抗HIV p24多肽单克隆抗体和抗血清的制备及初步应用

检测p2 4抗原的诊断试剂在AIDS研究和防治方面具有重要意义。为研制国产HIVp2 4抗原诊断试剂 ,根据国内外文献 ,合成了HIVp2 4抗原的六个肽段 :G -A 12 ,D -R 16 ,P -S 18,A -G 2 3(HIV - 2ROD) ,P -S 18,N -I 15以及D -C 2 2。六个肽段中 ,N -I 15肽和D -C 2 2肽用于制备McAb ,其余 4个用于制备山羊抗血清。除D -R 16肽的滴度较低 (1∶2 0 0 0 )外 ,其余三个肽的抗血清滴度都在 1∶10 0 0 0以上。McAb中 ,N -I 15肽的反应比D -C 2 2肽要强 ,但两者的腹水滴度相同 ,均为 1∶10 0 0。单抗铺板检测病毒 p2 4抗原的非特异性反应比较强。用山羊抗血清铺板 ,1∶80 0 0的稀释度检测效果最好。用我们研制的抗体检测不出裂解HIV - 1ⅢB中 p2 4 ,却能够测出Abbott公司p2 4抗原诊断试剂盒的阳性对照 (HIVAG - 1)。用北海道大学免疫科学研究所研制的单抗和人阳性血清做的交叉对照实验表明 ,可能是由于合成肽免疫产生的抗体与裂解病毒p2 4的亲和力较差引...

抗HIV药物的体外筛选和研究方法

　合适的体内外药物研究方法是研究和开发新药中最关键的环节之一。由于至今尚无理想的艾滋病 (AIDS) 动物模型,目前抗人免疫缺陷病毒(HIV) 药物的筛选和研究仍然主要依赖于各种体外的筛选和研究方法。 本文简要综述了抗HIV 药物的体外筛选和研究方法。

三个非核苷类逆转录酶抑制剂S-DABO类衍生物的体外抗HIV作用(英文)

本文对3个新S-DABO类衍合物(RZK-4、RZK-5、RZK-6)的体外抗HIV活性进行了研究。化合物RZK-4、RZK-5和RZK-6在200μg·mL-1的浓度下均能完全抑制HIV-1逆转录酶的活性。3个化合物对多种细胞均呈现出低毒性,且均在较低浓度下具有抑制HIV-1病毒实验株、临床株和耐药株的作用,治疗指数为3704～38462。其中,化合物RZK-6对HIV-1耐药株HIV-1IIIBA17具有非常显著的抑制作用。结果表明,这3种S-DABO类衍生物有良好的体外抗HIV-1作用,具有开发成为抗HIV-1药物的前景。

我国中药来源的抗HIV天然化合物研究进展

由于迄今仍无艾滋病(acquired immunodeficiency syndrome,AIDS)疫苗问世,抗人类免疫缺陷病毒(human immunodeficiency virus,HIV)药物仍然是艾滋病治疗的主要手段。传统中药和药用植物来源的天然化合物具有结构多样性、毒性较低、来源广泛等特点,因而在防治艾滋病方面有着独特的优势和巨大的潜力。研究者已经对天然化合物抗HIV作用进行了大量研究,并取得了可喜的成绩,发现了一些生物碱、香豆素、木脂素、黄酮类、萜类、鞣质类、多糖类、蛋白质和多肽类等天然化合物具有抗HIV的活性。然而,多数研究都是在体外试验完成的,大多数天然化合物体外抗HIV活性偏低,而且抗HIV的靶点仍不十分清楚。本文结合笔者实验室研究工作,重点介绍近年来我国传统中药来源的抗HIV活性较强的天然化合物研究进展。

HIV感染中的细胞凋亡

HIV感染以后,病毒蛋白的持续性产出导致免疫系统的持续性激活,引起Th1细胞的丢失,Th1细胞通过合成Ⅰ型细胞因子,抑制淋巴细胞的自发凋亡。另外,病毒蛋白或其他因素能够使CD4~(+)、CD8~(+) T细胞和APC转化为凋亡的效应细胞,通过Fas/FasL或其他途径引起细胞凋亡。HIV感染人体后凋亡细胞不仅有CD4~(+) T细胞,还包括B细胞、NK细胞、粒细胞、神经细胞和单细胞。凋亡作为机体的自我防护措施,在清除感染细胞的同时,并没有抑制HIV在单细胞/巨噬细胞内的复制,反而造成大量未感染细胞的凋亡,导致对HIV复制的失控,发展为严重的免疫缺陷,引起AIDS相关的机会性感染。

Xanthohumol, a novel anti-HIV-1 agent purified from Hops Humulus lupulus

Xanthohumol, prenylchacone flavonoid, is a natural product with multi-biofunctions purified from Hops Humulus lupulus. Its anti-HIV-1 activity was tested in the present study. Results showed that xanthohumol inhibited HIV-1 induced cytopathic effects, the production of viral p24 antigen and reverse transcriptase in C8166 lymphocytes at non-cytotoxic concentration. The EC50 values were 0.82, 1.28 and 0.50 mug/ml, respectively. The therapeutic index (TI) was about 10.8. Xanthohumol also inhibited HIV-1 replication in PBMC with EC50 value of 20.74 mug/ml. The activity of recombinant HIV-1 reverse transcriptase and the HIV-1 entry were not inhibited by xanthohumol. The results from this study suggested that xanthohumol is effective against HIV-1 and might serve as an interesting lead compound. It may represent a novel chemotherapeutic agent for HIV-1 infection. However, the mechanism of its anti-HIV-1 effect needs to be further clarified. (C) 2004 Elsevier B.V. All rights reserved.

The anti-HIV-1 effect of scutellarin

Scutellarin was purified from the plant Erigeron breuiscapus (Vant.) Hand.-Mazz. The activity against 3 strains of human immunodeficiency virus (HIV) was determined in vitro in this study. These were laboratory-derived virus (HIV-I-IIIB), drug-resistant virus (HIV-I-74V), and low-passage clinical isolated virus (HIV-1(KM018)). From syncytia inhibition study, the EC50 of scutellarin against HIV-I-IIB direct infection in C8166 cells was 26 mu M with a therapeutic index of 36. When the mode of infection changed from acute infection to cell-to-cell infection, this compound became even more potent and the EC50 reduced to 15 mu M. This suggested that cell fusion might be affected by this compound. By comparing the inhibitory effects on p24 antigen, scutellarin was also found to be active against HIV-1(74V) (EC50 253 mu M) and HIV-1(KM018) (EC50 136 mu M) infection with significant difference in potency. The mechanism of its action was also explored in this study. At a concentration of 433 mu M, scutellarin inhibited 48% of the cell free recombinant HIV-1 RT activity. It also caused 82% inhibition of HIV-1 particle attachment and 45% inhibition of fusion at the concentrations of 54 mu M. In summary, scutellarin was found to inhibit several strains of HIV-1 replication with different potencies. It appeared to inhibit HIV-1 RT activity, HIV-1 particle attachment and cell fusion. These are essential activities for viral transmission and replication. (c) 2005 Elsevier Inc. All rights reserved.

Structure and anti-HIV activity of micrandilactones B and C, new nortriterpenoids possessing a unique skeleton from Schisandra micrantha

Two new highly oxygenated nortriterpenoids with a unique norcycloartane skeleton, micrandilactones B and C (1-2), were isolated from Schisandra micrantha; micrandilactone C ( 2) exhibited an EC50 value of 7.71 mu g/mL (SI > 25.94) against HIV-1 replication with minimal cytotoxicity, and the potent anti-HIV-1 activity and unique structural features of 2 make it a promising lead for therapeutic development of a new generation of anti-HIV drug.

Sifuvirtide, a potent HIV fusion inhibitor peptide

Enfuvirtide (ENF) is currently the only FDA approved HIV fusion inhibitor in clinical use. Searching for more drugs in this category with higher efficacy and lower toxicity seems to be a logical next step. In line with this objective, a synthetic peptide

Anti-HIV Activities of the Compounds Isolated from Polygonum cuspidatum and Polygonum multiflorum

The 70% EtOH extract of Polygonum cuspidatum showed inhibitory action against HIV-1-induced syncytium formation at non-cytotoxic concentrations in vitro with a 50% effective concentration (EC50) of 13.94 +/- 3.41 mu g/mL. Through bioactivity-guided fractionation, 20 phenolic compounds, including eight stilbenoids, were isolated from the roots of Polygonum cuspidatum, and their anti-HIV-1 activities were evaluated. Results showed that compounds 1, 13, 14, and 16 demonstrated fairly strong antiviral activity against HIV-1-induced cytopathic effects in C8166 lymphocytes at non-cytotoxic concentrations, with EC50 values of 4.37 +/- 1.96 mu g/mL, 19.97 +/- 5.09, 14.4 +/- 1.34 mu g/mL, and 11.29 +/- 6.26 mu g/mL and therapeutic index (TI) values of 8.12, > 10.02, > 13.89, and > 17.71, respectively. Other compounds showed either weak or no effects. Compound 6 also showed weak inhibition (153.42 +/- 19.25 mu g/mL); however, it possesses very good water solubility and showed almost no cytotoxicity (> 2000 mu g/mL), therefore achieving a fairly good TI (13.04). The activities of the two compounds (3 and 18) from Polygonum multiflorum were also assayed. The relationship between molecular structures and their bioactivities was also discussed.

Anti-human Immunodeficiency Virus-1 Constituents of the Bark of Poncirus trifoliata

A total of 36 compounds (1-36) were obtained from the stem bark of Poncirus trifoliata including three new prenylated flavonoids, (-)-5,4'-dihydroxy-7,8-[(3 '',4 ''-cis-dihydroxy-3 '',4 ''-dihydro)-2 '',2 ''-dimethylpyrano]-flavone (1), (-)-5,4'-dihydroxy-7,8-[(3 ''-hydroxy-4 ''-one)-2 '',2 ''-dimethylpyrano]-flavone (2), and (-)-5,4'-dihydroxy-7,8-[(cis-3 ''-hydroxy-4 ''-ethoxy-3 '',4 ''-dihydro)-2 '',2 ''-dimethylpyrano]-flavone (3). The new structures were elucidated by means of spectroscopic methods. Compounds 1-20 were evaluated for their anti-human immunodeficiency virus-1 (HIV-1) activity, in which 2 showed significant anti-HIV-1 activity with high therapeutic index (T1) of 143.65.

Anti Human Immunodeficiency Virus Type 1 (HIV-1) Agents 4. Discovery of 5,5 '-(p-Phenylenebisazo)-8-hydroxyquinoline Sulfonates as New HIV-1 Inhibitors in Vitro

To search for compounds with superior anti-human immunodeficiency virus type 1 (HIV-1) activity, ten 5,5'-(p-phenylenebisazo)-8-hydroxyquinoline sulfonates (4a-j) were synthesized and preliminarily evaluated as HIV-1 inhibitors in vitro for the first time. Some compounds demonstrated anti-HIV-1 activity, especially 5,5'-(p-phenylenebisazo)-8-hydroxyquinoline p-ethylbenzenesulfonate (4g) and 5,5'-(p-phenylenebisazo)-8-hydroxyquinoline p-chlorobenzenesulfonate (41) showed the more potent anti-HIV-1 activity with 50% effective concentration (EC50) values of 2.59 and 4.01 mu g/ml, and therapeutic index (TI) values of 31.77 and 24.51, respectively.

Dibenzocyclooctadiene Lignans from Schisandra wilsoniana and Their Anti-HIV-1 Activities

Twelve new dibenzocyclooctadiene lignans, marlignans A-L (1-12), together with 16 known compounds, were isolated from the leaves and stems of Schisandra wilsoniana. The structures of 1-12 were elucidated by spectroscopic methods including 1D- and 2D-NMR techniques. Compounds 1-12 were evaluated for their anti-HIV activities, of which compounds 3, 6, 8, and 12 showed modest activities with therapeutic index values of 13.2, 15.6, 17.6, and 16.4, respectively.

动物模型在HIV-1杀微生物剂有效性和安全性评价中的应用

HIV/AIDS的流行趋势没有减弱的迹象,人们迫切需要新的预防HIV传播的手段。杀微生物剂旨在通过局部用药于阴道或直肠,从而阻止HIV的传播。鉴于目前有大量的杀微生物剂候选物,亟待能够有效评价其有效性及安全性的动物模型。通过比较非灵长类小型动物模型与非人灵长类动物模型在评价HIV杀微生物剂的有效性及安全性上的重要作用,该文总结了评价杀微生物剂有效性及安全性的动物模型的优缺点,同时指出了杀微生物剂研究与发展的方向和建议,希望能够对杀微生物剂的研发有所帮助。

An improved microtiter assay for evaluating anti-HIV-I neutralizing antibodies from sera or plasma

Background: The anti-HIV-1 neutralizing antibody assay is widely used in AIDS vaccine research and other experimental and clinical studies. The vital dye staining method applied in the detection of anti-HIV-1 neutralizing antibody has been used in many laboratories. However, the unknown factor(s) in sera or plasma affected cell growth and caused protection when the tested sera or plasma was continuously maintained in cell culture. In addition, the poor solubility of neutral red in medium (such as RPMI-1640) also limited the use of this assay. Methods: In this study, human T cell line C8166 was used as host cells, and 3-(4,5-Dimethyl-2-thiazolyl)- 2,5-diphenyl-2H-tetrazolium bromide (MTT) instead of neutral red was used as vital dye. In order to avoid the effect of the unknown factor( s), the tested sera or plasma was removed by a washout procedure after initial 3 - 6 h culture in the assay. Result: This new assay eliminated the effect of the tested sera or plasma on cell growth, improved the reliability of detection of anti-HIV-1 neutralizing antibody, and showed excellent agreement with the p24 antigen method. Conclusion: The results suggest that the improved assay is relatively simple, highly duplicable, cost-effective, and well reliable for evaluating anti-HIV-1 neutralizing antibodies from sera or plasma.