单克隆抗体：生命科学的革命、治疗肿瘤的希望

一、单克隆抗体的概念及制备 免疫反应是人类对疾病具有抵抗力的重要因素。当动物体受抗原刺激后可产生抗体。抗体的特异性取决于抗原分子的决定簇，各种抗原分子具有很多抗原决定簇，因此，免疫动物所产生的抗体实为多种抗体的混合物。用这种传统方法制备抗体效率低、产量有限，且动物抗体注入人体可产生严重的过敏反应。     

应用酶联免疫吸附法(ELISA)诊断家蚕微粒子病的研究

感染微粒子病的家蚕中肠壁匀浆作抗原,注射于家兔制备抗血清。用戊二醛简易法将辣根过氧化物酶与抗体蛋白交联,制成酶联免疫抗体。用ELISA直接法或双抗体(夹心)法均可将未发现孢子的患病中肠检出阳性反应。用分光光度计对聚苯乙烯塑料多孔板吸附免疫反应的读数(OD值)表示更则为准确。 酶联抗体—抗原点滴法试验(AST),将抗原滴于硝酸纤维薄膜,再加入酶标抗体,可获得肉眼判别阳性的斑点。     

丙型肝炎患者血清的自身抗体检测及临床分析

目的：探讨自身免疫反应在丙型肝炎中的作用。方法：应用间接免疫荧光法，以生物薄片马赛克技术制备的冰冻组织细胞切片为抗原与待测血清结合，加入荧光素标记物抗人免疫球蛋白，检测94例丙型肝炎(丙肝)患者血清中抗核抗体(ANA)、抗线粒体抗体(AMA)、抗平滑肌抗体(SMA)、抗肝膜抗体(LMA)和抗肝特异性脂蛋白抗体(LSP)等自身抗体，并根据荧光反映模式判定结果。结果：(1)急性丙肝组患者14例中有6例出现自身抗体阳性，检出率为42．8％；慢性丙肝组80例中自身抗体阳性者30例，检出率37．5％。两组比较差别无统计学意义。(2)抗—HCV阳性的64例中自身抗体阳性者28例，检出率43．75％；HCVRNA阳性的30例中8例抗体阳性，检出率26．67％。结论：HCV感染过程中机体可出现自身免疫反应，HCV患者在常规诊疗过程中应检测自身抗体。     

试谈肾小球肾炎的治标与治本

Comlron就肾炎的治疗提出了针对免疫反应的三个环节的治疗方案:(1)针对抗原:避免接触抗原,排除抗原。(2)针对抗体:抑制、排除抗体或增加抗体。(3)针对免疫效应:如降低补体的活性,抑制纤维蛋白的形成,抑制白细胞趋化因子和抑制激肽系统等。     

鲨鱼肝再生因子(SHRF)半抗原偶联与多抗制备

探讨鲨鱼肝细胞再生因子(Shark Hepatic Regenerative Factor,SHRF)完全抗原的偶联方法和兔抗体的制备。分别考察了戊二醛(GA)一步法,戊二醛二步法和1-乙基-3-(3-二甲氨丙基)碳化二亚胺(EDC)法3 种偶联方法对SHRF与BSA的偶联效果,并将制备的完全抗原免疫新西兰白兔。实验表明碳化二亚胺法具有反应条件温和,偶联效率高等优点,并得到了高效价的兔抗体,为SHRF的ELISA方法的建立奠定了基础。     

Sabin株脊髓灰质炎病毒灭活疫苗的制备及免疫原性评价

目的:建立Sabin株毒种制备脊髓灰质炎灭活疫苗(inactivated poliovirus vaccine, IPV)生产工艺并评价其免疫原性。方法:采用Vero细胞培养Sabin株脊髓灰质炎Ⅰ、Ⅱ、Ⅲ型病毒,制备3价Sabin株IPV(Sabin strain IPV, sIPV)。通过ELISA检测D抗原含量。...     

人体对流感活和灭活疫苗的抗体及细胞毒性T淋巴细胞应答

本文比较了 HI、单向辐射状溶血(SRH)和蚀斑中和(PN)试验对血凝素(HA)抗原的抗体应答结果。分析了这些方法测定的抗体应答与 HLA 限制的病毒特异性细胞毒性 T淋巴细胞(CTL)测定结果的相互关系。同时检测了对病毒核蛋白和内膜蛋白抗原的抗体应答。     

妊娠妇女血清柯萨奇B组病毒抗体测定

本文利用检测抗体的方法，对在本院体检的一组孕妇柯萨奇B组病毒（CVB）感染情况进行了调查，并对妊娠结果进行随访，发现某些异常妊娠可能与该组病毒感染有关。材料与方法一、检测对象本院门诊产前体检孕妇130人，受检时孕期平均26十周。同期体检非孕妇女100人。两组均取血清进行病毒抗体测定。二、试剂与方法用间接ELISA法测定CVBI～6型IgG、IgM多价抗体。所用病毒抗原，酶标抗人IgG、酶标抗人lgM均由本室自制，病毒抗原用Hela细胞制备。1．IgG抗体测定CVBI～6型病毒抗原按比例混合，经适当稀释包被微孔板，同样包被未感染细胞…     

脂质体亚单位疫苗预防EB病毒诱发的恶性淋巴瘤

最近报道与脂质体结合的纯化的病毒膜抗原gp340能预防EB病毒诱发的淋巴瘤,因而再次强调了对有效的免疫佐剂的需要。抗原gp340的免疫原性弱,但其脂质体形式能在小鼠体内诱生高效价的特异性病毒中和抗体。然而,在狮猴体内产生能足以提供保护作用的抗体,所需时间较长,并需多次腹腔内注射。Epstein等最近报道,较大剂量的脂质体gp340能在狮猴体内迅速诱生高效价的抗体,从而可通过较少的接种次数和较小剂量的dg340以及通过改变免疫程序达到保护性免疫应答。脂质体的大小、表面特征、脂质组分和调节抗原的方式并不相同。目前用的是冻干     

体外白细胞介素2的产生:一种评价狂犬病疫苗不同形式糖蛋白免疫原性的新方法

狂犬病病毒糖蛋白(GP)是唯一诱生病毒中和抗体和提供保护作用的抗原。GP的保护活性与其在疫苗中的形式有关,这对于制备狂犬病病毒亚单位疫苗相当重要。作者制备了纯化的灭活病毒(IPV)、脂质体结合的GP(GPL)、自动聚集的GP(GPR)及与脂质结合的糖蛋白GP1、GP2和GP3,将它们分别注射Balb/c小鼠,观察不同形式GP抗原诱生保护性免疫应答的各种指标的变化。结果表明,IPV和GPL可被中和性单克隆抗体(McAb)所识别,酶免疫试验(EIA)滴度均达1万任意单位(Au)/mg GP以上,     

Nasal priming with immunobiotic lactobacilli improves the adaptive immune response against influenza virus

The nasal priming with Lactobacillus rhamnosus CRL1505 modulates the respiratory antiviral innate immune response and improves protection against influenza virus (IFV) challenge in mice. However, the potential beneficial effect of the CRL1505 strain on the adaptive immune response triggered by IFV infection or vaccination was not evaluated before. In this work, we demonstrated that nasally administered L. rhamnosus CRL1505 is able to improve both the humoral and cellular adaptive immune responses induced by IFV infection or vaccination. Higher levels of IFV-specific IgA and IgG as well as IFN-γ were found in the serum and the respiratory tract of CRL1505-treated mice after IFV challenge. Lactobacilli treated mice also showed reduced concentrations of IL-17 and improved levels of IL-10 during IFV infection. The differential balance of inflammatory and regulatory cytokines induced by L. rhamnosus CRL1505 contributed to the protection against IFV by favoring an effective effector immune response without inducing inflammatory-mediated lung damage. The optimal immunomodulatory effect of the CRL1505 strain was achieved with viable bacteria. However, non-viable L. rhamnosus CRL1505 was also efficient in improving the adaptive immune responses generated by IFV challenges and therefore, emerged as an interesting alternative for vaccination of immunocompromised hosts. Similar to other immunomodulatory properties of lactobacilli, it was shown here that the adjuvant effect in the context of IFV vaccination was a strain dependent ability, since differences were found when L. rhamnosus CRL1505 and the immunomodulatory strain L. rhamnosus IBL027 were compared. This investigation represents a thorough exploration of the role of immunobiotic lactobacilli in improving humoral and cellular adaptive immune responses against IFV in the context of both infection and vaccination.     

Oral administration of heat-killed Lactobacillus pentosus strain b240 augments protection against influenza virus infection in mice

Host-defense mechanisms against influenza virus (IFV) infection involve both innate and acquired immunities. Among other components, secretory immunoglobulin A (SIgA) in the airway mucosa plays a particularly pivotal role in preventing IFV infection. Among 150 strains of lactic acid bacteria, Lactobacillus pentosus strain b240 (b240) has the highest IgA-inducing potency in mouse Peyer's patch cells. We previously reported a practical new finding that oral ingestion of nonviable heat-killed b240 elevates salivary IgA secretion in humans. The present study aimed to determine if nonviable b240 can prevent IFV infection in mice. In a BALB/c mouse model infected with lethal levels of IFV A/PR8/34 (H1N1), oral administration of b240 for 3 weeks by gavage prior to IFV infection significantly prolonged the survival period. For IFV infection at nonlethal levels, the infectious titers of IFV in the lungs 7 days after infection were significantly reduced after similar b240 administration. Both anti-IFV IgA and immunoglobulin G titers in bronchoalveolar lavage fluid and plasma on day 7 were significantly higher in the b240-treated group than the control group. The augmentation of the anti-IFV immune response by b240 application was preliminarily confirmed by the elevated production of IFV-driven T-cell factors during mixed lymphocyte reactions with b240-primed splenocytes. These results suggest that oral nonviable heat-killed b240 intake can facilitate protection against IFV infection.     

Oral administration of patchouli alcohol isolated from Pogostemonis Herba augments protection against influenza viral infection in mice

Seasonal influenza A infection results in considerable morbidity and mortality. The limited efficacy of available therapeutic strategies stresses the need for development and study of new molecules against influenza virus (IFV). Patchouli alcohol (PA), the major chemical constituent of Pogostemonis Herba, was previously found to strongly inhibit influenza H1N1 replication in vitro. In the present study, the in vivo anti-IFV effect of PA was investigated. In a mouse model infected with lethal levels of FM1, oral administration of PA (20 mg/kg to 80 mg/kg) for 7 d post IFV infection significantly increased the survival rate and survival time. For IFV infection at nonlethal levels, the quantity of IFV in the lungs 5 d after infection was significantly reduced after PA (20 mg/kg to 80 mg/kg) administration. Anti-IFV IgA, IgM, and IgG titers in serum on day 6 were significantly higher in the PA-treated group than the IFV-control group. Anti-IFV immune response augmentation was further confirmed by the elevated production of CD3+, CD4+, and CD8+ T cell levels in blood. Furthermore, the levels of inflammatory cytokines, including TNF-alpha, IL-10 and IFN-gamma in serum of mice, were regulated. Lung inflammation was reduced significantly after PA administration, and the effect may be mediated, at least in part, by regulating the lung levels of inflammatory cytokines. Thus, oral administration of PA appears to be able to augment protection against IFV infection in mice via enhancement of host immune responses, and attenuation of systemic and pulmonary inflammatory responses.     

Oral administration of Bifidobacterium longum ameliorates influenza virus infection in mice

We investigated whether the oral administration of Bifidobacterium longum BB536 could ameliorate influenza virus (IFV) infection in a mice model. Mice were orally administrated BB536 or saline for 2 weeks and then infected with IFV. Orally administered BB536 significantly alleviated symptoms, reduced the loss of body weight, and inhibited viral proliferation in the lungs relative to the control group findings. Histopathological findings in the lungs were improved in the BB536 group compared to control group findings. There was no significant difference in the levels of interleukin-6 (IL-6), interferon-γ (IFN-γ), IL-10 and IL-12p40 in the lungs between the groups, but the levels of IL-6 and IFN-γ were lower (p=0.076, 0.103, respectively) in the BB536 group compared with those of control group. The levels of IL-6 and IL-10 correlated significantly with the values of weight loss, and the levels of IFN-γ correlated with the virus titers in the lungs. These results suggested the potential of the oral administration of BB536 in ameliorating IFV infection and the possible involvement of anti-inflammatory effects of BB536 in the anti-infection effects against IFV.     

Lactobacillus plantarum strain YU from fermented foods activates Th1 and protective immune responses

Lactic acid bacteria (LAB) are known to have effects on immune function. From 203 strains of LAB isolated from fermented foods, we selected a beneficial strain, Lactobacillus plantarum strain YU (LpYU), which has high interleukin (IL)-12-inducing activity in mouse peritoneal macrophages. This activity of LpYU was partially mediated by Toll-like receptor (TLR) 2, but not TLR4 or TLR9. Oral administration of LpYU to ovalbumin (OVA)-immunized mice caused suppression of serum OVA-specific immunoglobulin E (IgE) levels, enhancing interferon (IFN)-γ production from spleen cells in response to OVA. Furthermore, LpYU enhanced natural killer cell activity in spleen cells and the production of IgA from Peyer's patch cells. Because activation of Th1 immune responses and IgA production induce antiviral effects, we evaluated the inhibitory effects of LpYU against the influenza A virus (A/NWS/33, H1N1) (IFV). Oral administration of LpYU suppressed viral proliferation in the lungs and in bronchoalveolar lavage fluids (BALFs). Both levels of IFV-specific secretory IgA in BALF and feces and titers of IFV-specific neutralizing antibody in BALFs and sera were increased. These results indicate that LpYU has a protective effect against IFV replication. We conclude that this strain has a beneficial effect in activating Th1 immune responses and preventing viral infection.     

Lactobacillus plantarum strain YU from fermented foods activates Th1 and protective immune responses

Lactic acid bacteria (LAB) are known to have effects on immune function. From 203 strains of LAB isolated from fermented foods, we selected a beneficial strain, Lactobacillus plantarum strain YU (LpYU), which has high interleukin (IL)-12-inducing activity in mouse peritoneal macrophages. This activity of LpYU was partially mediated by Toll-like receptor (TLR) 2, but not TLR4 or TLR9. Oral administration of LpYU to ovalbumin (OVA)-immunized mice caused suppression of serum OVA-specific immunoglobulin E (IgE) levels, enhancing interferon (IFN)-γ production from spleen cells in response to OVA. Furthermore, LpYU enhanced natural killer cell activity in spleen cells and the production of IgA from Peyer's patch cells. Because activation of Th1 immune responses and IgA production induce antiviral effects, we evaluated the inhibitory effects of LpYU against the influenza A virus (A/NWS/33, H1N1) (IFV). Oral administration of LpYU suppressed viral proliferation in the lungs and in bronchoalveolar lavage fluids (BALFs). Both levels of IFV-specific secretory IgA in BALF and feces and titers of IFV-specific neutralizing antibody in BALFs and sera were increased. These results indicate that LpYU has a protective effect against IFV replication. We conclude that this strain has a beneficial effect in activating Th1 immune responses and preventing viral infection.     

太原地区急性呼吸道感染病毒病原谱研究

目的了解太原地区急性呼吸道感染(ARI)病例的病毒病原学特性及分布情况,为ARI疾病的预防控制和治疗提供科学依据。方法利用荧光定量聚合酶链式反应(PCR)方法,对381份上呼吸道和下呼吸道ARI病例的呼吸道样本进行了流感病毒(IFV-A、H1、H3、甲型H1N1、B型、C型)、呼吸道合胞病毒(RSV-A、B)、人副流感病毒(HPIV-1～4型)、人冠状病毒(HCoV-229E、OC43、NL63、HKU1型)、人偏肺病毒(hMPV-A、B)、人博卡病毒(HBoV)、呼吸道腺病毒(R-ADV)等7种病毒(共计20个型别)的检测,并对结果数据进行了统计分析。结果 381份样本中病毒检测阳性结果有164份,阳性率为43.0%,分别为IFV(22.8%)、RSV(10.8%)、hMPV(3.7%)、HPIV(2.9%)、HCoV(2.6%)、R-ADV(1.8%)、HBoV(0.8%),其中同时2种以上病毒混合感染8份,占4.9%。全年中均有病毒感染病例被检出,主要集中在冬春季,IFV、RSV、hMPV阳性率最高月份分别为12月(71.4%)、2月(84.2%)、1月(33.3%)。另外,上呼吸道ARI病例样本323份,病毒阳性率为34.4%,以IFV为主(占76.6%);下呼吸道ARI病例样本58份,病毒阳性率为91.4%,以RSV为主(占75.5%);ARI病例中15岁以下儿童占有较大比例,其中3岁以下患儿的病毒感染率为85.7%。结论 IFV、RSV、hMPV为太原地区2009-2011年ARI病例的主要致病原,并且各病毒随年龄、流行季节中月份、感染部位等不同而有各自的流行特性。     

婴幼儿病毒性肺炎病原学及临床特点

目的探讨引起小儿病毒性肺炎病原学情况及其临床表现,为临床诊治提供依据。方法应用间接免疫荧光（IIF）方法检测267例婴幼儿肺炎患儿急性期血清中11种病毒（呼吸道合胞病毒RSV;腺病毒ADV;流感病毒IFV-A,B;副流感病毒PIY1-4;柯萨奇B1病毒CB1V;柯萨奇A,病毒;CA7V;埃可病毒）血清特异性IgM抗体。结果267例婴幼儿肺炎中共检出病毒感染102例,病毒感染率38．20％,单一病毒感染84例（82．35％）。其中RSV27株（26．47％）,IFV20株（19．61％）,ADV18株（17．65％）,PIV19株（18．62％）,混合病毒感染共18例（17．65％）。RSV肺炎患儿喘息重,缺氧明显;ADV肺炎患儿以高热和中毒症状表现为主;IFV和PIV肺炎患儿主要表现为咳嗽重、喘息轻。结论病毒性肺炎以RSV为主要病原,婴儿期病毒感染率最高,好发于冬、春季节。各类病毒性肺炎各有其临床特点。     

Lactobacillus fermentum CJL-112 protects mice against influenza virus infection by activating T-helper 1 and eliciting a protective immune response

We have previously reported that nasally administered Lactobacillus fermentum CJL-112 (CJL-112) efficiently improves resistance against lethal influenza infection in both mice and chicken. The aim of the present study was to understand the underlying mechanisms of the significant anti-influenza activity of this lactobacilli strain. In vitro, co-culturing of the chicken macrophage cell line HD-11 with CJL-112 significantly increased nitric oxide (NO) production. In vivo, CJL-112 was nasally administered to BALB/c mice for 21 days prior to influenza A/NWS/33 (H1N1) virus (IFV) infection. Significant up-regulation of T-helper 1 (Th1) cytokines (IL-2, IFN-γ) was observed, while the levels of T-helper 2 (Th2) cytokines (IL-4, IL-5, IL-10) was either reduced or unchanged than that in control mice were. Furthermore, IgA and specific anti-influenza IgA levels increased significantly in the treated mice than those in untreated mice. Therefore, CJL-112 likely protects the mice against lethal IFV infection via stimulation of macrophages, activation of Th1 and augmentation of IgA production, when directly delivered into the respiratory tract.     

Efficacy of oral administration of heat-killed probiotics from Mongolian dairy products against influenza infection in mice: alleviation of influenza infection by its immunomodulatory activity through intestinal immunity

Some probiotics possess immunomodulatory activities and have been used as complementary and alternative medicines. We previously found that 10 lactic acid bacteria (LAB) strains isolated from traditional Mongolian dairy products showed probiotic potential in vitro. In this study, we assessed the immunomodulatory activity of 10 LABs on influenza virus (IFV) infection in relation to their efficacies in IFV-infected mice. In an intranasal IFV infection model in mice, oral administration of boiled Lactobacillus plantarum 06CC2 strain (20 mg/mouse), one of the 10 LABs, twice daily for 10 days starting two days before infection was significantly effective in protecting the body weight loss of infected mice, reducing virus yields in the lungs on days 2, 4, and 6 after infection, and prolonging survival times without toxicity. The total numbers of infiltrated cells in the bronchoalveolar lavage fluid (BALF), especially macrophages and neutrophils, were significantly reduced by 06CC2 administration on day 2. On day 2, tumor necrosis factor (TNF)-α production in BALF was also reduced significantly, but interferon-α, interleukin-12, and interferon-γ productions were augmented and natural killer (NK) cell activity was significantly elevated. Furthermore, the gene expressions of interleukin-12 receptor and interferon-γ in Peyer's patches were augmented by 06CC2 administration on day 2. Thus, 06CC2 was suggested to alleviate influenza symptoms in mice in correlation with the augmentation of NK cell activity associated with the enhancement of interferon-α and Th1 cytokine productions through intestinal immunity and the reduction of TNF-α in the early stage of infection.