分子佐剂C3d增强人绒毛膜促性腺激素β基因避孕疫苗的免疫效应及转变免疫应答模式的研究

目的　研究分子佐剂C3d能否增强人绒毛膜促性腺激素 (hCG) βDNA疫苗的免疫原性 ,并转变Th1/Th2型免疫应答模式。方法　抽提纯化pcDNA3、pcDNA3 hCGβ、pcDNA3 hCGβ C3d3、pCMV4 hCGβ C3d34种质粒。将 12只 6周龄BALB/c雌性小鼠分为 12组 ,A1～ 3组各 6只以pcDNA3免疫 ,作为空白对照组 ;B1～ 3组各 6只以pcDNA3hCGβ免疫 ,作为阴性对照组 ;C1～ 3组各 6只以pcDNA3 hCGβ C3d3免疫 ,作为实验 1组 ;D1～ 3组各 5只以pCMV4 hCGβ C3d3免疫 ,作为实验 2组。免疫前 1周 ,于小鼠左后腿肌肉注射 0 2 5 %普鲁卡因 0 1ml。 1周后 ,在相同部位分别肌注前述 4种质粒 ,剂量分别为 :5pmol(A1～D1组 )、10pmol(A2～D2组 )、2 0pmol(A3～D3组 )。 3周后 ,再次给予相同剂量质粒加强免疫。于第 2次免疫后 3周 ,处死小鼠。用ELISA法测定外周血抗hCGβ抗体效价和经hCG抗原体外刺激脾细胞培养上清中Th1型 (IL 2、INF γ、TNF α) /Th2型 (IL 4、IL 10 )细胞因子分泌水平。结果　C3d分子佐剂能明显提高抗hCGβ抗体滴度。当免疫剂量为 2 0pmol时 ,与B3组比较 ,C3组抗hCG抗体最大效价达到 1∶4 5 0 ,提高了 9倍。D3组抗hCG抗体最大效价达到 1∶12 15 0 ,提高了 2 4 3倍。D3组与C3组相比较 ,抗hCG抗体水平提高了 2 7倍。受     

GM-CSF表达质粒对pcDNA3/MDC-VP1 DNA疫苗的增强作用

目的:观察粒-巨噬细胞集落刺激因子(GM-CSF)对pcDNA3/MDC-VP1DNA疫苗免疫作用的影响,为柯萨奇病毒B组3型(CVB3)疫苗的研制提供理论和实验依据。方法:4～6周龄雄性BALB/c小鼠随机分成pcDNA3组、pcDNA3/MDC-VP1组、pcDNA3/mGM-CSF组、pcDNA3/MDC-VP1与pcDNA3/mGM-CSF混合注射组,每组10只。每3周接种1次,共3次。每次接种的第20天眼眶采血,用微量中和试验(固定病毒-稀释血清法)检测血清中和抗体效价。第3次免疫后3周,每组取3只小鼠脾脏制备淋巴细胞悬液,检测淋巴细胞增殖活性与特异性细胞毒性T淋巴细胞(CTL)杀伤活性。结果:pcDNA3/MDC-VP1+pcDNA3/mGM-CSF组的血清中和抗体滴度明显提高,小鼠脾脏淋巴细胞增殖活性和特异性CTL杀伤活性均有增强。结论:GM-CSF可以增强pcDNA3/MDC-VP1DNA疫苗的特异性免疫应答。     

Breakthrough of immune self-tolerance to calreticulin induced by CpG-oligodeoxynucleotides as adjuvant

Reportedly, bacterial DNA containing unmethylated cytosine-guanosine dinucleotide motif-containing oligodeoxynucleotides (CpG-ODNs) can induce Th1-type adjuvant effects. We produced autoantibodies and induced hepatitis in mice using extracted proteins from human hepatocytes with CpG-ODNs as adjuvant. Western blot analysis was performed of sera from immunized mice and two patients with autoimmune hepatitis (AIH). When a common band was detected, N-terminal amino acid sequencing was performed to determine its site. For detection of antibodies against the identified protein (calreticulin), ELISA was performed of sera of 50 patients with AIH: 45 with primary biliary cirrhosis (PBC), 24 with chronic hepatitis C (CH), and 24 healthy controls. Mice were immunized with calreticulin protein with CpG-ODNs as adjuvant. Several reacted bands were detected in their sera; in addition, a common band to the sera of patients with AIH was detected at 60 kDa. Subsequent N-terminal amino acid sequencing revealed that the protein was human calreticulin. ELISA showed that, of patients with AIH, PBC, and CH, 30.0% (15/50), 17.8% (8/45), and 12.5% (3/24), respectively, were positive for anti calreticulin antibodies. Splenocytes from immunized mice produced IFN-gamma after they were pulsed with calreticulin protein. Histological analyses of liver specimens taken from mice immunized with calreticulin protein together with CpG-ODNs showed spotty and focal necrosis. Immunofluorescence analysis showed increased expression of calreticulin in the liver treated with CpG-ODNs. These results suggest that a breakthrough of immune self tolerance to calreticulin is induced with CpG-ODNs as adjuvant and that calreticulin protein might be a target antigen in this model.     

Cellular and humoral immune response of foals to vaccination with Corynebacterium equi

Transformation of peripheral blood lymphocytes from pony foals vaccinated and subsequently infected with Corynebacterium equi was studied. Three foals were vaccinated on two occasions using a formalinized C. equi vaccine with aluminum hydroxide as an adjuvant. Three nonvaccinated foals served as controls. Foals were challenged intratracheally with 9 x 10(9) C. equi six weeks after the initial vaccination.Foals survived this infection for one to two weeks. Significant lymphocyte transformation in response to C. equi antigens was detected in two vaccinated foals at the third week after initial vaccination and in all vaccinated animals at the fifth week. No statistically significant transformation was seen in nonvaccinated foals before infection. Vaccinated and nonvaccinated foals showed responsive lymphocytes following challenge. Vaccination offered no obvious protection against experimental challenge but this failure was probably due to an excessive infective dose of organisms. Low levels of humoral antibodies were detected in some challenged foals. The pathological changes in the lungs of infected animals were comparable with, but more fulminating than, changes observed in the natural disease.     

恶性疟MSP-2、CSP多价DNA疫苗免疫小鼠应答类型研究

目的：探讨恶性疟原虫FCC-1/HN株裂殖子表面蛋白-2(MPS-2)和环子孢子蛋白(CSP)组成的DNA多价疫苗在小鼠体内诱导的免疫应答的特性及抗感染的保护性免疫机制。方法：将重组真核表达质粒pBK/CSP和pBK/MSP-2经骨骼肌注射BALB/c小鼠，小鼠经多价疫苗免疫8wk后，用流式细胞仪分析脾脏T淋巴细胞的分化，并体外培养脾脏细胞，用夹心ELISA法测定IFN-γ和IL-2的产生；用血清学方法测定免疫鼠IgG抗体的动态变化。结果：与对照组相比，疫苗组CD^4 cd^8 淋巴细胞有显的增高，体外培养的脾脏细胞IFN-γ有一个高浓度的分泌。同时，免疫鼠血清对疟原虫抗原都表现了一个较高水平的IgG类抗体反应。结论：恶性疟原虫FCC-1/HN，pBK/MSP-2和pBK/CSP多价疫苗诱导了一个以细胞免疫为主的免疫应答类型。     

约氏疟原虫感染早期小鼠免疫应答的实验研究

目的:探讨约氏疟原虫感染早期小鼠免疫应答的差异性.方法:约氏疟原虫腹腔感染DBA/2、BALB/c和C57BL/6小鼠,制备薄血膜,姬姆萨染色,计数红细胞感染率;通过ELISA和Griess实验分别检测脾细胞培养上清中γ干扰素(IFN-γ)和一氧化氮(NO)水平.结果:感染第3 d之后,DBA/2小鼠的虫体血症水平显著低于BALB/c和C57BL/6小鼠;将疟原虫寄生的红细胞(PRBC)和脂多糖(LPS)或IFN-γ分别与3种感染第3 d小鼠脾细胞共同培养后,DBA/2小鼠脾细胞培养上清中的IFN-γ和NO水平明显高于BALB/c和C57BL/6小鼠.结论:约氏疟原虫感染早期,不同遗传背景小鼠在Th1型细胞免疫应答的建立和巨噬细胞活化方面存在显著差异.     

乙型肝炎疫苗免疫应答宿主相关遗传因素研究进展

乙型肝炎病毒（hepatitis B virus,HBV）感染是引起肝脏疾病最主要的原因之一。据估计,全球有20亿人曾感染过HBV,3.5亿~4亿人为HBV慢性感染者,其中75%的感染者为亚洲人[1]。乙型肝炎（乙肝）疫苗是预防HBV感染最有效的手段,但首次全程接种乙肝疫苗后,仍有5%~10%的人群HBV表面抗体（抗-HBs）达不到保护水平（抗-HBs〈10mIU/ml）[2-3]。     

Humoral immune response to HCV core DNA vaccine in mice

DNAvaccine,alsocallednucleicacidvaccine,isarecombinanteukatyoticexpressionplasmidencodingcertainantigenofpathogen.DNAvaccinecanexpressantigenandinducecellularandhumoralimmuneresponsesaftervaccinationbyintramuscularinjectionorplasmid--coatedmicroparticlebombardment.Itshightemperaturestability,lowexpenseofmassproductionandtheabilitytoinducestrongimmuneresponsesevenfortheuseintherapyhavemadeDNAvaccineanexcitingwayfornewvaccinedevelopment['J.InordertofurnishevidenceforHCVDNAvaccinesuitablefo…     

Significance of isolated antibody to hepatitis B core antigen determined by immune response to hepatitis B vaccination

The immune response to hepatitis B vaccine was studied in 14 individuals with isolated, high-titer antibody to hepatitis B core antigen (anti-HBc) and examined as an indicator of this serologic pattern's significance. Four subjects demonstrated a low-titer antibody to hepatitis B surface antigen (anti-HBs) on repeated testing, and three in this subgroup had anamnestic responses (anti-HBs, 82 to 140 ratio units) after vaccination. Compared with 22 seronegative controls, the remaining ten had significantly higher anti-HBs response rates (78% vs 22%, P = .003) and median anti-HBs titers (4 vs 0 ratio units, P = .008) two weeks after vaccination. One of ten subjects had an anamnestic response, while another exhibited no response. The general pattern of anti-HBs responsiveness observed in those subjects with isolated, high-titer anti-HBc was intermediate between seronegative and anti-HBs-positive groups and may indicate a state of waning immunity after natural infection. Hepatitis B vaccination with follow-up anti-HBs testing should be done for those patients with isolated, high-titer anti-HBc to help exclude chronic infection and boost protective immunity.     

不同术式再次剖宫产腹壁愈合腹腔粘连情况比较分析

目的:分析3种剖宫产术式术后腹壁愈合及腹腔粘连的情况。方法:采用回顾性分析的方法,对剖宫产术后再次剖宫产时患者腹壁愈合腹腔粘连的情况进行观察。按第1次剖宫产术式不同分为3组:改良腹部横切口式剖宫产者为A组92例,Stark式腹部横切口剖宫产者为B组88例,下腹纵切口式剖宫产者为C组112例。比较3组患者腹壁愈合以及腹直肌、腹膜、膀胱腹膜反折粘连的情况。结果:A、B组腹壁愈合情况好,愈合佳的比率分别为76.09%(70/92)和77.27%(68/88),明显高于C组的25.89%(29/112),A、B组比较差异无统计学意义(P>0.05),与C组比较各级差异均有统计学意义(P<0.001);腹直肌粘连严重的比率A组为40.22%(37/92),C组为11.61%(13/112),明显低于B组的71.59%(63/88),3组比较差异均有统计学意义(P<0.001);腹膜粘连率A组为42.39%(39/92),C组为33.04%(37/112),与B组的61.36%(54/88)相比,差异有统计学意义(P<0.05),A组与C组比较差异无统计学意义(P>0.05);3组膀胱腹膜反折的粘连率差异无统计学意义(P>0.05)。结论:剖宫产术后腹壁愈合、腹腔粘连情况与手术术式有关,改良腹部横切口式剖宫产者再次剖宫产时腹壁愈合佳,腹直肌、腹膜粘连少。为预防再次手术粘连的问题应慎重选择手术术式。     

Tramadol as an adjuvant to intravenous regional anesthesia with lignocaine

OBJECTIVE: To assess the effect of different doses of tramadol when added to lignocaine during intravenous regional anesthesia (IVRA). METHODS: Sixty patients, scheduled for hand surgery under IVRA in King Fahd University Hospital, Al-Khobar, Saudi Arabia from January 2006 to January 2007 were randomly allocated into 3 groups (20 patients each) in a double blind controlled study. All patients received 0.5% lignocaine, 40ml plus 2ml of a study solution containing either isotonic saline control group, or tramadol 50mg (group T50) or tramadol 100 mg (group T100). Hemodynamic changes, sensory and motor block onset and recovery times, tourniquet tolerance time, the quality of intraoperative anesthesia and the duration of postoperative analgesia were assessed. RESULTS: All patients, 20 in each group completed the study period. Patients who received tramadol had earlier onset of sensory block (5.2 +/= 1.2; 4.9 +/= 1.2 min in the T50; and T100 groups) compared with the control group (7.6 +/= 1.4 min). Patients who received 100mg of tramadol had better tolerance of tourniquet (p=0.011), and less intraoperative fentanyl supplementation (p=0.042). They had also a longer time to the first postoperative analgesic request (p=0.001) compared with the control group. CONCLUSION: Tramadol 100 mg is a beneficial additive to lignocaine for IVRA since it shortened the onset of sensory block, enhanced the tourniquet tolerance and improved the perioperative analgesia.     

三种常见眼病视力康复效果观察

目的：了解严重影响视力的3种眼疾的视力康复效果。方法：统计分析118例159眼的白内障、角膜炎、青光眼的视力康复情况。结果：白内障、角膜炎、青光眼的视力康复率分别为44．0％、68．8％、14．7％，白内障、角膜炎的视力康复率无显著性差异（P〉0．05）；青光眼视力康复率与前两者有显著性差异（P〈0．001）。结论：白内障、角膜炎临床疗效好，而青光眼重在早期发现、早期治疗。     

丙肝核酸免疫效果初步观察

目的 观察丙肝核酸疫苗的免疫效果。方法 用丙肝病毒Ｃ＋Ｅ１区基因的真核表达重组质粒ｐＳＶＬ－ＨＣＶ／Ｃ＋Ｅ１免疫蛇毒预处理的ＢＡＬＢ／Ｃ和Ｃ５７ＢＬ小鼠，两周后采用ＥＬＩＳＡ法开始检测抗体滴度。结果 真核表达重组质粒ｐＳＶＬ－ＨＣＶ／Ｃ＋Ｅ１免疫能诱导习免疫应答，产生高水平的特异抗体。特别是Ｃ５７ＢＬ小鼠抗体反应最好，初次免疫就能产生高水平的特异抗体，加强后能持续数周，最高滴度ＯＤ值达１．４５。结     

HCV核心蛋白DNA疫苗诱导的小鼠体液免疫应答

目的：观察丙型肝炎病毒（ＨＣＶ）核心蛋白ＤＮＡ疫苗诱导ＢＡＬＢ／ｃ小鼠体液免疫应答的效力，为研制应用于人类的ＨＣＶ ＤＮＡ疫苗提供实验依据。方法：将全长ＨＣＶ核心蛋白基因插入真核表达质粒载体ｐｃＤＮＡ３．１，构建ＨＣＶ核心蛋白重组表达质粒ｐｃＤＮＡ３．１ＨＣＶｃｏｒｅ，肌注ＢＡＬＢ／ｃ小鼠，以ＥＬＩＳＡ法检测小鼠血清ＨＣＶ抗体。以此重组质粒转染小鼠ＮＩＨ３Ｔ３细胞，以ＨＣＶ抗体阳性小鼠血清为捕获抗     

HBsAg阳性母亲所生婴儿全程接种乙肝疫苗免疫应答分析

目的：对HBsAg阳性母亲所生婴儿进行全程接种乙肝疫苗免疫应答进行分析。方法：观察对象为7月龄和1～6岁,每年静脉采血,观察两组免疫应答及乙肝免疫后不同年龄的HBsAb阳性率。结果：HBsAg阳性母亲所生婴儿全程接种乙肝疫苗经统计学分析,两组免疫应答有显著性差异（P〈0.05）。结论：被动加主动免疫对乙肝病毒感染具有良好的保护效果和促进免疫应答反应。对乙肝免疫后不同年龄的HBsAb阳性率观察,随年龄增长HBsAb阳性率呈进行性下降,免疫后3～5年应加强免疫一次,这样将会起到更加持久、稳定的保护作用。     

酶联免疫法检测伤寒杆菌抗原的结果分析

目的 评价伤寒杆菌抗原酶联免疫法(ELISA)对伤寒的临床诊断价值。方法 对413例疑似伤寒患血液用ELISA和细菌培养法平行检查，然后对检测结果进行对比分析。结果 ELISA检测法敏感性为100％。特异性为81．1％。结论 ELISA检测法对伤寒的诊断具有灵敏度高、反应时间短、操作简便等特点，可用于伤寒的辅助性诊断，具有早期诊断的价值，值得基层医疗单位进一步推广。     

胃黏膜局部免疫反应在以壳聚糖为佐剂的幽门螺杆菌疫苗免疫保护中的作用

目的探讨以壳聚糖为佐剂的幽门螺杆菌(Hp)疫苗的免疫保护作用及其机理。方法BALB/C小鼠随机分为9组:①空白对照组;②壳聚糖酸溶液组;③壳聚糖颗粒组;④Hp抗原组;⑤Hp抗原+壳聚糖酸溶液组;⑥Hp抗原+壳聚糖颗粒组;⑦Hp抗原+霍乱毒素(CT)组;⑧Hp抗原+壳聚糖酸溶液+霍乱毒素组;⑨Hp抗原+壳聚糖颗粒+霍乱毒素组,各组于第0、7、14、21天灌胃各免疫1次,免疫后4周给予1×109/ml的SS1Hp菌液0.5ml/只进行攻击,隔日1次,共2次。4周后,采用定量Hp培养和病理改良Giemsa染色法检测胃黏膜内Hp感染。用酶联免疫吸附实验(ELISA)检测唾液和胃黏膜内抗HpIgA,用定量ELISA法检测胃黏膜内Th1和Th2细胞因子,用SP免疫组织化学法检测胃黏膜内分泌型IgA(sIgA)。结果(1)以壳聚糖为佐剂的Hp疫苗的免疫保护率达60%,与以霍乱毒素为佐剂的Hp疫苗的免疫保护率(58.33%)相似,显著高于单纯Hp抗原组及其他不含Hp抗原组(P<0.01或P<0.05),同时以霍乱毒素+壳聚糖为佐剂的Hp疫苗的保护率为84.62%、85.71%,其Hp的定植评分显著低于无佐剂组及以霍乱毒素为佐剂组(P<0.01或P<0.05)。(2)胃黏膜内sIgA及特异性抗HpIgA水平在壳聚糖为佐剂组与以霍乱毒素为佐剂组无差别(P>0.05),显著高于无佐剂组,而壳聚糖与霍乱毒素联合应用组显著高于单以霍乱毒素为佐剂组(P<0.01或P<0.05)。(3)胃黏膜内白细胞介素(IL)2含量在攻击前各组间差异无统计学意义(P>0.05),攻击后含佐剂组显著高于对照组(P<0.01或P<0.05),且攻击后含Hp抗原各组均显著高于攻击前(P<0.05)。(4)胃黏膜内IL10含量攻击前以壳聚糖为佐剂组显著高于无佐剂组(P<0.01或P<0.05),攻击后各组之间差异无统计学意义(P>0.05),且攻击后含佐剂组均显著低于攻击前(P<0.01)。(5)胃黏膜内IL4含量攻击前以壳聚糖为佐剂组显著高于无佐剂组(P<0.05),攻击后以壳聚糖颗粒为佐剂组显著高于以霍乱毒素为佐剂组(P<0.05),以壳聚糖溶液为佐剂组显著高于对照组、无佐剂组及佐剂中含霍乱毒素组(P<0.01或P<0.05),且攻击后含佐剂组均显著低于攻击前(P<0.01或P<0.05)。结论(1)以壳聚糖为佐剂的Hp疫苗对Hp感染具有免疫保护作用。并可成功的诱导黏膜局部特异性的体液免疫反应,这可能在其免疫防御中起作用;(2)以壳聚糖为佐剂的Hp疫苗可促进Th1和Th2的混合免疫反应,并可逆转攻击后Th2反应的抑制,恢复Hp感染所致的Th1/Th2失衡,从而发挥其免疫保护作用。