照组(P<0.05)。说明甘草多糖能够提高小鼠腹腔巨噬细胞的吞噬能力。     

猪繁殖与呼吸综合征弱毒活疫苗免疫猪脾脏转移因子的制备及其活性分析

从猪繁殖与呼吸综合征(PRRS)弱毒活疫苗免疫猪脾制备转移因子(TF),检测其免疫活性,为开发新型免疫增强剂和有效防制PRRS提供科学依据。用PRRSV弱毒活疫苗免疫健康育成猪,无菌采集脾,通过透析法从脾提取TF。应用磺基水杨酸法、茚三酮反应和硝酸-钼酸铵试验等方法检测TF的主要成分,经高效液相色谱法测定TF中氨基酸种类及其含量;通过鸡红细胞吞噬试验和MTT法,检测TF对小鼠腹腔巨噬细胞吞噬功能和外周血淋巴细胞增殖活性的调节作用。进而将9头4周龄健康仔猪随机分为A、B和C组,每组3头。A组每头猪左、右侧耳后分别肌内注射1头份PRRSV疫苗和1mL TF;B组每头猪注射1头份PRRSV疫苗与1mL TF的混合液;C组每头猪注射1头份PRRSV疫苗与1mL生理盐水混合液。免疫后20d,通过MTT法检测每组猪外周血淋巴细胞(PBLC)的增殖活性。制备的TF内无蛋白质成分,含有核酸和18种氨基酸,每毫升TF含核酸21.5μg、总氨基酸3.479mg。TF能显著增强小鼠腹腔巨噬细胞对鸡红细胞的吞噬能力和PBLC增殖活性。当TF与PRRSV疫苗分开或混合注射给4周龄仔猪后,能够显著提高仔猪PBLC的增殖能力。从PRRS弱毒活疫苗免疫猪的脾制备了TF,其能显著提高巨噬细胞的吞噬功能和外周血淋巴细胞的增殖活性。     

Interleukin-2 corrects defective NK activity of patients with leukemia

NK cells of patients with leukemia display low cytotoxic potential. Since the NK cells have been suggested to play a role in natural resistance to leukemia, we considered it of importance to investigate the approaches leading to the correction of NK defect of leukemic patients. Our studies demonstrate that culture of effector cells with interleukin-2 (IL-2) resulted in restoration of cytotoxic defect. This was indicated by normalization of tumor-binding as well as lytic NK activity, by normal frequency of cytotoxic cells and their ability to recycle. The NK cell nature of cytotoxic cells was shown by abrogation or depletion of cytotoxicity by antibody directed against NK cell-associated, but not T cell-associated antigen. The generation of NK cell activity against fresh leukemic cells suggests that adoptive transfer of IL-2 activated NK cells may be a new approach to leukemic treatment.     

Characterization of NCR1+ cells residing in lymphoid tissues in the gut of lambs indicates that the majority are NK cells

Natural killer (NK) cells are important for immune protection of the gut mucosa. Previous studies have shown that under pathologic conditions NK cells, T cells and dendritic cells are found co-localised in secondary lymphoid organs where their interaction coordinates immune responses. However, in the gut-associated lymphoid tissues (GALTs), there are few detailed reports on the distribution of NK cells. Sheep harbour several types of organised lymphoid tissues in the gut that have different functions. The ileal Peyer’s patch (IPP) functions as a primary lymphoid tissue for B cell generation, while the jejunal Peyer’s patches (JPPs) and colon patches (CPs) are considered secondary lymphoid tissues. In the present study, we analysed tissues from healthy lambs by flow cytometry and in situ multicolour immunofluorescence, using recently described NCR1 antibodies to identify ovine NK cells. Most NCR1+ cells isolated from all tissues were negative for the pan T cell marker CD3, and thus comply with the general definition of NK cells. The majority of NCR1+ cells in blood as well as secondary lymphoid organs expressed CD16, but in the GALT around half of the NCR1+ cells were negative for CD16. A semi-quantitative morphometric study on tissue sections was used to compare the density of NK cells in four compartments of the IPPs, JPP and CPs. NCR1+ cells were found in all gut segments. Statistical analysis revealed significant differences between compartments of the primary lymphoid organ IPP and the secondary lymphoid organs of the JPPs and CP. NK cells co-localised and made close contact with T cells, dendritic cells and other NK cells, but did not show signs of proliferation. We conclude that NK cells are present in all investigated segments of the sheep gut, but that presence of other innate lymphoid cells expressing NCR1 cannot be excluded.     

奶牛乳房炎白色念珠菌的分离鉴定及生物学特性研究

试验旨在了解宁夏地区奶牛真菌性乳房炎的发生情况,为诊断和治疗真菌性奶牛乳房炎提供有效依据。采集宁夏周边地区各奶牛场使用抗生素治疗无效的乳房炎奶样,对奶样中的真菌进行分离鉴定,并对分离菌进行生物学特性研究。通过选择性培养基从乳房炎奶样中获得真菌,进一步应用生物化学和分子生物学手段鉴定出样品中的真菌为白色念珠菌。通过溶血性、磷脂酶活性及产膜等方面的研究,证实其具有不同毒力,分离菌株对试验动物有较强的致病性。同时,采用药敏纸片法对分离株进行药敏试验,结果显示,菌株对临床常用抗生素耐药,对临床常用抗真菌药物敏感。结果表明,宁夏地区真菌性乳房炎奶样中分离出的致病菌均为白色念珠菌,感染率为43%,分离株具有较强的毒力,对常用抗生素耐药,对抗真菌药物和中药敏感。     

Isolation,Identification and Analysis of Biological Characteristics of Candida albicans in Bouine Mastitis

The study was aimed to learn the infection of dairy cows with fungi in Ningxia and take effective measures to prevent and control the fungi infection. Milk samples were collected from dairy cows that did not respond to antibiotics in Ningxia, fungi in milk samples were separated and identified, and the biological characteristics of isolated bacteria were analyzed. Milk samples infected by fungus were screened out using fungal selective medium. The yeast pathogen was shown to be Candida albicans by biochemistry and molecular biology methods. Determination of enzyme activity and biofilm-forming tests confirmed that it had different levels of virulence with strong pathogenicity to experimental animals. Drug sensitive test was carried out by K-B programe, the results showed that the isolates were resistant to clinical commonly used antibiotics, and were sensitive to clinical common antifungal drugs. The fungal mastitis was caused by Candida albicans in Ningxia, the ratio of fungal infection was 43%. Candida albicans had strong virulence and were resistant to common antibiotic and sensitivity to antifungal drugs and traditional Chinese medicine.     

Isolation and characterization of porcine natural killer (NK) cells

The morphologic and biological properties of porcine cells mediating natural killer (NK) activity were determined. In a previous study, we demonstrated that lymphocytes from the peripheral blood of pigs greater than 1 week of age possessed NK activity to K562 tumor cells and that lymphocytes from the blood and spleen of pigs greater than 1 day of age were able to mediate natural cytotoxicity against parainfluenza-3 (PI3) virus-infected Vero cells (Yang and Schultz, 1986a). Discontinuous density gradients were used to enrich NK cells. NK cytotoxicity was mainly present in high-density Percoll fractions (50 to 55% and 55 to 60%); little or no NK activity was present in lower density fractions. The NK cell enriched lymphocytes responded to the mitogens PHA, ConA and PWM. NK cells were sensitive to the suppressive effect of corticosteroid, but Protein A did not affect NK activity. The amount of cytotoxicity directly corresponded to the degree of binding that occurred between the NK enriched lymphocyte population and the target cells. Cytochemical and morphological studies demonstrated that these bond cells which are believed to be responsible for the NK activities, were mainly small to medium lymphocytes lacking azurophilic cytoplasmic granules. These findings were confirmed by ultrastructural studies of effector and "target-binding" cells. The results of the present study suggested that the cells mediating NK activity in pigs have the morphological and density characteristics of small and medium sized lymphocytes; findings that differ from those described for NK cells in human and other animal species.     

山羊、水牛感染肝片吸虫后淋巴细胞自然杀伤活性动态变化

自然杀伤 (NK)活性是在细胞水平反映机体天然免疫力的重要指标之一。本实验以SP2 / 0细胞为靶细胞 (即效应细胞 ) ,采用乳酸脱氢酶(LDH)短程释放法测定人工感染肝片吸虫的水牛和山羊的外周血液中淋巴细胞的NK活性动态变化。从测定结果的相对值来看 ,发现山羊在感染肝片吸虫后NK活性有 2次高峰 ,推测与虫体的发育或感染山羊的免疫状态有关 ;2个感染组 (Ⅰ ,2 0 0个囊蚴 /只 ;Ⅱ ,50 0个囊蚴 /只 )的变化趋势表明 ,感染量与NK活性无相关关系。感染水牛的NK值仅略高于对照组 ,无明显的高峰。而绝对值结果又反映出不同动物的NK活性有差异 ,水牛的NK活性比山羊高 ,说明水牛对肝片吸虫的天然免疫力比山羊强     

Pyrimidinones: inducers of NK cell activity and antitumor immunity

We have shown that pyrimidinone molecule, 2 amino-5-iodo-6-phenyl-4-pyrimidinone (AIPP) after regional administration, displayed both prophylactic and therapeutic effect on ascitic mammary adenocarcinoma, ACA-755. The antitumor effect was mediated by natural killer (NK) cells, since depletion of this lymphocyte population resulted in abrogation of AIPP-induced tumor resistance. This observation confirms the role of NK cells in antitumor immunity and suggests that AIPP may be therapeutically beneficial also in man.     

Effects of intravenous administration of neurokinin receptor subtype-selective agonists on gonadotropin-releasing hormone pulse generator activity and luteinizing hormone secretion in goats

Recent evidence suggests that neurokinin B (NKB), a member of the neurokinin (tachykinin) peptide family, plays a pivotal role in gonadotropin-releasing hormone (GnRH) pulse generation. Three types of neurokinin receptors (NKRs), NK1R, NK2R and NK3R, are found in the brain. Although NKB preferentially binds to NK3R, other NKRs are possibly also involved in NKB action. The present study examined the effects of intravenous administration of the NKR subtype-selective agonists GR73632 (NK1R), GR64349 (NK2R), and senktide (NK3R) on GnRH pulse generator activity and luteinizing hormone (LH) secretion. Multiple-unit activity (MUA) was monitored in ovariectomized goats (n = 5) implanted with recording electrodes. Characteristic increases in MUA (MUA volleys) were considered GnRH pulse generator activity. Although three NKR agonists dose-dependently induced an MUA volley and an accompanying increase in LH secretion, the efficacy in inducing the volley markedly differed. As little as 10 nmol of senktide induced an MUA volley in all goats, whereas a dose of 1000 nmol was only effective for the NK1R and NK2R agonists in two and four goats, respectively. When the treatment failed to evoke an MUA volley, no apparent change was observed in the MUA or LH secretion. Similar effects of the NK2R and NK3R agonists were observed in the presence of estradiol. The results demonstrated that NK3R plays a predominant role in GnRH pulse generation and suggested that the contributions of NK1R and NK2R to this mechanism may be few, if any, in goats.     

Immunology of fungal infections in animals

The nature of immunity to fungal infection is discussed predominantly for mammals and birds. T-cell-mediated immunity seems essential for recovery both from cutaneous and mucosal infections (Candida, Malassezia and dermatophytes) and from infections of systemic fungal pathogens (Cryptococcus, Blastomyces, Histoplasma, and Coccidioides). Often chronic progressive disease caused by these fungi is associated with a depression or absence of T-cell-mediated immunity to antigens of the infecting fungus. In contrast recovery from disease, or absence of clinical disease after exposure to these fungi, is associated with the presence of strong T-cell-mediated immune responses to the fungus. The activation of macrophages and the stimulation of epidermal growth and keratinization are the processes induced by T-cell-mediated immunity which result in the resolution of systemic or cutaneous and mucosal disease. Other cell types, for example NK cells and PMNs (polymorphonuclear leucocytes), may be important in these diseases in reducing the effective amount of inoculum to which an animal is exposed and thereby reducing the likelihood of disseminated disease. Invasive opportunistic fungi (Candida, Aspergillus, Mucorales) are resisted by PMNs which attach to the hyphae or pseudohyphae and damage them via an extracellular mechanism. Other host cell types may be important in natural resistance, fungal spores being handled by the macrophages which, under conditions when animals are not immunosuppressed, are likely to be an effective first line of defense. Subcutaneous pathogens and miscellaneous other fungal diseases are discussed from a point of view of host immunity and immunodiagnosis. Vaccine development for ringworm and for other mycoses is discussed.