升血汤促进辐射损伤小鼠造血与免疫功能恢复的实验研究

本文研究了中药升血汤对辐射损伤小鼠造血与免疫功能恢复的促进作用,结果证明,升血汤能显著地增加辐射损伤小鼠外周血中各类血细胞数以及骨髓及脾脏中的有核细胞数,明显促进小鼠巨噬细胞,Ｔ及Ｂ淋巴细胞功能的恢复,表明升血汤可以明显促进辐射损伤小鼠造血与免疫功能的恢复。     

升血汤促进辐射后骨髓移植小鼠造血与免疫重建的研究

本文用多项血液学及免疫学指标研究了中药升血汤对骨髓移植小鼠造血与免疫重建的影响,结果证明,升血汤有明显促进骨髓移植小鼠造血与免疫功能恢复的功效,表现在小鼠骨髓造血干细胞数量的恢复,小鼠外周血象的改善,脾及骨髓中有核细胞数的增加,明显促进小鼠细胞免疫和体液免疫功能的恢复,研究结果为升血汤的临床应用提供了实验依据。     

小蘖胺免疫抑制作用和抗氧化实验研究

对小蘖胺（Ｂｅｒｂａｍｉｎｅ）的抗氧化作用和免疫抑制作用进行了研究,发现小蘖胺（２５ｍｇ／ｋｇ／ｄ）腹腔给药一周能明显降低小鼠肝脏过氧化脂质的含量,提高血及心肌内超氧化物歧化酶的活性,并能明显减低同种异型抗原诱导的迟发型超敏反应和混合淋巴细胞反应。皮肤移植试验表明小蘖胺５０ｍｇ／ｋｇ／ｄ可以延长小鼠对异基因皮肤移植的排斥时间。用流式细胞仪对小鼠脾脏Ｔ淋巴细胞亚群分析结果表明：小蘖胺能降低小鼠脾脏ＣＤ＋４／ＣＤ８＋的比例。说明小蘖胺对小鼠免疫功能的抑制作用可能在于改变Ｔｈ／Ｔｓ的细胞平衡,使抑制活性增强,从而发挥其对免疫反应的负调节作用。     

脾虚证小鼠胸腺细胞周期和免疫细胞化学的实验研究

本文用ＦＣＭ和免疫细胞化学技术检测了小鼠胸腺细胞的活动周期,Ｂｒｄｕ＋细胞和表达Ｔｈｙｌ．２抗原的Ｔ细胞。小鼠包括对照组,利血平诱发的脾虚组,自然恢复组和健脾汤治疗后复健组。ＦＣＭ和免疫细胞化学结果显示,脾虚组小鼠Ｇ１期细胞堆积,Ｓ期和Ｇ２＋Ｍ期细胞减少。同时胸腺内Ｂｒｄｕ＋细胞和Ｔ细胞也相应减少。经过健脾汤治疗后,除Ｇ１期细胞外,Ｓ期和Ｇ２＋Ｍ期细胞,Ｂｒｄｕ＋细胞和Ｔ细胞均显著增加,并且几乎达正常水平。但是自然恢复组在恢复细胞活动周期正常化,以及恢复Ｂｒｄｕ＋细胞和Ｔ细胞方面比复健组缓慢。说明健脾汤对脾虚小鼠的胸腺细胞有促进ＤＮＡ合成和细胞增殖功能。     

双歧杆菌及中药对小鼠血中抗氧化酶,T3,TSH和吞噬细胞功能的影响

本文用双歧杆菌、中药及复方双歧杆菌制剂饲喂ＢＡＬＢ／ｃ小鼠,发现中药及复方双歧杆菌制剂对老龄鼠血中ＳＯＤ、ＧＳＨ—Ｐｘ活性有明显增高作用,而对ＬＰＯ含量有明显减低作用,双歧杆菌及复方双歧杆菌能增强ＢＡＬＢ／ｃ小鼠内分泌功能及免疫功能,本试验中尚发现双歧杆菌与中药制成的复方双歧杆菌制剂能明显协同促进ＧＳＨ—Ｐｘ及吞噬细胞功能。说明双歧杆菌与一些传统补益类中药具有协同抗衰老作用。     

神经束路电镜追踪中影响HRP－TMB反应产物因素的分析

采用ＨＲｐ－ＴＭＢ－ＳＴ法,对小鼠延髓下橄榄核下端外侧网状核细胞的ＨＲＰ－ＴＭＢ反应产物影响因素分析表明：（１）较高的ｐＨ值（７．０以上）是反应产物消失的关键因素;０．２ｍｏｌ／ＬＰＢ,ｐＨ５．０—６．０对ＨＲＰ－ＴＭＢ反应灵敏度、反应产物的稳定性及组织超微结构的保存是适宜的;当ｐＨ值相同时,缓冲液浓度不同,反应产物消失的程度和速度也不相同。（２）酒精有稳定反应产物的作用。（３）反应前的预浸、灌注液的成分、浓度和固定时间等则与反应产物的丢失无关。此外,用ｐＨ５．５和ｐＨ７．０的缓冲液处理的切片经ＤＡＢ强化、锇化后,组织的超微结构无明显的差异。     

“思壮”虫草菌丝体胶囊免疫功能评价*

将“思壮”虫草菌丝体胶囊（商品名）经口给予小鼠,在４５、４５０（相当于成人每日每公斤推荐摄入量的１０倍）和９００ｍｇ／ｋｇ体重剂量下,在１５～３０ｄ时,通过二硝基氟苯（ＤＮＦＢ）诱导小鼠迟发性变态反应（ＤＴＨ）、血清溶血素试验和小鼠碳廓清试验,表明中、高剂量“思壮”虫草菌丝体胶囊具有增强小鼠细胞免疫功能,而３种剂量均能明显增强小鼠体液免疫功能和单核巨噬细胞吞噬功能,且有剂量－反应关系,由此证明“思壮”虫草菌丝体胶囊有明显的免疫调节作用。     

"思壮"虫草菌丝体胶囊免疫功能评价

将“思壮”虫草菌丝体胶囊（商品名）经口给予小鼠,在４５、４５０（相当于成人每日每公斤推荐摄入量的１０倍）和９００ｍｇ／ｋｇ体重剂量下,在１５￣３０ｄ时,通过二硝基氟苯（ＤＮＦＢ）诱导小鼠迟发性变态反应（ＤＴＨ）、血清溶血素试验和小鼠碳廓清试验,表明中、高剂量“思壮”虫草菌丝体胶囊具有增强小鼠细胞免疫功能,而３种剂量均能明显增强小鼠体液免疫功能和单核巨噬细胞功能,且有剂量－反应关系,由此证明“思壮”     

UV照射与非UV照射FITC致敏小鼠淋巴结树突状细胞的光电镜免疫组织化学特性

本工作观察了小鼠淋巴结树突状细胞（ＤＣ）的光、电镜免疫组织化学特性,并探讨其功能。从ＦＩＴＩＣ致敏小鼠淋巴结分离的ＤＣ在体外可刺激ＦＩＴＣ特异性Ｔ－细胞株增殖。注射经ＵＶ照射、ＦＩＴＣ致敏的小鼠淋巴结细胞（ＤＬＮＣ）至受体小鼠,可导致该小鼠的免疫耐受;这些细胞在体外仍可促进ＦＩＴＣ特异性Ｔ－细胞株增殖,但明显弱于非ＵＶ照射小鼠ＤＬＮＣ的作用。免疫光镜下显示ＵＶ－ＦＩＴＣ致敏小鼠淋巴结的ＦＩＴＣ阳性ＤＣ与非ＵＶ照射ＦＩＴＣ致敏组一样也表达了ＭＡＣ－１、２、３和Ｆ４／８０等巨噬细胞标志,唯其ＦＩＴＣ阳性细胞率明显高于非照射ＦＩＴＣ致敏组动物。免疫电镜下显示这些细胞呈Ｉａ阳性和ＦＩＴＣ阳性,ＦＩＴＣ主要定位于线粒体和溶酶体结构等处。研究表明这些与ＦＩＴＣ致敏小鼠ＤＬＮＣ有关的细胞活性的差异与Ｉａ阳性ＤＣ数量减少、表面Ｉａ的表达、ＦＩＴＣ在ＤＣ内的分布变化无关。某些Ｉａ阳性ＤＣ胞质内可见Ｂｉｒｂｅｃｋ颗粒样结构,提示Ｉａ阳性ＤＣ的不同群体可迁移至ＵＶ照射小鼠的淋巴结。     

NIDDM发病前后中国地鼠胰岛和肝脏细胞GLUT的变化研究

用胶原酶法分离胰岛细胞、超速离心法提取中国地鼠胰岛和肝脏细胞膜蛋白,ＳＤＳ电泳、ＷｅｓｔｅｒｎＢｌｏｔｔｉｎｇ分离ＧＬＵＴ后,应用ＧＬＵＴ２抗体免疫结合法测定ＧＬＵＴ的含量与分子量,同时测定了动物血糖、尿糖和血浆胰岛素的含量、显微镜下直接计数了胰岛细胞总数。结果表明：ＮＩＤＤＭ发病后,自发ＮＩＤＤＭ中国地鼠胰岛细胞ＧＬＵＴ含量减少,与ＧＬＵＴ２抗体反应明显减弱,但肝脏ＧＬＵＴ无明显变化。胰岛细胞总数发病前后大体一致。而血糖、尿精及血浆胰岛素浓度明显升高。     

戴氏虫草菌丝体水提物对放射损伤小鼠体液免疫的作用

为了研究戴氏虫草菌丝体水提物对γ射线辐射损伤小鼠的脾细胞产生抗羊红细胞抗体、血清溶血素水平、淋巴细胞增殖等的影响,以＾６０Ｃｏ γ射线放射损伤小鼠为动物模型,采用脾细胞ＳＲＢＣ溶血反应（ＱＨＳ）比色法测定脾细胞产生抗羊红细胞抗体,溶血素测定（ＨＣ５０）比色法测定血清溶血素水平,ＭＴＴ比色法测定淋巴细胞增殖。结果表明在一定剂量条件下,戴氏虫草菌丝体水提物可以恢复γ射线辐射损伤小鼠脾细胞产生抗ＳＲＢＣ抗体的能力和血清溶血素的水平,同时也能够恢复脾Ｂ淋巴细胞的增殖能力和脾淋巴细胞数目。因此我们初步认为戴氏虫草菌丝体水提物对放射损伤小鼠体液免疫具有保护作用。     

螺旋藻保健功能的研究(Ⅰ)——免疫调节保健功能的检验与评价

本研究对螺旋藻产品的免疫调节保健功能进行了检验和评价。小鼠实验结果显示 ,饲喂螺旋藻可以增强其体液免疫、细胞免疫、自然杀伤细胞和巨噬细胞等免疫系统的重要组成部分的活力。研究表明 ,螺旋藻能协调地改善免疫系统的功能 ,它不愧是优秀的保健食品。     

HIV-1 p24 DNA和P24蛋白联合免疫小鼠的效果

DNA疫苗能够诱导机体产生特异的细胞免疫和体液免疫反应,在肿瘤和感染性疾病的疫苗开发中显示出巨大的潜能。以HIV-1核心蛋白P24为抗原基因,构建pVAX1-p24 DNA,经Western blotting和动物活体成像检测证明,pVAX1 DNA携带的外源基因可以在293T 细胞和小鼠肌肉组织有效表达。采用不同的免疫策略免疫BALB/c小鼠 (DNA/DNA,DNA/Protein),实验结果表明：pVAX1-p24单独免疫BALB/c小鼠,可诱导明显的体液免疫及细胞免疫反应;pVAX1-p24与P24蛋白联合免疫诱导的体液免疫反应高于pVAX1-p24单独免疫,所获得的抗体滴度是单独免疫的7.3~8.0倍,但细胞免疫反应则不及单独免疫组。研究结果表明采取不同的免疫策略可以诱导产生不同的免疫反应,根据具体情况调整免疫策略将获得更好的免疫效果。这些研究为艾滋病疫苗的研发提供了实验依据。     

汉滩病毒84Fli株DNA疫苗诱导小鼠免疫应答的初步研究

为了加强我国病毒性出血热的防治,本研究将汉滩病毒84Fli株核蛋白S和糖蛋白M编码片段分别克隆至pcDNA3.0载体,构建了pcDNA3/84S和pcDNA3/84M重组质粒,等量混合采用肌肉注射途径免疫C57BL/6小鼠,免疫3次,每次间隔2周,同时与双价出血热病毒灭活疫苗进行对比。ELISA及免疫荧光(IFA)分别检测小鼠血清中汉滩病毒核蛋白及糖蛋白特异性抗体,流式细胞仪和ELISPOT方法分析小鼠免疫后的细胞免疫水平。微量中和试验检测小鼠血清抗体的的中和活性。结果显示,DNA疫苗免疫组C57BL/6小鼠在初次免疫2周后即能检测到汉滩病毒核蛋白与糖蛋白的特异性抗体,与灭活疫苗组相比,重组质粒诱导的抗体滴度高,产生时间早,产生的抗体具有中和活性;同时可诱导产生特异性细胞免疫应答。研究表明,汉滩病毒pcDNA3/84S和pcDNA3/84M重组质粒能有效刺激小鼠产生特异性体液免疫和细胞免疫应答。     

Hypogammaglobulinemia in BLT Humanized Mice – An Animal Model of Primary Antibody Deficiency

Primary antibody deficiencies present clinically as reduced or absent plasma antibodies without another identified disorder that could explain the low immunoglobulin levels. Bone marrow-liver-thymus (BLT) humanized mice also exhibit primary antibody deficiency or hypogammaglobulinemia. Comprehensive characterization of B cell development and differentiation in BLT mice revealed other key parallels with primary immunodeficiency patients. We found that B cell ontogeny was normal in the bone marrow of BLT mice but observed an absence of switched memory B cells in the periphery. PC-KLH immunizations led to the presence of switched memory B cells in immunized BLT mice although plasma cells producing PC- or KLH- specific IgG were not detected in tissues. Overall, we have identified the following parallels between the humoral immune systems of primary antibody deficiency patients and those in BLT mice that make this in vivo model a robust and translational experimental platform for gaining a greater understanding of this heterogeneous array of humoral immunodeficiency disorders in humans: (i) hypogammaglobulinemia; (ii) normal B cell ontogeny in bone marrow; and (iii) poor antigen-specific IgG response to immunization. Furthermore, the development of strategies to overcome these humoral immune aberrations in BLT mice may in turn provide insights into the pathogenesis of some primary antibody deficiency patients which could lead to novel clinical interventions for improved humoral immune function.     

Progressive depletion of peripheral B lymphocytes in 4-1BB (CD137) ligand/I-Ealpha)-transgenic mice

Interaction of 4-1BB (CD137) and its ligand (4-1BBL) is thought to positively regulate cell-mediated and humoral immune responses. We have prepared transgenic mouse strains that express 4-1BBL cDNA under the control of MHC class II I-Ealpha promoter. The 4-1BBL-transgenic mice show progressive splenomegaly and selective depletion of B220(+) B cells accompanied with low levels of circulating IgG and defective humoral responses to Ag challenge. In addition, splenocytes from the transgenic mice fail to provide stimulation for allogeneic T cells in both lymphoproliferative and CTL responses in vitro, whereas their T cells remain functionally normal. Our results reveal unexpected functions of 4-1BBL in the regulation of humoral immune responses and Ag presentation.     

携带HBV包膜大蛋白DNA疫苗的减毒沙门菌在小鼠诱导免疫应答

探索一种简便、有效的乙型肝炎病毒DNA疫苗免疫方法。将编码绿色荧光蛋白的真核表达质粒pEGFPN1转化到减毒鼠伤寒沙门菌SL7207,灌胃饲服BALB/c小鼠,流式细胞术检测出小鼠脾细胞内表达的绿色荧光蛋白;构建编码HBV包膜大蛋白的DNA疫苗pCIS1S2S,分别以SL7207为载体的口服途径或直接肌肉注射途径免疫BALB/c小鼠,检测小鼠的血清抗体、T细胞增殖和细胞毒性T淋巴细胞反应,结果表明两种免疫途径均能在小鼠体内诱生细胞和体液免疫应答,但口服途径诱导免疫应答的强度明显强于肌肉注射途径。口服携带HBV DNA疫苗的减毒伤寒沙门菌可能代表一种简便、有效的治疗乙型肝炎的新方法。      

Murine encephalitozoonosis: the effect of age and mode of transmission on occurrence of infection

Experiments were conducted to determine whether neonatal mice are more susceptible to E. cuniculi than adult mice, and whether vertical and/or horizontal transmission occur in murine encephalitozoonosis. E. cuniculi infection in neonates did not cause mortality or clinical signs, but did result in chronic infection. Despite initial age-related immunodeficiency, mice infected as neonates eventually developed humoral and cell-mediated immune responses against the parasite comparable to those seen in adult mice. The results suggested that neonatal mice are not more susceptible to E. cuniculi than adult mice. Pups from either infected or normal parents did not differ in humoral and cell-mediated immune responses after challenge, suggesting that pups from infected parents were not infected with E. cuniculi during gestation. In contrast, mice became infected by caging with infected mice demonstrating that horizontal infection does occur.     

The role of T-lymphocytes in the development of a humoral immune response to the corpuscular antigen of Staphylococcus aureus

The dependence of humoral immune response and the formation of immunological memory to corpuscular staphylococcal antigen (CSA) on the T-system of immunity was studied in experiments on B-mice and on mice with the congenital absence of the thymus (nude). Primary and secondary immune response to CSA in athymic mice was found to be considerably less than in normal animals. After the repeated immunization of genetically athymic mice the pronounced secondary reaction of the formation of antibodies to CSA was observed. As shown in this investigation carried out with the use of adoptive transfer techniques, the induction of memory B-cells to CSA may occur in animals with congenital or experimentally induced T-immunodeficiency. The conclusion was made on the T-dependence of humoral immune response to CSA, the formation of immunological memory to this antigen being relatively T-independent.     

Platycodin D2 Improves Specific Cellular and Humoral Responses to Hepatitis B Surface Antigen in Mice

Platycodin D2 ( 1 ), a less hemolytic saponin from the root of Platycodon grandiflorum than platycodin D ( 2 ), was evaluated for the potential to enhance specific cellular and humoral immune responses to hepatitis B surface antigen (HBsAg) in mice. It significantly increased the concanavalin A (Con A)‐, lipopolysaccharide (LPS)‐, and HBsAg‐induced splenocyte proliferation in HBsAg‐immunized mice (P<0.05, P<0.01, and P<0.001, resp.). HBsAg‐specific IgG, IgG1, IgG2a, and IgG2b antibody titers in the serum were also markedly enhanced by 1 compared to the HBsAg control group (P<0.01 or P<0.001). Moreover, 1 significantly promoted the production of Th1 (IL‐2 and IFN‐γ) and Th2 (IL‐4 and IL‐10) cytokines from splenocytes in the HBsAg‐immunized mice (P<0.001). The adjuvant potential of 1 on splenocyte proliferation, serum HBsAg‐specific IgG2a and IgG2b antibody response, as well as Th1‐cytokine secretion from splenocytes in the HBsAg‐immunized mice was higher than that of Alum. The results suggest that 1 could improve both cellular and humoral immune responses to HBsAg in mice. Hence, 1 might be a promising adjuvant for hepatitis B vaccine with dual Th1‐ and Th2‐potentiating activity.