Prof Sir Andrew J McMichael
|Keywords:||HIV, Vaccines, T cell immunity and Influenza|
Our HIV vaccines have completed five small phase I clinical trials and one large phase I/II clinical trials in London and Oxford. The vaccine is DNA encoding HIV clade A gag p24 and p17 plus a string of epitopes, and the same inserted into recombinant Modified Vaccinia Virus Ankara (MVA). Both stimulate strong CD8+ T cell responses in mice and macaques. In humans both have stimulated measurable CD8+ T cell responses in HIV low risk, uninfected volunteers. Current work is focussing on improving immunogenicity and working out assays for measuring immune responses that are likely to be protective against HIV infection. We tested the effect of those vaccines on boosting T cell responses in HIV infected patients who are on anti-retroviral drugs, with the aim of interrupting drug treatment when the T cell response is enhanced. Encouraging boosts are seen using the MVA-HIVA vaccine.
Nef (key virus protein in virus pathogenesis) initiates a programme of transcription closely similar to that triggered by T cell receptor activation. This favours HIV virus replication. Nef also alters the composition of proteins in cell signalling site, and exclusion of certain regulators favours the activation state.
We continue to explore selection of HIV-1 mutants by the immune response. T cells are very sensitive to small changes in the composition, orientation or flexibility of the exposed peptide bound to HLA. We are studying the immunodominant SLYNTVATL epitope presented by HLA-A2, showing selection of a sequence of mutations that impair T cell recognition. Why the T cells do not respond by new primary responses to the variants is also under investigation.
|Prof Bart Haynes||Duke University, North Carolina||USA|
Cytotoxic T lymphocytes (CTLs) are critical for the control of human immunodeficiency virus, but containment of virus replication can be undermined by mutations in CTL epitopes that lead to virus escape. We analyzed the evolution in vivo of an immunodominant, HLA-A2-restricted CTL epitope and found two principal, diametrically opposed evolutionary pathways that exclusively affect T cell-receptor contact residues. One pathway was characterized by acquisition of CTL escape mutations and the other by selection for wild-type amino acids. The pattern of CTL responses to epitope variants shaped which variant(s) prevailed in the virus population. The pathways notably influenced the amount of plasma virus, as patients with efficient CTL selection had lower plasma viral loads than did patients without efficient selection. Thus, viral escape from CTL responses does not necessarily correlate with disease progression. Hide abstract
Lentiviral Nef increases T cell signaling activity, but the molecular nature of the stimulus involved is incompletely described. We explored CD4 T cell lipid raft composition in the presence and absence of Nef. Here, the E2 ubiquitin-conjugating enzyme UbcH7, which acts in conjunction with c-Cbl, is absent from lipid rafts. This Nef-mediated exclusion is associated with failure of ubiquitination of activated Vav. In the presence of Nef, lipid raft Cdc42 is activated and forms a ternary complex between the c-Cbl-interacting protein p85Cool-1/betaPix and c-Cbl, displacing UbcH7 from rafts. Suppression of p85Cool-1/betaPix expression restores UbcH7 raft localization and Vav ubiquitination and diminishes Cdc42 activity. Moreover, p85Cool-1/betaPix knockdown attenuates HIV replication. Thresholds for activation of signaling involve the intricate balance of positive and negative regulators. Here we provide evidence for Nef disruption of a negative regulator of T cell signaling in promoting HIV replication. Hide abstract
All thymically selected T cells are inherently cross-reactive, yet many data indicate a fine specificity in antigen recognition, which enables virus escape from immune control by mutation in infections such as the human immunodeficiency virus (HIV). To address this paradox, we analyzed the fine specificity of T cells recognizing a human histocompatibility leukocyte antigen (HLA)-A2-restricted, strongly immunodominant, HIV gag epitope (SLFNTVATL). The majority of 171 variant peptides tested bound HLA-A2, but only one third were recognized. Surprisingly, one recognized variant (SLYNTVATL) showed marked differences in structure when bound to HLA-A2. T cell receptor (TCR) recognition of variants of these two peptides implied that they adopted the same conformation in the TCR-peptide-major histocompatibility complex (MHC) complex. However, the on-rate kinetics of TCR binding were identical, implying that conformational changes at the TCR-peptide-MHC binding interface occur after an initial permissive antigen contact. These findings have implications for the rational design of vaccines targeting viruses with unstable genomes. Hide abstract
J Immunol, 172 (12), pp. 7350-7358. Read abstract2004. CD4(-)CD8alphaalpha subset of CD1d-restricted NKT cells controls T cell expansion.
Valpha24 invariant (Valpha24i) CD1d-restricted NKT cells are widely regarded to have immune regulatory properties. They are known to have a role in preventing autoimmune diseases and are involved in optimally mounted immune responses to pathogens and tumor cells. We were interested in understanding how these cells provide protection in autoimmune diseases. We first observed, using EBV/MHC I tetrameric complexes, that expansion of Ag-specific cells in human PBMCs was reduced when CD1d-restricted NKT cells were concomitantly activated. This was accompanied by an increase in a CD4(-)CD8alphaalpha(+) subset of Valpha24i NKT cells. To delineate if a specific subset of NKT cells was responsible for this effect, we generated different subsets of human CD4(-) and CD4(+) Valpha24i NKT clones and demonstrate that a CD4(-)CD8alphaalpha(+) subset with highly efficient cytolytic ability was unique among the clones in being able to suppress the proliferation and expansion of activated T cells in vitro. Activated clones were able to kill CD1d-bearing dendritic or target cells. We suggest that one mechanism by which CD1d-restricted NKT cells can exert a regulatory role is by containing the proliferation of activated T cells, possibly through timely lysis of APCs or activated T cells bearing CD1d. Hide abstract
The anti-influenza CD8+ T cell response in HLA-A2-positive adults is almost exclusively directed at residues 58-66 of the virus matrix protein (MP(58-66)). V(beta)17V(alpha)10.2 T cell receptors (TCRs) containing a conserved arginine-serine-serine sequence in complementarity determining region 3 (CDR3) of the V(beta) segment dominate this response. To investigate the molecular basis of immunodominant selection in an outbred population, we have determined the crystal structure of V(beta)17V(alpha)10.2 in complex with MP(58-66)-HLA-A2 at a resolution of 1.4 A. We show that, whereas the TCR typically fits over an exposed side chain of the peptide, in this structure MP(58-66) exposes only main chain atoms. This distinctive orientation of V(beta)17V(alpha)10.2, which is almost orthogonal to the peptide-binding groove of HLA-A2, facilitates insertion of the conserved arginine in V(beta) CDR3 into a notch in the surface of MP(58-66)-HLA-A2. This previously unknown binding mode underlies the immunodominant T cell response. Hide abstract
Gene expression profiling was used to explore the role of Nef in HIV. Nef induces a transcriptional program in T cells that is 97% identical to that of anti-CD3 T cell activation. This program is inhibited in the presence of cyclosporin. A requirement for TCR zeta and ZAP-70 is demonstrated for formation of the complete profile. Among eight factors particular to the anti-CD3 activation profile are IL16 and YY1, negative regulators of HIV transcription. In contrast, Nef exclusively upregulates factors positively regulating HIV, including Tat-SF1, U1 SNRNP, and IRF-2. New genes associated with Nef include CDK9, the induction of which enhances Tat function. Thus, Nef acts as a master switch early in the viral life cycle, forcing an environment conducive to dynamic viral production. Hide abstract
HIV-1-specific cytotoxic T-lymphocyte (CTL) responses have been detected at a low frequency in many HIV-1-exposed, persistently seronegative (HEPS) subjects. However, it is unclear how CTLs could protect against HIV acquisition in HEPS subjects, when high levels of circulating CTL fail to prevent disease progression in most seropositive subjects. To address this issue we studied CD8(+) lymphocyte responses to a panel of HIV-1 CTL epitopes in 91 HEPS and 87 HIV-1-infected Nairobi sex workers. HIV-specific responses in seropositive women focused strongly on epitopes rarely or never recognized in HEPS subjects, who targeted epitopes that were subdominant or unrecognized in infected women. These differences in epitope specificity were restricted by only those HLA class I alleles that are associated with a reduced risk of HIV-1 infection in this cohort. Late seroconversion in HEPS donors was associated with a switch in epitope specificity and/or immunodominance to those epitopes preferentially recognized by HIV-1-infected women. The likelihood of detecting HIV-1-specific responses in HEPS women increased with the duration of viral exposure, suggesting that HIV-1-specific CD8(+) responses are acquired over time. The association between differential recognition of distinct CTL epitopes and protection from HIV-1 infection may have significant implications for vaccine design. Hide abstract
Naive and Memory CD8 T cells specific for HIV-1
We have recently found that in HIV negative and uninfected blood donors, there are HIV specific naive and memory CD4 T cells. The latter have been primed by cross reacting antigens, most likely in the microbiome. We now plan to examine CD8 T cells with the aim of determining whether the hierarchy of immunodominant T cells seen after infection with HIV is determined by the relative frequencies of naive or memory T cells present before infection. We will also identify dominant clonotypes by ...
Mechanistic basis of the decline in HIV-specific CD8 T cell function during chronic infection
Virus-specific CD8 T cell responses are induced during the acute phase of HIV-1 infection and play an important role in containment of the acute viral burst and subsequent control of ongoing viral replication. Prophylactic and therapeutic strategies to combat HIV infection are therefore being designed to target this arm of the immune response. However, moderate-high levels of HIV replication continue in the majority of chronically-infected individuals despite the presence of high frequencies of ...
The impact of IFTIM3 genetic variation on Influenza virus infection, immune responses and disease outcome
Interferon inducible transmembrane 3 (IFITM3) protein is a virus restriction factor mediating cellular resistance to influenza viruses and other viruses. However, the role of IFTIM3 in antiviral immunity is largely unknown. Everitt et al (1) that showed the severity of influenza virus infection was increased in IFITM3 knock out mice compared to wild type animals. They also showed that In northern European patients infected and hospitalized with seasonal influenza or pandemic influenza ...