63 research outputs found
P05-14 LB. Analysis of virus swarms to identify vaccine immunogens with improved exposure of epitopes recognized by broadly neutralizing antibodies
P04-47. High-titer autologous and heterologous neutralizing antibody activity in acute SIV-infection
P04-50 LB. Prevalence of cross-reactive HIV-1 neutralizing activity in HIV-1 infected patients with rapid or slow disease progression
Transduction of SIV-Specific TCR Genes into Rhesus Macaque CD8+ T Cells Conveys the Ability to Suppress SIV Replication
The SIV/rhesus macaque model for HIV/AIDS is a powerful system for examining the contribution of T cells in the control of AIDS viruses. To better our understanding of CD8(+) T-cell control of SIV replication in CD4(+) T cells, we asked whether TCRs isolated from rhesus macaque CD8(+) T-cell clones that exhibited varying abilities to suppress SIV replication could convey their suppressive properties to CD8(+) T cells obtained from an uninfected/unvaccinated animal.We transferred SIV-specific TCR genes isolated from rhesus macaque CD8(+) T-cell clones with varying abilities to suppress SIV replication in vitro into CD8(+) T cells obtained from an uninfected animal by retroviral transduction. After sorting and expansion, transduced CD8(+) T-cell lines were obtained that specifically bound their cognate SIV tetramer. These cell lines displayed appropriate effector function and specificity, expressing intracellular IFNγ upon peptide stimulation. Importantly, the SIV suppression properties of the transduced cell lines mirrored those of the original TCR donor clones: cell lines expressing TCRs transferred from highly suppressive clones effectively reduced wild-type SIV replication, while expression of a non-suppressing TCR failed to reduce the spread of virus. However, all TCRs were able to suppress the replication of an SIV mutant that did not downregulate MHC-I, recapitulating the properties of their donor clones.Our results show that antigen-specific SIV suppression can be transferred between allogenic T cells simply by TCR gene transfer. This advance provides a platform for examining the contributions of TCRs versus the intrinsic effector characteristics of T-cell clones in virus suppression. Additionally, this approach can be applied to develop non-human primate models to evaluate adoptive T-cell transfer therapy for AIDS and other diseases
CTL Escape Mediated by Proteasomal Destruction of an HIV-1 Cryptic Epitope
Cytotoxic CD8+ T cells (CTLs) play a critical role in controlling viral
infections. HIV-infected individuals develop CTL responses against epitopes
derived from viral proteins, but also against cryptic epitopes encoded by viral
alternative reading frames (ARF). We studied here the mechanisms of HIV-1 escape
from CTLs targeting one such cryptic epitope, Q9VF, encoded by an
HIVgag ARF and presented by HLA-B*07. Using PBMCs of
HIV-infected patients, we first cloned and sequenced proviral DNA encoding for
Q9VF. We identified several polymorphisms with a minority of proviruses encoding
at position 5 an aspartic acid (Q9VF/5D) and a majority encoding an asparagine
(Q9VF/5N). We compared the prevalence of each variant in PBMCs of
HLA-B*07+ and HLA-B*07- patients. Proviruses encoding Q9VF/5D were
significantly less represented in HLA-B*07+ than in HLA-B*07-
patients, suggesting that Q9FV/5D encoding viruses might be under selective
pressure in HLA-B*07+ individuals. We thus analyzed ex
vivo CTL responses directed against Q9VF/5D and Q9VF/5N. Around
16% of HLA-B*07+ patients exhibited CTL responses targeting Q9VF
epitopes. The frequency and the magnitude of CTL responses induced with Q9VF/5D
or Q9VF/5N peptides were almost equal indicating a possible cross-reactivity of
the same CTLs on the two peptides. We then dissected the cellular mechanisms
involved in the presentation of Q9VF variants. As expected, cells infected with
HIV strains encoding for Q9VF/5D were recognized by Q9VF/5D-specific CTLs. In
contrast, Q9VF/5N-encoding strains were neither recognized by Q9VF/5N- nor by
Q9VF/5D-specific CTLs. Using in vitro proteasomal digestions
and MS/MS analysis, we demonstrate that the 5N variation introduces a strong
proteasomal cleavage site within the epitope, leading to a dramatic reduction of
Q9VF epitope production. Our results strongly suggest that HIV-1 escapes CTL
surveillance by introducing mutations leading to HIV ARF-epitope destruction by
proteasomes
Proteomic Modeling for HIV-1 Infected Microglia-Astrocyte Crosstalk
Background: HIV-1-infected and immune competent brain mononuclear phagocytes (MP; macrophages and microglia) secrete cellular and viral toxins that affect neuronal damage during advanced disease. In contrast, astrocytes can affect disease by modulating the nervous system’s microenvironment. Interestingly, little is known how astrocytes communicate with MP to influence disease. Methods and Findings: MP-astrocyte crosstalk was investigated by a proteomic platform analysis using vesicular stomatitis virus pseudotyped HIV infected murine microglia. The microglial-astrocyte dialogue was significant and affected microglial cytoskeleton by modulation of cell death and migratory pathways. These were mediated, in part, through F-actin polymerization and filament formation. Astrocyte secretions attenuated HIV-1 infected microglia neurotoxicity and viral growth linked to the regulation of reactive oxygen species. Conclusions: These observations provide unique insights into glial crosstalk during disease by supporting astrocytemediated regulation of microglial function and its influence on the onset and progression of neuroAIDS. The results open new insights into previously undisclosed pathogenic mechanisms and open the potential for biomarker discovery an
The glial growth factors deficiency and synaptic destabilization hypothesis of schizophrenia
BACKGROUND: A systems approach to understanding the etiology of schizophrenia requires a theory which is able to integrate genetic as well as neurodevelopmental factors. PRESENTATION OF THE HYPOTHESIS: Based on a co-localization of loci approach and a large amount of circumstantial evidence, we here propose that a functional deficiency of glial growth factors and of growth factors produced by glial cells are among the distal causes in the genotype-to-phenotype chain leading to the development of schizophrenia. These factors include neuregulin, insulin-like growth factor I, insulin, epidermal growth factor, neurotrophic growth factors, erbB receptors, phosphatidylinositol-3 kinase, growth arrest specific genes, neuritin, tumor necrosis factor alpha, glutamate, NMDA and cholinergic receptors. A genetically and epigenetically determined low baseline of glial growth factor signaling and synaptic strength is expected to increase the vulnerability for additional reductions (e.g., by viruses such as HHV-6 and JC virus infecting glial cells). This should lead to a weakening of the positive feedback loop between the presynaptic neuron and its targets, and below a certain threshold to synaptic destabilization and schizophrenia. TESTING THE HYPOTHESIS: Supported by informed conjectures and empirical facts, the hypothesis makes an attractive case for a large number of further investigations. IMPLICATIONS OF THE HYPOTHESIS: The hypothesis suggests glial cells as the locus of the genes-environment interactions in schizophrenia, with glial asthenia as an important factor for the genetic liability to the disorder, and an increase of prolactin and/or insulin as possible working mechanisms of traditional and atypical neuroleptic treatments
In vivo analysis of Nef’s role in HIV-1 replication, systemic T cell activation and CD4+ T cell loss
Prime-boost immunization of rabbits with HIV-1 gp120 elicits potent neutralization activity against a primary viral isolate
<div><p>Development of a vaccine for HIV-1 requires a detailed understanding of the neutralizing antibody responses that can be experimentally elicited to difficult-to-neutralize primary isolates. Rabbits were immunized with the gp120 subunit of HIV-1 JR-CSF envelope (Env) using a DNA-prime protein-boost regimen. We analyzed five sera that showed potent autologous neutralizing activity (IC50s at ∼10<sup>3</sup> to 10<sup>4</sup> serum dilution) against pseudoviruses containing Env from the primary isolate JR-CSF but not from the related isolate JR-FL. Pseudoviruses were created by exchanging each variable and constant domain of JR-CSF gp120 with that of JR-FL or with mutations in putative N-glycosylation sites. The sera contained different neutralizing activities dependent on C3 and V5, C3 and V4, or V4 regions located on the glycan-rich outer domain of gp120. All sera showed enhanced neutralizing activity toward an Env variant that lacked a glycosylation site in V4. The JR-CSF gp120 epitopes recognized by the sera are generally distinct from those of several well characterized mAbs (targeting conserved sites on Env) or other type-specific responses (targeting V1, V2, or V3 variable regions). The activity of one serum requires specific glycans that are also important for 2G12 neutralization and this serum blocked the binding of 2G12 to gp120. Our findings show that different fine specificities can achieve potent neutralization of HIV-1, yet this strong activity does not result in improved breadth.</p> </div
Evaluating the proper administration and counseling by community pharmacists of intranasal mupiricin for methicillin-resistant Stapholococcus aureus decolonization
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