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.

In vitro anti-HIV and -HSV activity and safety of sodium rutin sulfate as a microbicide candidate

Sodium rutin sulfate (SRS) is a sulfated rutin modified from the natural flavonol glycoside rutin. Here, we investigated its in vitro anti-HIV and -HSV activities and its cytotoxic profile. Fifty percent inhibitory concentration (IC50) values of SRS against HIV-1 X4 virus IIIB, HIV-1 R5 isolates Ada-M and Ba-L were 2.3 +/- 0.2, 4.5 +/- 2.0 and 8.5 +/- 3.8 mu M with a selectivity index (SI) of 563, 575 and 329, respectively. Its IC50 against primary R5 HIV-1 isolate from Yunnan province in China was 13.1 +/- 5.5 mu M, with a Sl of 197. In contrast, unsulfated rutin had no activity against any of the HIV-1 isolates tested. Further study indicated that SRS blocked viral entry and virus-cell fusion likely through interacting with the HIV- I envelope glycoprotein. SRS also demonstrated some activity against human herpes simplex virus (HSV) with an IC50 of 88.3 +/- 0.1 mu M and a Sl of 30. The 50% cytotoxicity concentration (CC50) of SRS was >3.0 mM, as determined in human genital ME 180, HeLa and primary human foreskin fibroblast cells. Minimum inhibitory concentration of SRS for vaginal lactobacilli was >3.0 mM. These results collectively indicate that SRS represents a novel candidate for anti-HIV-1/HSV microbicide development. (C) 2007 Elsevier B.V. All rights reserved.

Effects of anti-bursin monoclonal antibody on immunosuppression in the duck (Cherry Valley duck)

To study the immunologic function of bursin, we analyzed the effects of anti-bursin monoclonal antibody (mAb) on the immunosuppression in ducks (Cherry Valley duck) by injecting various doses of the anti-bursin mAb into 13-d duck embryos. After hatch, cell-mediated immune activity and humoral responses were studied using lymphocyte proliferation test, tube agglutination test, and indirect enzyme-linked immuno-sorbent assay to detect anti-Escherichia coli antibodies and antibodies to Riemerella anatipester, respectively. Simultaneously, relative weights (BW-adjusted) of bursa of Fabricius (BF), spleen, and thymus were determined. Additionally, the morphology of BF, spleen, and thymus was examined at various ages using conventional histology. Follicle morphology of BF was analyzed by image analysis. The results indicated that anti-bursin mAb markedly decreased duck lymphocyte proliferation, the antibody-producing ability to bacteria, as well as the relative BF weight. Moreover, the anti-bursin mAb hindered the development of BF follicles.

Weak anti-localization in InAlAs/InGaAs/InAlAs high mobility two-dimensional electron gas systems

Magneto-transport measurements have been carried out on three heavily Si delta-doped In-0.52 Al-0.48 As/In-0.53 Ga-0.47 As/In-0.52 A(10.48) As single quantum well samples in which two subbands were occupied by electrons. The weak anti-localization (WAL) has been found in such high electron mobility systems. The strong Rashba spin-orbit (SO) coupling is due to the high structure inversion asymmetry (SIA) of the quantum wells. Since the WAL theory model is so complicated in fitting our experimental results, we obtained the Rashba SO coupling constant alpha and the zero-field spin splitting Delta(0) by an approximate approach. The results are consistent with that obtained by the Shubnikov-de Haas (SdH) oscillation analysis. The WAL effect in high electron mobility system suggests that finding a useful approach for deducing alpha and Delta(0) is important in designing future spintronics devices that utilize the Rashba SO coupling.

Anti-Stokes photoluminescence in ZnO microcrystal

Low temperature (10 K) strong anti-Stokes photoluminescence (ASPL) of ZnO microcrystal excited by low power cw 532 nm laser is reported here. Energy upconversion of 1.1 eV is obtained in our experiment with no conventional nonlinear effect. Through the study of the normal photoluminescence and temperature dependence of ASPL we conclude that the green band luminescence in ZnO is related to deep donor to valance band transition. Using the two-step two-photon absorption model, we provide a plausible mechanism leading to the ASPL phenomenon in our experiment. (c) 2006 American Institute of Physics.

Preparation and optical property of a novel hydrophobic anti-reflection coating for KDP crystal

A novel inorganic-organic hybrid hydrophobic anti-reflection silica film used for laser crystal was obtained by sol-gel process. The film consisted of silica sols mixed with a small amount of polymethyl methacrylate (PMMA) or polystyrene (PS). The optical transparency, hydrophobic property and surface morphology of the film were characterized by UV-VIS-NIR spectrophotometer; contact angle instrument and Scanning Electron Microscopy (SEM), respectively. The results showed that the anti-reflection coating had good hydrophobility and optical transparency from 400 nm to 1200 nm. The contact angle reached to 130-140 degrees. SEM images indicated the hydrophobic films modified with PMMA or PS had compact structure compared to the pure silica sol film. (C) 2008 Elsevier B.V. All rights reserved.

Recombinant protein of heptad-repeat HR212, a stable fusion inhibitor with potent anti-HIV action in vitro

HR212, a recombinant protein expressed in Escherichia coli, has been previously reported to inhibit HIV-1 membrane fusion at low nanomolar level. Here we report that HR212 is effective in blocking laboratory strain HIV-1IIIB entry and replication with EC50 values of 3.92±0.62 and 6.59±1.74 nM, respectively, and inhibiting infection by clinic isolate HIV-1KM018 with EC50 values of 44.44±10.20 nM, as well as suppressing HIV-1- induced cytopathic effect with an EC50 value of 3.04±1.20 nM. It also inhibited HIV-2ROD and HIV-2CBL-20 entry and replication in the μM range. Notably, HR212 was highly effective against T20-resistant strains with EC50 values ranging from 5.09 to 7.75 nM. Unlike T20, HR212 showed stability sufficient to inhibit syncytia formation in a time-of-addition assay, and was insensitive to proteinase K digestion. These results suggest that HR212 has great potential to be further developed as novel HIV-1 fusion inhibitor for treatment of HIV/ AIDS patients, particularly for those infected by T20-resistant variants.