Envelope Region Research Articles

Cs-GRP78 Recognition Site on Dengue Virus Envelope Protein: in silico perspective

Aim: To understand the binding of the dengue virus (DENV) envelope and the host cell factor, GRP78. Materials & methods: In this study, we simulate the binding of the DENV envelope against GRP78 using structural bioinformatics tools. Results: The sequence similarity of the DENV envelope C3–C30 and C302–C333 regions against the Pep42 cyclic peptide suggest these regions are possible recognition sites for GRP78. C3–C30 has a more similar grand average hydrophobicity index to that of Pep42 and a more negative binding affinity toward GRP78. Conclusion: We predict this region (C3–C30) of the DENV envelope to be the recognition site of GRP78. Further experimental validation will be important to future studies.

Relativistic stellar modeling with perfect fluid core and anisotropic envelope fluid

We investigate the effect of density perturbations and local anisotropy on the stability of stellar matter structures in general relativity using the concept of cracking. Adopting a core-envelope model of a super-dense star, we examine the properties and stability conditions by introducing anisotropic pressure to the envelope region. Furthermore, we propose self-bound compact stars with an anisotropic envelope as a potential progenitor for starquakes. We show how the difference between sound propagation in radial and tangential directions would be used to identify potentially stable regions within a configuration. Due to an increase in the anisotropic parameter, strain energy accumulates in the envelope region and becomes a potential candidate for building-up quake like situation. This stress-energy stored in the envelope region that would be released during a starquake of a self-bound compact star is computed as a function of the magnitude of anisotropy at the core-envelope boundary. Numerical studies for spherically asymmetric compact stars indicate that the stress energy can be as high as $$10^{50}$$ erg if the tangential pressure is slightly more significant than the radial pressure. It is happened to be of the same order as the energy associated with giant $$\gamma $$ -ray bursts. Thus, the present study will be useful for the correlation studies between starquakes and GRBs.

Substitution of gp120 C4 region compensates for V3 loss-of-fitness mutations in HIV-1 CRF01_AE co-receptor switching

ABSTRACT HIV-1 infection is mediated by a viral envelope subsequently binding to CD4 receptor and two main coreceptors, CCR5 (R5) for primary infection and CXCR4 (X4) in chronic infection. Switching from R5 to X4 tropism in HIV-1 infection is associated with increased viral pathogenesis and disease progression. The coreceptor switching is mainly due to variations in the V3 loop, while the mechanism needs to be further elucidated. We systematically studied the determinant for HIV-1 coreceptor switching by substitution of the genes from one R5 and one X4 pseudoviruses. The study results in successfully constructing two panels of chimeric viruses of R5 to X4 forward and X4 to R5 reverse switching. The determinants for tropism switching are the combined substitution of the V3 loop and C4 region of the HIV-1 envelope. The possible mechanism of the tropism switching includes two components, the V3 loop to enable the viral envelope binding to the newly switched coreceptor and the C4 region, to compensate for the loss of fitness caused by deleterious V3 loop mutations to maintain the overall viral viability. The combined C4 and V3 substitution showed at least an eightfold increase in replication activity compared with the pseudovirus with only V3 loop substitution. The site-directed mutations of N425R and S440-I442 with charged amino acids could especially increase viral activity. This study could facilitate HIV-1 phenotype surveillance and select right entry inhibitor, CCR5 or CXCR4 antagonists, for antiviral therapy.

Structure-Based Identification of Natural-Product-Derived Compounds with Potential to Inhibit HIV-1 Entry.

Broadly neutralizing antibodies (bNAbs) are potent in neutralizing a wide range of HIV strains. VRC01 is a CD4-binding-site (CD4-bs) class of bNAbs that binds to the conserved CD4-binding region of HIV-1 envelope (env) protein. Natural products that mimic VRC01 bNAbs by interacting with the conserved CD4-binding regions may serve as a new generation of HIV-1 entry inhibitors by being broadly reactive and potently neutralizing. This study aimed to identify compounds that mimic VRC01 by interacting with the CD4-bs of HIV-1 gp120 and thereby inhibiting viral entry into target cells. Libraries of purchasable natural products were virtually screened against clade A/E recombinant 93TH057 (PDB: 3NGB) and clade B (PDB ID: 3J70) HIV-1 env protein. Protein-ligand interaction profiling from molecular docking and dynamics simulations showed that the compounds had intermolecular hydrogen and hydrophobic interactions with conserved amino acid residues on the CD4-binding site of recombinant clade A/E and clade B HIV-1 gp120. Four potential lead compounds, NP-005114, NP-008297, NP-007422, and NP-007382, were used for cell-based antiviral infectivity inhibition assay using clade B (HXB2) env pseudotype virus (PV). The four compounds inhibited the entry of HIV HXB2 pseudotype viruses into target cells at 50% inhibitory concentrations (IC50) of 15.2 µM (9.7 µg/mL), 10.1 µM (7.5 µg/mL), 16.2 µM (12.7 µg/mL), and 21.6 µM (12.9 µg/mL), respectively. The interaction of these compounds with critical residues of the CD4-binding site of more than one clade of HIV gp120 and inhibition of HIV-1 entry into the target cell demonstrate the possibility of a new class of HIV entry inhibitors.

Измерение расстояния по максимальной частоте интерференционного сигнала при гармонической девиации длины волны лазерного автодина

A method is proposed for measuring absolute distances from the low-frequency spectrum of the interference signal of a frequency-modulated laser diode. The method of modulation of the self-mixing laser signal according to the harmonic law is used, in the spectrum of which a wide range of frequency components is observed. The connection between the maximum frequency of change of the interference signal and the absolute distance to the reflector is shown. A linear dependence of the frequency corresponding to the inflection region of the decay envelope of the interference signal spectrum on the distance is shown, which can be used to implement a non-contact distance measurement method with harmonic modulation of the self-mixing laser supply current.

Measurement of distance by the maximum frequency of the interference signal with harmonic deviation of the wavelength of the self-mixing laser

A method is proposed for measuring absolute distances from the low-frequency spectrum of the interference signal of a frequency-modulated laser diode. The method of modulation of the self-mixing laser signal according to the harmonic law is used, in the spectrum of which a wide range of frequency components is observed. The connection between the maximum frequency of change of the interference signal and the absolute distance to the reflector is shown. A linear dependence of the frequency corresponding to the inflection region of the decay envelope of the interference signal spectrum on the distance is shown, which can be used to implement a non-contact distance measurement method with harmonic modulation of the self-mixing laser supply current. Keywords: laser interferometry, self-mixing laser, semiconductor laser, laser radiation modulation, distance measurement, signal spectral analysis.

A generalised double layered model with polytropic and quadratic equations of state

This study generates a new exact model for a massive stellar spheres by utilising the Einstein–Maxwell field equations. The model has two interior regions with distinct equations of state (EoS): the polytropic EoS at the core region, and quadratic EoS at the envelope region. Variation of the physical features in the interior of star affirms that the matter variables, gravitational potentials and other physical quantities are consistent with massive stellar bodies. Interestingly, our model can regain solutions of the two-layered model generated by Mardan, Noureen and Khalid. Matching of the layers in junctions is done with help of the Reissner–Nordstrom exterior spacetime. Using our model, stellar masses, and radii consistent with observations and well known stars are generated.

Investigating Anomalous Photochemistry in the Inner Wind of IRC+10216 through Interferometric Observations of HC3N

In recent years, many questions have arisen regarding the chemistry of photochemical products in the carbon-rich winds of evolved stars. To address them, it is imperative to constrain the distributions of such species through high-angular-resolution interferometric observations covering multiple rotational transitions. We used archival Atacama Large Millimeter/submillimeter Array observations to map rotational lines involving high energy levels of cyanoacetylene (HC3N) toward the inner envelope (radius < 8″/1000 au) of the carbon star IRC+10216. The observed lines include the J = 28 − 27, J = 30 − 29, and J = 38 − 37 transitions of HC3N in its ground vibrational state. In contrast to previous observations of linear carbon chains toward this asymptotic giant branch (AGB) star that show extended, hollow emission at 15″–20″ radii (e.g., C4H, C6H, and HC5N), the maps of the HC3N lines here show compact morphologies comprising various arcs and density enhancements, with significant emission from gas clumps at an angular distance of ∼3″ (350 au) from the central AGB star. We compared visibility sampled non-LTE radiative transfer models with the observed brightness distributions, and derive a fractional abundance with respect to H2 of 10−8 for HC3N at the radii probed by these lines. These results are consistent with enhanced photochemistry occurring in warm (∼200 K) regions of the circumstellar envelope. After application of a specialized chemical model for IRC+10216, we find evidence that the enhanced HC3N abundances in the inner wind are most likely due to a solar-type binary companion initiating photochemistry in this region.

Membrane-bound mRNA immunogens lower the threshold to activate HIV Env V2 apex-directed broadly neutralizing B cell precursors in humanized mice.

Eliciting broadly neutralizing antibodies (bnAbs) is the core of HIV vaccine design. bnAbs specific to the V2-apex region of the HIV envelope acquire breadth and potency with modest somatic hypermutation, making them attractive vaccination targets. To evaluate Apex germline-targeting (ApexGT) vaccine candidates, we engineered knockin (KI) mouse models expressing the germline B cell receptor (BCR) of the bnAb PCT64. We found that high affinity of the ApexGT immunogen for PCT64-germline BCRs was necessary to specifically activate KI B cells at human physiological frequencies, recruit them to germinal centers, and select for mature bnAb mutations. Relative to protein, mRNA-encoded membrane-bound ApexGT immunization significantly increased activation and recruitment of PCT64 precursors to germinal centers and lowered their affinity threshold. We have thus developed additional models for HIV vaccine research, validated ApexGT immunogens for priming V2-apex bnAb precursors, and identified mRNA-LNP as a suitable approach to substantially improve the B cell response.

Anisotropic model of super dense star with linearized core and Van der Waals envelope

In this research, core envelope model of a super dense spherically symmetric compact star is developed by considering anisotropic matter configuration. The core is represented by a linear equation of state (EOS), whereas the Van der Waals EOS is used in the envelope region. In the core and envelope of the star, all geometrical and physical factors are viable. The three regions, i.e. the core, envelope and outer space satisfy the junction conditions. The proposed model validates with the properties of Vela X-1, Her X-1 and SMC X-1. It is concluded that in the model presented, the core of the star compresses as the mass increases justifying the domination of gravitational effects on massive astronomical objects.

Relationship between clinical and laboratory features with infecting dengue virus serotypes in a sample of dengue suspected adult patients from 2015-2017 in Sri Lanka

Dengue is a major viral disease affecting the tropics. Although previous research has focused on the relationship between the infecting dengue virus (DENV) serotypes and disease severity, less work has been done on the relationship between the clinical and laboratory features and the infecting DENV serotypes in Sri Lanka. We evaluated the relationship between the clinical and laboratory features and the infecting DENV serotypes in adult patients with clinically suspected dengue admitted to the Base Hospital, Mawanella, Sri Lanka from December 2015 to March 2017.Blood samples of 200 dengue suspected patients were tested for the infecting DENV serotypes using a reverse transcription polymerase chain reaction with primers targeting the envelope region of the virus. Relationship between the infecting DENV serotypes with clinical and laboratory features was assessed using Z score and paired t tests.Of the 200 patients tested, 39 (19.5%) were positive for DENV, any of the four DENV serotypes alone or in combination. The highest number of infections was noted with DENV-2 (n=18, 46.1%). Fever (P=0.000) and rash (P=0.017) were frequently noted in DENV negative patients while bleeding (P=0.012) was more frequently noted in DENV serotype positive patients. Platelet count of <100,000 μl−1 was significantly associated with DENV serotype positivity (P=0.000). Platelet count of <100,000 μl−1 (P=0.035) and haemoglobin (Hb) of >13mgdl−1 (P=0.016) were noted in 15 of the 18 DENV-2 positive patients.Clinical and laboratory features of severe dengue with bleeding manifestations, low platelet counts and high Hb were noted in DENV-2 infections.

Glycyrrhizic Acid Derivatives Bearing Amino Acid Residues in the Carbohydrate Part as Dengue Virus E Protein Inhibitors: Synthesis and Antiviral Activity.

Dengue virus (DENV) is one of the most geographically distributed mosquito-borne flaviviruses, like Japanese encephalitis virus (JEV), and Zika virus (ZIKV). In this study, a library of the known and novel Glycyrrhizic acid (GL) derivatives bearing amino acid residues or their methyl/ethyl esters in the carbohydrate part were synthesized and studied as DENV inhibitors in vitro using the cytopathic effect (CPE), viral infectivity and virus yield assays with DENV1 and DENV-2 in Vero E6 and A549 cells. Among the GL conjugates tested, compound hits GL-D-ValOMe 3, GL-TyrOMe 6, GL-PheOEt 11, and GL-LysOMe 21 were discovered to have better antiviral activity than GL, with IC50 values ranging from <0.1 to 5.98 μM on the in vitro infectivity of DENV1 and DENV2 in Vero E6 and A549 cells. Compound hits 3, 6, 11, and 21 had a concentration-dependent inhibition on the virus yield in Vero E6, in which GL-D-ValOMe 3 and GL-PheOEt 11 were the most active inhibitors of DENV2 yield. Meanwhile, the time-of-addition assay indicated that conjugates GL-D-ValOMe 3 and GL-PheOEt 11 exhibited a substantial decrease in the DENV2 attachment stage. Subsequently, chimeric single-round infectious particles (SRIPs) of DENV2 C-prM-E protein/JEV replicon and DENV2 prM-E/ZIKV replicon were utilized for the DENV envelope I protein-mediated attachment assay. GL conjugates 3 and 11 significantly reduced the attachment of chimeric DENV2 C-prM-E/JEV and DENV2 prM-E/ZIKV SRIPs onto Vero E6 cells in a concentration-dependent manner but did not impede the attachment of wild-type JEV CprME/JEV and ZIKV prM-E/ZIKV SRIPs, indicating the inhibition of Compounds 3 and 11 on DENV2 E-mediated attachment. Molecular docking data revealed that Compounds 3 and 11 have hydrophobic interactions within a hydrophobic pocket among the interfaces of Domains I, II, and the stem region of the DENV2 envelope (E) protein. These results displayed that Compounds 3 and 11 were the lead compounds targeting the DENV E protein. Altogether, our findings provide new insights into the structure–activity relationship of GL derivatives conjugated with amino acid residues and can be the new fundamental basis for the search and development of novel flavivirus inhibitors based on natural compounds.

Dependence on a variable residue limits the breadth of an HIV MPER neutralizing antibody, despite convergent evolution with broadly neutralizing antibodies

Broadly neutralizing antibodies (bNAbs) that target the membrane-proximal external region (MPER) of HIV gp41 envelope, such as 4E10, VRC42.01 and PGZL1, can neutralize >80% of viruses. These three MPER-directed monoclonal antibodies share germline antibody genes (IGHV1-69 and IGKV3-20) and form a bNAb epitope class. Furthermore, convergent evolution within these two lineages towards a 111.2GW111.3 motif in the CDRH3 is known to enhance neutralization potency. We have previously isolated an MPER neutralizing antibody, CAP206-CH12, that uses these same germline heavy and light chain genes but lacks breadth (neutralizing only 6% of heterologous viruses). Longitudinal sequencing of the CAP206-CH12 lineage over three years revealed similar convergent evolution towards 111.2GW111.3 among some lineage members. Mutagenesis of CAP206-CH12 from 111.2GL111.3 to 111.2GW111.3 and the introduction of the double GWGW motif into CAP206-CH12 modestly improved neutralization potency (2.5–3-fold) but did not reach the levels of potency of VRC42.01, 4E10 or PGZL1. To explore the lack of potency/breadth, viral mutagenesis was performed to map the CAP206-CH12 epitope. This indicated that CAP206-CH12 is dependent on D674, a highly variable residue at the solvent-exposed elbow of MPER. In contrast, VRC42.01, PGZL1 and 4E10 were dependent on highly conserved residues (W672, F673, T676, and W680) facing the hydrophobic patch of the MPER. Therefore, while CAP206-CH12, VRC42.01, PGZL1 and 4E10 share germline genes and show some evidence of convergent evolution, their dependence on different amino acids, which impacts orientation of binding to the MPER, result in differences in breadth and potency. These data have implications for the design of HIV vaccines directed at the MPER epitope.

Design, Analysis and Experimental Investigations of a Double-Arm Based Micro-Gripper for Thin and Flexible Metal Wires Manipulation.

A robotic system for the automatic wire pulling of coreless motor winding is designed, including the design of an opening-closing control system and a micro-gripper’s tip structure with a double-armed elastic-beam structure for the support part and an enveloping clamping structure for the tip part. The micro-gripper captures the electrode wire from the root, encircles the wire after the envelope region is closed, and the thin and flexible electrode wire is pulled to the top of the electrode pad by the movement of the micro-gripper and released. The mechanical index of the micro-gripper is simulated to obtain the optimal structural parameters. The experimental results show that the electrode wire’s maximum bearing force is about 0.3 N. Under this reaction force, the deformation of the tip-envelope region of the micro-gripper is about 27.5 μm, which is sufficient for electrode wire pulling micro-manipulation. By comparison with the steel micro-gripper, the silicon micro-gripper has more advantages in shape integrity, machinability and mechanical properties.

Immunogenicity of a germline-targeting nanoparticle in knock-in mice expressing human B cell receptors of the HIV gp41 neutralizing antibody, DH511

Abstract The Membrane Proximal External Region (MPER) of HIV Envelope represents a key target for vaccine development due to high neutralization breadth and potency of MPER-specific broadly neutralizing antibodies (bnAbs). However, neutralizing antibody responses to MPER epitopes are restricted by tolerance control and the MPER epitope is absent from many HIV immunogens under clinical investigation. Using computational design and yeast display, a candidate germline-targeting (GT5) immunogen was developed that bound strongly to the inferred human unmutated common ancestor (UCA) of the distal MPER bnAb DH511, as well as to several human DH511-like potential precursor antibodies. We studied the immunogenicity of a multimeric nanoparticle of the GT5 immunogen in a knock-in mouse line expressing human DH511.UCA B cell receptors (BCRs). Naïve DH511.UCA knock-in mice exhibited a reduction in overall B cell numbers, and DH511.UCA-bearing B cells expressed low levels of surface IgM and IgD, suggesting that DH511.UCA expression is subject to immune tolerance control. Nonetheless, following immunization with GT5 nanoparticles mixed with a saponin/monophosphoryl lipid A adjuvant, knock-in mice mounted robust anti-GT5 humoral responses, including anti-GT5 IgG in serum and GT5-specific germinal center B cells and T follicular helper cells in lymphoid tissues. Sequencing analysis of IgG+ GT5-specific B cells revealed improbable mutations in knock-in immunoglobulin genes of DH511.UCA. These studies will guide further optimization of immunogens with potential to select for development of bnAbs against MPER epitopes. Supported by a grant from NIH (P01-AI138211)

Identification of Major SARS CoV-2 Variants in India using in silico PCR-RFLP analysis

PurposeAs of June 30, 2021, WHO reported 181,344,224 confirmed cases worldwide and 3,934,252 deaths due to SARS CoV-2. India accounts almost 10% of the total mortality. The mutated Variants of Concern (VoC) and Variants of Interest (VoI) has acquired non-synonymous amino acid substitutions in Spike (S), ORF1a, ORF1b, Nucleocapsid (N), Membrane (M), Envelope (E), ORF6, ORF7a, ORF3a and ORF8 regions exhibiting more virulence and higher transmission rate. The S gene displaying nucleotide polymorphisms are studied for identification of major SARS CoV-2 variants circulating in India using in silico PCR-RFLP analysis.Methods & MaterialsDNA sequences of major SARS CoV-2 variants [alpha (B.1.1.7), beta (B.1.351), delta (B.1.617.2), epsilon (B.1.429+B.1.427), eta (B.1.525) and zeta (P.2)] prevailing in India retrieved from GISAID database were annotated using VIPR-VIGOR4 genome annotator tool. The S gene of variants were aligned using CLUSTALW algorithm in MEGA-X with default parameters and analyzed for sequence identity using Geneious Prime v2019.2.1. Primers were designed using Primer-BLAST tool, proof-read using FastPCR v6.7.46 and PCR-amplified using Snapgene v.3.2.1. Unique restriction sites in S amplicons of each variant were subjected to online Restriction Analyzer tool. The S amplicons were digested with restriction endonucleases and the band profile of each variant were visualized in the gel simulation tool using Snapgene v.3.2.1.ResultsThe percentage of identical sites present in S region among SARS CoV-2 variants was found to be 98.6%. The amplified products were in the length of 3,689 and 3,698 bp. Out of 400 restriction endonucleases identified, 14 buffer-compatible enzymes were selected for single-step restriction digestion to generate unique RFLP profiles for each variant. The BsaXI-XcmI-AcuI triple digest showed unique banding pattern identifiers on 2% agarose gel for individual variants including alpha, eta, delta and zeta. The ApaLI-BsaI double digest produced distinct band profiles for beta and epsilon variants.ConclusionOur study strongly suggests PCR-RFLP analysis of Spike region can differentiate major SARS CoV-2 variants that are circulating in India. Further, quadruple digestion based wet lab experiments are underway to explore the possibility of surveillance of the major variants using a single-tube reaction followed by agarose gel-based profiling.

Heat Transfer and Residence Time in Lean Direct Injection Fuel Galleries

Abstract In radially staged lean direct injection (LDI) systems, pilot fuel plays an important role in cooling the mains fuel gallery in regions of the flight envelope where the mains fuel is stagnant. Under these conditions, managing wetted wall temperatures is vital to avoid the formation of carbonaceous particles, which become deposited on the surfaces of the fuel gallery and can lead to a deterioration in system performance. The prediction of wetted wall temperatures therefore represents an important aspect of the injector design phase. Such predictions are often based on injector thermal models, which tend to rely on the application of convective boundary conditions from empirical heat transfer correlations. The use of these correlations leads to questions over the accuracy of predicted wetted wall temperatures and therefore uncertainty over the probability of deposition. This paper seeks to improve on the current situation by applying the inverse conduction technique to determine heat transfer coefficients (HTCs) specific to the pilot fuel gallery. These HTCs are crucial for determining wetted wall temperatures in the pilot and mains fuel galleries and principally govern the risk of deposition in the stagnant mains. The pilot heat transfer data are further examined alongside measurements of the pilot residence time distribution, which together control the risk of pilot deposition at low fuel flow rates. Both the heat transfer and residence time measurements demonstrate the opportunity for future fuel gallery design refinement and provide the supporting data to facilitate this.

Differential V2-directed antibody responses in non-human primates infected with SHIVs or immunized with diverse HIV vaccines

V2p and V2i antibodies (Abs) that are specific for epitopes in the V1V2 region of the HIV gp120 envelope (Env) do not effectively neutralize HIV but mediate Fc-dependent anti-viral activities that have been correlated with protection from, or control of HIV, SIV and SHIV infections. Here, we describe a novel molecular toolbox that allows the discrimination of antigenically and functionally distinct polyclonal V2 Ab responses. We identify different patterns of V2 Ab induction by SHIV infection and three separate vaccine regimens that aid in fine-tuning an optimized immunization protocol for inducing V2p and V2i Abs. We observe no, or weak and sporadic V2p and V2i Abs in non-vaccinated SHIV-infected NHPs, but strong V2p and/or V2i Ab responses after immunization with a V2-targeting vaccine protocol. The V2-focused vaccination is superior to both natural infection and to immunization with whole Env constructs for inducing functional V2p- and V2i-specific responses. Strikingly, levels of V2-directed Abs correlate inversely with Abs specific for peptides of V3 and C5. These data demonstrate that a V1V2-targeting vaccine has advantages over the imprecise targeting of SIV/SHIV infections and of whole Env-based immunization regimens for inducing a more focused functional V2p- and V2i-specific Ab response.

Position Effect Variegation: Role of the Local Chromatin Context in Gene Expression Regulation

Position effect variegation (PEV) is a phenomenon wherein the expression level of a gene strongly depends on its genomic position. PEV can be observed when a gene is moved via a chromosome rearrangement or identical genetic constructs are inserted into different regions of the genome. The eukaryotic genome has a domain organization, and gene activity within a domain depends not only on the nucleotide sequence of a gene, but also on the state of surrounding chromatin, thus being regulated epigenetically. Chromatin is a complex of DNA, RNA, and associated structural and regulatory proteins. The epigenetic status of chromatin depends on the replication time of a given genomic region, particular regulatory DNA motifs, and contacts with the inner nuclear envelope (lamina) and other chromosome regions (topologically associated domains). PEV results from the changes in the epigenetic state of a gene and provides a unique tool to study the molecular and biochemical processes that underlie the establishment and switching of epigenetic states. Understanding the molecular mechanisms of PEV in human is of clinical importance, in particular, for the detection and treatment of retroviral infections because the local chromatin state may determine the latent/active state transition of an infection, such as HIV. In addition, a large number of human neurodegenerative diseases are caused by epigenetic gene inactivation due to expansion of short repeats. Finally, to apply gene therapy methods, it is important to develop approaches that ensure a necessary level of transgene expression with sufficient accuracy.

A recombinant adenoviral vector with a specific tropism to CD4-positive cells: a new tool for HIV-1 inhibition.

Gene therapy can be an option to overcome the side effects of chemotherapy and prevent the development of drug-resistant HIV viruses in HIV-infected patients. The need to develop a safe and efficient vector for gene transfer is always necessary and an appropriate option might be adenovirus (Ad). The use of Ad vectors in gene delivery applications is limited due to the semi-specific tropism. A strategy to overcome this tropism limitation may be the modification of the fiber protein domain involved in the viral binding to cells. Therefore, we designed an Ad5 vector with a specific tropism to CD4 + cells containing an expression system limited to HIV-infected cells. We replaced the knob region of Ad5 fiber protein with the extracellular region of the HIV-1 envelope. We also used a specific Tat-inducible promoter to express two anti-HIV-1 shRNAs. Tropism of recombinant Ad5 was assayed by a comparison of the shRNA expression level in CEM and PBMC cells (as CD4 + cells) and HEK293 cells (as CD4 cells). HIV-1 inhibition was assayed by the determination of p24 antigen in the HIV-infected CEM cells transduced with the recombinant Ad5 vector. Our results showed that the shRNA expression was significantly higher in CEM and PBMC cells than HEK293 cells when transduced with recombinant Ad5 vector. This new Ad5 vector also inhibited HIV-1 proliferation in a Tat-inducible manner. Our new recombinant Ad5 vector has a specific tropism to CD4-positive cells that can effectively suppress the HIV-1 replication.