The HR2 polymorphism N140I in the HIV-1 gp41 combined with the HR1 V38A mutation is associated with a less cytopathic phenotype

<p>Abstract</p> <p>Background</p> <p>Resistance to the fusion inhibitor enfuvirtide (ENF) is achieved by changes in the gp41 subunit of the HIV envelope glycoprotein (Env). Specific ENF-associated mutational pathways correlate with immunological recovery, even after virological failure, suggesting that the acquisition of ENF resistance alters gp41 pathogenicity. To test this hypothesis, we have characterized the expression, fusion capability, induction of CD4⁺T cell loss and single CD4⁺T cell death of 48 gp41 proteins derived from three patients displaying different amino acids (N, T or I) at position 140 that developed a V38A mutation after ENF-based treatment.</p> <p>Results</p> <p>In all cases, intra-patient comparison of Env isolated pre- or post-treatment showed comparable values of expression and fusogenic capacity. Furthermore, Env with either N or T at position 140 induced comparable losses of CD4⁺T-cells, irrespective of the residue present at position 38. Conversely, Env acquiring the V38A mutation in a 140I background induced a significantly reduced loss of CD4⁺T cells and lower single-cell death than did their baseline controls. No altered ability to induce single-cell death was observed in the other clones.</p> <p>Conclusions</p> <p>Overall, primary gp41 proteins with both V38A and N140I changes showed a reduced ability to induce single cell death and deplete CD4⁺T cells, despite maintaining fusion activity. The specificity of this phenotype highlights the relevance of the genetic context to the cytopathic capacity of Env and the role of ENF-resistance mutations in modulating viral pathogenicity <it>in vivo</it>, further supporting the hypothesis that gp41 is a critical mediator of HIV pathogenesis.</p

Susceptibility of Human Lymphoid Tissue Cultured ex vivo to Xenotropic Murine Leukemia Virus-Related Virus (XMRV) Infection

Xenotropic murine leukemia virus-related virus (XMRV) was generated after a recombination event between two endogenous murine leukemia viruses during the production of a prostate cancer cell line. Although the associations of the XMRV infection with human diseases appear unlikely, the XMRV is a retrovirus of undefined pathogenic potential, able to replicate in human cells in vitro. Since recent studies using animal models for infection have yielded conflicting results, we set out an ex vivo model for XMRV infection of human tonsillar tissue to determine whether XMRV produced by 22Rv1 cells is able to replicate in human lymphoid organs. Tonsil blocks were infected and infection kinetics and its pathogenic effects were monitored XMRV, though restricted by APOBEC, enters and integrates into the tissue cells. The infection did not result in changes of T or B-cells, immune activation, nor inflammatory chemokines. Infectious viruses could be recovered from supernatants of infected tonsils by reinfecting DERSE XMRV indicator cell line, although these supernatants could not establish a new infection in fresh tonsil culture, indicating that in our model, the viral replication is controlled by innate antiviral restriction factors. Overall, the replication-competent retrovirus XMRV, present in a high number of laboratories, is able to infect human lymphoid tissue and produce infectious viruses, even though they were unable to establish a new infection in fresh tonsillar tissue. Hereby, laboratories working with cell lines producing XMRV should have knowledge and understanding of the potential biological biohazardous risks of this virus

Susceptibility of Human Lymphoid Tissue Cultured ex vivo to Xenotropic Murine Leukemia Virus-Related Virus (XMRV) Infection

BACKGROUND: Xenotropic murine leukemia virus-related virus (XMRV) was generated after a recombination event between two endogenous murine leukemia viruses during the production of a prostate cancer cell line. Although the associations of the XMRV infection with human diseases appear unlikely, the XMRV is a retrovirus of undefined pathogenic potential, able to replicate in human cells in vitro. Since recent studies using animal models for infection have yielded conflicting results, we set out an ex vivo model for XMRV infection of human tonsillar tissue to determine whether XMRV produced by 22Rv1 cells is able to replicate in human lymphoid organs. Tonsil blocks were infected and infection kinetics and its pathogenic effects were monitored RESULTS: XMRV, though restricted by APOBEC, enters and integrates into the tissue cells. The infection did not result in changes of T or B-cells, immune activation, nor inflammatory chemokines. Infectious viruses could be recovered from supernatants of infected tonsils by reinfecting DERSE XMRV indicator cell line, although these supernatants could not establish a new infection in fresh tonsil culture, indicating that in our model, the viral replication is controlled by innate antiviral restriction factors. CONCLUSIONS: Overall, the replication-competent retrovirus XMRV, present in a high number of laboratories, is able to infect human lymphoid tissue and produce infectious viruses, even though they were unable to establish a new infection in fresh tonsillar tissue. Hereby, laboratories working with cell lines producing XMRV should have knowledge and understanding of the potential biological biohazardous risks of this virus

Changes in the HIV-1 env gene: Implications at the RNA and protein structure levels

La glicoproteïna de l’envolta (Env) és una de les proteïnes claus del VIH-1 en patogènesi. La seqüència del gen env és important per a codificar l’Env però també per les estructures secundàries dels RREs que contenen els seus trànscrits. La rellevància dels dos elements s’ha comprovat extensament, tot i que assajos funcionals són requerits en ambdós casos per estudiar l’impacte de canvis específics. El tractament amb l’inhibidor de fusió T-20 selecciona virus resistents després d’adquirir canvis al gen env. Per estudiar variants funcionalment rellevants d’env que es trobessin in vivo vam decidir utilitzar mutacions associades a T-20. Les prediccions de les estructures secundàries dels RRE van resultar alterades en els canvis que codificaven per G36V/D, V38A, Q40H i L45M, però no per N43D o Q40H-L45M. Funcionalment vam demostrar que només els mutants que presenten el canvi L45M a la seva seqüència pateixen un impacte en la capacitat d’unir-se a la proteïna viral Rev quan aquesta es troba en baixes concentracions, suggerint que els canvis nucleotídics que afecten a la formació de l’aminoàcid 45 de gp41 juguen un paper en la unió a Rev. De totes maneres, ninguna variant d’RRE va ser afectada en la seva capacitat per a ser transportada al citoplasma. Per tant, alteracions en les prediccions d’estructures secundàries a partir de seqüències nucleotídiques no necessàriament impliquen impactes funcionals i la mutació L45M no s’incorpora com a mutació secundària degut a un restaurament de les funcions del RRE. Com que és important caracteritzar funcionalment Envs derivades de pacients per a estudiar la seva patogenicitat, vam establir una metodologia completa per a estudiar-les basant-nos en el paper principal que hi juga la subunitat gp41. Els resultats demostren que la selecció de la línia cel·lular efectora és essencial per optimitzar la sensibilitat dels assajos. La línia cel·lular 293T hauria de ser utilitzada per experiments fusogènics i la HeLa per analitzar paràmetres relacionats amb la mort. Estudis clínics havien suggerit que virus que contenien la mutació V38A i que havien aparegut sota un context que tinguessin el polimorfisme viral N140I eren menys patogènics degut a que estaven associats amb fallada virològica i beneficis immunològics. Per tal d’entendre aquestes dades discordants vam basar-nos en la nostra metodologia per a analitzar Envs derivades de pacients que tinguessin aquests canvis. Aquestes Envs V38A-N140I van resultar tenir menys capacitat per induir la mort per cèl·lula individual tot i no veure’s alterada la seva capacitat fusogènica, remarcant la importància del context de l’Env i el paper central de gp41 a la patogènesi del VIH-1.La glicoproteína de la envuelta (Env) es una de les proteínas claves del VIH-1 en patogénesis. La secuencia del gen env es importante para codificar la Env pero también para las estructuras secundarias de los RREs presentes en sus tránscritos. La relevancia de estos dos elementos ha sido comprobada extensamente, pese a que ensayos funcionales aún son requeridos en ambos casos para estudiar el impacto de cambios específicos. El tratamiento con el inhibidor de fusión T-20 selecciona virus resistentes después de adquirir cambios en el gen env. Para estudiar variantes funcionalmente relevantes de env que se encontraran in vivo decidimos utilizar mutaciones asociadas a T-20. Las predicciones de las estructuras secundarias de los RRE resultaron alteradas en los cambios que codificaban para G36V/D, V38A, Q40H y L45M, pero no para N43D o Q40H-L45M. Funcionalmente demostramos que sólo los mutantes que presentan el cambio L45M en su secuencia sufren un impacto de unión a la proteína viral Rev cuando ésta se encuentra en bajas concentraciones, sugiriendo que los cambios nucleotídicos que afectan a la formación del aminoácido 45 de gp41 juegan un papel en la unión a Rev. No obstante, la capacidad de transporte al citoplasma de estos RRE no está afectada. Por lo tanto, alteraciones en las estructuras secundarias predictivas a partir de secuencias nucleotídicas no necesariamente implican impactos funcionales y la mutación L45M tampoco se incorpora como mutación secundaria debido a la restauración de las funciones del RRE. Como es importante caracterizar funcionalmente Envs derivadas de pacientes para estudiar su patogenicidad, establecimos una metodología completa basándonos en el papel principal de la subunidad gp41. Los resultados demostraron que la línea celular efectora es esencial para optimizar la sensibilidad de los ensayos. La línea celular 293T debería utilizarse para experimentos de fusión y la HeLa per analizar parámetros de citopaticidad. Estudios clínicos habían sugerido que virus que contenían la mutación V38A y el polimorfismo viral N140I estaban asociados a una fallo virológico y beneficios inmunológicos del paciente debido a que eran menos patogénicos. Para entender estos datos discordantes analizamos Envs derivadas de pacientes y vimos que cuando éstas tenían los cambios V38A-N140I tenían menos capacidad para matar células diana de forma individual pese a no tener alterada su capacidad fusogénica. Estos datos remarcan la importancia del contexto de la Env y el papel central de gp41 en la patogénesis del VIH-1.The Env glycoprotein is one of the key proteins used by HIV-1 to mediate its pathogenicity. The sequence of the env gene is important to encode the Env glycoprotein but also for the secondary structure of the RRE that harbor its transcripts. The relevance of both elements has been extensively proved, although functional assays were needed in both cases to study the impact of specific changes. Treatment with the fusion inhibitor T-20 in patients infected with HIV-1 select resistant viruses to this drug after acquiring changes in their env gene. Therefore, we decided to use T-20-associated changes in order to functionally study relevant in vivo env variants. Predictions of the RRE secondary structures showed alterations when they were encoding for the changes G36V/D, V38A, Q40H and L45M, but not when harboring N43D and Q40H-L45M. Functional data showed that only the mutants harboring the L45M mutation had impairments in their binding capacity to Rev when this protein was at low concentrations, suggesting that the nucleotide changes affecting the encoding of the amino acid 45 in gp41 plays a role in Rev binding. However, none of the RRE variants were affected in their ability to being transported to the cytoplasm. Thus, it was found that alterations in RRE secondary structures predicted from nucleotide sequences might not necessary imply functional impairments and that the L45M change was not incorporated as a secondary mutation due to a restoration of the RRE functions. As it is important to functionally characterize patient-derived Envs to understand their in vivo pathogenesis, we established a complete methodology to study them by focusing on the main role of the subunit gp41. Based on our results, the correct selection of the effector cell line is essential to optimize the sensitivity of the assays. The 293T cell line might be used for fusogenic experiments and the HeLa for analyzing death parameters. Clinical trials had suggested that V38A viral mutants arising in an Env context containing the N140I polymorphism, were low pathogenic due to they were associated with virological failure and immunologic benefits. In order to understand these in vivo discordant data, we used our complete methodology to analyze patient-derived Envs that harbored these changes. We found that the V38A mutant Envs that were arisen in a N140I context were less able to induce single-cell death to target cells despite not having an altered fusogenic capacity. Thus, these data supports the importance of the Env context and the main role of gp41 in HIV-1 pathogenesis

Colony-Stimulating Factor 1 Receptor Antagonists Sensitize Human Immunodeficiency Virus Type 1-Infected Macrophages to TRAIL-Mediated Killing

Strategies aimed at eliminating persistent viral reservoirs from HIV-1-infected individuals have focused on CD4(+) T-cell reservoirs. However, very little attention has been given to approaches that could promote elimination of tissue macrophage reservoirs. HIV-1 infection of macrophages induces phosphorylation of colony-stimulating factor 1 receptor (CSF-1R), which confers resistance to apoptotic pathways driven by tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), thereby promoting viral persistence. In this study, we assessed whether CSF-1R antagonists (PLX647, PLX3397, and PLX5622) restored apoptotic sensitivity of HIV-1-infected macrophages in vitro. PLX647, PLX3397, and PLX5622 at clinically relevant concentrations blocked the activation of CSF-1R and reduced the viability of infected macrophages, as well as the extent of viral replication. Our data show that strategies targeting monocyte colony-stimulating factor (MCSF) signaling could be used to promote elimination of HIV-1-infected myeloid cells and to contribute to the elimination of persistent viral reservoirs. IMPORTANCE As the HIV/AIDS research field explores approaches to eliminate HIV-1 in individuals on suppressive antiviral therapy, those approaches will need to eliminate both CD4(+) T-cell and myeloid cell reservoirs. Most of the attention has focused on CD4(+) T-cell reservoirs, and scant attention has been paid to myeloid cell reservoirs. The distinct nature of the infection in myeloid cells versus CD4(+) T cells will likely dictate different approaches in order to achieve their elimination. For CD4(+) T cells, most strategies focus on promoting virus reactivation to promote immune-mediated clearance and/or elimination by viral cytopathicity. Macrophages resist viral cytopathic effects and CD8(+) T-cell killing. Therefore, we have explored clearance strategies that render macrophages sensitive to viral cytopathicity. This research helps inform the design of strategies to promote clearance of the macrophage reservoir in infected individuals on suppressive antiviral therapy

Changes in the HIV-1 env gene : implications at the RNA and protein structure levels /

La glicoproteïna de l'envolta (Env) és una de les proteïnes claus del VIH-1 en patogènesi. La seqüència del gen env és important per a codificar l'Env però també per les estructures secundàries dels RREs que contenen els seus trànscrits. La rellevància dels dos elements s'ha comprovat extensament, tot i que assajos funcionals són requerits en ambdós casos per estudiar l'impacte de canvis específics. El tractament amb l'inhibidor de fusió T-20 selecciona virus resistents després d'adquirir canvis al gen env. Per estudiar variants funcionalment rellevants d'env que es trobessin in vivo vam decidir utilitzar mutacions associades a T-20. Les prediccions de les estructures secundàries dels RRE van resultar alterades en els canvis que codificaven per G36V/D, V38A, Q40H i L45M, però no per N43D o Q40H-L45M. Funcionalment vam demostrar que només els mutants que presenten el canvi L45M a la seva seqüència pateixen un impacte en la capacitat d'unir-se a la proteïna viral Rev quan aquesta es troba en baixes concentracions, suggerint que els canvis nucleotídics que afecten a la formació de l'aminoàcid 45 de gp41 juguen un paper en la unió a Rev. De totes maneres, ninguna variant d'RRE va ser afectada en la seva capacitat per a ser transportada al citoplasma. Per tant, alteracions en les prediccions d'estructures secundàries a partir de seqüències nucleotídiques no necessàriament impliquen impactes funcionals i la mutació L45M no s'incorpora com a mutació secundària degut a un restaurament de les funcions del RRE. Com que és important caracteritzar funcionalment Envs derivades de pacients per a estudiar la seva patogenicitat, vam establir una metodologia completa per a estudiar-les basant-nos en el paper principal que hi juga la subunitat gp41. Els resultats demostren que la selecció de la línia cel·lular efectora és essencial per optimitzar la sensibilitat dels assajos. La línia cel·lular 293T hauria de ser utilitzada per experiments fusogènics i la HeLa per analitzar paràmetres relacionats amb la mort. Estudis clínics havien suggerit que virus que contenien la mutació V38A i que havien aparegut sota un context que tinguessin el polimorfisme viral N140I eren menys patogènics degut a que estaven associats amb fallada virològica i beneficis immunològics. Per tal d'entendre aquestes dades discordants vam basar-nos en la nostra metodologia per a analitzar Envs derivades de pacients que tinguessin aquests canvis. Aquestes Envs V38A-N140I van resultar tenir menys capacitat per induir la mort per cèl·lula individual tot i no veure's alterada la seva capacitat fusogènica, remarcant la importància del context de l'Env i el paper central de gp41 a la patogènesi del VIH-1.La glicoproteína de la envuelta (Env) es una de les proteínas claves del VIH-1 en patogénesis. La secuencia del gen env es importante para codificar la Env pero también para las estructuras secundarias de los RREs presentes en sus tránscritos. La relevancia de estos dos elementos ha sido comprobada extensamente, pese a que ensayos funcionales aún son requeridos en ambos casos para estudiar el impacto de cambios específicos. El tratamiento con el inhibidor de fusión T-20 selecciona virus resistentes después de adquirir cambios en el gen env. Para estudiar variantes funcionalmente relevantes de env que se encontraran in vivo decidimos utilizar mutaciones asociadas a T-20. Las predicciones de las estructuras secundarias de los RRE resultaron alteradas en los cambios que codificaban para G36V/D, V38A, Q40H y L45M, pero no para N43D o Q40H-L45M. Funcionalmente demostramos que sólo los mutantes que presentan el cambio L45M en su secuencia sufren un impacto de unión a la proteína viral Rev cuando ésta se encuentra en bajas concentraciones, sugiriendo que los cambios nucleotídicos que afectan a la formación del aminoácido 45 de gp41 juegan un papel en la unión a Rev. No obstante, la capacidad de transporte al citoplasma de estos RRE no está afectada. Por lo tanto, alteraciones en las estructuras secundarias predictivas a partir de secuencias nucleotídicas no necesariamente implican impactos funcionales y la mutación L45M tampoco se incorpora como mutación secundaria debido a la restauración de las funciones del RRE. Como es importante caracterizar funcionalmente Envs derivadas de pacientes para estudiar su patogenicidad, establecimos una metodología completa basándonos en el papel principal de la subunidad gp41. Los resultados demostraron que la línea celular efectora es esencial para optimizar la sensibilidad de los ensayos. La línea celular 293T debería utilizarse para experimentos de fusión y la HeLa per analizar parámetros de citopaticidad. Estudios clínicos habían sugerido que virus que contenían la mutación V38A y el polimorfismo viral N140I estaban asociados a una fallo virológico y beneficios inmunológicos del paciente debido a que eran menos patogénicos. Para entender estos datos discordantes analizamos Envs derivadas de pacientes y vimos que cuando éstas tenían los cambios V38A-N140I tenían menos capacidad para matar células diana de forma individual pese a no tener alterada su capacidad fusogénica. Estos datos remarcan la importancia del contexto de la Env y el papel central de gp41 en la patogénesis del VIH-1.The Env glycoprotein is one of the key proteins used by HIV-1 to mediate its pathogenicity. The sequence of the env gene is important to encode the Env glycoprotein but also for the secondary structure of the RRE that harbor its transcripts. The relevance of both elements has been extensively proved, although functional assays were needed in both cases to study the impact of specific changes. Treatment with the fusion inhibitor T-20 in patients infected with HIV-1 select resistant viruses to this drug after acquiring changes in their env gene. Therefore, we decided to use T-20-associated changes in order to functionally study relevant in vivo env variants. Predictions of the RRE secondary structures showed alterations when they were encoding for the changes G36V/D, V38A, Q40H and L45M, but not when harboring N43D and Q40H-L45M. Functional data showed that only the mutants harboring the L45M mutation had impairments in their binding capacity to Rev when this protein was at low concentrations, suggesting that the nucleotide changes affecting the encoding of the amino acid 45 in gp41 plays a role in Rev binding. However, none of the RRE variants were affected in their ability to being transported to the cytoplasm. Thus, it was found that alterations in RRE secondary structures predicted from nucleotide sequences might not necessary imply functional impairments and that the L45M change was not incorporated as a secondary mutation due to a restoration of the RRE functions. As it is important to functionally characterize patient-derived Envs to understand their in vivo pathogenesis, we established a complete methodology to study them by focusing on the main role of the subunit gp41. Based on our results, the correct selection of the effector cell line is essential to optimize the sensitivity of the assays. The 293T cell line might be used for fusogenic experiments and the HeLa for analyzing death parameters. Clinical trials had suggested that V38A viral mutants arising in an Env context containing the N140I polymorphism, were low pathogenic due to they were associated with virological failure and immunologic benefits. In order to understand these in vivo discordant data, we used our complete methodology to analyze patient-derived Envs that harbored these changes. We found that the V38A mutant Envs that were arisen in a N140I context were less able to induce single-cell death to target cells despite not having an altered fusogenic capacity. Thus, these data supports the importance of the Env context and the main role of gp41 in HIV-1 pathogenesis

Nef Is Dispensable for Resistance of Simian Immunodeficiency Virus-Infected Macrophages to CD8 +

Simian immunodeficiency virus (SIV)-specific CD8(+) T cells kill SIV-infected CD4(+) T cells in an major histocompatibility complex class I (MHC-I)-dependent manner. However, they are reportedly less efficient at killing SIV-infected macrophages. Since the viral accessory protein Nef has been shown to downregulate MHC-I molecules and enhance cytotoxic T lymphocyte (CTL) evasion in human immunodeficiency virus type 1 (HIV-1)-infected CD4(+) T cells, we examined whether Nef played a role in protecting SIV-infected macrophages from killing by SIV-specific CD8(+) T cells. To explore the role of Nef in CD8(+) T cell evasion, we compared the ability of freshly sorted SIV-specific CD8(+) T cells to readily suppress viral replication or eliminate CD4(+) T cells or monocyte-derived macrophages infected with SIV variants containing wild-type (WT) or mutated nef genes. As expected, SIV-specific CD8(+) T cells suppressed viral replication and eliminated the majority of SIV-infected CD4(+) T cells, and this killing was enhanced in CD4(+) T cells infected with the nef variants. However, macrophages infected with nef variants that disrupt MHC-I downregulation did not promote rapid killing by freshly isolated CD8(+) T cells. These results suggest that mechanisms other than Nef-mediated MHC-I downregulation govern the resistance of SIV-infected macrophages to CD8(+) T cell-mediated killing. This study has implications for viral persistence and suggests that macrophages may afford primate lentiviruses some degree of protection from immune surveillance. IMPORTANCE Myeloid cells are permissive for HIV/SIV replication in vitro and may contribute to viral persistence in vivo. While many studies have been geared to understanding how CD8(+) T cells control viral replication in CD4(+) T cells, the role of these cells in controlling viral replication in macrophages is less clear. Primary, unstimulated CD8(+) T cells insignificantly suppress viral replication or eliminate SIV-infected macrophages. Since the viral Nef protein downregulates MHC-I and provides infected cells some degree of protection from CD8(+) T cell-mediated effector functions, we evaluated whether Nef may be contributing to the resistance of macrophages to CD8(+) T cell suppression. Our results suggest that Nef is not involved in protecting infected macrophages from CD8(+) T cell killing and suggest that other mechanisms are involved in macrophage evasion from CD8 surveillance

<i>In vitro</i> Rev-RRE binding assay.

<p>Electro-mobility shift assay (EMSA) in the absence or in the presence of 20 ng, 40 ng or 80 ng of Rev protein with the different RRE RNA variants (RREWT, sRRE40, sRRE45 and RRE40-45). The reaction products were separated in a polyacrylamide gel and the quantification of the gel shifts are displayed.</p

Functional Analyses Reveal Extensive RRE Plasticity in Primary HIV-1 Sequences Selected under Selective Pressure

<div><p>Background</p><p>HIV-1 Rev response element (RRE) is a functional region of viral RNA lying immediately downstream to the junction of gp120 and gp41 in the <i>env</i> coding sequence. The RRE is essential for HIV replication and binds with the Rev protein to facilitate the export of viral mRNA from nucleus to cytoplasm. It has been suggested that changes in the predicted secondary structure of primary RRE sequences impact the function of the RREs; however, functional assays have not yet been performed. The aim of this study was to characterize the genetic, structural and functional variation in the RRE primary sequences selected <i>in vivo</i> by Enfuvirtide pressure.</p><p>Results</p><p>Multiple RRE variants were obtained from viruses isolated from patients who failed an Enfuvirtide-containing regimen. Different alterations were observed in the predicted RRE secondary structures, with the abrogation of the primary Rev binding site in one of the variants. In spite of this, most of the RRE variants were able to bind Rev and promote the cytoplasmic export of the viral mRNAs with equivalent efficiency in a cell-based assay. Only RRE45 and RRE40-45 showed an impaired ability to bind Rev in a gel-shift binding assay. Unexpectedly, this impairment was not reflected in functional capacity when RNA export was evaluated using a reporter assay, or during virus replication in lymphoid cells, suggesting that <i>in vivo</i> the RRE would be highly malleable.</p><p>Conclusions</p><p>The Rev-RRE functionality is unaffected in RRE variants selected in patients failing an ENF-containing regimen. Our data show that the current understanding of the Rev-RRE complex structure does not suffice and fails to rationally predict the function of naturally occurring RRE mutants. Therefore, this data should be taken into account in the development of antiviral agents that target the RRE-Rev complex.</p></div

Novel anti-HIV molecules produced in transgenic BY-2 suspension cultures

International audienc