bluuebunny/crossref_metadata_2025_split huggingface dataset:
Efficacy of Vaccination of Calves against Hemorrhagic Septicemia with a Live
aroA
Derivative of
Pasteurella multocida
B:2 by Two Different Routes of Administration
Authors: Hodgson J. Christopher , Finucane Anna , Dagleish Mark P. , Ataei Saeed , Parton Roger , Coote John G. 
Published: February 2005

ABSTRACT

Two groups of four calves each were immunized either intramuscularly (i.m. vaccinated) or intranasally (i.n. vaccinated) at 2 and 6 weeks of age with ca. 10
9
CFU of a derivative of
P. multocida
serotype B:2 strain 85020 containing a deletion in the
aroA
gene (strain JRMT12). Both groups of calves and three unvaccinated control calves were challenged subcutaneously at 8 weeks of age with ca. 10
7
CFU of the wild-type 85020 strain. The first and second vaccinations caused a significant pyrexia and increase in the mean demeanor score (
P
< 0.05) in i.m. but not i.n. vaccinated calves. Serum agglutinating activity against whole cells of
P. multocida
strain 85020 and immunoglobulin G antibody concentrations increased after the second vaccination in i.m. but not in i.n. vaccinated animals, and this difference was statistically significant (
P
< 0.05). Concentrations of serum amyloid A (SAA) increased significantly 3 h after both the primary (
P
< 0.05) and booster (
P
< 0.001) i.m. vaccinations, but not in i.n. vaccinated calves. All four i.m. vaccinated calves were solidly immune to challenge with wild-type
P. multocida
B:2. However, the mean rectal temperatures, demeanor scores, and serum SAA concentrations of i.n. vaccinated and control calves increased significantly (
P
< 0.01). Three i.n. vaccinated and two control calves were killed for humane reasons within 14 h postchallenge, and postmortem examination revealed pathological lesions consistent with hemorrhagic septicemia. These data showed that the
aroA
mutant strain, given i.m. as two doses 4 weeks apart, acted as an effective live-attenuated vaccine strain to protect calves against challenge with the virulent parent strain.
Helicobacter pyloriInduces Interleukin-8 Secretion by Toll-Like Receptor 2- and Toll-Like Receptor 5-Dependent and -Independent Pathways
Authors: Torok Anastasia M. , Bouton Amy H. , Goldberg Joanna B. 
Published: February 2005

ABSTRACTHelicobacter pyloriis an important human pathogen that causes gastritis and is strongly associated with gastric ulcers, gastric adenocarcinomas, and mucosa-associated lymphoid tissue lymphomas. In response toH. pylori, interleukin-8 (IL-8) is secreted from host cells to attract components of the innate and adaptive immune systems to the site of infection. Toll-like receptor 2 (TLR2) and TLR5 have been shown to recognizeH. pyloriand to initiate signaling pathways that result in enhanced activation of NF-κB. Here, we evaluated the contribution of mitogen-activated protein kinase signaling pathways to TLR2-dependent and TLR5-dependent secretion of IL-8. Secretion of IL-8 fromH. pylori-infected HEK293 cells was augmented by the expression of TLR2 or TLR5. WhileH. pyloriinfection resulted in the activation of ERK, JNK, and p38, the enhanced IL-8 secretion from TLR2- and TLR5-expressing cells coincided with increased p38 activation and phosphorylation of the transcription factor ATF2. When p38 activity was inhibited in TLR2- or TLR5-expressing cells,H. pylori-dependent IL-8 secretion returned to the level observed in infected parental HEK293 cells that did not express TLR2 or TLR5; inhibition of p38 had no effect on IL-8 secretion from infected parental HEK cells. In contrast, inhibition of JNK and/or ERK resulted in substantially less IL-8 secretion from infected cells, independent of TLR2 or TLR5 expression. Based on these data, we propose thatH. pyloriinduces IL-8 secretion through a dual mechanism that includes a TLR2/5-independent component involving the activities of JNK and ERK and a TLR2/5-dependent component that requires p38 activity.
Antibody against V Antigen Prevents Yop-Dependent Growth ofYersinia pestis
Authors: Philipovskiy Alexander V. , Cowan Clarissa , Wulff-Strobel Christine R. , Burnett Sandra H. , Kerschen Edward J. , Cohen Donald A. , Kaplan Alan M. , Straley Susan C. 
Published: February 2005

ABSTRACTThe V antigen (LcrV) of the plague bacteriumYersinia pestisis a potent protective antigen that is under development as a vaccine component for humans. LcrV is multifunctional. On the bacterial surface it mediates delivery of a set of toxins called Yops into host cells, and as a released protein it can cause production of the immunosuppressive cytokine interleukin-10 (IL-10) and can inhibit chemotaxis of polymorphonuclear neutrophils. It is not known how these mechanisms of LcrV operate, what their relative importance is, when they function during plague, and which are critical to protection by antibody. This study investigated several of these issues. C57BL/6 mice, mice unable to express IL-10, or mice with the macrophage lineage eliminated were treated with a protective anti-LcrV antibody or a nonprotective antibody against YopM and infected intravenously byY. pestisKIM5 or a strain that lacked the genes encoding all six effector Yops. Viable bacterial numbers were determined at various times. The data indicated that Yops were necessary forYersiniagrowth after the bacteria had seeded liver and spleen. Anti-LcrV antibody prevented this growth, even in IL-10−/−mice, demonstrating that one protective mechanism for anti-LcrV antibody is independent of IL-10. Anti-LcrV antibody had no effect on persistence in organs ofY. pestislacking effector Yops, even though the yersiniae could strongly express LcrV, suggesting that Yops are necessary for building sufficient bacterial numbers to produce enough LcrV for its immunosuppressive effects. In vitro assays showed that anti-LcrV antibody could partially block delivery of Yops and downstream effects of Yops in infected macrophage-like J774A.1 cells. However, cells of the macrophage lineage were found to be dispensable for protection by anti-LcrV antibody in spleen, although they contributed to protection in liver. Taken together, the data support the hypothesis that one protective effect of the antibody is to block delivery of Yops to host cells and prevent early bacterial growth. The findings also identified the macrophage lineage as one host cell type that mediates protection.
Characterization of a Novel Leucine-Rich Repeat Protein Antigen from Group B Streptococci That Elicits Protective Immunity
Authors: Seepersaud Ravin , Hanniffy Sean B. , Mayne Peter , Sizer Phil , Le Page Richard , Wells Jerry M. 
Published: February 2005

ABSTRACTGroup B streptococci (GBS) usually behave as commensal organisms that asymptomatically colonize the gastrointestinal and urogenital tracts of adults. However, GBS are also pathogens and the leading bacterial cause of life-threatening invasive disease in neonates. While the events leading to transmission and disease in neonates remain unclear, GBS carriage and level of colonization in the mother have been shown to be significant risk factors associated with invasive infection. Surface antigens represent ideal vaccine targets for eliciting antibodies that can act as opsonins and/or inhibit colonization and invasion. Using a genetic screen for exported proteins in GBS, we identified a gene, designatedlrrG, that encodes a novel LPXTG anchored surface antigen containing leucine-rich repeat (LRR) motifs found in bacterial invasins and other members of the LRR protein family. Southern blotting showed thatlrrGwas present in all GBS strains tested, representing the nine serotypes, and revealed the presence of anlrrGhomologue inStreptococcus pyogenes. Recombinant LrrG protein was shown in vitro to adhere to epithelial cells in a dose-dependent manner, suggesting that it may function as an adhesion factor in GBS. More importantly, immunization with recombinant LrrG elicited a strong immunoglobulin G response in CBA/ca mice and protected against lethal challenge with virulent GBS. The data presented in this report suggest that this conserved protein is a highly promising candidate antigen for use in a GBS vaccine.
Salmonella entericaSerovar Typhimurium Infection of Dendritic Cells Leads to Functionally Increased Expression of the Macrophage-Derived Chemokine
Authors: Fu Guo , Wijburg Odilia L. C. , Cameron Paul U. , Price Jason D. , Strugnell Richard A 
Published: February 2005

ABSTRACTGene expression in murine dendritic cells (DCs) infected with green fluorescent protein-expressingSalmonella entericaserovar Typhimurium BRD509 was studied by mRNA differential display. Infected DCs were sorted from uninfected cells by flow cytometry. The mRNA expression patterns of infected and uninfected cells revealed a number of differentially expressed transcripts, which included the macrophage-derived chemokine (MDC). Up-regulation ofMDCtranscription in infected DCs was confirmed by Northern blotting, and the kinetics ofMDCexpression was examined by real-time reverse transcription-PCR, with which 31- and 150-fold increases were detected at 2 and 6 h postinfection, respectively. The increased release by DCs of MDC into culture media was detected by an enzyme-linked immunosorbent assay. The biological activity of MDC was investigated in in vitro and in vivo assays. In vitro, supernatants fromS. entericaserovar Typhimurium-infected DCs were chemoattractive to T cells, and neutralization of MDC in these supernatants inhibited T-cell migration. Passive transfer of anti-MDC antibody to mice infected with BRD509 revealed that neither growth of the bacterium nor resistance of the mice to reinfection was affected and that in vivo inhibition of MDC did not affect T-cell responses, as measured by the gamma interferon ELISPOT method 3 days after challenge infection.
Use of Green Fluorescent Protein and Reverse Transcription-PCR To Monitor
Candida albicans
Agglutinin-Like Sequence Gene Expression in a Murine Model of Disseminated Candidiasis
Authors: Green Clayton B. , Zhao Xiaomin , Hoyer Lois L. 
Published: February 2005

ABSTRACT

Candida albicans
P
ALS
-green fluorescent protein (GFP) reporter strains were inoculated into mice in a disseminated candidiasis model, and GFP production was monitored by immunohistochemistry and reverse transcription-PCR (RT-PCR). GFP production from the
ALS1
and
ALS3
promoters was detected immunohistochemically.
ALS1
,
ALS2
,
ALS3
,
ALS4
, and
ALS9
transcription was detected by RT-PCR, further identifying ALS genes expressed in this model.
Production of a Proteolytically Active Protein, Chlamydial Protease/Proteasome-Like Activity Factor, by Five Different
Chlamydia
Species
Authors: Dong Feng , Zhong Youmin , Arulanandam Bernard , Zhong Guangming 
Published: February 2005

ABSTRACT

We have previously identified a chlamydial protein, chlamydial protease/proteasome-like activity factor (CPAF), for degrading host transcription factors in cells infected with the human chlamydial species
Chlamydia trachomatis
or
Chlamydia pneumoniae
. We now report that functional CPAF was also produced during infection with the species
Chlamydia muridarum
,
Chlamydia psittaci
, and
Chlamydia caviae
, which primarily infect nonhuman hosts.
Glutathione and Nitrosoglutathione in Macrophage Defense againstMycobacterium tuberculosis
Authors: Venketaraman Vishwanath , Dayaram Yaswant K. , Talaue Meliza T. , Connell Nancy D. 
Published: February 2005

ABSTRACTWe demonstrate thatMycobacterium tuberculosisgrown in vitro is sensitive to glutathione and its derivativeS-nitrosoglutathione. Furthermore, our infection studies with J774.1 macrophages indicate that glutathione is essential for the control of the intracellular growth ofM. tuberculosis. This study indicates the important role of glutathione in the control of macrophages byM. tuberculosis.
Genetic Vaccination against Murine Cysticercosis by Using a Plasmid Vector CarryingTaenia soliumParamyosin
Authors: Solís Carlos F. , Ostoa-Saloma Pedro , Lugo-Martínez Verónica H. , Johnston Stephen Albert , Laclette Juan Pedro 
Published: February 2005

ABSTRACTA plasmid vector carrying the immunoprotective amino-terminal fragment ofTaenia soliumparamyosin (VW2-1) was designed for genetic vaccination studies. Mice that were genetically immunized with VW2-1 and challenged by intraperitoneal inoculation ofTaenia crassicepscysticerci showed 43 to 48% reductions in the parasite burden, values which were similar to values obtained previously when the recombinant protein was used.
Amino Acids 1055 to 1192 in the S2 Region of Severe Acute Respiratory Syndrome Coronavirus S Protein Induce Neutralizing Antibodies: Implications for the Development of Vaccines and Antiviral Agents
Authors: Keng Choong-Tat , Zhang Aihua , Shen Shuo , Lip Kuo-Ming , Fielding Burtram C. , Tan Timothy H. P. , Chou Chih-Fong , Loh Chay Boon , Wang Sifang , Fu Jianlin , Yang Xiaoming , Lim Seng Gee , Hong Wanjin , Tan Yee-Joo 
Published: February 2005

ABSTRACT

The spike (S) protein of the severe acute respiratory syndrome coronavirus (SARS-CoV) interacts with cellular receptors to mediate membrane fusion, allowing viral entry into host cells; hence it is recognized as the primary target of neutralizing antibodies, and therefore knowledge of antigenic determinants that can elicit neutralizing antibodies could be beneficial for the development of a protective vaccine. Here, we expressed five different fragments of S, covering the entire ectodomain (amino acids 48 to 1192), as glutathione
S
-transferase fusion proteins in
Escherichia coli
and used the purified proteins to raise antibodies in rabbits. By Western blot analysis and immunoprecipitation experiments, we showed that all the antibodies are specific and highly sensitive to both the native and denatured forms of the full-length S protein expressed in virus-infected cells and transfected cells, respectively. Indirect immunofluorescence performed on fixed but unpermeabilized cells showed that these antibodies can recognize the mature form of S on the cell surface. All the antibodies were also able to detect the maturation of the 200-kDa form of S to the 210-kDa form by pulse-chase experiments. When the antibodies were tested for their ability to inhibit SARS-CoV propagation in Vero E6 culture, it was found that the anti-SΔ10 antibody, which was targeted to amino acid residues 1029 to 1192 of S, which include heptad repeat 2, has strong neutralizing activities, suggesting that this region of S carries neutralizing epitopes and is very important for virus entry into cells.
Protection of Rhesus Monkeys against Infection with Minimally Pathogenic Simian-Human Immunodeficiency Virus: Correlations with Neutralizing Antibodies and Cytotoxic T Cells
Authors: Quinnan Gerald V. , Yu Xiao-Fang , Lewis Mark G. , Zhang Peng Fei , Sutter Gerd , Silvera Peter , Dong Ming , Choudhary Anil , Sarkis Phuong T. N. , Bouma Peter , Zhang Zhiqiang , Montefiori David C. , VanCott Thomas C. , Broder Christopher C. 
Published: February 2005

ABSTRACT

We studied the capacity of active immunization of rhesus monkeys with HIV-1 envelope protein (Env) to induce primary virus cross-reactive neutralizing antibodies to prevent infection following intravenous challenge with simian-human immunodeficiency virus (SHIV). Monkeys were immunized with the human immunodeficiency type 1 (HIV-1) strain R2 Env. Initially, the Env was expressed in vivo by an alphavirus replicon particle system, and then it was administered as soluble oligomeric gp140. Concurrently, groups of monkeys received expression vectors that encoded either simian immunodeficiency virus (SIV)
gag
/
pol
genes or no SIV genes in vivo to test the additional protective benefit of concurrent induction of virus-specific cell-mediated immune (CMI) responses. Groups of control monkeys received either the
gag
/
pol
regimen or sham immunizations. The antibodies induced by the Env immunization regimen neutralized diverse primary HIV-1 strains. Similarly, potent CMI responses were induced by the
gag
/
pol
regimen, as measured by gamma interferon enzyme-linked immunospot assays. Differences in the responses among groups of monkeys strongly suggested that there was interference between the Env and g
ag
/
pol
immunization regimens. Complete protection of some of the monkeys against infection after intravenous challenge with the partially pathogenic SHIV
DH12R (Clone 7)
was associated independently with both neutralizing antibody and CMI responses. Protection was associated with SHIV
DH12 (Clone 7)
serum neutralizing antibody titers of ≥1:80 or with cellular immune responses corresponding to >2,000 spot forming cells per 10
6
peripheral blood mononuclear cells. Immunization was also associated with a reduction in the magnitude and duration of virus load. Induction of cross-reactive, primary HIV-1-neutralizing antibodies is feasible and, when potent, may result in complete protection against infection with a heterologous challenge virus strain.
A Noninfectious Simian/Human Immunodeficiency Virus DNA Vaccine That Protects Macaques against AIDS
Authors: Singh Dinesh K. , Liu Zhenqian , Sheffer Darlene , Mackay Glenn A. , Smith Marilyn , Dhillon Sukhbir , Hegde Ramakrishna , Jia Fenglan , Adany Istvan , Narayan Opendra 
Published: February 2005

ABSTRACT

Simian/human immunodeficiency virus SHIV
KU2
replicates with extremely high titers in macaques. In order to determine whether the DNA of the viral genome could be used as a vaccine if the DNA were rendered noninfectious, we deleted the reverse transcriptase gene from SHIV
KU2
and inserted this DNA (Δ
rt
SHIV
KU2
) into a plasmid that was then used to test gene expression and immunogenicity. Transfection of Jurkat and human embryonic kidney epithelial (HEK 293) cells with the DNA resulted in production of all of the major viral proteins and their precursors and transient export of a large quantity of the Gag p27 into the supernatant fluid. As expected, no infectious virus was produced in these cultures. Four macaques were injected intradermally with 2 mg of the DNA at 0, 8, and 18 weeks. The animals developed neutralizing antibodies and low enzyme-linked immunospot assay (E-SPOT) titers against SHIV
KU2
. These four animals and two unvaccinated control animals were then challenged with heterologous SHIV89.6P administered into their rectums. The two control animals developed viral RNA titers exceeding 10
6
copies/ml of plasma, and these titers were accompanied by the loss of CD4
+
T cells by 2 weeks after challenge. The two control animals died at weeks 8 and 16, respectively. All four of the immunized animals became infected with the challenge virus but developed lower titers of viral RNA in plasma than the control animals, and the titers decreased over time in three of the four macaques. The fourth animal remained viremic and died at week 47. Whereas the control animals failed to develop E-SPOT responses, all four of the immunized animals developed anamnestic E-SPOT responses after challenge. The animal that died developed the highest E-SPOT response and was the only one that produced neutralizing antibodies against the challenge virus. These results established that noninfectious DNA of pathogenic SHIV could be used as a vaccine to prevent AIDS, even though the immunological assays used did not predict the manner in which the challenge virus would replicate in the vaccinated animals.
Glycoprotein M Is an Essential Lytic Replication Protein of the Murine Gammaherpesvirus 68
Authors: May Janet S. , Colaco Susanna , Stevenson Philip G. 
Published: February 2005

ABSTRACT
All herpesviruses encode a homolog of glycoprotein M (gM), which appears to function in virion morphogenesis. Despite its conservation, gM is inessential for the lytic replication of alphaherpesviruses. In order to address the importance of gM in gammaherpesviruses, we disrupted it in the murine gammaherpesvirus 68 (MHV-68). The mutant virus completely failed to propagate in normally permissive fibroblasts. The defective genome was rescued by either homologous recombination to restore the wild-type gM in situ or the insertion of an ectopic, intergenic expression cassette encoding gM into the viral genome. Thus, gM was essential for the lytic replication of MHV-68.
Complex Formation among Murine Cytomegalovirus US22 Proteins Encoded by Genes M139, M140, and M141
Authors: Karabekian Zaruhi , Hanson Laura K. , Slater Jacquelyn S. , Krishna Neel K. , Bolin Lisa L. , Kerry Julie A. , Campbell Ann E. 
Published: February 2005

ABSTRACT

The murine cytomegalovirus (MCMV) proteins encoded by US22 genes M139, M140, and M141 function, at least in part, to regulate replication of this virus in macrophages. Mutant MCMV having one or more of these genes deleted replicates poorly in macrophages in culture and in the macrophage-dense environment of the spleen. In this report, we demonstrate the existence of stable complexes formed by the products of all three of these US22 genes, as well as a complex composed of the products of M140 and M141. These complexes form in the absence of other viral proteins; however, the pM140/pM141 complex serves as a requisite binding partner for the M139 gene products. Products from all three genes colocalize to a perinuclear region of the cell juxtaposed to or within the
cis
-Golgi region but excluded from the
trans
-Golgi region. Interestingly, expression of pM141 redirects pM140 from its predominantly nuclear residence to the perinuclear, cytoplasmic locale where these US22 proteins apparently exist in complex. Thus, complexing of these nonessential, early MCMV proteins likely confers a function(s) independent of each individual protein and important for optimal replication of MCMV in its natural host.
Large-Scale Molecular Characterization of Adeno-Associated Virus Vector Integration in Mouse Liver
Authors: Nakai Hiroyuki , Wu Xiaolin , Fuess Sally , Storm Theresa A. , Munroe David , Montini Eugenio , Burgess Shawn M. , Grompe Markus , Kay Mark A. 
Published: February 2005

ABSTRACT
Recombinant adeno-associated virus (rAAV) vector holds promise for gene therapy. Despite a low frequency of chromosomal integration of vector genomes, recent studies have raised concerns about the risk of rAAV integration because integration occurs preferentially in genes and accompanies chromosomal deletions, which may lead to loss-of-function insertional mutagenesis. Here, by analyzing 347 rAAV integrations in mice, we elucidate novel features of rAAV integration: the presence of hot spots for integration and a strong preference for integrating near gene regulatory sequences. The most prominent hot spot was a harmless chromosomal niche in the rRNA gene repeats, whereas nearly half of the integrations landed near transcription start sites or CpG islands, suggesting the possibility of activating flanking cellular disease genes by vector integration, similar to retroviral gain-of-function insertional mutagenesis. Possible cancer-related genes were hit by rAAV integration at a frequency of 3.5%. In addition, the information about chromosomal changes at 218 integration sites and 602 breakpoints of vector genomes have provided a clue to how vector terminal repeats and host chromosomal DNA are joined in the integration process. Thus, the present study provides new insights into the risk of rAAV-mediated insertional mutagenesis and the mechanisms of rAAV integration.