bluuebunny/crossref_metadata_2025_split huggingface dataset:
PCR-Based Method for Isolation and Detection of
Chlamydia pneumoniae
DNA in Cerebrospinal Fluids
Authors: Ikejima Hideaki , Haranaga Shusaku , Takemura Hiromu , Kamo Tsutomu , Takahashi Youichi , Friedman Herman , Yamamoto Yoshimasa 
Published: July 2002

ABSTRACT

Since current studies indicate the possible involvement of
Chlamydia pneumoniae
in the pathogenesis of multiple sclerosis (MS), demonstration of
C. pneumoniae
in the cerebrospinal fluid (CSF) of patients with MS is highly desirable. However, there is controversy concerning the detection of
C. pneumoniae
in CSFs from MS patients due to the lack of a standard protocol for extraction and detection of
C. pneumoniae
DNA. In this regard, we attempted to establish a highly effective extraction protocol for
C. pneumoniae
DNA from CSFs utilizing a commercial kit and a PCR detection method. The extraction and PCR detection protocol established in this study succeeded in detecting as few as 20
C. pneumoniae
organisms in 200 μl of mock CSF. The use of this protocol to detect
C. pneumoniae
DNA in CSFs revealed that 68% of CSF samples obtained from patients with MS were positive (11 out of 16 samples) for chlamydia DNA. Thus, the protocol established here is sensitive enough to detect chlamydia DNA from CSFs and can be used by other laboratories for evaluation of the presence of chlamydiae in CSFs because the protocol is based on the use of a commercial kit.
Borna Disease Virus and Human Disease
Authors: Carbone Kathryn M. 
Published: July 2002

SUMMARYThe biology of Borna disease virus (BDV) strongly supports the likelihood of human infection with BDV or a variant of BDV. Thus far, the evidence supporting BDV infection in humans has initiated much controversy among basic and clinical scientists; only time and additional research will support or refute the hypothesis of human BDV infection. Until an assay of acceptable specificity and sensitivity has been developed, validated, and used to document human BDV infection, scientists cannot reasonably begin to associate BDV infection with specific disease syndromes. Clinical studies seeking causal associations between BDV infection and specific diseases must ensure the proper identification of the BDV infection status of patients and control subjects by using a validated, highly sensitive, and highly specific assay (or series of assays). For clinical studies, a highly sensitive “screening” test followed by a highly specific confirmatory test will be of significant benefit. Although it is possible to formulate hypotheses about the clinical outcomes of human BDV infection based on animal model work, to date no human disease has been causally linked to human BDV infection. Scientists all over the world are actively pursuing these issues, and with continuing advances in clinical and basic BDV research, the answers cannot be far away.
Genetic Immunization of BALB/c mice with a Plasmid Bearing the Gene Coding for a Hybrid Merozoite Surface Protein 1-Hepatitis B Virus Surface Protein Fusion Protects Mice against LethalPlasmodium chabaudi chabaudiPC1 Infection
Authors: Wunderlich Gerhard , Moura Ivan C. , del Portillo Hernando A. 
Published: July 2002

ABSTRACTThe genetic immunization of rodents with a plasmid coding for aPlasmodium chabaudimerozoite surface protein 1 (C terminus)-hepatitis B virus surface fusion protein (pPcMSP119-HBs) provided protection of mice against subsequent lethal challenge withP. chabaudi chabaudiPC1-infected red blood cells. The percentage of survivor mice was higher in DNA-immunized mice than in animals immunized with a recombinant rPcMSP119– glutathioneS-transferase fusion protein administered in Freund adjuvant. In all mice immunized with the pPcMSP119-HBs, a Th1-specific response, including the production of anti-MSP119-specific immunoglobulins predominantly of the immunoglobulin G2a subtype and reacting almost exclusively against discontinuous epitopes, was elicited. The coinjection of Th1-type cytokine-expressing plasmids (gamma interferon, interleukin-2, and granulocyte-macrophage colony-stimulating factor) mostly abolished protection and boosting of MSP119-specific antibodies. The inclusion of a lymph node-targeting signal did not significantly increase protection. These data provide further evidence that MSP119-HBs DNA constructs might be useful as components of a genetic vaccine against the asexual blood stages ofPlasmodium.
Formation of Methyl Mercaptan from
l
-Methionine by
Porphyromonas gingivalis
Authors: Yoshimura Mamiko , Nakano Yoshio , Yamashita Yoshihisa , Oho Takahiko , Saito Toshiyuki , Koga Toshihiko 
Published: July 2002

ABSTRACT

Methyl mercaptan production by oral bacteria is thought to be one of the main causes of oral malodor. We examined the ability of periodontopathic
Porphyromonas gingivalis
to produce methyl mercaptan from
l
-methionine and found that the invasive strains W83 and W50 produced large amounts of methyl mercaptan. We cloned and sequenced the
mgl
gene encoding
l
-methionine-α-deamino-γ-mercaptomethane-lyase (METase) from
P. gingivalis
W83. The structural
mgl
gene consisted of 1,200 bp and encoded a 43.3-kDa protein. To examine the role of methyl mercaptan in the pathogenesis of
P. gingivalis
, a METase-deficient mutant of
P. gingivalis
W83 was constructed. The methionine degradation activity and virulence of the mutant (M1217) and the parent strain (W83) in mice were compared. M1217 showed a marked decrease in the formation of methyl mercaptan from
l
-methionine and decreased virulence compared with the wild-type strain W83. These results suggest that methyl mercaptan not only is one of the sources of oral malodor, but may also play a role in the pathogenicity of
P. gingivalis.
ESAT-6 Subunit Vaccination againstMycobacterium tuberculosis
Authors: Brandt Lise , Elhay Martin , Rosenkrands Ida , Lindblad Erik B. , Andersen Peter 
Published: July 2002

ABSTRACTThe ESAT-6 antigen fromMycobacterium tuberculosisis a dominant target for cell-mediated immunity in the early phase of tuberculosis (TB) in TB patients as well as in various animal models. The purpose of our study was to evaluate the potential of ESAT-6 in an experimental TB vaccine. We started out using dimethyl dioctadecylammonium bromide (DDA), an adjuvant which has been demonstrated to be efficient for the induction of cellular immune responses and has been used successfully before as a delivery system for TB vaccines. Here we demonstrate that, whereas immune responses to both short-term-culture filtrate and Ag85B are efficiently induced with DDA, this adjuvant was inefficient for the induction of immune responses to ESAT-6. Therefore, we investigated the modulatory effect of monophosphoryl lipid A (MPL), an immunomodulator which in different combinations has demonstrated strong adjuvant activity for both cellular and humoral immune responses. We show in the present study that vaccination with ESAT-6 delivered in a combination of MPL and DDA elicited a strong ESAT-6-specific T-cell response and protective immunity comparable to that achieved withMycobacterium bovisBCG.
Shigella flexneri
2a Strain CVD 1207, with Specific Deletions in
virG
,
sen
,
set
, and
guaBA
, Is Highly Attenuated in Humans
Authors: Kotloff Karen L. , Noriega Fernando R. , Samandari Taraz , Sztein Marcelo B. , Losonsky Genevieve A. , Nataro James P. , Picking William D. , Barry Eileen M. , Levine Myron M. 
Published: July 2002

ABSTRACT

A phase 1 clinical trial was conducted among 35 healthy adult volunteers to evaluate the safety, immunogenicity, and shedding of different doses of CVD 1207, a live attenuated
Shigella flexneri
2a vaccine candidate with specific deletion mutations in
virG
,
sen
,
set
, and
guaBA
. CVD 1207 retains the ability to invade epithelial cells but cannot effectively spread intercellularly after invasion (Δ
virG
), does not produce enterotoxin (Δ
sen
and Δ
set
), and has limited proliferation in vivo (Δ
guaBA
). In a consecutive fashion, groups of three to seven subjects ingested a single oral dose of CVD 1207 at an inoculum of either 10
6
, 10
7
, 10
8
, 10
9
, or 10
10
CFU. CVD 1207 was remarkably well-tolerated at inocula as high as 10
8
CFU. In comparison, one of 12 subjects who received 10
9
CFU experienced mild diarrhea and another experienced a single episode of emesis. One of five subjects who received 10
10
CFU experienced watery diarrhea and emesis. All subjects who ingested doses of 10
8
to 10
10
CFU excreted the vaccine; in 23 of 25, the duration of excretion was ≤3 days. A dose-related, immunoglobulin A antibody-secreting cell (ASC) response to
S. flexneri
2a O-specific lipopolysaccharide was seen, with geometric mean peak values of 6.1 to 35.2 ASCs/10
6
peripheral blood mononuclear cells (PBMC) among recipients of 10
7
to 10
10
CFU. The cytokine response to
Shigella
-specific antigens observed in volunteers' PBMC following vaccination suggested a Th1 pattern with stimulation of gamma interferon and absence of interleukin 4 (IL-4) or IL-5. CVD 1207 represents a
Shigella
live oral vaccine strain prepared from wild-type
S. flexneri
2a by rational use of recombinant DNA technology that achieves a remarkable degree of attenuation compared with earlier recombinant strains, even when administered at high dosage.
Cable-Piliated
Burkholderia cepacia
Binds to Cytokeratin 13 of Epithelial Cells
Authors: Sajjan Umadevi S. , Sylvester Francisco A. , Forstner Janet F. 
Published: July 2002

ABSTRACT

Although the
Burkholderia cepacia
complex consists of several genomovars, one highly transmissible strain of
B. cepacia
has been isolated from the sputa of cystic fibrosis (CF) patients throughout the United Kingdom and Canada. This strain expresses surface cable (Cbl) pili and is thought to be the major strain associated with the fatal “cepacia syndrome.” In the present report we characterize the specific 55-kDa buccal epithelial cell (BEC) protein that binds cable pilus-positive
B. cepacia
. N-terminal sequences of CNBr-generated internal peptides identified the protein as cytokeratin 13 (CK13). Western blots of BEC extracts probed with a specific monoclonal antibody to CK13 confirmed the identification. Mixed epidermal cytokeratins (which contain CK13), cytokeratin extract from BEC (which consists essentially of CK13 and CK4), and a polyclonal antibody to mixed cytokeratins inhibited
B. cepacia
binding to CK13 blots and to normal human bronchial epithelial (NHBE) cells. Preabsorption of the antikeratin antibody with the BEC cytokeratin fraction reversed the inhibitory effect of the antibody. A cytokeratin mixture lacking CK13 was ineffective as an inhibitor of binding. Colocalization of CK13 and
B. cepacia
by confocal microscopy demonstrated that intact nonpermeabilized NHBE cells express small amounts of surface CK13 and bind Cbl-positive
B. cepacia
in the same location. Binding to intact NHBE cells was dependent on bacterial concentration and was saturable, whereas a Cbl-negative isolate exhibited negligible binding. These findings raise the possibility that surface-accessible CK13 in respiratory epithelia may be a biologically relevant target for the binding of cable piliated
B. cepacia
.
Effects of Estradiol and Progesterone on Susceptibility and Early Immune Responses toChlamydia trachomatisInfection in the Female Reproductive Tract
Authors: Kaushic Charu , Zhou Fan , Murdin Andrew D. , Wira Charles R. 
Published: July 2002

ABSTRACTWe have used a previously described rodent model to examine the influence of hormonal environment on susceptibility and immune responses to genitalChlamydiainfection. Ovariectomized rats were administered estradiol, progesterone, or a combination of both, infected withChlamydia trachomatisvia the intrauterine route, and sacrificed 5 days later. Histopathological examination showed severe inflammation in the uteri and vaginae of progesterone-treated animals, whereas animals receiving estradiol or a combination of both hormones showed no inflammation. Large numbers of chlamydiae were found in vaginal secretions of progesterone-treated and combination-treated animals, while estradiol-treated animals had none. Tissue localization showed that numerous chlamydial inclusions were present in the uterine epithelium of the progesterone group and the cervicovaginal epithelium of the combination group. Examination of the acute immune responses of the infected animals showed that maximum activation was present in the draining lymph node cells from the progesterone-treated group, and these cells were producing large amounts of interleukin-10 and gamma interferon compared to other hormone-treated groups. In contrast, spleen cell proliferation was suppressed in progesterone-treated animals compared to other hormone-treated groups. We conclude that progesterone increases and estradiol decreases susceptibility to intrauterine chlamydial infection in this rat model. Our data demonstrate that hormone environment, at the time of infection, has a profound effect on the outcome of microbial infection in the female reproductive tract.
β-1,2-Linked Oligomannosides fromCandida albicansBind to a 32-Kilodalton Macrophage Membrane Protein Homologous to the Mammalian Lectin Galectin-3
Authors: Fradin Chantal , Poulain Daniel , Jouault Thierry 
Published: July 2002

ABSTRACTβ-1,2-linked oligomannoside residues are present, associated with mannan and a glycolipid, the phospholipomannan, at theCandida albicanscell wall surface. β-1,2-linked oligomannoside residues act as adhesins for macrophages and stimulate these cells to undergo cytokine production. To characterize the macrophage receptor involved in the recognition ofC. albicansβ-1,2-oligomannoside we used the J774 mouse cell line, which is devoid of the receptor specific for α-linked mannose residues. A series of experiments based on affinity binding on eitherC. albicansyeast cells or β-1,2-oligomannoside-conjugated bovine serum albumin (BSA) and subsequent disclosure with biotinylated conjugated BSA repeatedly led to the detection of a 32-kDa macrophage protein. An antiserum specific for this 32-kDa protein inhibitedC. albicansbinding to macrophages and was used to immunoprecipitate the molecule. Two high-pressure liquid chromatography-purified peptides from the 32-kDa tryptic digest showed complete homology to galectin-3 (previously designated Mac-2 antigen), an endogenous lectin with pleiotropic functions which is expressed in a wide variety of cell types with whichC. albicansinteracts as a saprophyte or a parasite.
Protection against
Streptococcus pneumoniae
Elicited by Immunization with Pneumolysin and CbpA
Authors: Ogunniyi Abiodun David , Woodrow Matthew C. , Poolman Jan T. , Paton James C. 
Published: July 2002

ABSTRACT

The need for the development of cheap and effective vaccines against pneumococcal disease has necessitated the evaluation of common virulence-associated proteins of
Streptococcus pneumoniae
as potential vaccine antigens. In this study, we examined the capacity of active immunization with a genetic toxoid derivative of pneumolysin (PdB) and/or a fragment of choline binding protein A (CbpA; also known as PspC, Hic, and SpsA) to protect mice from intraperitoneal challenge with medium to very high doses of a highly virulent capsular type 2 pneumococcal strain, D39. The median survival times for mice immunized with the individual protein antigens in different adjuvant combinations were significantly longer than those for mice that received the respective adjuvants alone. Mice immunized with CbpA alone were significantly better protected than mice immunized with PdB alone. Correspondingly, the median survival times for mice that were immunized with a combination of PdB and CbpA were significantly longer than those for mice that received PdB alone but not significantly different from those that received CbpA alone. Mice immunized with the protein antigens in a mixture of monophospholipid A (MPL) and aluminium phosphate (AlPO
4
) adjuvants had higher antibody titers than mice that received the antigens in AlPO
4
alone. Mice immunized with PdB in MPL plus AlPO
4
were also significantly better protected than mice that received PdB in AlPO
4
alone.
Cryptosporidium parvum-Specific Mucosal Immune Response in C57BL/6 Neonatal and Gamma Interferon-Deficient Mice: Role of Tumor Necrosis Factor Alpha in Protection
Authors: Lacroix Sonia , Mancassola Roselyne , Naciri Muriel , Laurent Fabrice 
Published: July 2002

ABSTRACTBoth neonatal and C57BL/6 gamma interferon (IFN-γ) knockout (C57BL/6-GKO) mice are susceptible toCryptosporidium parvum, but the course of infection is different. Neonatal mice are able to clear the parasite within 3 weeks, whereas C57BL/6-GKO mice, depending on age, die rapidly or remain chronically infected. The mechanism by which IFN-γ leads to a protective immunity is yet poorly understood. In order to investigate the effect of IFN-γ on other cytokines expressed in the intestinal mucosa duringC. parvuminfection, we studied cytokine mRNA expression in the neonates and GKO (neonatal and adult) mice by quantitative reverse transcription-PCR (RT-PCR) at 4 and 9 days after infection. IFN-γ mRNA levels were quickly and strongly up-regulated in the mucosa of neonatal mice. In GKO mice, the Th1-type response was dramatically altered during the infection, whereas the mRNA expression levels of the Th2-type cytokines interleukin 4 (IL-4) and IL-10 were increased in both mouse models. In the absence of IFN-γ, the adult knockout mice up-regulated the mRNA levels of inflammatory cytokines, such as IL-1β, IL-6, and granulocyte-macrophage colony-stimulating factor, in the mucosa, but not tumor necrosis factor alpha (TNF-α), whereas all these cytokines were up-regulated in the infected neonatal mice. Further experiments indicated that injections of TNF-α into GKO adult mice significantly reduced oocyst shedding. The results of the present study indicate that the resolution of infection is dependent on the expression of Th1-type cytokines in the mucosa of C57BL/6 mice and that TNF-α may participate in the control of parasite development.
Rhoptry-Associated Protein 1-Binding Monoclonal Antibody Raised against a Heterologous Peptide Sequence InhibitsPlasmodium falciparumGrowth In Vitro
Authors: Moreno Rafael , Pöltl-Frank Friederike , Stüber Dietrich , Matile Hugues , Mutz Michael , Weiss Niklaus A. , Pluschke Gerd 
Published: July 2002

ABSTRACTMonoclonal antibodies (MAbs) specific forPlasmodium falciparumrhoptry-associated protein 1 (RAP-1) were generated and tested for inhibition of parasite growth in vitro. The majority of indirect immunofluorescence assay (IFA)-positive MAbs raised against recombinant RAP-1 positions 23 to 711 (rRAP-123–711) recognized epitopes located in the immunodominant N-terminal third of RAP-1. MAbs specific for the building block 35.1 of the synthetic peptide malaria vaccine SPf66 also yielded an IFA staining pattern characteristic for rhoptry-associated proteins and reacted specifically with rRAP-1 and parasite-derived RAP-1 molecules p67 and p82. Cross-reactivity with RAP-1 was blocked by the 35.1 peptide. Epitope mapping with truncated rRAP-1 molecules and overlapping peptides identified the linear RAP-1 sequence Y218KYSL222as a target of the anti-35.1 MAbs. This sequence lacks primary sequence similarity with the 35.1 peptide (YGGPANKKNAG). Cross-reactivity of the anti-35.1 MAbs thus appears to be associated with conformational rather than sequence homology. While the anti-35.1 MAb SP8.18 exhibited parasite growth-inhibitory activity, none of the tested anti-rRAP-123–711MAbs inhibited parasite growth, independently of their fine specificity for the RAP-1 sequences at positions 33 to 42, 213 to 222, 243 to 247, 280 to 287, or 405 to 446. The growth-inhibitory activity of MAb SP8.18 was, however, accelerated by noninhibitory anti-RAP-1 MAbs. Results demonstrate that in addition to fine specificity, other binding parameters are also crucial for the inhibitory potential of an antibody.
Boosting Vaccine for Tuberculosis
Authors: Brooks Jason V. , Frank Anthony A. , Keen Marc A. , Bellisle John T. , Orme Ian M. 
Published: July 2002

ABSTRACT

An effective new vaccine for the control of tuberculosis is badly needed. While current research attempts to improve vaccination are concentrating on new prophylactic or immunotherapeutic vaccines, virtually no emphasis has been placed on boosting individuals already inoculated with
Mycobacterium bovis
BCG. It is shown here that mice vaccinated with BCG gradually lose their capacity to resist an aerosol challenge infection as they age. However, if these mice are inoculated with the 30-kDa mycolyl transferase A protein in midlife, after challenge when aged they express levels of protection in the lungs comparable to those of young mice, associated with minimal pathological damage.
Intranasal Immunization Enhances Clearance of NontypeableHaemophilus influenzaeand Reduces Stimulation of Tumor Necrosis Factor Alpha Production in the Murine Model of Otitis Media
Authors: Sabirov Albert , Kodama Satoru , Hirano Takashi , Suzuki Masashi , Mogi Goro 
Published: July 2002

ABSTRACTNontypeableHaemophilus influenzae(NTHi) is a major pathogen causing otitis media (OM). One of the outer membrane proteins of NTHi, P6, is a common antigen to all strains and is considered a candidate for mucosal vaccine. We have previously reported that intranasal immunization with P6 and cholera toxin (CT) could induce P6-specific immunoglobulin A (IgA) antibodies in the middle ear. In the present study, we assessed the effect of intranasal immunization for the protection against NTHi-induced OM. Mice were immunized intranasally with P6 and CT as an adjuvant on days 0, 7, and 14. Control mice were given phosphate-buffered saline (PBS) without antigen. One week after the final immunization, a suspension of live NTHi (107CFU) was injected into the tympanic cavity to induce experimental OM. On days 3 and 7 after bacterial challenge, mice were killed and middle ear effusions (MEEs) were collected. All immunized mice showed elevated titers of P6-specific antibodies in MEEs. The rank order of specific antibody included, from highest to lowest levels, IgG, IgA, and IgM. In addition, immunized mice showed enhanced clearance of NTHi from the middle ear and the number of NTHi in MEEs of immunized mice was reduced by 97% on day 3 and by 92% on day 7 after bacterial challenge relative the number in the MEEs of control mice. The protective effect of intranasal immunization on the incidence of NTHi-induced experimental OM was evident on day 7 after challenge. By day 7, the number of MEEs in immunized mice was 64% less than that in control mice and the incidence of NTHi culture-positive MEEs in immunized mice was 56% less than that in control mice. Less stimulation of tumor necrosis factor alpha (TNF-α) production in the middle ear was evident on day 3 after challenge. Immunized mice showed lower concentrations of TNF-α in MEEs. These results indicate that intranasal immunization affords protection against experimental OM as evidenced by enhanced clearance of NTHi and less stimulation of TNF-α production in the middle ear. These findings suggest that a nasal vaccine might be useful for preventing OM.