bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of the Cell Division Protein FtsZ as a Means of Differentiating among
Bartonella
Species
Authors: Kelly Timothy M. , Padmalayam Indira , Baumstark Barbara R.
Published: December 2019

Genes coding for homologs of the highly conserved cell division protein FtsZ were isolated from
Bartonella henselae
and
Bartonella quintana
, the causative agents of cat scratch disease and trench fever, respectively. DNA fragments coding for the
ftsZ
open reading frames (ORFs) were cloned into
Escherichia coli
following PCR amplification with primers based on the
ftsZ
sequence of the closely related species
Bartonella bacilliformis
. The amino acid sequences predicted from the cloned
B. henselae
and
B. quintana ftsZ
ORFs are 81 to 83% identical to the corresponding protein in
B. bacilliformis
. Like the FtsZ protein of
B. bacilliformis
, the
B. henselae
and
B. quintana
homologs are about twice as large as the FtsZ proteins reported in most other organisms. Localized sequence differences within the C-terminal coding regions of the
Bartonella ftsZ
genes were used as the basis for species-specific identification of these organisms at both the DNA and protein levels. Oligonucleotide primers which permit the amplification of an
ftsZ
fragment from each of the
Bartonella
species without amplifying DNA from the other two species were designed. Anti-FtsZ antisera raised in rabbits against synthetic peptides corresponding to the relatively divergent C-terminal regions were shown via Western blot analysis to react only with the FtsZ protein from the cognate
Bartonella
species. These observations raise the possibility that the differences in
ftsZ
sequences can be used as the basis for diagnostic tests to differentiate among these closely related pathogens.
Extracellular and Cytosolic Iron Superoxide Dismutase from
Mycobacterium bovis
BCG
Authors: Kang Sung-Koo , Jung Yong-Jae , Kim Cheorl-Ho , Song Chul-Yong
Published: December 2019

Two forms of iron superoxide dismutase (SOD) were purified from cell extract (CE) and culture filtrate (CF) of
Mycobacterium bovis
BCG, respectively. The molecular weight of both enzymes was estimated to be approximately 84,000 by gel filtration, whereas that of their subunits was 21,500, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, indicating that each of purified enzymes is composed of four identical subunits. The specific activities of CE SOD and CF SOD were 3,850 and 4,040, respectively. The purified enzymes were not joined by disulfide bonds and were, to some extent, resistant to sodium dodecyl sulfate. Their activities were lost by H
2
O
2
, but not by azide and cyanide, indicating iron SODs. Enzyme activities were detectable over a broad range of pHs, from 5.0 to 9.0, and were stable for 6 months at −20°C. Each value of pI was 4.5. In Western blots, both enzymes reacted with sera of tuberculosis patients, but not with normal sera. The N-terminal amino acid sequences of CE SOD and CF SOD were the same, suggesting that there is no N-terminal signal sequence.
Human T-Cell Lymphotropic Virus Type 1 Tax among American Blood Donors
Authors: Zucker-Franklin Dorothea , Pancake Bette A.
Published: December 2019

In the United States, all blood used for transfusion is tested for the presence of antibodies to the structural components of the human T-cell lymphotropic viruses types 1 and 2 (HTLV-1 and -2). Based on such serologic tests, the prevalence of HTLV-1 infection is estimated to range from 0.016 to 0.1%. As a consequence of studies of patients with mycosis fungoides and some of their healthy relatives who are antibody negative but were found to carry thetaxsequence of HTLV-1 in their lymphocytes and who had antibodies to the p40taxprotein, a study was undertaken to determine the prevalence of the “tax-only” state in 250 healthy blood donors and other volunteers. Using PCR and Southern analysis for cell lysates and using Western blotting for plasmas, 8.6% of the blood donors proved to betaxsequence positive and antibody positive. Sequence analysis of specimens from 22 individuals proved that 20 of the sequences were homologous with that of HTLV-1 while 2 resembled the HTLV-2 sequence. The latter were obtained from volunteers of Indian origin. The possible clinical significance of thetax-only carrier state is discussed.
Effects of Ginseng Treatment on Neutrophil Chemiluminescence and Immunoglobulin G Subclasses in a Rat Model of ChronicPseudomonas aeruginosaPneumonia
Authors: Song Zhijun , Kharazmi Arsalan , Wu Hong , Faber Viggo , Moser Claus , Krogh Johansen Helle , Rygaard Jørgen , Høiby Niels
Published: December 2019

ChronicPseudomonas aeruginosalung infection in cystic fibrosis (CF) patients is almost impossible to eradicate with antibiotic treatment. In the present study, the effects of treatment with the Chinese herbal medicine ginseng on blood polymorphonuclear leukocyte (PMN) chemiluminescence and serum specific antibody responses were studied in a rat model of chronicP. aeruginosapneumonia mimicking CF. An aqueous extract of ginseng was administered by subcutaneous injection at a dosage of 25 mg/kg of body weight/day for 2 weeks. Saline was used as a control. Two weeks after the start of ginseng treatment, significantly increased PMN chemiluminescence (P≤ 0.001) and a decreased level in serum of immunoglobulin G (IgG) againstP. aeruginosa(P< 0.05) were found. Furthermore, a higher IgG2a level (P< 0.04) but lower IgG1 level (P< 0.04) were found in the ginseng-treated infected group than in the control group. In the ginseng-treated group the macroscopic lung pathology was milder (P= 0.0003) and the percent PMNs in the cells collected by bronchoalveolar lavage was lower (P= 0.0006) than in the control group. However, the alveolar macrophage (AM) chemiluminescence values were not significantly different in the two groups infected withP. aeruginosa. The differences between the ginseng-treated noninfected rats and the control group (withoutP. aeruginosalung infection) for the PMN chemiluminescence and AM chemiluminescence were not significant. These results suggest that ginseng treatment leads to an activation of PMNs and modulation of the IgG response toP. aeruginosa, enhancing the bacterial clearance and thereby reducing the formation of immune complexes, resulting in a milder lung pathology. The changes in IgG1 and IgG2a subclasses indicate a possible shift from a Th-2-like to a Th-1-like response. These findings indicate that the therapeutic effects of ginseng may be related to activation of a Th-1 type of cellular immunity and down-regulation of humoral immunity.
Comparison of Three Different Methods for Measuring Classical Pathway Complement Activity
Authors: Jaskowski Troy D. , Martins Thomas B. , Litwin Christine M. , Hill Harry R.
Published: December 2019

The complement system plays an important role in host defense against infection and in most inflammatory processes. The standard 50% hemolytic complement (CH
50
) assay is the most commonly used method of screening patient sera for functional activity of the classical complement pathway. Our objective in this study was to compare two newer methods (the enzyme immunoassay and the liposome immunoassay) to a commercial CH
50
assay for measuring total classical complement activity. We conclude that both newer methods compare well with a CH
50
assay and are equally sensitive in screening routine clinical sera.
Expression and Self-Assembly of Grimsby Virus: Antigenic Distinction from Norwalk and Mexico Viruses
Authors: Hale Antony D. , Crawford Sue E. , Ciarlet Max , Green Jonathan , Gallimore Christopher , Brown David W. G. , Jiang Xi , Estes Mary K.
Published: December 2019

A cDNA obtained from Grimsby virus (GRV), a Norwalk-like virus, purified from a stool sample of a symptomatic adult associated with a gastroenteritis outbreak in the United Kingdom, was used to obtain the complete nucleotide sequence of the second open reading frame (ORF2). The ORF2 sequence of GRV predicts a capsid of 539 amino acids (aa) which exhibits aa identities of 96% to Lordsdale virus, 67% to Mexico virus (MXV), and 43% to Norwalk virus (NV). The GRV capsid protein was expressed in insects cells by using a recombinant baculovirus, and the resulting virus-like particles (VLPs) possessed a protein with an apparent molecular weight of 58,000. Hyperimmune antisera raised against purified GRV, MXV, and NV VLPs were tested in an indirect enzyme-linked immunosorbent assay (ELISA) against GRV, NV, and MXV VLPs, revealing that GRV is antigenically distinct from both NV and MXV. The antigenic specificity of the GRV-hyperimmune antiserum was confirmed in an antigen capture ELISA using GRV-, NV-, or MXV-containing fecal specimens. The expression of the GRV capsid protein has, for the first time, allowed the antigenic comparison of three distinct recombinant Norwalk-like viruses.
Transplacental Transmission of Serotype-Specific Pneumococcal Antibodies in a Brazilian Population
Authors: Carvalho Beatriz T. Costa , Carneiro-Sampaio Magda M. , Solé Dirceu , Naspitz Charles , Leiva Lily E. , Sorensen Ricardo U.
Published: December 2019

The highest incidence of severe pneumococcal infections in children occurs in the first 6 months of life; however, immunization of infants with the existing polysaccharide vaccines is ineffective. We wished to determine the prevalence of immunoglobulin G (IgG) pneumococcal antibodies in unimmunized Brazilian mothers and their transplacental transmission to term and preterm infants. Total IgG, IgG1 and -2 subclass levels, and IgG antibodies against
Streptococcus pneumoniae
serotypes 1, 3, 6B, 9V, and 14 were determined in 15 pairs of mothers and term newborns (gestational age, ≥37 weeks) and in 18 pairs of mothers and preterm newborns (gestational age, 32 to 36 weeks). Serotype-specific anti-pneumococcal antibodies were detected by a recently standardized enzyme-linked immunosorbent assay calibrated with the 89-SF reference serum. Varying percentages of the mothers had antibody concentrations below arbitrarily defined protective levels: 33% for serotype 1, 67% for serotype 3, 30% for serotype 6B, 52% for serotype 9V, and 22% for serotype 14. In term newborns, IgG1 concentrations were slightly higher than maternal concentrations; in preterm newborns, the concentrations were much lower. Concentrations of IgG2 in term and preterm infants were significantly lower than in the mothers. Transplacental transmission of antibodies to serotypes 3 and 14 was clearly different from that of antibodies to serotypes 1, 6B, and 9V. Concentrations of IgG antibodies against serotypes 3 and 14 were similar to or higher than those of the mothers; against serotypes 1, 6B, and 9V they ranged from 77 to 83% of maternal concentrations in term newborns and also in preterm infants, although transplacental transmission of antibodies was proportionally lower for each specific serotype in preterm than in term infants. These data are relevant for developing strategies to protect infants against pneumococcal infections in the first months of life. Our findings and a review of existing information stress the importance of understanding the relationships among pneumococcal immunization, IgG subclass antibodies to individual serotypes, transplacental transport, half-life, and antibody function and their protective values against infection.
Contributory and Exacerbating Roles of Gaseous Ammonia and Organic Dust in the Etiology of Atrophic Rhinitis
Authors: Hamilton T. D. C. , Roe J. M. , Hayes C. M. , Jones P. , Pearson G. R. , Webster A. J. F.
Published: December 2019

Pigs reared commercially indoors are exposed to air heavily contaminated with particulate and gaseous pollutants. Epidemiological surveys have shown an association between the levels of these pollutants and the severity of lesions associated with the upper respiratory tract disease of swine atrophic rhinitis. This study investigated the role of aerial pollutants in the etiology of atrophic rhinitis induced by
Pasteurella multocida
. Forty, 1-week-old Large White piglets were weaned and divided into eight groups designated A to H. The groups were housed in Rochester exposure chambers and continuously exposed to the following pollutants: ovalbumin (groups A and B), ammonia (groups C and D), ovalbumin plus ammonia (groups E and F), and unpolluted air (groups G and H). The concentrations of pollutants used were 20 mg m
−3
total mass and 5 mg m
−3
respirable mass for ovalbumin dust and 50 ppm for ammonia. One week after exposure commenced, the pigs in groups A, C, E, and G were infected with
P. multocida
type D by intranasal inoculation. After 4 weeks of exposure to pollutants, the pigs were killed and the extent of turbinate atrophy was assessed with a morphometric index (MI). Control pigs kept in clean air and not inoculated with
P. multocida
(group H) had normal turbinate morphology with a mean MI of 41.12% (standard deviation [SD], ± 1.59%). In contrast, exposure to pollutants in the absence of
P. multocida
(groups B, D, and F) induced mild turbinate atrophy with mean MIs of 49.65% (SD, ±1.96%), 51.04% (SD, ±2.06%), and 49.88% (SD, ±3.51%), respectively. A similar level of atrophy was also evoked by inoculation with
P. multocida
in the absence of pollutants (group G), giving a mean MI of 50.77% (SD, ±2.07%). However, when
P. multocida
inoculation was combined with pollutant exposure (groups A, C, and E) moderate to severe turbinate atrophy occurred with mean MIs of 64.93% (SD, ±4.64%), 59.18% (SD, ±2.79%), and 73.30% (SD, ±3.19%), respectively. The severity of atrophy was greatest in pigs exposed simultaneously to dust and ammonia. At the end of the exposure period, higher numbers of
P. multocida
bacteria were isolated from the tonsils than from the nasal membrane, per gram of tissue. The severity of turbinate atrophy in inoculated pigs was proportional to the number of
P. multocida
bacteria isolated from tonsils (
r
2
= 0.909,
P
< 0.05) and nasal membrane (
r
2
= 0.628,
P
< 0.05). These findings indicate that aerial pollutants contribute to the severity of lesions associated with atrophic rhinitis by facilitating colonization of the pig’s upper respiratory tract by
P. multocida
and also by directly evoking mild atrophy.
Human Herpesviruses in Chronic Fatigue Syndrome
Authors: Wallace Howard L. , Natelson Benjamin , Gause William , Hay John
Published: December 2019

We have conducted a double-blind study to assess the possible involvement of the human herpesviruses (HHVs) HHV6, HHV7, Epstein-Barr virus (EBV), and cytomegalovirus in chronic fatigue syndrome (CFS) patients compared to age-, race-, and gender-matched controls. The CFS patient population was composed of rigorously screened civilian and Persian Gulf War veterans meeting the Centers for Disease Control and Prevention’s CFS case definition criteria. Healthy control civilian and veteran populations had no evidence of CFS or any other exclusionary medical or psychiatric condition. Patient peripheral blood mononuclear cells were analyzed by PCR for the presence of these HHVs. Using two-tailed Fisher’s exact test analyses, we were unable to ascertain any statistically significant differences between the CFS patient and control populations in terms of the detection of one or more of these viruses. This observation was upheld when the CFS populations were further stratified with regard to the presence or absence of major axis I psychopathology and patient self-reported gradual versus acute onset of disease. In tandem, we performed serological analyses of serum anti-EBV and anti-HHV6 antibody titers and found no significant differences between the CFS and control patients.
Production, Characterization, and Uses of Monoclonal Antibodies against Recombinant Nucleoprotein of Elk Coronavirus
Authors: Daginakatte Girish C. , Chard-Bergstrom Cindy , Andrews Gordon A. , Kapil Sanjay
Published: December 2019

This is the first report of the production of monoclonal antibodies against elk coronavirus. The nucleoprotein gene of elk coronavirus was amplified by PCR and was cloned and expressed in a prokaryotic expression vector. Recombinant nucleocapsid protein was used to immunize mice for the production of hybridomas. Twelve hybridomas that produced monoclonal antibodies against the nucleocapsid protein of elk coronavirus were selected by an indirect fluorescent-antibody test, an enzyme-linked immunosorbent assay, and a Western blot assay. Ten of the monoclonal antibodies were of the immunoglobulin G1 (IgG1) isotype, one was IgG2a, and one was IgM. All had kappa light chains. By immunohistochemistry four monoclonal antibodies detected bovine coronavirus and elk coronavirus in formalin-fixed intestinal tissues. Antinucleoprotein monoclonal antibodies were found to be better at ruminant coronavirus detection than the anti-spike protein monoclonal antibodies. Because nucleoprotein is a more abundant antigen than spike protein in infected cells, this was not an unexpected finding.
Comparative Effects of Antilactoferrin Antibodies and Tumor Necrosis Factor on Neutrophil Adherence to Matrix Proteins
Authors: Zweiman Burton , von Allmen Carolyn
Published: December 2019

Neutrophil adherence to matrix proteins likely plays an important role in inflammatory responses. Antineutrophil cytoplasm antibodies may activate neutrophils in certain diseases. Using an in vitro method that allows simultaneous quantitation of neutrophil adherence and superoxide secretion, we compared the effects of antibodies against neutrophil granule proteins and tumor necrosis factor alpha (TNF-α), a known neutrophil agonist. Antilactoferrin antibodies but not antielastase or antimyeloperoxidase antibodies stimulated increased adherence to fibronectin and laminin similar in degree to that induced by TNF-α. This, but not the simultaneous superoxide secretion, was inhibited in the presence of anti-CD18 antibodies. Humoral immune responses to lactoferrin, likely expressed on the neutrophil surface, can activate neutrophils in proinflammatory responses that may be pathogenic.
Helicobacter pylori
Heat Shock Protein A: Serologic Responses and Genetic Diversity
Authors: Ng Enders K. W. , Thompson Stuart A. , Pérez-Pérez Guillermo I. , Kansau Imad , van der Ende Arie , Labigne Agnès , Sung Joseph J. Y. , Chung S. C. Sydney , Blaser Martin J.
Published: December 2019

Helicobacter pylori
synthesizes an unusual GroES homolog, heat shock protein A (HspA). The present study was aimed at an assessment of the serological response to HspA in a group of Chinese patients with defined gastroduodenal pathologies and determination of whether diversity is present in the nucleotide sequences encoding HspA in isolates from these patients. Serum samples collected from 154 patients who had an upper gastrointestinal pathology and the presence of
H. pylori
defined by biopsy were tested for an immunoglobulin G (IgG) serologic response to
H. pylori
HspA by an enzyme linked immunosorbant assay. HspA-encoding nucleotide sequences in
H. pylori
isolates from 14 patients (7 seropositive and 7 seronegative for HspA) were analyzed by PCR and direct sequencing of the PCR products. The sequencing results were compared to those of 48 isolates from other parts of the world. Of the 154 known
H. pylori
-positive patients, 54 (35.1%) were seropositive for HspA. The A domain (GroES homology) of HspA was highly conserved in the 14 isolates tested. Although the B domain (metal-binding site unique to
H. pylori
) resembled that in the known major variant, particular amino acid substitutions allowed definition of an HspA variant associated with isolates from East Asia. There were no associations between patient characteristics and HspA seropositivity or amino acid sequences. We confirmed in this study that the clinical outcomes of
H. pylori
infection are not related to HspA antigenicity or to sequence variation. However, B-domain sequence variation may be a marker for the study of the genetic diversity of
H. pylori
strains of different geographic origins.
Interactions of
Streptococcus mutans
Fimbria-Associated Surface Proteins with Salivary Components
Authors: Ray Chad A. , Gfell Linda E. , Buller Tiffany L. , Gregory Richard L.
Published: December 2019

Streptococcus mutans
has been implicated as the major causative agent of human dental caries.
S. mutans
binds to saliva-coated tooth surfaces, and previous studies suggested that fimbriae may play a role in the initial bacterial adherence to salivary components. The objectives of this study were to establish the ability of an
S. mutans
fimbria preparation to bind to saliva-coated surfaces and determine the specific salivary components that facilitate binding with fimbriae. Enzyme-linked immunosorbent assay (ELISA) established that the
S. mutans
fimbria preparation bound to components of whole saliva. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot techniques were used to separate components of whole saliva and determine fimbria binding. SDS-PAGE separated 15 major protein bands from saliva samples, and Western blot analysis indicated significant binding of the
S. mutans
fimbria preparation to a 52-kDa salivary protein. The major fimbria-binding salivary protein was isolated by preparative electrophoresis. The ability of the
S. mutans
fimbria preparation to bind to the purified salivary protein was confirmed by Western blot analysis and ELISA. Incubation of the purified salivary protein with the
S. mutans
fimbria preparation significantly neutralized binding of the salivary protein-fimbria complex to saliva-coated surfaces. The salivary protein, whole saliva, and commercial amylase reacted similarly with antiamylase antibody in immunoblots. A purified 65-kDa fimbrial protein was demonstrated to bind to both saliva and amylase. These data indicated that the
S. mutans
fimbria preparation and a purified fimbrial protein bound to whole-saliva-coated surfaces and that amylase is the major salivary component involved in the binding.
Intranasal Immunization against Dental Caries with aStreptococcus mutans-Enriched Fimbrial Preparation
Authors: Fontana Margherita , Dunipace Ann J. , Stookey George K. , Gregory Richard L.
Published: December 2019

Streptococcus mutanshas been identified as the major etiological agent of human dental caries. The first step in the initiation of infection by this pathogenic bacterium is its attachment (i.e., through bacterial surface proteins such as glucosyltransferases, P1, glucan-binding proteins, and fimbriae) to a suitable receptor. It is hypothesized that a mucosal vaccine against a combination ofS. mutanssurface proteins would protect against dental caries by inducing specific salivary immunoglobulin A (IgA) antibodies which may reduce bacterial pathogenesis and adhesion to the tooth surface by affecting several adhesins simultaneously. Conventional Sprague-Dawley rats, infected withS. mutansat 18 to 20 days of age, were intranasally immunized with a mixture ofS. mutanssurface proteins, enriched for fimbriae and conjugated with cholera toxin B subunit (CTB) plus free cholera toxin (CT) at 13, 15, 22, 29, and 36 days of age (group A). Control rats were either not immunized (group B) or immunized with adjuvant alone (CTB and CT [group C]). At the termination of the study (when rats were 46 days of age), immunized animals (group A) had significantly (P< 0.05) higher salivary IgA and serum IgG antibody responses to the mixture of surface proteins and to whole bacterial cells than did the other two groups (B and C). No significant differences were found in the average numbers of recoveredS. mutanscells among groups. However, statistically fewer smooth-surface enamel lesions (buccal and lingual) were detected in the immunized group than in the two other groups. Therefore, a mixture ofS. mutanssurface proteins, enriched with fimbria components, appears to be a promising immunogen candidate for a mucosal vaccine against dental caries.
Evaluation of Newly Developed Immunoperoxidase Monolayer Assays for Detection of Antibodies against Bovine Herpesvirus 4
Authors: Wellenberg G. J. , van Rooij E. M. A. , Maissan J. , Van Oirschot J. T.
Published: December 2019

This study describes the evaluation of immunoperoxidase monolayer assays (IPMAs) for detection of antibodies against bovine herpesvirus 4 (BHV4) DN-599 or BHV4 LVR 140 in sera of cattle. We compared the quality of these IPMAs with the quality of a BHV4 indirect enzyme-linked immunosorbent assay (ELISA). In addition, a preliminary serological survey of BHV4 antibodies was carried out to estimate the seroprevalence of BHV4 in Dutch cattle at different ages. The specificities of both BHV4 IPMAs were 1.00. The geometrical mean titers (detection limit) of the BHV4 IPMAs were twice as high as that of the BHV4 indirect ELISA. In experimentally infected cattle, BHV4 antibodies were detectable by IPMAs 16 to 18 days postinfection, which was almost 2 weeks earlier than in the indirect ELISA. The reproducibility of the BHV4 DN-599 IPMA (
κ
D value, 0.92) and of the BHV4 LVR 140 IPMA (
κ
D value, 0.87) were good. For field sera the overall agreement between the BHV4 indirect ELISA and the two BHV4 IPMAs, DN-599 and LVR 140, was 95 and 96%, respectively. The serological-survey study showed that the estimated seroprevalence of BHV4 in Dutch cattle was 16 to 18% and that the percentage of BHV4-positive animals varied by age category (between 6 and 43%). In summary, the two BHV4 IPMA formats have several advantages that make IPMA a useful alternative to the BHV4 indirect ELISA for detecting BHV4 antibodies in cattle.
Cytokine mRNA Expression in Lesions in Cats with Chronic Gingivostomatitis
Authors: Harley R. , Helps C. R. , Harbour D. A. , Gruffydd-Jones T. J. , Day M. J.
Published: December 2019

Semiquantitative reverse transcription-PCR assays were developed to measure feline interleukin-2 (IL-2), IL-4, IL-5, IL-6, IL-10, and IL-12 (p35 & p40); gamma interferon (IFN-γ); and glyceraldehyde-3-phosphate dehydrogenase mRNA concentrations in biopsies of feline oral mucosa. Biopsies were collected from 30 cats with chronic gingivostomatitis (diseased) prior to each cat receiving one of four treatments. In 23 cases replicate biopsies were collected 3 months after treatment commenced. Biopsies were also analyzed from 11 cats without clinical disease (nondiseased). Expression of IL-2, IL-10, IL-12 (p35 and p40), and IFN-γ was detected in most nondiseased biopsies, while IL-6 was detected in a minority, and IL-4 and IL-5 were both undetectable. Compared to nondiseased cats, the diseased population showed a significant increase in the relative mRNA expression of IL-2, IL-4, IL-6, IL-10, IL-12 (p35 and p40), and IFN-γ. In contrast, IL-5 mRNA expression was unchanged and was only detected in one case. No significant relationship was demonstrable between the change in relative expression of specific cytokine mRNA and the change in clinical severity of the local mucosal lesions over the treatment period. The results demonstrate that the normal feline oral mucosa is biased towards a predominantly (Th) type 1 profile of cytokine expression and that during the development of lesions seen in feline chronic gingivostomatitis there is a shift in the cytokine profile from a type 1 to a mixed type 1 and type 2 response.
Decreased Superoxide Production, Degranulation, Tumor Necrosis Factor Alpha Secretion, and CD11b/CD18 Receptor Expression by Adherent Monocytes from Preterm Infants
Authors: Kaufman David , Kilpatrick Laurie , Hudson R. Guy , Campbell Donald E. , Kaufman Ann , Douglas Steven D. , Harris Mary C.
Published: December 2019

Preterm infants have an increased incidence of infection, which is principally due to deficiencies in neonatal host defense mechanisms. Monocyte adherence is important in localizing cells at sites of infection and is associated with enhanced antimicrobial functions. We isolated cord blood monocytes from preterm and full-term infants to study their adhesion and immune functions, including superoxide (O2−) generation, degranulation, and cytokine secretion and their adhesion receptors. O2−production and degranulation were significantly diminished, by 28 and 37%, respectively, in adherent monocytes from preterm infants compared to full-term infants (P< 0.05); however, these differences were not seen in freshly isolated cells. We also observed a significant decrease of 35% in tumor necrosis factor alpha secretion by lipopolysaccharide-stimulated adherent monocytes from preterm infants compared to full-term infants (P< 0.05); however, this difference was not observed in interleukin-1β or interleukin-6 production by the monocytes. The cell surface expression of the CD11b/CD18 adhesion receptor subunits was significantly decreased (by 60 and 52%, respectively) in monocytes from preterm infants compared to full-term infants (P< 0.01). The cascade of the immune response to infection involves monocyte upregulation and adherence via CD11b/CD18 receptors followed by cell activation and the release of cytokines and bactericidal products. We speculate that monocyte adherence factors may be important in the modulation of immune responses in preterm infants.
Outcomes ofBordetellaInfections in Vaccinated Children: Effects of Bacterial Number in the Nasopharynx and Patient Age
Authors: He Qiushui , Arvilommi Heikki , Viljanen Matti K. , Mertsola Jussi
Published: December 2019

Five outbreaks of infection (three pertussis, one parapertussis, and one mixed) in schools were studied prospectively. Nasopharyngeal swabs were obtained from a total of 697 children for culture ofBordetellaorganisms. Of 50 vaccinated children with culture-confirmedBordetellainfections (29 with pertussis and 21 parapertussis), 40 were symptomatic and 10 remained symptom-free. Smaller numbers of colonies were recovered from the nasopharyngeal swabs of the asymptomatic children than from those of the symptomatic children. Older children had longer durations of illness than younger ones. Our results indicate that during outbreaks children who do not develop disease may have small amounts ofBordetellaorganisms in their nasopharynges and/or better immune defenses against the disease.
Development of an Antigen Spot Test for Detection of Coronavirus in Bovine Fecal Samples
Authors: Gaber Fathy , Kapil Sanjay
Published: December 2019

We have developed a rapid and sensitive microimmunodot blot assay, the antigen spot test (AST), for the detection of bovine coronavirus (BCV) antigen from neonatal calf fecal samples. The AST procedure can be completed in 3.5 h, whereas the previously reported immunodot blot assays require 10 to 12 h. Ninety-six samples can be tested per membrane, and 10 membranes (960 samples) may be processed by a single technologist in 1 working day. The effects of detergents, oxidizing chemicals, chaotropic agents, and enzyme substrates in improving the sensitivity and signal-to-noise ratio of the AST were studied. Finally, the sensitivity and specificity of AST for the detection of BCV antigen were compared to those of a sandwich enzyme-linked immunosorbant assay (ELISA) and a hemagglutination assay (HA). Of 347 field samples tested by all three methods, 94.2% were positive by AST, 91.4% were positive by ELISA, and 86.7% were positive by HA. The sensitivity of the AST was determined to be 100% compared to the results of the ELISA reference method. The specificity of the AST was 67%, which reflects a lower limit of detection of 10
4
viral particles per ml in a 10% fecal suspension.
A Flow Cytometric Opsonophagocytic Assay for Measurement of Functional Antibodies Elicited after Vaccination with the 23-Valent Pneumococcal Polysaccharide Vaccine
Authors: Martinez Joseph E. , Romero-Steiner Sandra , Pilishvili Tamara , Barnard Suzanne , Schinsky Joseph , Goldblatt David , Carlone George M.
Published: December 2019

Opsonophagocytosis is the primary mechanism for clearance of pneumococci from the host, and the measurement of opsonophagocytic antibodies appears to correlate with vaccine-induced protection. We developed a semiautomated flow cytometric opsonophagocytosis assay using HL-60 granulocytes as effector cells and nonviable 5,6-carboxyfluorescein, succinimidyl ester-labeledStreptococcus pneumoniae(serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F) as bacterial targets. The flow cytometric opsonophagocytosis assay was highly reproducible (for 87% of repetitive assays the titers were within 1 dilution of the median titer) and serotype specific, with ≥97% inhibition of opsonophagocytic titer by addition of homologous serotype-specific polysaccharide. In general, opsonophagocytic titers were not significantly inhibited by the presence of either heterologous pneumococcal polysaccharide or penicillin in the serum. The flow cytometric assay could reproducibly measure functional antibody activity in prevaccination (n= 28) and postvaccination (n= 36) serum specimens from healthy adult volunteers vaccinated with the 23-valent pneumococcal polysaccharide vaccine. When compared with a standardized manual viable opsonophagocytic assay, a high correlation (r= 0.89;P≤ 0.01) was found between the two assays for the seven serotypes tested. The flow cytometric assay is rapid (∼4 h) with high throughput (∼50 serum samples per day per technician) and provides a reproducible measurement of serotype-specific functional antibodies, making it a highly suitable assay for the evaluation of the immune responses elicited by pneumococcal vaccines.
Heterophile Antibodies to Bovine and Caprine Proteins Causing False-Positive Human Immunodeficiency Virus Type 1 and Other Enzyme-Linked Immunosorbent Assay Results
Authors: Willman Joseph H. , Martins Thomas B. , Jaskowski Troy D. , Hill Harry R. , Litwin Christine M.
Published: December 2019

Heterophile antibodies are a well-recognized cause of erroneous results in immunoassays. We describe here a 22-month-old child with heterophile antibodies reactive with bovine serum albumin and caprine proteins causing false-positive results to human immunodeficiency virus type 1 and other infectious serology testing.
Total Immunoglobulin E Levels and Dengue Infection on San Andrés Island, Colombia
Authors: Míguez-Burbano María José , Jaramillo Carlos A. , Palmer Carol J. , Shor-Posner Gail , Velásquez Luz Stella , Lai Hong , Baum Marianna K.
Published: December 2019

An evaluation of total immunoglobulin E (IgE) and dengue serostatus in 168 subjects from San Andrés Island, Colombia, revealed altered levels of IgE in 89% of the population. IgE levels were higher in patients with a history of dengue or with a current secondary or current primary infection than in subjects with no exposure (
P
= 0.01). Dengue infection accounted for 23% of the variation in IgE levels.
Brucella
Outer Membrane Lipoproteins Share Antigenic Determinants with Bacteria of the Family
Rhizobiaceae
Authors: Cloeckaert Axel , Tibor Anne , Zygmunt Michel S.
Published: December 2019

Brucellae have been reported to be phylogenetically related to bacteria of the family
Rhizobiaceae
. In the present study, we used a panel of monoclonal antibodies (MAbs) to
Brucella
outer membrane proteins (OMPs) to determine the presence of common OMP epitopes in some representative bacteria of this family, i.e.,
Ochrobactrum anthropi
,
Phyllobacterium rubiacearum
,
Rhizobium leguminosarum
, and
Agrobacterium tumefaciens
, and also in bacteria reported to serologically cross-react with brucella, i.e.,
Yersinia enterocolitica
O:9,
Escherichia coli
O:157, and
Salmonella urbana
. In particular, most MAbs to the
Brucella
outer membrane lipoproteins Omp10, Omp16, and Omp19 cross-reacted with
O. anthropi
and
P. rubiacearum
, which are actually the closest relatives of brucellae. Some of them also cross-reacted, but to a lower extent, with
R. leguminosarum
and
A. tumefaciens
. The putative Omp16 and Omp19 homologs in these bacteria showed the same apparent molecular masses as their
Brucella
counterparts. None of the antilipoprotein MAbs cross-reacted with
Y. enterocolitica
O:9,
E. coli
O:157, or
S. urbana
.
Mastitis and Immunological Factors in Breast Milk of Lactating Women in Malawi
Authors: Semba Richard D. , Kumwenda Newton , Taha Taha E. , Hoover Donald R. , Lan Yin , Eisinger Ward , Mtimavalye Laban , Broadhead Robin , Miotti Paolo G. , Van Der Hoeven Len , Chiphangwi John D.
Published: December 2019

Although an elevated sodium concentration in human milk is suggested to be an indicator of mastitis, it is unclear whether elevated sodium concentrations are associated with immunological and inflammatory mediators in human milk. We conducted a cross-sectional study to evaluate the relationships between elevated breast milk sodium concentrations and levels of lactoferrin, lysozyme, secretory leukocyte protease inhibitor (SLPI), interleukin-8 (IL-8), and RANTES (regulated on activation normal T cell expressed and secreted) in human milk at 6 weeks postpartum in 96 lactating women in Blantyre, Malawi. Mastitis, as indicated by an elevated breast milk sodium concentration, was present in 15.6% of the women. Women with and without mastitis had respective median levels of other factors as follows: lactoferrin, 1,230 versus 565 mg/liter (P< 0.0007); lysozyme, 266 versus 274 mg/liter (P= 0.55); SLPI, 76 versus 15 μg/liter, (P< 0.0002); IL-8, 339 versus 25 ng/liter (P< 0.0001); and RANTES, 82 versus 3 ng/liter (P< 0.0001). Elevated sodium concentrations in breast milk are associated with an increase in levels of some immunological and inflammatory factors in breast milk.
Development of Diagnostic Reagents To Differentiate betweenMycobacterium bovisBCG Vaccination andM. bovisInfection in Cattle
Authors: Vordermeier H. M. , Cockle P. C. , Whelan A. , Rhodes S. , Palmer N. , Bakker D. , Hewinson R. G.
Published: December 2019

In Great Britain a recent independent scientific review for the government has concluded that the development of a cattle vaccine againstMycobacterium bovisholds the best long-term prospect for tuberculosis control in British herds. A sine qua non for vaccination is the development of a complementary diagnostic test to differentiate between vaccinated animals and those infected withM. bovisso that test-and-slaughter-based control strategies can continue alongside vaccination. In order to assess the feasibility of developing a differential diagnostic test for a live vaccine, we choseM. bovisBCG Pasteur as a model system. Recombinant forms of antigens which are expressed inM. bovisbut not, or only at low levels, in BCG Pasteur (ESAT-6, MPB64, MPB70, and MPB83) were produced. These reagents were tested either alone or in combination by using peripheral blood mononuclear cells fromM. bovis-infected, BCG-vaccinated, andMycobacterium avium-sensitized calves. All four antigens induced in vitro proliferation and gamma interferon responses only inM. bovis-infected animals. A cocktail composed of ESAT-6, MPB64, and MPB83 identified infected animals but not those vaccinated with BCG. In addition, promiscuous T-cell epitopes of ESAT-6, MPB64, and MPB83 were formulated into a peptide cocktail. In T-cell assays with this peptide cocktail, infected animals were identified with frequencies similar to those obtained in assays with the protein cocktail, while BCG-vaccinated orM. avium-sensitized animals did not respond. In summary, our results suggest that peptide and protein cocktails can be designed to discriminate betweenM. bovisinfection and BCG vaccination.
Urinary Excretion of Thiol Compounds in Patients with Rheumatoid Arthritis
Authors: Rojkovich Bernadette , Nagy Eszter , Pröhle Tamás , Poór Gyula , Gergely Péter
Published: December 2019

The objective of the present study was to assess the excretion of urinary thiol compounds in patients with active and inactive rheumatoid arthritis (RA). Urinary thiol compounds were measured by the method of Kokonov (M. T. Kokonov, Lab. Delo 5:273–276, 1965) in 51 outpatients with active and inactive RA. Those with active disease had significantly higher levels of urinary thioamine excretion.
Detection of Borna Disease Virus-Reactive Antibodies from Patients with Psychiatric Disorders and from Horses by Electrochemiluminescence Immunoassay
Authors: Yamaguchi Kazunari , Sawada Takashi , Naraki Tohru , Igata-Yi Ruriko , Shiraki Hiroshi , Horii Yoichiro , Ishii Toshinori , Ikeda Kazuhiko , Asou Norio , Okabe Hiroaki , Mochizuki Manabu , Takahashi Kazuo , Yamada Shogo , Kubo Kaori , Yashiki Shinji , Waltrip Royce W. , Carbone Kathryn M.
Published: December 2019

The prevalence of Borna disease virus (BDV)-specific antibodies among patients with psychiatric disorders and healthy individuals has varied in several reports using several different serological assay methods. A reliable and specific method for anti-BDV antibodies needs to be developed to clarify the pathological significance of BDV infections in humans. We developed a new electrochemiluminescence immunoassay (ECLIA) for the antibody to BDV that uses two recombinant proteins of BDV, p40 and p24 (full length). Using this ECLIA, we examined 3,476 serum samples from humans with various diseases and 917 sera from blood donors in Japan for the presence of anti-BDV antibodies. By ECLIA, 26 (3.08%) of 845 schizophrenia patients and 9 (3.59%) of 251 patients with mood disorders were seropositive for BDV. Among 323 patients with other psychiatric diseases, 114 with neurological diseases, 75 with chronic fatigue syndrome, 85 human immunodeficiency virus-infected patients, 50 with autoimmune diseases including rheumatoid arthritis and systemic lupus erythematosis and 17 with leprosy, there was no positive case except one case each with alcohol addiction, AIDS, and dementia. Although 19 (1.36%) of 1,393 patients with various ocular diseases, 10 (1.09%) of 917 blood donors, and 3 (4.55%) of 66 multitransfused patients were seropositive for BDV-specific antigen, high levels of seroprevalence in schizophrenia patients and young patients (16 to 59 years old) with mood disorders were statistically significant. The immunoreactivity of seropositive sera could be verified for specificity by blocking with soluble p40 and/or p24 recombinant protein. Anti-p24 antibody was more frequent than p40 antibody in most cases, and in some psychotic patients antibody profiles showed only p40 antibody. Although serum positive for both p40 and p24 antibodies was not found in this study, the p40 ECLIA count in schizophrenia patients was higher than that of blood donors. Furthermore, we examined 90 sera from Japanese feral horses. Antibody profiles of control human samples are similar to that of naturally BDV-infected feral horses. We concluded that BDV infection was associated in some way with psychiatric disorders.
Cell Culture of Sporadic Hepatitis E Virus in China
Authors: Huang Rutong , Li Derong , Wei Shaojing , Li Qinghong , Yuan Xitong , Geng Liqing , Li Xiaoyu , Liu Minxia
Published: December 2019

The isolation and identification of the 87A strain of epidemic hepatitis E virus (HEV) by means of cell culturing have been described previously. This paper reports the successful isolation of a sporadic HEV strain (G93-2) in human lung carcinoma cell (A549) cultures. The etiology, molecular and biological properties, and serological relationship of this new strain to other, epidemic HEV strains are described. The propagation of both sporadic and epidemic HEV strains in a cell culture system will facilitate vaccine research.
Hepatitis B Virus DNA in Blood Samples Positive for Antibodies to Core Antigen and Negative for Surface Antigen
Authors: Gutiérrez C. , León G. , Loureiro C. L. , Uzcátegui N. , Liprandi F. , Pujol F. H.
Published: December 2019

Anti-hepatitis B core antigen (HBcAg)-positive hepatitis B surface antigen (HBsAg)-negative plasma samples from blood donors were tested by nested PCR. DNA positivity was more significantly associated with high levels of anti-HBcAg than with low levels of anti-HBsAg antibodies. Analysis of a dilution of anti-HBcAg antibodies might result in a more rational exclusion of anti-HBcAg-positive HBsAg-negative samples, reducing the number of donations discarded and enabling more countries to incorporate anti-HBcAg testing.
Antibody to Human Endogenous Retrovirus Peptide in Urine of Human Immunodeficiency Virus Type 1-Positive Patients
Authors: Stevens Roy W. , Baltch Aldona L. , Smith Raymond P. , McCreedy Bruce J. , Michelsen Phyllis B. , Bopp Lawrence H. , Urnovitz Howard B.
Published: December 2019

Human endogenous retrovirus (HERV)-like sequences are normal inherited elements that constitute several hundredths of the human genome. The expression of genes located within these elements can occur as a consequence of several different events, including persistent inflammation or genotoxic events. Antibodies to endogenous retroviral gene products have been found in a number of infectious, chronic, and malignant diseases, suggesting a role in disease initiation and progression. We studied human immunodeficiency virus type 1 (HIV-1)-infected patients for evidence of urine antibody to a HERV peptide and investigated correlates with clinical and laboratory parameters. Forty-three HIV-1-infected patients in documented asymptomatic, symptomatic, or AIDS stages of disease and 21 age- and gender-matched, uninfected controls were tested for antibody to HERV-related peptide 4.1. Urine specimens were examined in a blinded fashion with the Calypte Biomedical Corp. experimental enzyme immunoassay for antibody to peptide 4.1. Results were compared with demographic data, medical history, clinical state of disease, and results of other laboratory tests. Thirty-six percent of the asymptomatic (Centers for Disease Control and Prevention [CDC] category A) and 81.3% of both the symptomatic (CDC category B) and AIDS (CDC category C) patients were positive for antibody to HERV-related peptide 4.1. None of the controls were positive. In this study, antibodies to HERV-related peptide 4.1 were found more frequently in patients with advanced stages (categories B and C) of HIV-1 disease than in those patients with an earlier stage (category A) of HIV disease. In HIV patients, severe immunosuppression, defined as having had at least one opportunistic infection, correlated with the expression of antibody to a HERV-related peptide.
Cloning and Expression of a DNA Sequence Encoding a 41-Kilodalton
Cryptosporidium parvum
Oocyst Wall Protein
Authors: Jenkins Mark C. , Trout Jim , Murphy Charles , Harp James A. , Higgins Jim , Wergin William , Fayer Ron
Published: December 2019

This study was conducted to produce a recombinant species-specific oocyst wall protein of
Cryptosporidium parvum
. Antigens unique to
C. parvum
were identified by gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting of oocyst proteins from several different
Cryptosporidium
species. Antiserum was then prepared against a 41-kDa antigen unique to
C. parvum
and used to identify a recombinant DNA clone, designated rCP41. Expression of CP41 mRNA in
C. parvum
oocysts was confirmed by reverse transcriptase PCR (RT-PCR). Although the CP41 sequence was shown by PCR to be present in the genome of
C. baileyi
, CP41 mRNA was not detected in this species by RT-PCR. Immunofluorescence staining with antiserum against recombinant CP41 detected native CP41 antigen on the surface of
C. parvum
oocysts but failed to detect CP41 on
C. baileyi
oocysts. Immunoelectron microscopy demonstrated that native CP41 was distributed unevenly on the
C. parvum
oocyst surface and was associated with amorphous oocyst wall material. In an enzyme-linked immunosorbent assay, purified rCP41 performed as well as native
C. parvum
oocyst protein in measuring the serological responses of young calves and adult cows to experimental and natural
C. parvum
infections. These results indicate that recombinant CP41 antigen may have potential in the immunodiagnosis of cryptosporidiosis.