%0 Journal Article %~ PubMed %A Harman, Andrew N %A Bye, Chris R %A Nasr, Najla %A Sandgren, Kerrie J %A Kim, Min %A Mercier, Sarah K %A Botting, Rachel A %A Lewin, Sharon R %A Cunningham, Anthony L %A Cameron, Paul U %T Identification of Lineage Relationships and Novel Markers of Blood and Skin Human Dendritic Cells. %B Journal of Immunology %D 2013 %C United States %I American Association of Immunologists %V 190 %N 1 %P 66-79 %@ 1550-6606 %X %Z FOR Codes: 60103 60502 110704 %0 Journal Article %~ PubMed %A Cunningham, Anthony %A Griffiths, Paul %A Leone, Peter %A Mindel, Adrian %A Patel, Rajul %A Stanberry, Lawrence %A Whitley, Richard %T Current management and recommendations for access to antiviral therapy of herpes labialis. %B Journal of Clinical Virology %D 2012 %C Netherlands %I Elsevier BV %V 53 %N 1 %P 6-11 %@ 1873-5967 %X Herpes labialis is a common skin infective condition, worldwide, which is primarily caused by HSV-1. Recurrent episodes of herpes labialis, also known as cold sores, can be frequent, painful, long-lasting and disfiguring for infected patients. At present, there are two types of antivirals for the treatment of herpes labialis, topical and oral, which are available over the counter or as prescription-only. The aim of antiviral therapy is to block viral replication to enable shortening the duration of symptoms and to accelerate healing of the lesions associated with herpes labialis. This review examines the evidence for the effectiveness of current topical and oral antivirals in the management of recurrent episodes of herpes labialis. In most countries, oral antivirals for herpes labialis are available as prescription-only. However, in early 2010, the oral antiviral famciclovir was reclassified from prescription-only medicine to pharmacist-controlled status in New Zealand. The benefits and risks associated with moving an antiviral therapy for herpes labialis from prescription-only to pharmacist-controlled status are reviewed here, and the implications for patients, general physicians and pharmacists are considered. %Z FOR Codes: 110309 110304 %0 Journal Article %~ PubMed %A Nasr, Najla %A Maddocks, Susan %A Turville, Stuart G %A Harman, Andrew N %A Woolger, Natalie %A Helbig, Karla J %A Wilkinson, John %A Bye, Chris R %A Wright, Thomas K %A Rambukwelle, Dharshini %A Donaghy, Heather %A Beard, Michael R %A Cunningham, Anthony L %T HIV-1 infection of human macrophages directly induces viperin which inhibits viral production. %B Blood %D 2012 %C United States %I American Society of Hematology %V 120 %N 4 %P 778-788 %@ 0006-4971 %X Macrophages are key target cells for HIV-1. HIV-1BaL induced a subset of interferon-stimulated genes (ISGs) in monocyte-derived macrophages (MDMs), which differed to that in monocyte derived-dendritic cells and CD4 T cells, without inducing any interferons. Inhibition of type I IFN induction was mediated by HIV-1 inhibition of interferon regulated factor (IRF3) nuclear translocation. In MDMs, viperin was the most upregulated ISG and it significantly inhibited HIV-1 production. HIV-1 infection disrupted lipid rafts via viperin induction and redistributed viperin to CD81 compartments, the site of HIV-1 egress by budding in MDMs. Exogenous farnesol, which enhances membrane protein prenylation, reversed viperin mediated inhibition of HIV-1 production. Mutagenesis analysis in transfected cell lines showed that the internal S-adenosyl methionine (SAM) domains of viperin were essential for its antiviral activity. Thus viperin may contribute to persistent non-cytopathic HIV-1 infection of macrophages and possibly to biological differences with HIV-1 infected T cells. %Z FOR Codes: 110707 110804 60106 %0 Journal Article %~ PubMed %A Kim, Min %A Osborne, Naomi R %A Zeng, Weiguang %A Donaghy, Heather %A McKinnon, Kay %A Jackson, David C %A Cunningham, Anthony L %T Herpes simplex virus antigens directly activate NK cells via TLR2, thus facilitating their presentation to CD4 T lymphocytes. %B Journal of Immunology %D 2012 %C United States %I American Association of Immunologists %V 188 %N 9 %P 4158-4170 %@ 1550-6606 %X NK cells infiltrate human herpetic lesions, but their role has been underexplored. HSV can stimulate innate immune responses via surface TLR2, which is expressed on monocyte-derived dendritic cells (DCs) and NK cells. In this study, UV-inactivated HSV1/2 and immunodominant HSV2 glycoprotein D peptides conjugated to the TLR2 agonist dipalmitoyl-S-glyceryl cysteine stimulated CD4 T lymphocyte IFN-?? responses within PBMCs or in coculture with monocyte-derived DCs. NK cells contributed markedly to the PBMC responses. Furthermore, NK cells alone were activated directly by both Ags, also upregulating HLA-DR and HLA-DQ and then they activated autologous CD4 T lymphocytes. Using Transwells, Ag-stimulated NK cells and CD4 T lymphocytes were shown to interact through both cell-to-cell contact and cytokines, differing in relative importance in different donors. A distinct immunological synapse between Ag-stimulated NK cells and CD4 T lymphocytes was observed, indicating the significance of their cell-to-cell contact. A large proportion (57%) of NK cells was also in contact with CD4 T lymphocytes in the dermal infiltrate of human recurrent herpetic lesions. Thus, NK cells stimulated by TLR2-activating HSV Ags can present Ag alone or augment the role of DCs in vitro and perhaps in herpetic lesions or draining lymph nodes. In addition to DCs, NK cells should be considered as targets for adjuvants during HSV vaccine development. %Z FOR Codes: 1107 1108 %0 Journal Article %~ PubMed %A Kelly, Barbara J %A Mijatov, Branka %A Fraefel, Cornel %A Cunningham, Anthony L %A Diefenbach, Russell J %T Identification of a single amino acid residue which is critical for the interaction between HSV-1 inner tegument proteins pUL36 and pUL37. %B Virology %D 2012 %C United States %I Academic Press %V 422 %N 2 %P 308-16 %@ 0042-6822 %X The herpes simplex virus type 1 (HSV-1) structural tegument proteins pUL36 and pUL37 are essential for secondary envelopment during the egress of viral particles. For this study, scanning alanine mutagenesis of HSV-1 pUL37, in combination with yeast two-hybrid, identified pUL37 residue D631 as a major determinant for binding of pUL36. Further analysis of the binding of this pUL37 mutant to pUL36 by coimmunoprecipitation assay confirmed the role of pUL37 D631 in mediating binding of pUL36. A trans-complementation assay using pUL37 deletion virus FR??UL37 was then carried out, where pUL37 wild type or D631A were provided in trans. For pUL37 D631A, a significant reduction in virus titer was observed compared to that seen when pUL37 wild type was present. The results presented here underline the crucial role of the pUL36/pUL37 interaction in replication of HSV-1 and indicate a critical role for pUL37 D631 in mediating this interaction. %Z FOR Codes: 110804 %0 Journal Article %~ PubMed %A Aggarwal, Anupriya %A Iemma, Tina L %A Shih, Ivy %A Newsome, Timothy P %A McAllery, Samantha %A Cunningham, Anthony L %A Turville, Stuart G %T Mobilization of HIV Spread by Diaphanous 2 Dependent Filopodia in Infected Dendritic Cells. %B PLoS Pathogens %D 2012 %C United States %I Public Library of Science %V 8 %N 6 %P e1002762 %@ 1553-7374 %X Paramount to the success of persistent viral infection is the ability of viruses to navigate hostile environments en route to future targets. In response to such obstacles, many viruses have developed the ability of establishing actin rich-membrane bridges to aid in future infections. Herein through dynamic imaging of HIV infected dendritic cells, we have observed how viral high-jacking of the actin/membrane network facilitates one of the most efficient forms of HIV spread. Within infected DC, viral egress is coupled to viral filopodia formation, with more than 90% of filopodia bearing immature HIV on their tips at extensions of 10 to 20 ??m. Live imaging showed HIV filopodia routinely pivoting at their base, and projecting HIV virions at ??m.sec(-1) along repetitive arc trajectories. HIV filopodial dynamics lead to up to 800 DC to CD4 T cell contacts per hour, with selection of T cells culminating in multiple filopodia tethering and converging to envelope the CD4 T-cell membrane with budding HIV particles. Long viral filopodial formation was dependent on the formin diaphanous 2 (Diaph2), and not a dominant Arp2/3 filopodial pathway often associated with pathogenic actin polymerization. Manipulation of HIV Nef reduced HIV transfer 25-fold by reducing viral filopodia frequency, supporting the potency of DC HIV transfer was dependent on viral filopodia abundance. Thus our observations show HIV corrupts DC to CD4 T cell interactions by physically embedding at the leading edge contacts of long DC filopodial networks. %Z FOR Codes: 110704 %0 Journal Article %~ PubMed %A Aggarwal, Anupriya %A Miranda-Saksena, Monica %A Boadle, Ross A %A Kelly, Barbara J %A Diefenbach, Russell J %A Alam, Waafiqa %A Cunningham, Anthony L %T Ultrastructural Visualization of Individual Tegument Protein Dissociation during Entry of Herpes Simplex Virus 1 into Human and Rat Dorsal Root Ganglion Neurons. %B Journal of Virology %D 2012 %C United States %I American Society for Microbiology %V 86 %N 11 %P 6123-6137 %@ 1098-5514 %X Herpes simplex virus 1 (HSV-1) enters neurons primarily by fusion of the viral envelope with the host cell plasma membrane, leading to the release of the capsid into the cytosol. The capsid travels via microtubule-mediated retrograde transport to the nuclear membrane, where the viral DNA is released for replication in the nucleus. In the present study, the composition and kinetics of incoming HSV-1 capsids during entry and retrograde transport in axons of human fetal and dissociated rat dorsal root ganglia (DRG) neurons were examined by wide-field deconvolution microscopy and transmission immunoelectron microscopy (TIEM). We show that HSV-1 tegument proteins, including VP16, VP22, most pUL37, and some pUL36, dissociated from the incoming virions. The inner tegument proteins, including pUL36 and some pUL37, remained associated with the capsid during virus entry and transit to the nucleus in the neuronal cell body. By TIEM, a progressive loss of tegument proteins, including VP16, VP22, most pUL37, and some pUL36, was observed, with most of the tegument dissociating at the plasma membrane of the axons and the neuronal cell body. Further dissociation occurred within the axons and the cytosol as the capsids moved to the nucleus, resulting in the release of free tegument proteins, especially VP16, VP22, pUL37, and some pUL36, into the cytosol. This study elucidates ultrastructurally the composition of HSV-1 capsids that encounter the microtubules in the core of human axons and the complement of free tegument proteins released into the cytosol during virus entry. %Z FOR Codes: 110804 %0 Journal Article %~ PubMed %A Cashin, Kieran %A Roche, Michael %A Sterjovski, Jasminka %A Ellett, Anne %A Gray, Lachlan R %A Cunningham, Anthony L %A Ramsland, Paul A %A Churchill, Melissa J %A Gorry, Paul R %T Alternative coreceptor requirements for efficient CCR5- and CXCR4-mediated HIV-1 entry into macrophages. %B Journal of virology %D 2011 %C United States %I American Society for Microbiology %V 85 %N 20 %P 10699-10709 %@ 1098-5514 %X %Z FOR Codes: 110804 %0 Journal Article %~ PubMed %A Wang, Bin %A Dwyer, Dominic Edmund %A Soedjono, Maly %A Shi, Haijing %A Matlho, Kabo %A Ratnamohan, Mala %A Blyth, Christopher %A McPhie, Ken %A Cunningham, Anthony Lawrence %A Saksena, Nitin Kumar %T Evidence of the circulation of pandemic influenza (H1N1) 2009 with D222D/G/N/S hemagglutinin polymorphisms during the first wave of the 2009 influenza pandemic. %B Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology %D 2011 %C Netherlands %I Elsevier BV %V 52 %N 4 %P 304-6 %@ 1873-5967 %X The hemagglutinin HA1 D222G substitution may be associated with adverse outcomes in pandemic influenza A (H1N1) 2009 infections by enhancing the binding capacity of ??2-3 sialyl receptors to pandemic influenza (H1N1) 2009 viruses. %Z FOR Codes: 60506 60409 %0 Journal Article %~ PubMed %A Harman, Andrew N %A Lai, Joey %A Turville, Stuart %A Samarajiwa, Shamith %A Gray, Lachlan %A Marsden, Valerie %A Mercier, Sarah %A Jones, Kate %A Nasr, Najla %A Cumming, Helen %A Donaghy, Heather %A Mak, Johnson %A Churchill, Melissa %A Hertzog, Paul %A Cunningham, Anthony L %T HIV infection of dendritic cells subverts the interferon induction pathway via IRF1 and inhibits type 1 interferon production. %B Blood %D 2011 %C United States %I American Society of Hematology %V 118 %N 2 %P 298-308 %@ 0006-4971 %X Many viruses have developed mechanisms to evade the IFN response. Here, HIV-1 was shown to induce a distinct subset of IFN-stimulated genes (ISGs) in monocyte-derived dendritic cells (DCs), without detectable type I or II IFN. These ISGs all contained an IFN regulatory factor 1 (IRF-1) binding site in their promoters, and their expression was shown to be driven by IRF-1, indicating this subset was induced directly by viral infection by IRF-1. IRF-1 and -7 protein expression was enriched in HIV p24 antigen-positive DCs. A HIV deletion mutant with the IRF-1 binding site deleted from the long terminal repeat showed reduced growth kinetics. Early and persistent induction of IRF-1 was coupled with sequential transient up-regulation of its 2 inhibitors, IRF-8, followed by IRF-2, suggesting a mechanism for IFN inhibition. HIV-1 mutants with Vpr deleted induced IFN, showing that Vpr is inhibitory. However, HIV IFN inhibition was mediated by failure of IRF-3 activation rather than by its degradation, as in T cells. In contrast, herpes simplex virus type 2 markedly induced IFN?? and a broader range of ISGs to higher levels, supporting the hypothesis that HIV-1 specifically manipulates the induction of IFN and ISGs to enhance its noncytopathic replication in DCs. %Z FOR Codes: 110707 %0 Book Section %A Diefenbach, Russell %A Miranda Saksena, Monica %A Cunningham, Anthony %T Herpes simplex virus: Microtubuledependent transport and egress from neurons %B Viral Transport, Assembly and Egress %D 2011 %C India %I Research Signpost %V %N %P 49-78 %@ 9788130804330 %E Cunningham, Anthony %E Diefenbach, Russell %X %Z FOR Codes: 110804 %0 Book Section %A Cunningham, Anthony %A Garland, Suzanne M %A Donaghy, Heather %A Kim, Min %T Mucosal Immunity in Sexually Transmitted Infections %B Sexually Transmitted Infections and Sexually Transmitted Diseases %D 2011 %C Germany %I Springer %V %N %P 49-76 %@ 9783642146626 %E Gross, Gerd %E Tyring, Stephen K %X %Z FOR Codes: 60502 %0 Journal Article %~ PubMed %A Wang, Bin %A Steain, Megan C %A Dwyer, Dominic E %A Cunningham, Anthony L %A Saksena, Nitin K %T Synthetic long oligonucleotides to generate artificial templates for use as positive controls in molecular assays: drug resistance mutations in influenza virus as an example. %B Virology journal %D 2011 %C United Kingdom %I BioMed Central Ltd. %V 8 %N %P 405 %@ 1743-422X %X ABSTRACT: %Z FOR Codes: 60502 110303 60408 %0 Journal Article %~ PubMed %A Cunningham, Anthony L %A Booth, David %T The first common cold sore susceptibility gene. %B Journal of Infectious Diseases %D 2011 %C United States %I University of Chicago Press %V 204 %N 11 %P 1645-1647 %@ 0022-1899 %X %Z FOR Codes: 110309 60412 %0 Journal Article %A Leo, Oberdan %A Cunningham, Anthony %A Stern, Peter L %T Vaccine Immunology %B Understanding Modern Vaccines: Perspectives in Vaccinology %D 2011 %C United States %I Elsevier %V 1 %N 1 %P 25-59 %@ 2210-7622 %X %Z FOR Codes: 110804 %0 Journal Article %A Strugnell, Richard %A Zepp, Fred %A Cunningham, Anthony %A Tantawichien, Terapong %T Vaccine antigens %B Understanding Modern Vaccines: Perspectives in Vaccinology %D 2011 %C United States %I Elsevier %V 1 %N 1 %P 61-88 %@ 2210-7622 %X %Z FOR Codes: 110804 %0 Journal Article %~ PubMed %A Sandgren, Kerrie J %A Wilkinson, John %A Miranda-Saksena, Monica %A McInerney, Gerald M %A Byth-Wilson, Karen %A Robinson, Phillip J %A Cunningham, Anthony L %T A differential role for macropinocytosis in mediating entry of the two forms of vaccinia virus into dendritic cells. %B PLoS pathogens %D 2010 %C United States %I Public Library of Science %V 6 %N 4 %P e1000866 %@ 1553-7374 %X Vaccinia virus (VACV) is being developed as a recombinant viral vaccine vector for several key pathogens. Dendritic cells (DCs) are specialised antigen presenting cells that are crucial for the initiation of primary immune responses; however, the mechanisms of uptake of VACV by these cells are unclear. Therefore we examined the binding and entry of both the intracellular mature virus (MV) and extracellular enveloped virus (EV) forms of VACV into vesicular compartments of monocyte-derived DCs. Using a panel of inhibitors, flow cytometry and confocal microscopy we have shown that neither MV nor EV binds to the highly expressed C-type lectin receptors on DCs that are responsible for capturing many other viruses. We also found that both forms of VACV enter DCs via a clathrin-, caveolin-, flotillin- and dynamin-independent pathway that is dependent on actin, intracellular calcium and host-cell cholesterol. Both MV and EV entry were inhibited by the macropinocytosis inhibitors rottlerin and dimethyl amiloride and depended on phosphotidylinositol-3-kinase (PI(3)K), and both colocalised with dextran but not transferrin. VACV was not delivered to the classical endolysosomal pathway, failing to colocalise with EEA1 or Lamp2. Finally, expression of early viral genes was not affected by bafilomycin A, indicating that the virus does not depend on low pH to deliver cores to the cytoplasm. From these collective results we conclude that VACV enters DCs via macropinocytosis. However, MV was consistently less sensitive to inhibition and is likely to utilise at least one other entry pathway. Definition and future manipulation of these pathways may assist in enhancing the activity of recombinant vaccinia vectors through effects on antigen presentation. %Z FOR Codes: 605 1108 %0 Journal Article %~ PubMed %A Sterjovski, Jasminka %A Roche, Michael %A Churchill, Melissa J %A Ellett, Anne %A Farrugia, William %A Gray, Lachlan R %A Cowley, Daniel %A Poumbourios, Pantelis %A Lee, Benhur %A Wesselingh, Steven L %A Cunningham, Anthony L %A Ramsland, Paul A %A Gorry, Paul R %T An altered and more efficient mechanism of CCR5 engagement contributes to macrophage tropism of CCR5-using HIV-1 envelopes. %B Virology %D 2010 %C United States %I Academic Press %V 404 %N 2 %P 269-278 %@ 0042-6822 %X While CCR5 is the principal coreceptor used by macrophage (M)-tropic HIV-1, not all primary CCR5-using (R5) viruses enter macrophages efficiently. Here, we used functionally-diverse R5 envelope (Env) clones to characterize virus-cell interactions important for efficient CCR5-mediated macrophage entry. The magnitude of macrophage entry by Env-pseudotyped reporter viruses correlated with increased immunoreactivity of CD4-induced gp120 epitopes, increased ability to scavenge low levels of cell-surface CCR5, reduced sensitivity to the CCR5 inhibitor maraviroc, and increased dependence on specific residues in the CCR5 ECL2 region. These results are consistent with an altered and more efficient mechanism of CCR5 engagement. Structural studies revealed potential alterations within the gp120 V3 loop, the gp41 interaction sites at the gp120 C- and N-termini, and within the gp120 CD4 binding site which may directly or indirectly lead to more efficient CCR5-usage. Thus, enhanced gp120-CCR5 interactions may contribute to M-tropism of R5 HIV-1 strains through different structural mechanisms. %Z FOR Codes: 60506 %0 Journal Article %~ PubMed %A Jin, Fengyi %A Prestage, Garrett %A Imrie, John %A Kippax, Susan %A Donovan, Basil %A Templeton, David %A Cunningham, Anthony %A Mindel, Adrian %A Cunningham, Philip %A Kaldor, John %A Grulich, Andrew %T Anal Sexually Transmitted Infections and Risk of HIV Infection in Homosexual Men. %B Journal of Acquired Immune Deficiency Syndromes %D 2010 %C United States %I Lippincott Williams & Wilkins %V 53 %N 1 %P 144-149 %@ 1525-4135 %X BACKGROUND:: We examined a range of common bacterial and viral sexually transmitted infections as risk factors for HIV seroconversion in a community-based cohort of HIV-negative homosexual men in Sydney, Australia. METHODS:: Detailed information about HIV risk behaviors was collected by interview twice yearly. Participants were tested annually for HIV, anal and urethral gonorrhea and chlamydia, herpes simplex virus types 1 and 2, and syphilis. In addition, they reported annual diagnoses of these conditions and of genital and anal warts. RESULTS:: Among 1427 enrolled participants, 53 HIV seroconverters were identified, giving an incidence of 0.78 per 100 person-years. After controlling for number of episodes of insertive and receptive nonseroconcordant unprotected anal intercourse, there were independent associations with anal gonorrhea (adjusted hazard ratio = 7.12, 95% confidence interval: 2.05 to 24.79) and anal warts (hazard ratio = 3.63, 95% confidence interval: 1.62 to 8.14). CONCLUSIONS:: Anal gonorrhea and anal warts were independently associated with HIV acquisition. The added HIV prevention value of more frequent screening of the anus to allow early detection and treatment of anal sexually transmitted infections in homosexual men should be considered. %Z FOR Codes: 1103 1117 %0 Journal Article %~ PubMed %A Gowrishankar, Kavitha %A Steain, Megan %A Cunningham, Anthony L %A Rodriguez, Michael %A Blumbergs, Peter %A Slobedman, Barry %A Abendroth, Allison %T Characterization of the host immune response in human Ganglia after herpes zoster. %B Journal of virology %D 2010 %C United States %I American Society for Microbiology %V 84 %N 17 %P 8861-70 %@ 1098-5514 %X Varicella-zoster virus (VZV) causes varicella (chicken pox) and establishes latency in ganglia, from where it reactivates to cause herpes zoster (shingles), which is often followed by postherpetic neuralgia (PHN), causing severe neuropathic pain that can last for years after the rash. Despite the major impact of herpes zoster and PHN on quality of life, the nature and kinetics of the virus-immune cell interactions that result in ganglion damage have not been defined. We obtained rare material consisting of seven sensory ganglia from three donors who had suffered from herpes zoster between 1 and 4.5 months before death but who had not died from herpes zoster. We performed immunostaining to investigate the site of VZV infection and to phenotype immune cells in these ganglia. VZV antigen was localized almost exclusively to neurons, and in at least one case it persisted long after resolution of the rash. The large immune infiltrate consisted of noncytolytic CD8(+) T cells, with lesser numbers of CD4(+) T cells, B cells, NK cells, and macrophages and no dendritic cells. VZV antigen-positive neurons did not express detectable major histocompatibility complex (MHC) class I, nor did CD8(+) T cells surround infected neurons, suggesting that mechanisms of immune control may not be dependent on direct contact. This is the first report defining the nature of the immune response in ganglia following herpes zoster and provides evidence for persistence of non-latency-associated viral antigen and inflammation beyond rash resolution. %Z FOR Codes: 110804 %0 Journal Article %~ PubMed %A Wang, Bin %A Dwyer, Dominic E %A Blyth, Christopher C %A Soedjono, Maly %A Shi, Haijing %A Kesson, Alison %A Ratnamohan, Mala %A McPhie, Ken %A Cunningham, Anthony L %A Saksena, Nitin K %T Detection of the rapid emergence of the H275Y mutation associated with oseltamivir resistance in severe pandemic influenza virus A/H1N1 09 infections. %B Antiviral research %D 2010 %C Netherlands %I Elsevier BV %V 87 %N 1 %P 16-21 %@ 1872-9096 %X In 2009 a new swine-origin influenza virus A/H1N1 (A/H1N1 09) emerged, causing the century''s first pandemic. Most isolates of the new A/H1N1 09 virus are susceptible to neuraminidase inhibitors, but the H275Y mutation in the neuraminidase gene region associated with high-level oseltamivir resistance has been detected. Using rolling circle amplification (RCA) technology, 96 A/H1N1 09-specific RT-PCR positive clinical samples collected from 80 oseltamivir-treated and untreated patients were screened for the presence of the H275Y mutation. Samples positive for 275Y mutation by RCA were cloned and sequenced for confirmation. From 25 patients who had been treated with oseltamivir and remained A/H1N1 09 RT-PCR positive, we identified three (12%) individuals with the H275Y mutation: one immuno-suppressed adult, one immuno-competent adult and one child. Samples collected at multiple time points from the two adults showed a switch from wild-type oseltamivir-sensitive 275H to oseltamivir-resistant 275Y population after 9 days of treatment. The child had the 275Y mutation detected after being persistently A/H1N1 09 RT-PCR positive while receiving oseltamivir treatment. Resistance was not detected in 17 pre-treatment samples and 54 A/H1N1 09 RT-PCR positive outpatients. RCA demonstrates the rapid emergence of the H275Y resistance mutation in individuals with severe A/H1N1 09 infection receiving neuraminidase inhibitors. Rapid detection of oseltamivir resistance in severe infection is essential for patients to receive maximum therapeutic benefit. In the light of emerging resistance, close monitoring and understanding of the nature and dynamics of resistance mutations in newly emerging strains should be a priority. %Z FOR Codes: 110804 110309 %0 Journal Article %~ PubMed %A Cameron, Paul U %A Saleh, Suha %A Sallmann, Georgina %A Solomon, Ajantha %A Wightman, Fiona %A Evans, Vanessa A %A Boucher, Genevieve %A Haddad, Elias K %A Sekaly, Rafick-Pierre %A Harman, Andrew N %A Anderson, Jenny L %A Jones, Kate L %A Mak, Johnson %A Cunningham, Anthony L %A Jaworowski, Anthony %A Lewin, Sharon R %T Establishment of HIV-1 latency in resting CD4+ T cells depends on chemokine-induced changes in the actin cytoskeleton. %B Proceedings of the National Academy of Sciences of the United States of America %D 2010 %C United States %I National Academy of Sciences %V 107 %N 39 %P 16934-16939 %@ 0027-8424 %X Eradication of HIV-1 with highly active antiretroviral therapy (HAART) is not possible due to the persistence of long-lived, latently infected resting memory CD4(+) T cells. We now show that HIV-1 latency can be established in resting CD4(+) T cells infected with HIV-1 after exposure to ligands for CCR7 (CCL19), CXCR3 (CXCL9 and CXCL10), and CCR6 (CCL20) but not in unactivated CD4(+) T cells. The mechanism did not involve cell activation or significant changes in gene expression, but was associated with rapid dephosphorylation of cofilin and changes in filamentous actin. Incubation with chemokine before infection led to efficient HIV-1 nuclear localization and integration and this was inhibited by the actin stabilizer jasplakinolide. We propose a unique pathway for establishment of latency by direct HIV-1 infection of resting CD4(+) T cells during normal chemokine-directed recirculation of CD4(+) T cells between blood and tissue. %Z FOR Codes: 110707 %0 Journal Article %~ PubMed %A Puttur, Franz K %A Fernandez, Marian A %A White, Rose %A Roediger, Ben %A Cunningham, Anthony L %A Weninger, Wolfgang %A Jones, Cheryl A %T Herpes Simplex Virus Infects Skin {gamma}{delta} T Cells before Langerhans Cells and Impedes Migration of Infected Langerhans Cells by Inducing Apoptosis and Blocking E-Cadherin Downregulation. %B Journal of immunology (Baltimore, Md. : 1950) %D 2010 %C United States %I American Association of Immunologists %V 185 %N 1 %P 477-87 %@ 1550-6606 %X The role individual skin dendritic cell (DC) subsets play in the immune response to HSV remains unclear. We investigated the effect of HSV on DC virus uptake, viability, and migration after cutaneous infection in vitro and in vivo. HSV increased the emigration of skin DCs from whole skin explants over 3 d postinfection (p.i.) compared with mock controls, but the kinetics of emigration was influenced by the skin DC subset. Uninfected (bystander) Langerhans cells (LCs) were the major emigrant DC subset at 24 h p.i., but thereafter, large increases in infected CD103(+)langerin(+) dermal DC (dDC) and uninfected langerin(-) dDC emigration were also observed. LC infection was confirmed by the presence of HSV glycoprotein D (gD) and was associated with impaired migration from cultured skin. Langerin(+) dDC also expressed HSV gD, but infection did not impede migration. We then followed the virus in live MacGreen mice in which LCs express GFP using a fluorescent HSV-1 strain by time-lapse confocal microscopy. We observed a sequential infection of epidermal cells, first in keratinocytes and epidermal gammadelta T cells at 6 h p.i., followed by the occurrence of HSVgD(+) LCs at 24 h p.i. HSV induced CCR7 upregulation on all langerin(+) DC, including infected LCs, and increased production of skin TNF-alpha and IL-1beta. However, a large proportion of infected LCs that remained within the skin was apoptotic and failed to downregulate E-cadherin compared with bystander LCs or mock controls. Thus, HSV infection of LCs is preceded by infection of gammadelta T cells and delays migration. %Z FOR Codes: 110804 110701 110304 %0 Journal Article %~ PubMed %A Apcarian, Arin %A Cunningham, Anthony L %A Diefenbach, Russell J %T Identification of binding domains in the herpes simplex virus type 1 small capsid protein pUL35 (VP26). %B The Journal of general virology %D 2010 %C United Kingdom %I Society for General Microbiology %V 91 %N 11 %P 2659-63 %@ 1465-2099 %X In this study, fragments of the small capsid protein pUL35 (VP26) from herpes simplex virus type 1 (HSV-1) were generated to identify binding domains for a number of known ligands. Analysis of the binding of dynein light chain subunits, DYNLT1 and DYNLT3, as well the HSV-1 structural proteins pUL19 (VP5) and pUL37 was then undertaken using the LexA yeast two-hybrid assay. The N-terminal half of pUL35, in particular residues 30-43, was identified as a common region for the binding of DYNLT1 and DYNLT3. Additional distinct regions in the C terminus of pUL35 also contribute to the binding of DYNLT1 and DYNLT3. In contrast, only the C-terminal half of pUL35 was found to mediate the binding of pUL19 and pUL37 through distinct regions. The relevance of this information to the role of pUL35 in viral transport and assembly is discussed. %Z FOR Codes: 110804 %0 Journal Article %~ PubMed %A Huch, Jennifer H %A Cunningham, Anthony L %A Arvin, Ann M %A Nasr, Najla %A Santegoets, Saskia J A M %A Slobedman, Eric %A Slobedman, Barry %A Abendroth, Allison %T Impact of varicella-zoster virus on dendritic cell subsets in human skin during natural infection. %B Journal of Virology %D 2010 %C United States %I American Society for Microbiology %V 84 %N 8 %P 4060-4072 %@ 1098-5514 %X Varicella-zoster virus (VZV) causes varicella and herpes zoster, diseases characterized by distinct cutaneous rashes. Dendritic cells (DC) are essential for inducing antiviral immune responses; however, the contribution of DC subsets to immune control during natural cutaneous VZV infection has not been investigated. Immunostaining showed that compared to normal skin, the proportion of cells expressing DC-SIGN (a dermal DC marker) or DC-LAMP and CD83 (mature DC markers) were not significantly altered in infected skin. In contrast, the frequency of Langerhans cells was significantly decreased in VZV-infected skin, whereas there was an influx of plasmacytoid DC, a potent secretor of type I interferon (IFN). Langerhans cells and plasmacytoid DC in infected skin were closely associated with VZV antigen-positive cells, and some Langerhans cells and plasmacytoid DC were VZV antigen positive. To extend these in vivo observations, both plasmacytoid DC (PDC) isolated from human blood and Langerhans cells derived from MUTZ-3 cells were shown to be permissive to VZV infection. In VZV-infected PDC cultures, significant induction of alpha IFN (IFN-alpha) did not occur, indicating the VZV inhibits the capacity of PDC to induce expression of this host defense cytokine. This study defines changes in the response of DC which occur during cutaneous VZV infection and implicates infection of DC subtypes in VZV pathogenesis. %Z FOR Codes: 110804 %0 Journal Article %~ PubMed %A Morton, April M %A Cunningham, Anthony L %A Diefenbach, Russell J %T Kinesin-1 plays a role in transport of SNAP-25 to the plasma membrane. %B Biochemical and Biophysical Research Communications %D 2010 %C United States %I Academic Press %V 391 %N 1 %P 388-393 %@ 0006-291X %X The cellular molecular motor kinesin-1 mediates the microtubule-dependent transport of a range of cargo. We have previously identified an interaction between the cargo binding domain of kinesin-1 heavy chain KIF5B and the membrane-associated SNARE proteins SNAP-25 and SNAP-23. In this study we further defined the minimal SNAP-25 binding domain in KIF5B to residues 874-894. Overexpression of a fragment of KIF5B (residues 594-910) resulted in significant colocalization with SNAP-25 with resulting blockage of the trafficking of SNAP-25 to the periphery of cells. This indicates that kinesin-1 facilitates the transport of SNAP-25 containing vesicles as a prerequisite to SNAP-25 driven membrane fusion events. %Z FOR Codes: 60108 %0 Journal Article %~ PubMed %A Cunningham, Anthony L %A Donaghy, Heather %A Harman, Andrew N %A Kim, Min %A Turville, Stuart G %T Manipulation of dendritic cell function by viruses. %B Current opinion in microbiology %D 2010 %C United Kingdom %I Elsevier Ltd. %V 13 %N 4 %P 524-9 %@ 1879-0364 %X Viruses manipulate the function of dendritic cells (DCs) to enhance their entry, spread, survival and transmission. This review summarises recently published work identifying how viruses alter the expression of receptors, antiviral molecules, disrupt signalling pathways, subvert trafficking pathways and even affect DC function via interactions with second or third cell types. Different viruses such as human immunodeficiency virus (HIV) and herpes viruses may have widely divergent and even opposite effects on DC function, determined by the need for transfer to a primary target cell, replication within the DC or various immunoevasive mechanisms. %Z FOR Codes: 801 %0 Journal Article %~ PubMed %A Brazzale, Anthony G %A Russell, Darren B %A Cunningham, Anthony L %A Taylor, Janette %A McBride, William J H %T Seroprevalence of herpes simplex virus type 1 and type 2 among the Indigenous population of Cape York, Far North Queensland, Australia. %B Sexual Health %D 2010 %C Australia %I CSIRO Publishing %V 7 %N 4 %P 453-459 %@ 1448-5028 %X The objective of this study was to obtain representative seroprevalence data for the Indigenous population of Far North Queensland by measuring the age- and sex-specific seroprevalence of the herpes simplex viruses (HSV-1 and HSV-2) in Cape York. %Z FOR Codes: 111706 111701 %0 Journal Article %~ PubMed %A Ko, Debbie H %A Cunningham, Anthony L %A Diefenbach, Russell J %T The major determinant for addition of tegument protein pUL48 (VP16) to capsids in herpes simplex virus type 1 is the presence of the major tegument protein pUL36 (VP1/2). %B Journal of virology %D 2010 %C United States %I American Society for Microbiology %V 84 %N 3 %P 1397-405 %@ 1098-5514 %X In this study a number of herpes simplex virus type 1 (HSV-1) proteins were screened, using a yeast-two-hybrid assay, for interaction with the tegument protein pUL48 (VP16). This approach identified interactions between pUL48 and the capsid proteins pUL19 (VP5), pUL38 (VP19C), and pUL35 (VP26). In addition, the previously identified interaction of pUL48 with the major tegument protein pUL36 (VP1/2) was confirmed. All of these interactions, except that of pUL35 and pUL48, could be confirmed using an in vitro pulldown assay. A subsequent pulldown assay with intact in vitro-assembled capsids, consisting of pUL19, pUL38, and pUL18 (VP23) with or without pUL35, showed no binding of pUL48, suggesting that the capsid/pUL48 interactions initially identified were more then likely not biologically relevant. This was confirmed by using a series of HSV-1 mutants lacking the gene encoding either pUL35, pUL36, or pUL37. For each HSV-1 mutant, analysis of purified deenveloped C capsids indicated that only in the absence of pUL36 was there a complete loss of capsid-bound pUL48, as well as pUL37. Collectively, this study shows for the first time that pUL36 is a major factor for addition of both pUL48 and pUL37, likely through a direct interaction of both with nonoverlapping sites in pUL36, to unenveloped C capsids during assembly of HSV-1. %Z FOR Codes: 60506 %0 Journal Article %~ PubMed %A Cunningham, Anthony L %A Abendroth, Allison %A Jones, Cheryl %A Nasr, Najla %A Turville, Stuart %T Viruses and Langerhans cells. %B Immunology and cell biology %D 2010 %C United Kingdom, Australia %I Nature Publishing Group %V 88 %N 4 %P 416-23 %@ 0818-9641 %X Langerhans cells (LCs) are the resident dendritic cells (DCs) of epidermis in human mucosal stratified squamous epithelium and the skin. A phenotypically similar DC has recently been discovered as a minor population in the murine dermis. In epidermis, LCs function as sentinel antigen-presenting cells that can capture invading viruses such as herpes simplex virus (HSV), varicella-zoster virus (VZV) and human immunodeficiency virus (HIV). This interaction between LCs and viruses results in highly variable responses, depending on the virus as discussed in this review. For example, HSV induces apoptosis in LCs but HIV does not. LCs seem to be the first in a complex chain of antigen presentation to T cells in lymph nodes for HSV and possibly VZV, or they transport virus to T cells, as described for HIV and maybe VZV. Together with epidermal keratinocytes they may also have a role in the initial innate immune response at the site of infection in the epidermis, although this is not fully known. The full spectrum of biological responses of LCs even to these viruses has yet to be understood and will require complementary studies in human LCs in vitro and in murine models in vivo. %Z FOR Codes: 110804 110707 %0 Journal Article %~ PubMed %A Donaghy, Heather %A Bosnjak, Lidija %A Harman, Andrew N %A Marsden, Valerie %A Tyring, Stephen K %A Meng, Tze-Chiang %A Cunningham, Anthony L %T A role for Plasmacytoid Dendritic Cells in the immune control of human recurrent herpes simplex. %B Journal of virology %D 2009 %C United States %I American Society for Microbiology %V 83 %N 0 %P 1952-61 %@ 1098-5514 %X Plasmacytoid dendritic cells (pDC) are an important component of the innate immune response, producing large amounts of alpha interferon in response to viral stimulation in vitro. Under noninflammatory conditions, pDC are not found in the skin and are restricted in location to the blood and lymph nodes. Therefore, their role in mucosal and cutaneous herpes simplex virus (HSV) infection has not been well-defined. In this study we show a role for human pDC in the immune response to HSV infection. First, by confocal microscopy we showed that pDC infiltrate the dermis of recurrent genital herpes simplex lesions at early and late phases, often at the dermo-epidermal junction. We then showed that pDC in vitro are resistant to HSV infection despite expressing the entry receptors CD111, CD112, and HVE-A. Within the lesions, pDC were found closely associated with CD3(+) lymphocytes and NK cells, especially those which were activated (CD69(+)). Furthermore, these HSV-exposed pDC were able to stimulate virus-specific autologous T-lymphocyte proliferation. We conclude from this work that pDC may contribute to the immune control of recurrent herpes virus infection in vivo. %Z FOR Codes: 110701 %0 Journal Article %~ PubMed %A Templeton, David J %A Jin, Fengyi %A Prestage, Garrett P %A Donovan, Basil %A Imrie, John C %A Kippax, Susan C %A Cunningham, Phillip H %A Kaldor, John M %A Mindel, Adrian %A Cunningham, Anthony L %A Grulich, Andrew E %T Circumcision and Risk of Sexually Transmissible Infections in a Community-Based Cohort of HIV-Negative Homosexual Men in Sydney, Australia. %B The Journal of infectious diseases %D 2009 %C United States %I University of Chicago Press %V 200 %N 12 %P 1813-9 %@ 0022-1899 %X Circumcision status was examined as an independent risk factor for sexually transmissible infections (STIs) in the Health in Men cohort of homosexual men in Sydney. %Z FOR Codes: 1103 1117 %0 Journal Article %~ PubMed %A Steain, Megan C %A Dwyer, Dominic E %A Hurt, Aeron C %A Kol, Chenda %A Saksena, Nitin K %A Cunningham, Anthony L %A Wang, Bin %T Detection of influenza A H1N1 and H3N2 mutations conferring resistance to oseltamivir using rolling circle amplification. %B Antiviral research %D 2009 %C Netherlands %I Elsevier BV %V 84 %N 3 %P 242-8 %@ 1872-9096 %X In the event of an influenza pandemic, the use of oseltamivir (OTV) will undoubtedly increase and therefore it is more likely that OTV-resistant influenza strains will also arise. OTV-resistance genotyping using sequence-based testing on viruses isolated in cell culture is time consuming and less likely to detect the low-level presence of drug-resistant virus populations. We have developed a novel rolling circle amplification (RCA) method to achieve the sensitive detection of OTV-resistant viruses from clinical specimens. Using artificially created templates, RCA could detect the presence of OTV-resistant mutations (N2: 119V, 292K, N1: 274Y) even if the population carrying the mutations was <1% of the total. By applying RCA to clinical samples, we identified the emergence of the 274Y mutation in one OTV-treated patient, as well as in seven individuals who were treatment-na??ve (confirming community transmission of 274Y-containing resistant influenza A H1N1). These results were further confirmed by neuraminidase region sequencing. In conclusion, RCA technology can provide rapid (<24 h), high-throughput diagnosis of OTV resistance mutations with a high specificity and sensitivity. %Z FOR Codes: 60502 1103 %0 Journal Article %~ PubMed %A Kelly, Barbara J %A Fraefel, Cornel %A Cunningham, Anthony L %A Diefenbach, Russell J %T Functional roles of the tegument proteins of herpes simplex virus type 1. %B Virus research %D 2009 %C Netherlands %I Elsevier BV %V 145 %N 2 %P 173-86 %@ 0168-1702 %X Herpes virions consist of four morphologically distinct structures, a DNA core, capsid, tegument, and envelope. Tegument occupies the space between the nucleocapsid (capsid containing DNA core) and the envelope. A combination of genetic, biochemical and proteomic analysis of alphaherpes virions suggest the tegument contains in the order of 20 viral proteins. Historically the tegument has been described as amorphous but increasing evidence suggests there is an ordered addition of tegument during assembly. This review highlights the diverse roles, in addition to structural, that tegument plays during herpes viral replication using as an example herpes simplex virus type 1. Such diverse roles include: capsid transport during entry and egress; targeting of the capsid to the nucleus; regulation of transcription, translation and apoptosis; DNA replication; immune modulation; cytoskeletal assembly; nuclear egress of capsid; and viral assembly and final egress. %Z FOR Codes: 110804 %0 Journal Article %~ PubMed %A Maddocks, Susan %A Scandurra, Gabriella M %A Nourse, Craig %A Bye, Chris %A Williams, Rohan B %A Slobedman, Barry %A Cunningham, Anthony L %A Britton, Warwick J %T Gene expression in HIV-1/Mycobacterium tuberculosis co-infected macrophages is dominated by M. tuberculosis. %B Tuberculosis (Edinburgh, Scotland) %D 2009 %C United Kingdom %I Churchill Livingstone %V 89 %N 4 %P 285-93 %@ 1873-281X %X The resurgence of tuberculosis worldwide has closely mirrored the HIV pandemic. In regions like sub-Saharan Africa, a large proportion of individuals are co-infected with Mycobacterium tuberculosis and HIV. Macrophages are the reservoir host cells for both pathogens, however the interactions between both pathogens in co-infected cells remain poorly understood. Thus, the global gene responses of primary human macrophages following productive co-infection with highly purified HIV and M. tuberculosis were analyzed using cDNA microarrays. A broad range of genes was up-regulated in response to co-infection or M. tuberculosis infection of primary macrophages, including those encoding pro-inflammatory chemokines and cytokines, their receptors, signalling associated genes, type I IFN signalling genes and genes of the tryptophan degradation pathway. Real-time RT-PCR analysis confirmed up-regulation of a wide variety of genes including indoleamine 2,3 dioxygenase and Sp110 in M. tuberculosis and co-infected samples. Downstream analysis confirmed significant elevation of the chemokines CCL3, CCL4 and CCL8 in M. tuberculosis and co-infected culture supernatants. In contrast, the changes seen in gene expression following HIV infection alone were fewer in number and significantly less in magnitude. Thus, the effects of M. tuberculosis infection on global gene expression dominated the effects of HIV-1 in co-infected primary human macrophages. %Z FOR Codes: 60501 110707 60506 %0 Journal Article %~ PubMed %A Harman, Andrew N %A Kraus, Marianne %A Bye, Chris R %A Byth, Karen %A Turville, Stuart G %A Tang, Owen %A Mercier, Sarah K %A Nasr, Najla %A Stern, Josh L %A Slobedman, Barry %A Driessen, Christoph %A Cunningham, Anthony L %T HIV-1-infected dendritic cells show 2 phases of gene expression changes, with lysosomal enzyme activity decreased during the second phase. %B Blood %D 2009 %C United States %I American Society of Hematology %V 114 %N 1 %P 85-94 %@ 1528-0020 %X Dendritic cells (DCs) play a key role in the pathogenesis of HIV infection. HIV interacts with these cells through 2 pathways in 2 temporal phases, initially via endocytosis and then via de novo replication. Here the transcriptional response of human DCs to HIV-1 was studied in these phases and at different stages of the virus replication cycle using purified HIV-1 envelope proteins, and inactivated and viable HIV-1. No differential gene expression was detected in response to envelope. However, more than 100 genes were differentially expressed in response to entry of viable and inactivated HIV-1 in the first phase. A completely different set of genes was differentially expressed in the second phase, predominantly in response to viable HIV-1, including up-regulation of immune regulation genes, whereas genes encoding lysosomal enzymes were down-regulated. Cathepsins B, C, S, and Z RNA and protein decreased, whereas cathepsin L was increased, probably reflecting a concomitant decrease in cystatin C. The net effect was markedly diminished cathepsin activity likely to result in enhanced HIV-1 survival and transfer to contacting T lymphocytes but decreased HIV-1 antigen processing and presentation to these T cells. %Z FOR Codes: 60107 %0 Journal Article %~ PubMed %A Miranda-Saksena, Monica %A Boadle, Ross A %A Aggarwal, Anupriya %A Tijono, Bibing %A Rixon, Frazer J %A Diefenbach, Russell J %A Cunningham, Anthony L %T Herpes simplex virus utilizes the large secretory vesicle pathway for anterograde transport of tegument and envelope proteins and for viral exocytosis from growth cones of human fetal axons. %B Journal of Virology %D 2009 %C United States %I American Society for Microbiology %V 83 %N 7 %P 3187-3199 %@ 0022-538X %X Axonal transport of herpes simplex virus (HSV-1) is essential for viral infection and spread in the peripheral nervous system of the host. Therefore, the virus probably utilizes existing active transport and targeting mechanisms in neurons for virus assembly and spread from neurons to skin. In the present study, we used transmission immunoelectron microscopy to investigate the nature and origin of vesicles involved in the anterograde axonal transport of HSV-1 tegument and envelope proteins and of vesicles surrounding partially and fully enveloped capsids in growth cones. This study aimed to elucidate the mechanism of virus assembly and exit from axons of human fetal dorsal root ganglia neurons. We demonstrated that viral tegument and envelope proteins can travel in axons independently of viral capsids and were transported to the axon terminus in two types of transport vesicles, tubulovesicular membrane structures and large dense-cored vesicles. These vesicles and membrane carriers were derived from the trans-Golgi network (TGN) and contained key proteins, such as Rab3A, SNAP-25, GAP-43, and kinesin-1, involved in the secretory and exocytic pathways in axons. These proteins were also observed on fully and partially enveloped capsids in growth cones and on extracellular virions. Our findings provide further evidence to the subassembly model of separate transport in axons of unenveloped capsids from envelope and tegument proteins with final virus assembly occurring at the axon terminus. We postulate that HSV-1 capsids invaginate tegument- and envelope-bearing TGN-derived vesicles and utilize the large secretory vesicle pathway of exocytosis for exit from axons. %Z FOR Codes: 60506 60108 %0 Journal Article %~ PubMed %A Stein, Alicia N %A Britt, Helena %A Harrison, Christopher %A Conway, E Lynne %A Cunningham, Anthony %A Macintyre, C Raina %T Herpes zoster burden of illness and health care resource utilisation in the Australian population aged 50 years and older. %B Vaccine %D 2009 %C United Kingdom %I Elsevier Ltd %V 27 %N 4 %P 520-529 %@ 0264-410X %X Incidence of zoster and post-herpetic neuralgia (PHN) and associated health care resource utilisation were investigated in the Australian population aged > or =50 years, using general practice data from 2000 to 2006, and pharmaceutical prescribing, hospital morbidity and emergency department data from 1998 to 2005. Zoster and PHN incidence rates were estimated as approximately 10/1000 and 1.45/1000 persons, respectively, with antivirals prescribed for 73.5% of zoster cases. Estimated hospitalisation and emergency department visit rates were 0.67/1000 and 0.38/1000 persons, respectively. Management of zoster (including PHN) involved approximately 2.4 general practitioner consultations. Total costs to the health care system were estimated as approximately 32.8 million per year. The substantial burden of zoster and PHN highlights the potential benefit of zoster vaccination. %Z FOR Codes: 111706 %0 Journal Article %~ PubMed %A Zaunders, John J %A Munier, Mee Ling %A Seddiki, Nabila %A Pett, Sarah %A Ip, Susanna %A Bailey, Michelle %A Xu, Yin %A Brown, Kai %A Dyer, Wayne B %A Kim, Min %A de Rose, Robert %A Kent, Stephen J %A Jiang, Lele %A Breit, Samuel N %A Emery, Sean %A Cunningham, Anthony L %A Cooper, David A %A Kelleher, Anthony D %T High Levels of Human Antigen-Specific CD4+ T Cells in Peripheral Blood Revealed by Stimulated Coexpression of CD25 and CD134 (OX40). %B Journal of immunology (Baltimore, Md. : 1950) %D 2009 %C United States %I American Association of Immunologists %V 183 %N 4 %P 2827-36 %@ 1550-6606 %X Ag-specific human CD4(+) memory T lymphocytes have mostly been studied using assays of proliferation in vitro. Intracellular cytokine and ELISPOT assays quantify effector cell populations but barely detect responses to certain recall Ags that elicit strong proliferative responses, e.g., tetanus toxoid, that comprise non-Th1 CD4(+) cells. We have found that culturing whole blood with Ag for 40-48 h induces specific CD4(+) T cells to simultaneously express CD25 and CD134. This new technique readily detects responses to well-described CD4(+) T cell recall Ags, including preparations of mycobacteria, CMV, HSV-1, influenza, tetanus toxoid, Candida albicans, and streptokinase, as well as HIV-1 peptides, with high specificity. The assay detects much higher levels of Ag-specific cells than intracellular cytokine assays, plus the cells retain viability and can be sorted for in vitro expansion. Furthermore, current in vitro assays for human CD4(+) memory T lymphocytes are too labor-intensive and difficult to standardize for routine diagnostic laboratories, whereas the whole-blood CD25(+)CD134(+) assay combines simplicity of setup with a straightforward cell surface flow cytometry readout. In addition to revealing the true extent of Ag-specific human CD4(+) memory T lymphocytes, its greatest use will be as a simple in vitro monitor of CD4(+) T cell responses to Ags such as tuberculosis infection or vaccines. %Z FOR Codes: 110704 110309 %0 Journal Article %~ PubMed %A Lai, Joey %A Bernhard, Oliver K %A Turville, Stuart G %A Harman, Andrew N %A Wilkinson, John %A Cunningham, Anthony L %T Oligomerization of the macrophage mannose receptor enhances gp120-mediated binding of HIV-1. %B Journal of Biological Chemistry %D 2009 %C United States %I American Society for Biochemistry and Molecular B %V 284 %N 17 %P 11027-11038 %@ 0021-9258 %X C-type lectin receptors expressed on the surface of dendritic cells and macrophages are able to bind glycoproteins of microbial pathogens via mannose, fucose, and N-acetylglucosamine. Langerin on Langerhans cells, dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin on dendritic cells, and mannose receptor (MR) on dendritic cells and macrophages bind the human immunodeficiency virus (HIV) envelope protein gp120 principally via high mannose oligosaccharides. These C-type lectin receptors can also oligomerize to facilitate enhanced ligand binding. This study examined the effect of oligomerization of MR on its ability to bind to mannan, monomeric gp120, native trimeric gp140, and HIV type 1 BaL. Mass spectrometry analysis of cross-linked MR showed homodimerization on the surface of primary monocyte-derived dendritic cells and macrophages. Both monomeric and dimeric MR were precipitated by mannan, but only the dimeric form was co-immunoprecipitated by gp120. These results were confirmed independently by flow cytometry analysis of soluble monomeric and trimeric HIV envelope and a cellular HIV virion capture assay. As expected, mannan bound to the carbohydrate recognition domains of MR dimers mostly in a calcium-dependent fashion. Unexpectedly, gp120-mediated binding of HIV to dimers on MR-transfected Rat-6 cells and macrophages was not calcium-dependent, was only partially blocked by mannan, and was also partially inhibited by N-acetylgalactosamine 4-sulfate. Thus gp120-mediated HIV binding occurs via the calcium-dependent, non-calcium-dependent carbohydrate recognition domains and the cysteine-rich domain at the C terminus of MR dimers, presenting a much broader target for potential inhibitors of gp120-MR binding. %Z FOR Codes: 605 %0 Journal Article %~ PubMed %A Wang, Bin %A Dwyer, Dominic E %A Chew, Choo Beng %A Kol, Chenda %A He, Zhong Ping %A Joshi, Hemal %A Steain, Megan C %A Cunningham, Anthony L %A Saksena, Nitin K %T Sensitive detection of the K103N non-nucleoside reverse transcriptase inhibitor resistance mutation in treatment-na??ve HIV-1 infected individuals by rolling circle amplification. %B Journal of virological methods %D 2009 %C Netherlands %I Elsevier BV %V 161 %N 1 %P 128-35 %@ 0166-0934 %X Primary or transmitted antiretroviral drug resistance mutations pose a significant obstacle for optimizing antiviral treatment. When present at low-levels, resistance mutations are less likely to be detected by standard genotyping assays. This study utilizes a novel rolling circle amplification (RCA) method using padlock probes to achieve the sensitive, specific and low-level detection of the NNRTI resistance K103N from 59 HIV+ treatment-na??ve patients from Beijing, China. Using standard genotyping methods, primary drug resistance mutations to either protease or RT inhibitors were found in 25% (15/59) of patients attending hospital clinics in Beijing. Among these 15 patients with antiretroviral (ARV) resistance mutations, standard sequence-based genotyping revealed that most (10/15) had the 103N. Using a highly sensitive RCA assay, 5 more patients among the 59 treatment-na??ve cohort were found to have the 103N, but at low-levels, leading to an overall rate of 103N at 25.4% (15/59) in this population. The high prevalence of the 103N suggests that baseline resistance testing should be performed before treatment in this population. Importantly, the new RCA technology allows large-scale, sensitive detection of drug resistance mutations, including detection of minority populations with minimal equipment requirement. %Z FOR Codes: 110804 %0 Journal Article %~ PubMed %A Cheung, Allen K L %A Gottlieb, David J %A Plachter, Bodo %A Pepperl-Klindworth, Sandra %A Avdic, Selmir %A Cunningham, Anthony L %A Abendroth, Allison %A Slobedman, Barry %T The role of the human cytomegalovirus UL111A gene in down-regulating CD4+ T-cell recognition of latently infected cells: implications for virus elimination during latency. %B Blood %D 2009 %C United States %I American Society of Hematology %V 114 %N 19 %P 4128-4137 %@ 0006-4971 %X The capacity of human cytomegalovirus (HCMV) to establish and maintain a latent infection from which it can later reactivate ensures its widespread distribution in the population, but the mechanisms enabling maintenance of latency in the face of a robust immune system are poorly understood. We examined the role of the HCMV UL111A gene, which encodes homologs of the immunosuppressive cytokine interleukin-10 in the context of latent infection of myeloid progenitor cells. A UL111A deletion virus was able to establish, maintain, and reactivate from experimental latency in a manner comparable with parental virus, but major histocompatibility complex class II levels increased significantly on the surfaces of cells infected with the deletion virus. Importantly, there was an increase in both allogeneic and autologous peripheral blood mononuclear cells and CD4(+) T-cell responses to UL111A deletion virus-infected myeloid progenitors, indicating that loss of the capacity to express viral interleukin-10 during latency results in latently infected cells becoming more readily recognizable by a critical arm of the immune response. The detection of a viral gene that suppresses CD4(+) T-cell recognition of latently infected cells identifies an immune evasion strategy that probably enhances the capacity of HCMV to persist in a latent state within the human host. %Z FOR Codes: 110804 %0 Journal Article %~ Isi %A Gray, L. %A Roche, M. %A Churchill, M. J. %A Sterjovski, J. %A Ellett, A. %A Poumbourios, P. %A Sheffief, S. %A Wang, B. %A Saksena, N. %A Purcell, D. F. J. %A Wesselingh, S. %A Cunningham, A. L. %A Brew, B. J. %A Gabuzda, D. %A Gorry, P. R. %T Tissue-Specific Sequence Alterations in the Human Immunodeficiency Virus Type 1 Envelope Favoring CCR5 Usage Contribute to Persistence of Dual-Tropic Virus in the Brain %B Journal of Virology %D 2009 %C United States %I American Society for Microbiology %V 83 %N 11 %P 5430-5441 %@ 1098-5514 %X %Z FOR Codes: 110804 %0 Journal Article %~ Isi %A Page, A. %A Taylor, R. %A Richters, J. %A Shaw, J. %A Taylor, J. %A Cunningham, A. %A Mindel, A. %T Upstairs and Downstairs Socio-Economic and Gender Interactions in Herpes Simplex Virus Type 2 Seroprevalence in Australia %B Sexually Transmitted Diseases %D 2009 %C United States %I Lippincott Williams & Wilkins %V 36 %N 6 %P 344-349 %@ 0148-5717 %X %Z FOR Codes: 110324 %0 Journal Article %~ PubMed %A Lee, Jin H %A Vittone, Valerio %A Diefenbach, Eve %A Cunningham, Anthony L %A Diefenbach, Russell J %T Identification of structural protein-protein interactions of herpes simplex virus type 1. %B Virology %D 2008 %C United States %I Academic Press %V 378 %N 2 %P 347-54 %@ 0042-6822 %X In this study we have defined protein-protein interactions between the structural proteins of herpes simplex virus type 1 (HSV-1) using a LexA yeast two-hybrid system. The majority of the capsid, tegument and envelope proteins of HSV-1 were screened in a matrix approach. A total of 40 binary interactions were detected including 9 out of 10 previously identified tegument-tegument interactions (Vittone, V., Diefenbach, E., Triffett, D., Douglas, M.W., Cunningham, A.L., and Diefenbach, R.J., 2005. Determination of interactions between tegument proteins of herpes simplex virus type 1. J. Virol. 79, 9566-9571). A total of 12 interactions involving the capsid protein pUL35 (VP26) and 11 interactions involving the tegument protein pUL46 (VP11/12) were identified. The most significant novel interactions detected in this study, which are likely to play a role in viral assembly, include pUL35-pUL37 (capsid-tegument), pUL46-pUL37 (tegument-tegument) and pUL49 (VP22)-pUS9 (tegument-envelope). This information will provide further insights into the pathways of HSV-1 assembly and the identified interactions are potential targets for new antiviral drugs. %Z FOR Codes: 110309 %0 Journal Article %~ PubMed %A Kim, Min %A Taylor, Janette %A Sidney, John %A Mikloska, Zorka %A Bodsworth, Neil %A Lagios, Katerina %A Dunckley, Heather %A Byth-Wilson, Karen %A Denis, Martine %A Finlayson, Robert %A Khanna, Rajiv %A Sette, Alessandro %A Cunningham, Anthony L %T Immunodominant Epitopes in Herpes Simplex Virus Type 2 Glycoprotein D Are Recognized by CD4 Lymphocytes from Both HSV-1 and HSV-2 Seropositive Subjects. %B Journal of Immunology %D 2008 %C United States %I American Association of Immunologists %V 181 %N 9 %P 6604-6615 %@ 0022-1767 %X In human recurrent cutaneous herpes simplex, there is a sequential infiltrate of CD4 and then CD8 lymphocytes into lesions. CD4 lymphocytes are the major producers of the key cytokine IFN-gamma in lesions. They recognize mainly structural proteins and especially glycoproteins D and B (gD and gB) when restimulated in vitro. Recent human vaccine trials using recombinant gD showed partial protection of HSV seronegative women against genital herpes disease and also, in placebo recipients, showed protection by prior HSV1 infection. In this study, we have defined immunodominant peptide epitopes recognized by 8 HSV1(+) and/or 16 HSV2(+) patients using (51)Cr-release cytotoxicity and IFN-gamma ELISPOT assays. Using a set of 39 overlapping 20-mer peptides, more than six immunodominant epitopes were defined in gD2 (two to six peptide epitopes were recognized for each subject). Further fine mapping of these responses for 4 of the 20-mers, using a panel of 9 internal 12-mers for each 20-mers, combined with MHC II typing and also direct in vitro binding assay of these peptides to individual DR molecules, showed more than one epitope per 20-mers and promiscuous binding of individual 20-mers and 12-mers to multiple DR types. All four 20-mer peptides were cross-recognized by both HSV1(+)/HSV2(-) and HSV1(-)/HSV2(+) subjects, but the sites of recognition differed within the 20-mers where their sequences were divergent. This work provides a basis for CD4 lymphocyte cross-recognition of gD2 and possibly cross-protection observed in previous clinical studies and in vaccine trials. %Z FOR Codes: 110701 %0 Journal Article %~ PubMed %A Cunningham, Anthony L %A Carbone, Francis %A Geijtenbeek, Teunis B H %T Langerhans cells and viral immunity. %B European journal of immunology %D 2008 %C Germany %I Wiley-V C H Verlag Gmbh %V 38 %N 9 %P 2377-85 %@ 0014-2980 %X Langerhans cells (LC) are a unique dendritic cell subset that are located in mucosal stratified squamous epithelium and skin epidermis. Their location is ideally suited for their function as antigen presenting cells that capture invading viruses and induce anti-viral immunity. However, it is becoming evident that the interaction between LC and viruses can result in different responses, depending on the virus and the receptors involved. Here we will discuss the recent data on the similarities and differences in roles of LC in viral immunity to and infection with HIV, herpes simplex and varicella-zoster virus. Although all three viruses interact with LC during initial infection, the effects can be quite different, reflecting differences in biology and pathogenesis. %Z FOR Codes: 110701 %0 Journal Article %~ PubMed %A Lidbury, Brett A %A Rulli, Nestor E %A Suhrbier, Andreas %A Smith, Paul N %A McColl, Shaun R %A Cunningham, Anthony L %A Tarkowski, Andrej %A van Rooijen, Nico %A Fraser, Robert J %A Mahalingam, Suresh %T Macrophage-Derived Proinflammatory Factors Contribute to the Development of Arthritis and Myositis after Infection with an Arthrogenic Alphavirus. %B The Journal of infectious diseases %D 2008 %C United States %I University of Chicago Press %V 197 %N 11 %P 1585-93 %@ 0022-1899 %X Alphaviruses, such as chikungunya virus and Ross River virus (RRV), are associated with outbreaks of infectious rheumatic disease in humans worldwide. Using an established mouse model of disease that mimics RRV disease in humans, we showed that macrophage-derived factors are critical in the development of striated muscle and joint tissue damage. Histologic analyses of muscle and ankle joint tissues demonstrated a substantial reduction in inflammatory infiltrates in infected mice depleted of macrophages (i.e., "macrophage-depleted mice"). Levels of the proinflammatory factors tumor necrosis factor-alpha, interferon-gamma, and macrophage chemoattractant protein-1 were also dramatically reduced in tissue samples obtained from infected macrophage-depleted mice, compared with samples obtained from infected mice without macrophage depletion. These factors were also detected in the synovial fluid of patients with RRV-induced polyarthritis. Neutralization of these factors reduced the severity of disease in mice, whereas blocking nuclear factor kappaB by treatment with sulfasalazine ameliorated RRV inflammatory disease and tissue damage. To our knowledge, these findings are the first to demonstrate that macrophage-derived products play important roles in the development of arthritis and myositis triggered by alphavirus infection. %Z FOR Codes: 110309 %0 Journal Article %~ PubMed %A Cunningham, Anthony L %A Breuer, Judith %A Dwyer, Dominic E %A Gronow, David W %A Helme, Robert D %A Litt, John C %A Levin, Myron J %A Macintyre, C Raina %T The prevention and management of herpes zoster. %B The Medical Journal of Australia %D 2008 %C Australia %I Australasian Medical Publishing Company Pty. Ltd %V 188 %N 3 %P 171-176 %@ 0025-729X %X The burden of illness from herpes zoster (HZ) and postherpetic neuralgia (PHN) in the Australian community is high. The incidence and severity of HZ and PHN increase with age in association with a progressive decline in cell-mediated immunity to varicella-zoster virus (VZV). Antiviral medications (valaciclovir, famciclovir, aciclovir) have been shown to be effective in reducing much but not all of the morbidity associated with HZ and PHN, but are consistently underprescribed in Australia. Zoster-associated pain should be treated early and aggressively, as it is more difficult to treat once established. Clinicians should be proactive in their follow-up of individuals at high risk of developing PHN, and refer patients to a specialist pain clinic earlier, rather than later. A live, attenuated VZV vaccine (Oka/Merck strain, Zostavax [Merck Sharp & Dohme]) has proven to be efficacious in reducing the incidence of and morbidity associated with HZ and PHN in older adults. The vaccine''s efficacy has been shown to persist for at least 4 years, but is likely to last a lot longer. Ongoing surveillance will determine the duration of protection and whether a booster dose is required. Clinicians should consider recommending the vaccine, which can be safely administered at the same time as the inactivated influenza vaccine, to all immunocompetent patients aged 60 years or older. Clinicians should refer to the Australian immunisation handbook for advice on the use of the live vaccine in immunosuppressed individuals. %Z FOR Codes: 110701 %0 Journal Article %~ PubMed %A Wu, Jing Qin %A Wang, Bin %A Belov, Larissa %A Chrisp, Jeremy %A Learmont, Jenny %A Dyer, Wayne B %A Zaunders, John %A Cunningham, Anthony L %A Dwyer, Dominic E %A Saksena, Nitin K %T Antibody microarray analysis of cell surface antigens on CD4+ and CD8+ T cells from HIV+ individuals correlates with disease stages. %B Retrovirology %D 2007 %C England %I BioMed Central Ltd %V 4 %N 83 %P 1-13 %@ 1742-4690 %X BACKGROUND: Expression levels of cell surface antigens such as CD38 and HLA-DR are related to HIV disease stages. To date, the immunophenotyping of cell surface antigens relies on flow cytometry, allowing estimation of 3-6 markers at a time. The recently described DotScan antibody microarray technology enables the simultaneous analysis of a large number of cell surface antigens. This new technology provides new opportunities to identify novel differential markers expressed or co-expressed on CD4+ and CD8+ T cells, which could aid in defining the stage of evolution of HIV infection and the immune status of the patient. RESULTS: Using this new technology, we compared cell surface antigen expression on purified CD4+ and CD8+ T cells between 3 HIV disease groups (long-term non-progressors controlling viremia naturally; HIV+ patients on highly active antiretroviral therapy (HAART) with HIV plasma viral loads <50 copies/ml; and HIV+ patients with viremia during HAART) and uninfected controls. Pairwise comparisons identified 17 statistically differential cell surface antigens including 5 novel ones (CD212b1, CD218a, CD183, CD3 epsilon and CD9), not previously reported. Notably, changes in activation marker expression were more pronounced in CD8+ T cells, whereas changes in the expression of cell membrane receptors for cytokines and chemokines were more pronounced in CD4+ T cells. CONCLUSION: Our study not only confirmed cell surface antigens previously reported to be related to HIV disease stages, but also identified 5 novel ones. Of these five, three markers point to major changes in responsiveness to certain cytokines, which are involved in Th1 responses. For the first time our study shows how density of cell surface antigens could be efficiently exploited in an array manner in relation to HIV disease stages. This new platform of identifying disease markers can be further extended to study other diseases. %Z FOR Codes: %0 Journal Article %~ PubMed %A Sterjovski, Jasminka %A Churchill, Melissa J %A Ellett, Anne %A Gray, Lachlan R %A Roche, Michael J %A Dunfee, Rebecca L %A Purcell, Damian F J %A Saksena, Nitin %A Wang, Bin %A Sonza, Secondo %A Wesselingh, Steven L %A Karlsson, Ingrid %A Fenyo, Eva-Maria %A Gabuzda, Dana %A Cunningham, Anthony L %A Gorry, Paul R %T Asn 362 in gp120 contributes to enhanced fusogenicity by CCR5-restricted HIV-1 envelope glycoprotein variants from patients with AIDS. %B Retrovirology %D 2007 %C United Kingdom %I BioMed Central Ltd. %V 4 %N %P 89 %@ 1742-4690 %X CCR5-restricted (R5) human immunodeficiency virus type 1 (HIV-1) variants cause CD4+ T-cell loss in the majority of individuals who progress to AIDS, but mechanisms underlying the pathogenicity of R5 strains are poorly understood. To better understand envelope glycoprotein (Env) determinants contributing to pathogenicity of R5 viruses, we characterized 37 full-length R5 Envs from cross-sectional and longitudinal R5 viruses isolated from blood of patients with asymptomatic infection or AIDS, referred to as pre-AIDS (PA) and AIDS (A) R5 Envs, respectively. %Z FOR Codes: 110804 %0 Book Section %A Cunningham, Anthony %A Wilkinson, John %A Turville, Stuart %A Robbiani, Melissa %T Binding and Uptake of HIV by Dendritic Cells and Transfer to T Lymphocytes: Implications for Pathogenesis %B The Biology of Dendritic Cells and HIV Infection %D 2007 %C United States %I Springer %V %N %P 381-404 %@ 9780387337845 %E Gessani, Sandra %E Belardelli, Filippo %X %Z FOR Codes: 110804 %0 Book %A Cunningham, Anthony %A Mindel, A %A Dwyer, Dominic %T Clinician's Manual on Genital Herpes, 3rd Ed. %B %D 2007 %C United Kingdom %I Current Medicine Group Ltd. %V %N %P %@ 9781858734187 %X %Z FOR Codes: 110309 %0 Journal Article %~ PubMed %A Cunningham, Anthony L %A Harman, Andrew N %A Donaghy, Heather %T DC-SIGN 'AIDS' HIV immune evasion and infection. %B Nature immunology %D 2007 %C United States %I Nature Publishing Group %V 8 %N 6 %P 556-558 %@ 1529-2908 %X %Z FOR Codes: 110702 %0 Journal Article %~ PubMed %A Watson, Sarah %A Mercier, Sarah %A Bye, Chris %A Wilkinson, John %A Cunningham, Anthony L %A Harman, Andrew N %T Determination of suitable housekeeping genes for normalisation of quantitative real time PCR analysis of cells infected with human immunodeficiency virus and herpes viruses. %B Virology journal %D 2007 %C Netherlands, UK, US. %I BioMed Central Ltd. %V 4 %N 0 %P 130 %@ 1743-422X %X The choice of an appropriate housekeeping gene for normalisation purposes has now become an essential requirement when designing QPCR experiments. This is of particular importance when using QPCR to measure viral and cellular gene transcription levels in the context of viral infections as viruses can significantly interfere with host cell pathways, the components of which traditional housekeeping genes often encode. In this study we have determined the reliability of 10 housekeeping genes in context of four heavily studied viral infections; human immunodeficiency virus type 1, herpes simplex virus type 1, cytomegalovirus and varicella zoster virus infections using a variety of cell types and virus strains. This provides researchers of these viruses with a shortlist of potential housekeeping genes to use as normalisers for QPCR experiments. %Z FOR Codes: 110105 %0 Journal Article %~ PubMed %A Gorry, Paul R %A McPhee, Dale A %A Verity, Erin %A Dyer, Wayne B %A Wesselingh, Steven L %A Learmont, Jennifer %A Sullivan, John S %A Roche, Michael %A Zaunders, John J %A Gabuzda, Dana %A Crowe, Suzanne M %A Mills, John %A Lewin, Sharon R %A Brew, Bruce J %A Cunningham, Anthony L %A Churchill, Melissa J %T Pathogenicity and immunogenicity of attenuated, nef-deleted HIV-1 strains in vivo. %B Retrovirology %D 2007 %C United Kingdom %I BioMed Central Ltd. %V 4 %N 0 %P 66 %@ 1742-4690 %X In efforts to develop an effective vaccine, sterilizing immunity to primate lentiviruses has only been achieved by the use of live attenuated viruses carrying major deletions in nef and other accessory genes. Although live attenuated HIV vaccines are unlikely to be developed due to a myriad of safety concerns, opportunities exist to better understand the correlates of immune protection against HIV infection by studying rare cohorts of long-term survivors infected with attenuated, nef-deleted HIV strains such as the Sydney blood bank cohort (SBBC). Here, we review studies of viral evolution, pathogenicity, and immune responses to HIV infection in SBBC members. The studies show that potent, broadly neutralizing anti-HIV antibodies and robust CD8+ T-cell responses to HIV infection were not necessary for long-term control of HIV infection in a subset of SBBC members, and were not sufficient to prevent HIV sequence evolution, augmentation of pathogenicity and eventual progression of HIV infection in another subset. However, a persistent T-helper proliferative response to HIV p24 antigen was associated with long-term control of infection. Together, these results underscore the importance of the host in the eventual outcome of infection. Thus, whilst generating an effective antibody and CD8+ T-cell response are an essential component of vaccines aimed at preventing primary HIV infection, T-helper responses may be important in the generation of an effective therapeutic vaccine aimed at blunting chronic HIV infection. %Z FOR Codes: 110804 %0 Journal Article %~ PubMed %A Gray, Lachlan %A Churchill, Melissa J %A Sterjovski, Jasminka %A Witlox, Kristie %A Learmont, Jennifer C %A Sullivan, John S %A Wesselingh, Steven L %A Gabuzda, Dana %A Cunningham, Anthony L %A McPhee, Dale A %A Gorry, Paul R %T Phenotype and envelope gene diversity of nef-deleted HIV-1 isolated from long-term survivors infected from a single source. %B Virology journal %D 2007 %C United Kingdom %I BioMed Central Ltd. %V 4 %N %P 75 %@ 1743-422X %X BACKGROUND: The Sydney blood bank cohort (SBBC) of long-term survivors consists of multiple individuals infected with attenuated, nef-deleted variants of human immunodeficiency virus type 1 (HIV-1) acquired from a single source. Long-term prospective studies have demonstrated that the SBBC now comprises slow progressors (SP) as well as long-term nonprogressors (LTNP). Convergent evolution of nef sequences in SBBC SP and LTNP indicates the in vivo pathogenicity of HIV-1 in SBBC members is dictated by factors other than nef. To better understand mechanisms underlying the pathogenicity of nef-deleted HIV-1, we examined the phenotype and env sequence diversity of sequentially isolated viruses (n = 2) from 3 SBBC members. RESULTS: The viruses characterized here were isolated from two SP spanning a three or six year period during progressive HIV-1 infection (subjects D36 and C98, respectively) and from a LTNP spanning a two year period during asymptomatic, nonprogressive infection (subject C18). Both isolates from D36 were R5X4 phenotype and, compared to control HIV-1 strains, replicated to low levels in peripheral blood mononuclear cells (PBMC). In contrast, both isolates from C98 and C18 were CCR5-restricted. Both viruses isolated from C98 replicated to barely detectable levels in PBMC, whereas both viruses isolated from C18 replicated to low levels, similar to those isolated from D36. Analysis of env by V1V2 and V3 heteroduplex tracking assay, V1V2 length polymorphisms, sequencing and phylogenetic analysis showed distinct intra- and inter-patient env evolution. CONCLUSION: Independent evolution of env despite convergent evolution of nef may contribute to the in vivo pathogenicity of nef-deleted HIV-1 in SBBC members, which may not necessarily be associated with changes in replication capacity or viral coreceptor specificity. %Z FOR Codes: 110702 %0 Journal Article %~ PubMed %A Gowrishankar, Kavitha %A Slobedman, Barry %A Cunningham, Anthony L %A Miranda-Saksena, Monica %A Boadle, Ross A %A Abendroth, Allison %T Productive varicella-zoster virus infection of cultured intact human ganglia. %B Journal of virology %D 2007 %C US %I American Society for Microbiology %V 81 %N 12 %P 6752-6756 %@ 0022-538X %X Varicella-zoster virus (VZV) is a species-specific herpesvirus which infects sensory ganglia. We have developed a model of infection of human intact explant dorsal root ganglia (DRG). Following exposure of DRG to VZV, viral antigens were detected in neurons and nonneuronal cells. Enveloped virions were visualized by transmission electron microscopy in neurons and nonneuronal cells and within the extracellular space. Moreover, rather than remaining highly cell associated during infection of cultured cells, such as fibroblasts, cell-free VZV was released from infected DRG. This model enables VZV infection of ganglionic cells to be studied in the context of intact DRG. %Z FOR Codes: 1107 %0 Journal Article %~ PubMed %A Mijatov, Branka %A Cunningham, Anthony L %A Diefenbach, Russell J %T Residues F593 and E596 of HSV-1 tegument protein pUL36 (VP1/2) mediate binding of tegument protein pUL37. %B Virology %D 2007 %C United States %I Academic Press %V 368 %N 1 %P 26-31 %@ 0042-6822 %X The herpes simplex virus type 1 (HSV-1) structural tegument proteins pUL36 (VP1/2) and pUL37 are essential for secondary envelopment during the egress of viral particles. Our laboratory has previously shown that HSV-1 pUL36(512-767) fragment interacts with full-length pUL37. A number of single and double amino acid changes of conserved residues in the pUL36(512-767) fragment were generated using alanine-scanning site-directed mutagenesis. The interaction of pUL36(512-767) and pUL37 was then assessed using a combination of yeast two-hybrid and coimmunoprecipitation assays. Single changes to alanine of pUL36 residues F593 and E596 impaired binding of pUL37 with the greatest effect observed for the substitution E596A. Double mutations involving either of these residues in combination with the substitution E580A essentially blocked binding of pUL37. This information will provide the basis for generation of viral mutants to further define the importance of the pUL36/pUL37 interaction in assembly of HSV-1. %Z FOR Codes: 110804 %0 Journal Article %~ PubMed %A Diefenbach, Russell J %A Miranda-Saksena, Monica %A Douglas, Mark W %A Cunningham, Anthony L %T Transport and egress of herpes simplex virus in neurons. %B Reviews in medical virology %D 2007 %C United Kingdom %I John Wiley & Sons Ltd. %V 18 %N 1 %P 35-51 %@ 1052-9276 %X The mechanisms of axonal transport of the alphaherpesviruses, HSV and pseudorabies virus (PrV), in neuronal axons are of fundamental interest, particularly in comparison with other viruses, and offer potential sites for antiviral intervention or development of gene therapy vectors. These herpesviruses are transported rapidly along microtubules (MTs) in the retrograde direction from the axon terminus to the dorsal root ganglion and then anterogradely in the opposite direction. Retrograde transport follows fusion and deenvelopment of the viral capsid at the axonal membrane followed by loss of most of the tegument proteins and then binding of the capsid via one or more viral proteins (VPs) to the retrograde molecular motor dynein. The HSV capsid protein pUL35 has been shown to bind to the dynein light chain Tctex1 but is likely to be accompanied by additional dynein binding of an inner tegument protein. The mechanism of anterograde transport is much more controversial with different processes being claimed for PrV and HSV: separate transport of HSV capsid/tegument and glycoproteins versus PrV transport as an enveloped virion. The controversy has not been resolved despite application, in several laboratories, of confocal microscopy (CFM), real-time fluorescence with viruses dual labelled on capsid and glycoprotein, electron microscopy in situ and immuno-electron microscopy. Different processes for each virus seem counterintuitive although they are the most divergent in the alphaherpesvirus subfamily. Current hypotheses suggest that unenveloped HSV capsids complete assembly in the axonal growth cones and varicosities, whereas with PrV unenveloped capsids are only found travelling in a retrograde direction. %Z FOR Codes: 110804 %0 Journal Article %~ PubMed %A Verity, Erin E %A Zotos, Dimitra %A Wilson, Kim %A Chatfield, Catherine %A Lawson, Victoria A %A Dwyer, Dominic E %A Cunningham, Anthony %A Learmont, Jennifer %A Dyer, Wayne %A Sullivan, John %A Churchill, Melissa %A Wesselingh, Steven L %A Gabuzda, Dana %A Gorry, Paul R %A McPhee, Dale A %T Viral phenotypes and antibody responses in long term survivors infected with attenuated human immunodeficiency virus type 1 containing deletions in the nef and long terminal repeat region. %B Journal of Virology %D 2007 %C US %I American Society for Microbiology %V 81 %N 17 %P 9268-9278 %@ 0022-538X %X The Sydney Blood Bank Cohort (SBBC) consists of eight blood transfusion recipients infected with nef-attenuated human immunodeficiency virus type 1 (HIV-1) acquired from a single donor. Here, we show that viral phenotypes and antibody responses differ considerably between individual cohort members, despite the single source of infection. Replication of isolated virus varied from barely detectable to similar to that of the wild-type virus, and virus isolated from five SBBC members showed coreceptor usage signatures unique to each individual. Higher viral loads and stronger neutralizing antibody responses were associated with better-replicating viral strains, and detectable viral replication was essential for the development of strong and sustained humoral immune responses. Despite the presence of strong neutralizing antibodies in a number of SBBC members, disease progression was not prevented, and each cohort member studied displayed a unique outcome of infection with nef-attenuated HIV-1. %Z FOR Codes: 110705