ABSTRACT
UNLABELLED: Deletion of Gly-720 and Tyr-721 from a highly conserved GYxxØ trafficking signal in the SIVmac239 envelope glycoprotein cytoplasmic domain, producing a virus termed ΔGY, leads to a striking perturbation in pathogenesis in rhesus macaques (Macaca mulatta). Infected macaques develop immune activation and progress to AIDS, but with only limited and transient infection of intestinal CD4(+) T cells and an absence of microbial translocation. Here we evaluated ΔGY in pig-tailed macaques (Macaca nemestrina), a species in which SIVmac239 infection typically leads to increased immune activation and more rapid progression to AIDS than in rhesus macaques. In pig-tailed macaques, ΔGY also replicated acutely to high peak plasma RNA levels identical to those for SIVmac239 and caused only transient infection of CD4(+) T cells in the gut lamina propria and no microbial translocation. However, in marked contrast to rhesus macaques, 19 of 21 pig-tailed macaques controlled ΔGY replication with plasma viral loads of <15 to 50 RNA copies/ml. CD4(+) T cells were preserved in blood and gut for up to 100 weeks with no immune activation or disease progression. Robust antiviral CD4(+) T cell responses were seen, particularly in the gut. Anti-CD8 antibody depletion demonstrated CD8(+) cellular control of viral replication. Two pig-tailed macaques progressed to disease with persisting viremia and possible compensatory mutations in the cytoplasmic tail. These studies demonstrate a marked perturbation in pathogenesis caused by ΔGY's ablation of the GYxxØ trafficking motif and reveal, paradoxically, that viral control is enhanced in a macaque species typically predisposed to more pathogenic manifestations of simian immunodeficiency virus (SIV) infection. IMPORTANCE: The pathogenesis of human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) reflects a balance between viral replication, host innate and adaptive antiviral immune responses, and sustained immune activation that in humans and Asian macaques is associated with persistent viremia, immune escape, and AIDS. Among nonhuman primates, pig-tailed macaques following SIV infection are predisposed to more rapid disease progression than are rhesus macaques. Here, we show that disruption of a conserved tyrosine-based cellular trafficking motif in the viral transmembrane envelope glycoprotein cytoplasmic tail leads in pig-tailed macaques to a unique phenotype in which high levels of acute viral replication are followed by elite control, robust cellular responses in mucosal tissues, and no disease. Paradoxically, control of this virus in rhesus macaques is only partial, and progression to AIDS occurs. This novel model should provide a powerful tool to help identify host-specific determinants for viral control with potential relevance for vaccine development.
Subject(s)
Amino Acid Motifs , CD4-Positive T-Lymphocytes/immunology , Immunity, Mucosal , Macaca nemestrina/virology , Sequence Deletion , Simian Acquired Immunodeficiency Syndrome/immunology , Viral Envelope Proteins/immunology , Animals , CD4-Positive T-Lymphocytes/virology , Disease Progression , Female , Gene Expression , Intestines/immunology , Intestines/virology , Macaca mulatta/virology , Male , Molecular Sequence Data , Mucous Membrane/immunology , Mucous Membrane/virology , Protein Sorting Signals , Protein Structure, Tertiary , Protein Transport , Simian Acquired Immunodeficiency Syndrome/pathology , Simian Acquired Immunodeficiency Syndrome/virology , Simian Immunodeficiency Virus/genetics , Simian Immunodeficiency Virus/immunology , Species Specificity , Viral Envelope Proteins/deficiency , Viral Envelope Proteins/genetics , Viral Load/genetics , Viral Load/immunology , Viremia/immunology , Viremia/pathology , Virus Replication/genetics , Virus Replication/immunologyABSTRACT
Feline leukemia virus (FeLV) is a naturally transmitted gammaretrovirus that infects domestic cats. FeLV-945, the predominant isolate associated with non-T-cell disease in a natural cohort, is a member of FeLV subgroup A but differs in sequence from the FeLV-A prototype, FeLV-A/61E, in the surface glycoprotein (SU) and long terminal repeat (LTR). Substitution of the FeLV-945 LTR into FeLV-A/61E resulted in pathogenesis indistinguishable from that of FeLV-A/61E, namely, thymic lymphoma of T-cell origin. In contrast, substitution of both FeLV-945 LTR and SU into FeLV-A/61E resulted in multicentric lymphoma of non-T-cell origin. These results implicated the FeLV-945 SU as a determinant of pathogenic spectrum. The present study was undertaken to test the hypothesis that FeLV-945 SU can act in the absence of other unique sequence elements of FeLV-945 to determine the disease spectrum. Substitution of FeLV-A/61E SU with that of FeLV-945 altered the clinical presentation and resulted in tumors that demonstrated expression of CD45R in the presence or absence of CD3. Despite the evident expression of CD45R, a typical B-cell marker, T-cell receptor beta (TCRß) gene rearrangement indicated a T-cell origin. Tumor cells were detectable in bone marrow and blood at earlier times during the disease process, and the predominant SU genes from proviruses integrated in tumor DNA carried markers of genetic recombination. The findings demonstrate that FeLV-945 SU alters pathogenesis, although incompletely, in the absence of FeLV-945 LTR. Evidence demonstrates that FeLV-945 SU and LTR are required together to fully recapitulate the distinctive non-T-cell disease outcome seen in the natural cohort.
Subject(s)
Leukemia Virus, Feline/pathogenicity , Lymphoma/pathology , Membrane Glycoproteins/metabolism , Retroviridae Infections/virology , Terminal Repeat Sequences/genetics , Thymus Neoplasms/pathology , Tumor Virus Infections/virology , Amino Acid Sequence , Animals , Blotting, Southern , Cats , DNA, Viral/genetics , Disease Progression , Female , Immunoenzyme Techniques , Leukemia Virus, Feline/physiology , Lymphoma/genetics , Lymphoma/virology , Membrane Glycoproteins/genetics , Molecular Sequence Data , Polymerase Chain Reaction , Retroviridae Infections/metabolism , Retroviridae Infections/pathology , Sequence Homology, Amino Acid , Survival Rate , Thymus Neoplasms/genetics , Thymus Neoplasms/virology , Tumor Virus Infections/metabolism , Tumor Virus Infections/pathology , Viral Envelope Proteins/genetics , Viral Envelope Proteins/metabolismABSTRACT
Dimethyl fumarate (DMF), an antioxidant/anti-inflammatory drug approved for the treatment of multiple sclerosis, induces antioxidant enzymes, in part through transcriptional upregulation. We hypothesized that DMF administration to simian immunodeficiency virus (SIV)-infected rhesus macaques would induce antioxidant enzyme expression and reduce oxidative injury and inflammation throughout the brain. Nine SIV-infected, CD8+-T-lymphocyte-depleted rhesus macaques were studied. Five received oral DMF prior to the SIV infection and through to the necropsy day. Protein expression was analyzed in 11 brain regions, as well as the thymus, liver, and spleen, using Western blot and immunohistochemistry for antioxidant, inflammatory, and neuronal proteins. Additionally, oxidative stress was determined in brain sections using immunohistochemistry (8-OHdG, 3NT) and optical redox imaging of oxidized flavoproteins containing flavin adenine dinucleotide (Fp) and reduced nicotinamide adenine dinucleotide (NADH). The DMF treatment was associated with no changes in virus replication; higher expressions of the antioxidant enzymes NQO1, GPX1, and HO-1 in the brain and PRDX1 and HO-2 in the spleen; lower levels of 8-OHdG and 3NT; a lower optical redox ratio. The DMF treatment was also associated with increased expressions of cell-adhesion molecules (VCAM-1, ICAM-1) and no changes in HLA-DR, CD68, GFAP, NFL, or synaptic proteins. The concordantly increased brain antioxidant enzyme expressions and reduced oxidative stress in DMF-treated SIV-infected macaques suggest that DMF could limit oxidative stress throughout the brain through effective induction of the endogenous antioxidant response. We propose that DMF could potentially induce neuroprotective brain responses in persons living with HIV.
ABSTRACT
Cellular Al accumulation has been shown to alter iron metabolism and induce peroxidative injury. Therefore antioxidants could potentially reduce or prevent peroxidative injury in Al-loaded cells. To test this hypothesis we assessed the effect of the antioxidants N-acetyl cysteine (NAC), catalase, superoxide dismutase (SOD), and tetramethylpiperidine 1-oxyl (TEMPO) in abrogating Al-associated cell toxicity and melonyldialdehyde (MDA) accumulation in mouse hepatocytes. Mouse hepatocytes (MH) were grown in media containing the minimum toxic concentration of Al (100 microg/L as Al-transferrin). All antioxidants protected MH from injury as assessed by cell growth and enzyme leakage into media. The antioxidants did not affect Al uptake by MH, protect MH from lipid peroxidation or decrease the reactive iron content of MH. Although antioxidants protected Al loaded MH from injury the mechanisms of this effect are unknown.
Subject(s)
Aluminum/toxicity , Antioxidants/pharmacology , Hepatocytes/metabolism , Oxidative Stress/drug effects , Animals , Antioxidants/metabolism , Cell Proliferation/drug effects , Hepatocytes/pathology , Iron/metabolism , Lipid Peroxidation/drug effects , MiceABSTRACT
La infiltracion del sistema nervioso central (SNC) por celulas leucemicas es una complicacion que se presenta cada vez con mayor frecuencia. El diagnostico de infiltracion leucemica se confirma por la observacion de celulas malignas en el liquido cefalorraquideo (LCR). No obstante, la evaluacion morfologica de las celulas resulta dificil, ya que estas pierden frecuentemente su morfologia debido a las caracteristicas fisicas del LCR y porque en ocasiones la celulas leucemicas se encuentran confinadas unicamente al parenquima cerebral. Para demonstrar la infiltracion leucemica del SNC se midieron los niveles de ferritina en el LCR mediante una tecnica inmunorradiometrica. Los pacientes con infiltracion del SNC tuvieron niveles mas altos de ferritina en el LCR (11.2 +/- 1.4 ug/l) que los enfermos con leucemia aguda que no presentaban esta complicacion (p = 0.001). Los niveles de ferritina en LCR en el grupo control y en los enfermos con leucemia aguda sin infiltracion leucemica fueron casi identicos 4.4 +/- 0.6 y 4.5 +/- 0.7 ug/l respectivamente.Estos hallazgos sugieren que la dosificacion de ferritina en el LCR puede ser de utilidad en el diagnostico de esta grave complicacion