实验教学中颗粒性抗原与可溶性抗原制备抗血清的方法及其应用

目的 比较颗粒性抗原与可溶性抗原制备抗血清的效价,寻求获取符合实验教学需要的高效价抗血清的较佳方法.方法 牛血清白蛋白(Bovine Serum Albumin,BSA)作为可溶性抗原,伤寒"H"菌液作为颗粒性抗原,采用1周间隔的免疫程序,以BSA和伤寒杆菌"H"菌液免疫家兔,用对流免疫电泳和双向琼脂扩散试验、肥达试验检测并比较其血清抗体效价.结果 浓度为7×109个/ml伤寒杆菌"H"菌液免疫家兔后获取的伤寒"H"抗血清几何平均滴度(geometry mean titer,GMT)为1:9051.07,浓度为5mg/ml BSA免疫家兔后测得抗血清GMT分别为1:3.668和1:6.17.结论 颗粒性抗原与可溶性抗原1周间隔免疫程序制备的抗血清可获取符合实验教学的抗血清效价.     

两种免疫方法制备兔抗绵羊红细胞溶血素的效价观察

目的 比较两种免疫程序和途径制备抗血清的效价,并在免疫学溶血反应中的运用效果比较,寻求获取符合实验教学需要的高效价溶血素的较佳方法.方法 采用不同浓度的绵羊红细胞(Sheep red blood cells,SRBC)间隔1天和间隔3天腹腔、耳静脉免疫家兔,用血球凝集法浓度为1.5%SRBC悬液检测并比较两种免疫程序和...     

两种短程免疫方法制备伤寒抗血清的效价比较

目的通过对伤寒沙门菌抗体效价两种短程方法的比较,找出更为高效快速的免疫方法。方法采用鼠伤寒杆菌制备出伤寒沙门菌O抗原,将每天注射的免疫程序和每三天注射的免疫程序作比较,采用试管凝集法检测比较血清抗体效价,选出更为高效省时的制备免疫血清的方法。结果每天注射的免疫程序效价为1:2560,每三天注射的免疫程序效价为1:1280。结论将两种短效免疫方法进行比较,每天注射的免疫程序所制得的血清用时更短,效价更高。     

四氢呋喃与牛血清白蛋白相互作用的荧光光谱法研究

目的研究四氢呋喃（tetrahydrofuran,THF）与牛血清白蛋白（bovine serum albumin,BSA）相互作用的机制,探讨温度对其反应的影响。方法移取的BSA于石英比色皿中,搅拌混合均匀,避光静置,分别在25℃（298 K）、29℃（302 K）、33℃（306 K）、37℃（310 K）下以280 nm为激发波长,在LS-55荧光分光光度计上扫描280~500 nm波长范围内的发射光谱。结果不同温度下的结合猝灭常数（Ksv）为（3.466~6.317）×104L/mol,猝灭常数随温度的升高而减小。结论四氢呋喃对BSA的猝灭作用是属于静态猝灭。     

免疫比浊法测定人血清前白蛋白及其应用

用免疫比浊法测定人血清前白蛋白（ＰＡ）含量，对其测试条件进行了探讨。该方法的检测下限为０．２０ｍｇ／ｄｌ，批内ＣＶ为３．６４％，批间ＣＶ为５．３４％，回收率为９１．３０％～１０９．２２％。用本法与火箭电泳法同时测定了９０例健康成年人的血清ＰＡ含量，分别为３５．７１±６．１２ｍｇ／ｄｌ和３５．６２±６．４０ｍｇ／ｄｌ（ｘ±ｓ），两法结果无显著差异（Ｐ＞０．０５），相关系数为０．９２４０。本法具有简便、快速、灵敏等优点，适用于大批量样品测定。     

甲型肝炎减毒活疫苗不同免疫程序的长期免疫原性比较

为了探讨国产规范化甲型肝炎(甲肝)减毒活疫苗(LA-1株)不同免疫程序的长期免疫原性,在广西壮族自治区柳城县筛检出甲肝抗体IgG阴性的6～9岁儿童156名,分为3组,各组年龄和性别构成均衡.甲肝疫苗由长春生物制品研究所提供,滴度为106.75TCID50.A组接种1剂甲肝疫苗,B组按0、6个月程序,C组按0、12个月程序分别接种2剂甲肝疫苗.A组于免疫后6、12、24、36个月,B、C组于第2针免疫后1个月、首针免疫后12、24、36个月定人随访采血,用美国Abbott公司ImxmEIA试剂检测甲肝抗体IgG.结果显示A组免疫后6～24个月的抗体阳性率88.6%～91.4%,GMT105mIU/ml～106mIU/ml,第36个月时阳性率80.0%,GMT99.20mIU/ml.B、C组的抗体水平变化趋势大致相同.抗体GMT均在第2针免疫后1个月达到高峰,分别为1?204.63mIU/ml和3?463.21mIU/ml,从峰值到第24个月间,两组GMT分别迅速下降了59%和83%(为489.12mIU/ml和596.57mIU/ml);第36个月,分别缓慢降至459.68mIU/ml和506.23mIU/ml,较第24个月分别下降6%和15%.B、C组在2针免疫后,抗体水平始终显著高于A组,在第36个月,GMT分别是A组的4.6倍和5.1倍,阳性率(97%)亦高于同期A组(80%).甲肝减毒活疫苗0、6个月和0、12个月程序产生的高水平抗体免疫后呈先急后缓的下降趋势,但仍高于1针的抗体水平;免疫后3年抗体阳性率仍达97%和较高水平的GMT,预示着抗体有可能长期存在.     

不同效价组织培养人用狂犬病疫苗免疫后抗体水平的监测

本文对禹州市262例犬咬伤者接种不同效价的狂犬病疫苗,用ELISA法检测三组疫苗免疫后血清抗体。A组、B组和C组抗体阳转率分别是77.9％、85.06％和89.89％。三种不同的效价疫苗的免疫效果无显著性差异。免后抗体阳转率与年龄、性别无关。     

狂犬病暴露预防处置不同免疫程序比较

狂犬病(rabies)是由狂犬病毒引起的人兽共患急性传染病,死亡率近100％[1-2].近年来中国犬、猫数量快速增加,被犬、猫伤害的人数也不断增加.暴露后处置是预防狂犬病的唯一有效措施.WHO认为暴露后及时、科学和彻底的处置能够避免狂犬病的发生[3],其推荐的暴露后肌内接种程序有Essen 5针法和Zagreb 4针法(即2-1-1免疫程序)[4].以往在犬伤暴露后处置中,对狂犬疫苗的接种多采用Essen免疫程序.为进一步对比分析Essen免疫程序与2-1-1免疫程序在狂犬病暴露预防处置中的安全性、依从性及医疗成本,本研究以2010年10月-2012年10月浙江省嘉兴市疾病预防控制中心门诊就诊患者6 101例为研究对象,比较分析2种免疫程序效果,现将结果报告如下.     

抗人血白蛋白单克隆抗体的制备及其免疫金标试纸条的研制

目的:制备抗人血白蛋白单克隆抗体并对其抗体特性进行鉴定,将其标记胶体金,研制一种快速准确检测尿中人血白蛋白试纸条的方法。方法:用99.8%纯度的人血白蛋白免疫Balb/C小鼠,采用杂交瘤细胞技术制备单克隆抗体,并对其单克隆抗体的相关性质进行鉴定,制备胶体金,并标记所制备的单克隆抗体,按竞争法制备试纸条。结果:得到两株能够稳定分泌抗体,效价高的杂交瘤细胞系4C2,2F10。结论:建立的免疫层析试纸条,简便、可靠,有望通过进一步改进应用于临床检验。     

Intestinal absorption of dinitrophenyl-lysine and effect of immunization with dinitrophenylated bovine serum albumin

The intestinal absorption of dinitrophenyl-lysine (DNP-lys) was studied with a special interest on the role of the immune system in the absorption of small molecules which are recognized as nonself. [3H]-DNP-lys was rapidly absorbed by ligated intestinal loops in situ via a saturable and unique route. When [3H]-DNP-lys was preincubated with the immune serum obtained from rats immunized with dinitrophenylated bovine serum albumin (DNP-BSA), the [3H]-DNP-lys absorption was depressed. The absorption of [3H]-DNP-lys in DNP-BSA-immunized rats was depressed compared to the control. The results obtained suggest that the immune system play a role in avoiding the absorption of small molecules with antigenicity.     

Interaction of acrylamide with bovine serum albumin

The binding of acrylamide (ACR) with purified bovine serum albumin (BSA) was studied. Binding of ACR with BSA was characterized by equilibrium dialysis, fluorescence studies, and ultraviolet spectroscopy. ACR was quantitated by high-pressure liquid chromatography. Equilibrium dialysis studies on the binding of ACR with BSA showed that more than 25% of the added ligand was bound to the protein at equilibrium. ACR produced a concentration-dependent decrease in uv absorbance of BSA, indicating reactivity of ACR with BSA. Fluorescence quenching studies of ACR binding showed a concentration-dependent quenching of the fluorescence of BSA. ACR also caused a concentration-dependent decrease in the fluorescence of sulfhydryl (-SH) groups present on BSA, implicating a role of protein -SH groups in the binding of ACR. Prior blocking of -SH groups by N-ethylmaleimide resulted in 16% inhibition of the initial binding of ACR, suggesting involvement of -SH groups. Our results demonstrate that ACR binds to BSA through both aromatic amino acids and -SH groups and that such binding may play an important role in pharmacodynamics and toxicity of ACR.     

The binding of mercury to bovine serum albumin

Additional studies on the binding of mercury by serum albumin is presented. Data were obtained by radiotracer methods and the technique of equilibrium dialysis. The results showed that mercury is bound at other sites in addition to the carboxyl and sulfydryl groups reported previously.     

The binding of ascorbate to bovine serum albumin

Ultrafiltration has been used to investigate the interaction of ascorbate with bovine serum albumin in 0.1 M sodium phosphate buffer, pH 6.5. The results are interpreted in terms of the binding of ascorbate to four equivalent and independent protein sites, governed by an intrinsic association constant of 2 600 +/- 700 M-1 at 20 degrees C, thereby providing evidence for specificity of the interaction over a range of vitamin concentration (0.08-1.5 mM) pertinent to the physiological situation.     

Comparison of antibody titers after immunization with monovalent or tetravalent KLH conjugate vaccines.

Antigens such as ganglioside GD3, neutral glycolipid Lewis(y) (Le(y)) and mucins MUC1 and MUC2 are over-expressed on the cell surface of many tumors. We have shown previously that conjugation of antigens such as these to keyhole limpet hemocyanin (KLH) and the use of immunological adjuvant QS-21 is the optimal approach for inducing high titer IgM and IgG antibodies. These antibodies are able to bind with natural antigens on the tumor cell surface and mediate complement dependent cytotoxicity and/or antibody dependent cell mediated cytotoxicity. Immunization of patients with monovalent vaccines containing these and a variety of other antigens have demonstrated both the consistent immunogenicity and the safety of these vaccines. Now, in preparation for the use of polyvalent conjugate vaccines in the clinic, we have addressed for the first time with conjugate vaccines against cancer antigens several questions in the pre-clinical setting, including whether immunogenicity of the individual components is decreased in the polyvalent vaccine and issues relating to vaccine formulation and administration. We have immunized groups of mice with GD3-KLH, Le(y)-KLH, MUC1-KLH and MUC2-KLH conjugates and QS-21 separately or mixed and administered at one or four sites. High titer IgM and IgG antibodies were induced against each of the four antigens whether administered singly in separate mice, at separate sites in the same mice, or mixed and administered at a single site or at four sites, or administered subcutaneously (s.c.) or intraperitoneally (i.p.). These antibodies reacted specifically with the respective antigens and tumor cells expressing these antigens. There was no evidence of suppression of the antibody response against any one of the antigens by the presence of the other conjugates in the vaccine. Immunogenicity of the four individual antigens conjugated to KLH and QS-21 is not affected by mixing the four together and administering them at a single subcutaneous site.