bluuebunny/crossref_metadata_2025_split huggingface dataset:
Intranasal Delivery of an IgA Monoclonal Antibody Effective against Sublethal H5N1 Influenza Virus Infection in Mice
Authors: Ye Jianqiang , Shao Hongxia , Hickman Danielle , Angel Matthew , Xu Kemin , Cai Yibin , Song Haichen , Fouchier Ron A. M. , Qin Aijian , Perez Daniel R. 
Published: July 2010

ABSTRACTHighly pathogenic avian H5N1 influenza viruses are endemic in poultry in Asia and pose a pandemic threat to humans. Since the deployment of vaccines against a pandemic strain may take several months, adequate antiviral alternatives are needed to minimize the effects and the spread of the disease. Passive immunotherapy is regarded as a viable alternative. Here, we show the development of an IgA monoclonal antibody (DPJY01 MAb) specific to H5 hemagglutinin. The DPJY01 MAb showed a broad hemagglutination inhibition (HI) profile against Asian H5N1 viruses of clades 0, 1.0, 2.1, 2.2, and 2.3 and also against H5 wild bird influenza viruses of the North American and Eurasian lineages. DPJY01 MAb displayed also high neutralization activityin vitroandin vivo. In mice, DPJY01 MAb provided protection via a single dose administered intranasally before or after inoculation with a sublethal dose of H5N1 viruses of clades 1.0 and 2.2. Pretreatment with 50 mg of DPJY01 MAb kg of body weight at either 24, 48, or 72 h before highly pathogenic H5N1 virus (A/Vietnam/1203/2004 [H5N1]) inoculation resulted in complete protection. Treatment with 50 mg/kg at either at 24, 48, or 72 h after H5N1 inoculation provided 100%, 80%, and 60% protection, respectively. These studies highlight the potential use of DPJY01 MAb as an intranasal antiviral treatment for H5N1 influenza virus infections.
Mycobacterium avium
Complex Cervical Lymphadenitis in an Immunocompetent Adult
Authors: Christensen Joshua B. , Koeppe John 
Published: July 2010

ABSTRACT

Nontuberculosis mycobacterial cervical lymphadenitis is a relatively common disease in immunocompetent children but a rare disease in immunocompetent adults. We report the diagnosis and treatment of
Mycobacterium avium
complex cervical lymphadenitis in an adult female. Our evaluation of immune competence, including gamma interferon (IFN-γ) and interleukin-12 (IL-12) signaling, found no evidence of deficiency.
Screening of Predicted Secreted Antigens from
Mycobacterium bovis
Identifies Potential Novel Differential Diagnostic Reagents
Authors: Jones Gareth J. , Hewinson R. Glyn , Vordermeier H. Martin 
Published: July 2010

ABSTRACT

To date, the most promising vaccination strategies for the control of bovine tuberculosis (TB) focus on improving the efficacy of
M
ycobacterium
bovis
bacillus Calmette-Guérin (BCG). However, vaccination with BCG results in sensitization of animals to bovine tuberculin and compromises tests currently used for diagnosis of bovine TB infection. Thus, the development of specific diagnostic reagents capable of discriminating between infected and uninfected vaccinated animals (DIVA) is of high priority. To test the hypothesis that
M. bovis
-secreted proteins are likely to contain immunogenic antigens that can be used to increase the specificity of diagnostic tests, we screened 379 pools of overlapping peptides representing 119 antigens for their ability to stimulate a gamma inferferon (IFN-γ) response
in vitro
using whole blood from both TB reactor and BCG-vaccinated animals. Peptide pools representing antigens Rv3020c and Rv2346c induced responses in 61% and 57% of the TB reactor animals, respectively, without inducing responses in any BCG-vaccinated animal studied. Furthermore, individual peptides contained within pools recognized by BCG vaccinates were identified that were specific and induced IFN-γ responses in TB reactor animals. From these results, we constructed a cocktail of nine peptides representing multiple antigen targets that was recognized by 54% of TB reactor animals but also failed to induce responses in any BCG-vaccinated animal studied. In summary, we have identified three peptide cocktails for prioritization in larger trials to discriminate between
M. bovis
infection and BCG vaccination.
Multicenter Study Evaluating Performance of the Smart Group B Streptococcus (GBS) Assay Using an Enrichment Protocol for Detecting GBS Colonization in Patients in the Antepartum Period
Authors: Jordan Jeanne A. , Hall Geraldine , Davis Thomas 
Published: July 2010

ABSTRACT
Group B streptococcus (GBS) remains the leading cause of infectious morbidity and mortality in infants born in the United States, especially among black infants. Because a newborn can acquire GBS during and after delivery, the Centers for Disease Control and Prevention (CDC) recommends that pregnant women be screened for rectovaginal GBS colonization during the antepartum period between weeks 35 and 37 of gestation and, if they are colonized, that intrapartum antibiotic prophylaxis be administered. A prospective investigational study was undertaken from 2 May 2006 to 14 August 2006 at three sites to establish the performance characteristics of the Smart GBS LB assay on the SmartCycler II system for detecting GBS colonization in subjects in the antepartum period from combined vaginal/rectal swab-based specimens after broth enrichment. Results were compared to broth enrichment culture and to the predicate device, the BD GeneOhm StrepB direct assay. The collected specimens were randomized for swab testing order. Each swab sample was processed simultaneously by culture, Smart GBS LB assay, and the BD GeneOhm StrepB assay. A total of 310 subjects were enrolled, with 306 subject results included in the study. Compared to enrichment culture, the Smart GBS LB assay demonstrated a sensitivity, specificity, positive predictive value, and negative predictive value of 98.6%, 90.4%, 77.1%, and 99.5%, respectively. The Smart GBS LB assay demonstrated substantially equivalent or better performance than culture or the predicate device. Screening of broth enrichment fluids by nucleic acid amplification testing requires careful handling during sample processing to avoid possible contamination.
Agar-Based Disk Diffusion Assay for Susceptibility Testing of Dermatophytes
Authors: Nweze E. I. , Mukherjee P. K. , Ghannoum M. A. 
Published: July 2010

ABSTRACT
Currently, no agar-based susceptibility testing method has been standardized for testing dermatophytes. We describe a newly developed agar-based method employing disk diffusion assay to test the susceptibility of 47 isolates of dermatophytes against 8 antifungals. Our results show that the method is reproducible, is simple, and could be used to determine the antifungal susceptibility of dermatophytes.
Comparison of Citrated Human Blood, Citrated Sheep Blood, and Defibrinated Sheep Blood Mueller-Hinton Agar Preparations for Antimicrobial Susceptibility Testing of
Streptococcus pneumoniae
Isolates
Authors: Satzke Catherine , Seduadua Anna , Chandra Reginald , Carapetis Jonathan R. , Mulholland E. Kim , Russell Fiona M. 
Published: July 2010

ABSTRACT

The use of Mueller-Hinton agar supplemented with citrated human or citrated sheep blood was compared with the use of routinely used Mueller-Hinton agar supplemented with defibrinated sheep blood for antimicrobial susceptibility testing of
Streptococcus pneumoniae
. The alternate supplements were found to be unsatisfactory, particularly for testing resistant isolates, and therefore are not recommended.
Novel Phosphoprotein-Interacting Region in Nipah Virus Nucleocapsid Protein and Its Involvement in Viral Replication
Authors: Omi-Furutani Mio , Yoneda Misako , Fujita Kentaro , Ikeda Fusako , Kai Chieko 
Published: July 2010

ABSTRACT
The interaction of Nipah virus (NiV) nucleocapsid (N) protein with phosphoprotein (P) during nucleocapsid assembly is the essential process in the viral life cycle, since only the encapsidated RNA genome can be used for replication. To identify the region responsible for N-P interaction, we utilized fluorescent protein tags to visualize NiV N and P proteins in live cells and analyzed their cellular localization. N protein fused to monomeric enhanced cyan fluorescence protein (N-ECFP) exhibited a dotted pattern in transfected cells, while P protein fused to monomeric red fluorescent protein (P-mRFP) showed diffuse distribution. When the two proteins were coexpressed, P-mRFP colocalized with N-ECFP dots. N-ECFP mutants with serial amino acid deletions were generated to search for the region(s) responsible for this N-P colocalization. We found that, in addition to the 467- to 496-amino-acid (aa) region reported previously, aa 135 to 146 were responsible for the N-P colocalization. The residues crucial for N-P interaction were further investigated by introducing alanine substitutions into the untagged N protein. Alanine scanning in the region of aa 135 to 146 has revealed that there are distinct regions essential for the interaction of N-P and the function of N. This is the first study to visualize Nipah viral proteins in live cells and to assess the essential domain of N protein for the interaction with P protein.
P Body-Associated Protein Mov10 Inhibits HIV-1 Replication at Multiple Stages
Authors: Burdick Ryan , Smith Jessica L. , Chaipan Chawaree , Friew Yeshitila , Chen Jianbo , Venkatachari Narasimhan J. , Delviks-Frankenberry Krista A. , Hu Wei-Shau , Pathak Vinay K. 
Published: July 2010

ABSTRACT
Recent studies have shown that APOBEC3G (A3G), a potent inhibitor of human immunodeficiency virus type 1 (HIV-1) replication, is localized to cytoplasmic mRNA-processing bodies (P bodies). However, the functional relevance of A3G colocalization with P body marker proteins has not been established. To explore the relationship between HIV-1, A3G, and P bodies, we analyzed the effects of overexpression of P body marker proteins Mov10, DCP1a, and DCP2 on HIV-1 replication. Our results show that overexpression of Mov10, a putative RNA helicase that was previously reported to belong to the DExD superfamily and was recently reported to belong to the Upf1-like group of helicases, but not the decapping enzymes DCP1a and DCP2, leads to potent inhibition of HIV-1 replication at multiple stages. Mov10 overexpression in the virus producer cells resulted in reductions in the steady-state levels of the HIV-1 Gag protein and virus production; Mov10 was efficiently incorporated into virions and reduced virus infectivity, in part by inhibiting reverse transcription. In addition, A3G and Mov10 overexpression reduced proteolytic processing of HIV-1 Gag. The inhibitory effects of A3G and Mov10 were additive, implying a lack of functional interaction between the two inhibitors. Small interfering RNA (siRNA)-mediated knockdown of endogenous Mov10 by 80% resulted in a 2-fold reduction in virus production but no discernible impact on the infectivity of the viruses after normalization for the p24 input, suggesting that endogenous Mov10 was not required for viral infectivity. Overall, these results show that Mov10 can potently inhibit HIV-1 replication at multiple stages.
Introduction of the Six Major Genomic Deletions of Modified Vaccinia Virus Ankara (MVA) into the Parental Vaccinia Virus Is Not Sufficient To Reproduce an MVA-Like Phenotype in Cell Culture and in Mice
Authors: Meisinger-Henschel Christine , Späth Michaela , Lukassen Susanne , Wolferstätter Michael , Kachelriess Heike , Baur Karen , Dirmeier Ulrike , Wagner Markus , Chaplin Paul , Suter Mark , Hausmann Jürgen 
Published: July 2010

ABSTRACT

Modified vaccinia virus Ankara (MVA) has a highly restricted host range in cell culture and is apathogenic
in vivo
. MVA was derived from the parental chorioallantois vaccinia virus Ankara (CVA) by more than 570 passages in chicken embryo fibroblast (CEF) cells. During CEF cell passaging, six major deletions comprising 24,668 nucleotides occurred in the CVA genome. We have cloned both the MVA and the parental CVA genome as bacterial artificial chromosomes (BACs) and have sequentially introduced the six major MVA deletions into the cloned CVA genome. Reconstituted mutant CVA viruses containing up to six major MVA deletions showed no detectable replication restriction in 12 of 14 mammalian cell lines tested; the exceptions were rabbit cell lines RK13 and SIRC. In mice, CVA mutants with up to three deletions showed slightly enhanced virulence, suggesting that gene deletion in replicating vaccinia virus (VACV) can result in gain of fitness
in vivo
. CVA mutants containing five or all six deletions were still pathogenic, with a moderate degree of attenuation. Deletion V was mainly responsible for the attenuated phenotype of these mutants. In conclusion, loss or truncation of all 31 open reading frames in the six major deletions is not sufficient to reproduce the specific MVA phenotype of strong attenuation and highly restricted host range. Mutations in viral genes outside or in association with the six major deletions appear to contribute significantly to this phenotype. Host range restriction and avirulence of MVA are most likely a cooperative effect of gene deletions and mutations involving the major deletions.
Estimating Frequencies of Minority Nevirapine-Resistant Strains in Chronically HIV-1-Infected Individuals Naïve to Nevirapine by Using Stochastic Simulations and a Mathematical Model
Authors: Gadhamsetty Saikrishna , Dixit Narendra M. 
Published: July 2010

ABSTRACTNevirapine forms the mainstay of our efforts to curtail the pediatric AIDS epidemic through prevention of mother-to-child transmission of HIV-1. A key limitation, however, is the rapid selection of HIV-1 strains resistant to nevirapine following the administration of a single dose. This rapid selection of resistance suggests that nevirapine-resistant strains preexist in HIV-1 patients and may adversely affect outcomes of treatment. The frequencies of nevirapine-resistant strainsin vivo, however, remain poorly estimated, possibly because they exist as a minority below current assay detection limits. Here, we employ stochastic simulations and a mathematical model to estimate the frequencies of strains carrying different combinations of the common nevirapine resistance mutations K103N, V106A, Y181C, Y188C, and G190A in chronically infected HIV-1 patients naïve to nevirapine. We estimate the relative fitness of mutant strains from an independent analysis of previous competitive growth assays. We predict that single mutants are likely to preexist in patients at frequencies (∼0.01% to 0.001%) near or below current assay detection limits (>0.01%), emphasizing the need for more-sensitive assays. The existence of double mutants is subject to large stochastic variations. Triple and higher mutants are predicted not to exist. Our estimates are robust to variations in the recombination rate, cellular superinfection frequency, and the effective population size. Thus, with 107to 108infected cells in HIV-1 patients, even when undetected, nevirapine-resistant genomes may exist in substantial numbers and compromise efforts to prevent mother-to-child transmission of HIV-1, accelerate the failure of subsequent antiretroviral treatments, and facilitate the transmission of drug resistance.
Lipids and Proteins Act in Opposing Manners To Regulate Polyomavirus Infection
Authors: Qian Mengding , Tsai Billy 
Published: July 2010

ABSTRACT
How receptors control virus infection is poorly understood. Polyomavirus (Py) binds to the sialic acid-galactose moiety on receptors to gain entry into host cells and cause infection. We previously demonstrated that the sialic acid-galactose-containing glycolipids called gangliosides GD1a and GT1b promote Py infection, in part, by sorting the virus from the endolysosomes to the endoplasmic reticulum (ER), a critical infection route. Whether these glycolipids act as Py entry receptors, however, is not clear. Additionally, as the majority of glycoproteins also harbor terminal sialic acid-galactose residues, their roles in Py infection are also not well established. Using a ganglioside-deficient cell line, we show that GD1a is the functional entry receptor for Py. GD1a binds to Py on the plasma membrane, and the receptor-virus complex is internalized and transported to the late endosomes and then the ER to initiate infection. In contrast, our findings indicate that glycoproteins act as decoy receptors, restricting the ER transport and infection of Py. Thus, glycolipids and glycoproteins, two major constituents of the plasma membrane, execute opposing functions in regulating infection by a defined virus.
Comparison of Immune Restoration in Early versus Late Alpha Interferon Therapy against Hepatitis C Virus
Authors: Abdel-Hakeem Mohamed S. , Bédard Nathalie , Badr Gamal , Ostrowski Mario , Sékaly Rafick P. , Bruneau Julie , Willems Bernard , Heathcote E. Jenny , Shoukry Naglaa H. 
Published: July 2010

ABSTRACTEarly alpha interferon (IFN-α) therapy against hepatitis C virus (HCV) rescues polyfunctional, virus-specific memory CD8+T cells, but whether immune restoration is possible during late therapy remains controversial. We compared immune restoration of HCV-specific memory T cells in patients who cleared HCV infection spontaneously and following early or late IFN therapy. Multifunctional CD4+and CD8+memory T cells were detected in spontaneous resolvers and in individuals treated early following an acute infection. In contrast, limited responses were detected in patients treated during chronic infection, and the phenotype of HCV-specific cells was influenced by autologous viral sequences. Our data suggest that irreversible damage to the HCV-specific memory T-cell response is associated with chronic HCV infection.
Role of the Innate Immune Response and Tumor Immunity Associated with Simian Virus 40 Large Tumor Antigen
Authors: Lowe Devin B. , Shearer Michael H. , Aldrich Joel F. , Winn Richard E. , Jumper Cynthia A. , Kennedy Ronald C. 
Published: July 2010

ABSTRACTWe examined properties of the innate immune response against the tumor-specific antigen simian virus 40 (SV40) large tumor antigen (Tag) following experimental pulmonary metastasis in naive mice. Approximately 14 days after mKSA tumor cell challenge, expression of inflammatory mediators such as tumor necrosis factor alpha (TNF-α), interleukin-2 (IL-2), and RANTES was upregulated in splenocytes harvested from mice, as assessed by flow cytometry and antibody array assays. This response was hypothesized to activate and induce tumor-directed NK cell lysis since IL-2-stimulated NK cells mediated tumor cell destructionin vitro. The necessary function of NK cells was further validatedin vivothrough selected antibody depletion of NK cells, which resulted in an overwhelming lung tumor burden relative to that in animals receiving a control rabbit IgG depletion regimen. Interestingly, mice achieved increased protection from experimental pulmonary metastasis when NK cells were further activated indirectly throughin vivoadministration of poly(I:C), a Toll-like receptor 3 (TLR3) agonist. In a separate study, mice receiving treatments of poly(I:C) and recombinant SV40 Tag protein immunization mounted effective tumor immunity in an established experimental pulmonary metastasis setting. Initiating broad-based immunity with poly(I:C) was observed to induce a Th1 bias in the SV40 Tag antibody response that led to successful antitumor responses not observed in animals treated only with poly(I:C) or SV40 Tag. These data have direct implications for immunotherapeutic strategies incorporating methods to elicit inflammatory reactions, particularly NK cell-driven lysis, against malignant cell types that express a tumor-specific antigen such as SV40 Tag.
Analysis of A47, an Immunoprevalent Protein of Vaccinia Virus, Leads to a Reevaluation of the Total Antiviral CD8+T Cell Response
Authors: Yuen Tracy J. , Flesch Inge E. A. , Hollett Natasha A. , Dobson Bianca M. , Russell Tiffany A. , Fahrer Aude M. , Tscharke David C. 
Published: July 2010

ABSTRACTVaccinia virus (VACV) is the prototypic orthopoxvirus and was the live vaccine used to eradicate smallpox. In addition, VACV is a possible vector for recombinant vaccines. Despite these reasons for study, the roles of many VACV genes are unknown, and some fundamental aspects, such as the total size of immune responses, remain poorly characterized. VACV gene A47L is of interest because it is highly transcribed, has no sequence similarity to any nonpoxvirus gene, and contains a larger-than-expected number of CD8+T cell epitopes. Here it is shown that A47L is not required for growthin vitroand does not contribute to virulence in mice. However, we confirmed that this one protein primes CD8+T cells to three different epitopes in C57BL/6 mice. In the process, one of these epitopes was redefined and shown to be the most dominant in A47 and one of the more highly ranked in VACV as a whole. The relatively high immunogenicity of this epitope led to a reevaluation of the total CD8+T cell response to VACV. By the use of two methods, the true size of the response was found to be around double previous estimates and at its peak is on the order of 60% of all CD8+T cells. We speculate that more CD8+T cell epitopes remain to be mapped for VACV and that underestimation of responses is unlikely to be unique to VACV, so there would be merit in revisiting this issue for other viruses.
Widespread Infection with Homologues of Human Parvoviruses B19, PARV4, and Human Bocavirus of Chimpanzees and Gorillas in the Wild
Authors: Sharp Colin P. , LeBreton Matthew , Kantola Kalle , Nana Ahmadou , Diffo Joseph Le Doux , Djoko Cyrille F. , Tamoufe Ubald , Kiyang John A. , Babila Tafon G. , Ngole Eitel Mpoudi , Pybus Oliver G. , Delwart Eric , Delaporte Eric , Peeters Martine , Soderlund-Venermo Maria , Hedman Klaus , Wolfe Nathan D. , Simmonds Peter 
Published: July 2010

ABSTRACT

Infections with human parvoviruses B19 and recently discovered human bocaviruses (HBoVs) are widespread, while PARV4 infections are transmitted parenterally and prevalent specifically in injecting drug users and hemophiliacs. To investigate the exposure and circulation of parvoviruses related to B19 virus, PARV4, and HBoV in nonhuman primates, plasma samples collected from 73 Cameroonian wild-caught chimpanzees and gorillas and 91 Old World monkey (OWM) species were screened for antibodies to recombinant B19 virus, PARV4, and HBoV VP2 antigens by enzyme-linked immunosorbent assay (ELISA). Moderate to high frequencies of seroreactivity to PARV4 (63% and 18% in chimpanzees and gorillas, respectively), HBoV (73% and 36%), and B19 virus (8% and 27%) were recorded for apes, while OWMs were uniformly negative (for PARV4 and B19 virus) or infrequently reactive (3% for HBoV). For genetic characterization, plasma samples and 54 fecal samples from chimpanzees and gorillas collected from Cameroonian forest floors were screened by PCR with primers conserved within
Erythrovirus
,
Bocavirus
, and PARV4 genera. Two plasma samples (chimpanzee and baboon) were positive for PARV4, while four fecal samples were positive for HBoV-like viruses. The chimpanzee PARV4 variant showed 18% and 15% nucleotide sequence divergence in NS and VP1/2, respectively, from human variants (9% and 7% amino acid, respectively), while the baboon variant was substantially more divergent, mirroring host phylogeny. Ape HBoV variants showed complex sequence relationships with human viruses, comprising separate divergent homologues of HBoV1 and the recombinant HBoV3 species in chimpanzees and a novel recombinant species in gorillas. This study provides the first evidence for widespread circulation of parvoviruses in primates and enables future investigations of their epidemiology, host specificity, and (co)evolutionary histories.