bluuebunny/crossref_metadata_2025_split huggingface dataset:
Increased Levels of Alternatively Spliced Interleukin 4 (IL-4δ2) Transcripts in Peripheral Blood Mononuclear Cells from Patients with Systemic Sclerosis
Authors: Sakkas Lazaros I. , Tourtellotte Charles , Berney Steve , Myers Allen R. , Platsoucas Chris D. 
Published: December 2019

ABSTRACTRecent in vitro studies have shown that interleukin 4 (IL-4) induces and gamma interferon (IFN-γ) inhibits collagen production. To define the TH1(IFN-γ) and TH2(IL-4) cytokine profiles in systemic sclerosis (Sscl), a disease characterized by widespread fibrosis, we investigated IL-4 and IFN-γ transcripts in peripheral blood mononuclear cells and plasma protein levels in 13 patients with Sscl. Two previously identified IL-4 transcripts, a full-length transcript and an alternatively spliced (truncated) transcript (designated IL-4δ2), were identified in patients and normal controls. Significantly increased levels of total IL-4 transcripts (full-length plus IL-4δ2 transcripts) were found in patients with Sscl in comparison to those found in healthy controls (P= 0.003), and this increase was primarily due to an increase in the level of the alternatively spliced IL-4δ2 form. The IL-4δ2/full-length-IL-4 transcript ratio was significantly increased in Sscl patients (P< 0.0001, versus healthy controls). Sequencing analysis revealed that the frequency of IL-4 clones carrying the IL-4δ2 transcript was also substantially increased in patients with Sscl. Plasma IL-4 protein levels were increased in Sscl patients compared to those in healthy controls (P= 0.001) and correlated with total IL-4 transcript levels. The up-regulation of the fibrogenic IL-4 (a TH2 cytokine) in Sscl suggests a pathogenic role for IL-4 in this disease.
Comparison of PanBio Dengue Duo Enzyme-Linked Immunosorbent Assay (ELISA) and MRL Dengue Fever Virus Immunoglobulin M Capture ELISA for Diagnosis of Dengue Virus Infections in Southeast Asia
Authors: Cuzzubbo Andrea J. , Vaughn David W. , Nisalak Ananda , Solomon Tom , Kalayanarooj Siripen , Aaskov John , Dung Nguyen Minh , Devine Peter L. 
Published: December 2019

ABSTRACT

The performances of the MRL dengue fever virus immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (ELISA) and the PanBio Dengue Duo IgM capture and IgG capture ELISA were compared. Eighty sera from patients with dengue virus infections, 24 sera from patients with Japanese encephalitis (JE), and 78 sera from patients with nonflavivirus infections, such as malaria, typhoid, leptospirosis, and scrub typhus, were used. The MRL test showed superior sensitivity for dengue virus infections (94 versus 89%), while the PanBio test showed superior specificity for JE (79 versus 25%) and other infections (100 versus 91%). The PanBio ELISA showed better overall performance, as assessed by the sum of sensitivity and specificity (
F
value). When dengue virus and nonflavivirus infections were compared,
F
values of 189 and 185 were obtained for the PanBio and MRL tests, respectively, while when dengue virus infections and JE were compared,
F
values of 168 and 119 were obtained. The results obtained with individual sera in the PanBio and MRL IgM ELISAs showed good correlation, but this analysis revealed that the cutoff value of the MRL test was set well below that of the PanBio test. Comparing the sensitivity and specificity of the tests at different cutoff values (receiver-operator analysis) revealed that the MRL and PanBio IgM ELISAs performed similarly in distinguishing dengue virus from nonflavivirus infections, although the PanBio IgM ELISA showed significantly better distinction between dengue virus infections and JE. The implications of these findings for the laboratory diagnosis of dengue are discussed.
Cloning and Expression of an Immunogenic Membrane-Associated Protein of
Helicobacter hepaticus
for Use in an Enzyme-Linked Immunosorbent Assay
Authors: Livingston Robert S. , Riley Lela K. , Hook Reuel R. , Besch-Williford Cynthia L. , Franklin Craig L. 
Published: December 2019

ABSTRACT

Helicobacter hepaticus
is a bacterial pathogen that causes chronic active hepatitis and inflammatory bowel disease in mice. The purpose of this study was to develop a recombinant antigen-based enzyme-linked immunosorbent assay (ELISA) to detect
H. hepaticus
-infected mice. A genomic library of
H. hepaticus
was constructed and was screened with sera from
H. hepaticus
-infected mice. A 459-bp open reading frame that coded for an 18-kDa immunoreactive protein, MAP18, was identified. The gene had high identity with genes coding for outer membrane proteins of other bacteria, and the predicted amino acid sequence of MAP18 had a putative membrane-trafficking signal sequence and a putative signal peptidase II cleavage site. The recombinant protein was expressed in
Escherichia coli
as a glutathione
S
-transferase (GST) fusion protein, GST-MAP18, and purified by affinity chromatography. The 44-kDa fusion protein was detected on Western blots probed with sera from
H. hepaticus
-infected mice but was not detected on blots probed with sera from mice infected with
Helicobacter muridarum
or
Helicobacter bilis
or with sera from mice free of
Helicobacter
infection. The GST-MAP18 fusion protein was used as an antigen in an ELISA to detect anti-
H. hepaticus
antibodies in sera from infected mice. This ELISA was compared to an
H. hepaticus
-specific ELISA that uses a detergent extract of
H. hepaticus
as the antigen. Sera from mice naturally and experimentally infected with
H. hepaticus
,
H. bilis
, or
H. muridarum
and sera from mice free of
Helicobacter
infection were evaluated. Both ELISAs performed with a high specificity (98%); however, the detergent extract-based ELISA performed with a higher sensitivity (89%) than the recombinant protein-based ELISA (sensitivity, 66%). These data indicate that
H. hepaticus
carries a gene that encodes an immunogenic 18-kDa membrane-associated protein; however, antibodies to this protein are not detected in all infected mice.
Th1 Cytokine Patterns in Cervical Human Papillomavirus Infection
Authors: Scott Mark , Stites Daniel P. , Moscicki Anna-Barbara 
Published: December 2019

ABSTRACTThe host’s immune response to cervical human papillomavirus (HPV) infection is poorly understood. In a longitudinal cohort of women with cervical HPV infections, defined by PCR-based HPV DNA testing, we used exfoliated cervical cells and reverse transcription-PCR to examine the cervical mucosal mRNA expression of cytokines involved in regulating cell-mediated immunity. We identified seven HPV-positive subjects who were found to have cleared their HPV infections 4 months later. In all seven, a T-helper type 1 (Th1) cytokine pattern (expression of gamma interferon and absence of interleukin-4) preceded clearance. The more variable cytokine patterns seen in HPV-negative subjects suggest that the Th1 pattern in the women with subsequent clearance was a response to the HPV infection. This contention is supported by additional cross-sectional data showing a Th1 pattern in a majority of HPV-positive women. This study establishes a feasible means for assessing local cytokine expression in the cervical milieu and demonstrates that a Th1 cytokine response is associated with subsequent clearance of cervical HPV infection.
Serological Evidence of Human Infection with the ProtozoanNeospora caninum
Authors: Tranas Jennifer , Heinzen Robert A. , Weiss Louis M. , McAllister Milton M. 
Published: December 2019

ABSTRACTNeospora caninumis a protozoan parasite that is closely related toToxoplasma gondii. Dogs are a definitive host. Prior to its discovery in 1988,N. caninuminfection in animals was often mistakenly diagnosed as toxoplasmosis. Neosporosis in animals is characterized by encephalitis, abortion, and other conditions that clinically and pathologically resemble toxoplasmosis. The potential ofN. caninumto infect humans is unknown. Therefore, evidence of human exposure to this parasite was sought by screening for antibodies in blood donors by indirect fluorescent antibody (IFA) tests and immunoblotting. Of 1,029 samples screened, 69 (6.7%) had titers of 1:100 by IFA testing. Fifty of the 69 (72%) sera that were positive forN. caninumwere also negative for a closely related protozoan pathogen of humans,T. gondii. Immunoblot analysis confirmed the specificity of the positive sera forN. caninumantigens, with several sera recognizing multipleNeosporaantigens with molecular masses similar to those of antigens recognized by monkey anti-N. caninumserum. An immunodominant antigen of approximately 35 kDa was observed with 12 sera. These data provide evidence of human exposure toN. caninum, although the antibody titers in healthy donors were low. The significance of human exposure to, and possible infection with, this parasite is unknown and warrants further study.
Effects of Orally Administered Viable
Lactobacillus rhamnosus
GG and
Propionibacterium freudenreichii
subsp.
shermanii
JS on Mouse Lymphocyte Proliferation
Authors: Kirjavainen Pirkka V. , ElNezami Hani S. , Salminen Seppo J. , Ahokas Jorma T. , Wright Paul F. A. 
Published: December 2019

ABSTRACT

Immunomodulation by probiotics is a subject of growing interest, but the knowledge of dose response and time profile relationships is minimal. In this study we examined the effects of
Lactobacillus rhamnosus
GG (LGG) and
Propionibacterium freudenreichii
subsp.
shermanii
JS (PJS) on the proliferative activity of murine lymphocytes ex vivo. Dose dependency was assessed by treating animals perorally with a low or a high dose (i.e., 10
9
or 10
12
viable bacteria/kg of body weight) for 7 days. The lower dose levels of each strain appeared to enhance T-cell proliferation at the optimal concanavalin A (ConA) concentration (by 69 to 84%) and B-cell proliferation at the optimal and supraoptimal concentrations of lipopolysaccharide (by 57 to 82%). B-cell proliferation was also enhanced by the high LGG dose (by 32 to 39%) but was accompanied by a marginal decrease in T-cell proliferation (by 8%) at the optimal ConA concentration. The time profiles of the immune responses were assessed after daily treatment with the higher dose for 3, 7, and 14 days. A significant decrease in basal lymphoproliferation (by 32 to 42%) was observed with PJS treatment after the 3- and 7-day periods; however, this activity returned to control levels after 14 days of treatment, which also resulted in significantly enhanced T-cell proliferation at optimal and supraoptimal ConA concentrations (by 24 to 80%). The 14-day LGG treatment also enhanced the latter activity (by 119%). In conclusion, LGG and PJS have specific dose- and duration-dependent immunomodulatory effects on the proliferative activity of B and T lymphocytes and may also reduce lymphocyte sensitivity to the cytotoxic effects of lectin mitogens.
Lipopolysaccharide- and Cholera Toxin-Specific Subclass Distribution of B-Cell Responses in Cholera
Authors: Qadri Firdausi , Ahmed Firoz , Karim M. Manjurul , Wenneras Christine , Begum Yasmin Ara , Salam Mohammad Abdus , Albert M. John , McGhee Jerry R. 
Published: December 2019

ABSTRACTThe immunoglobulin subclass responses to homologous lipopolysaccharide (LPS) and to cholera toxin (CT) in adult patients infected withVibrio choleraeO1 andV. choleraeO139 were studied. LPS-specific antibody-secreting cells (ASC) of both the immunoglobulin A1 (IgA1) and IgA2 subclasses were seen, with the IgA1 ASC response predominating in bothV. choleraeO1- and O139-infected patients. For antibodies in plasma, by day 11 after onset of disease, allV. choleraeO1- infected patients responded to homologous LPS with the IgA1 subclass (P= 0.001), whereas fewer (68%) responded with the IgA2 subclass (P= 0.007). About 89% ofV. choleraeO139-infected patients responded with the IgA1 subclass (P= 0.003), and only 21% responded with the IgA2 subclass (not significant [NS]). Both groups of cholera patients showed significant increases in LPS-specific IgG1, IgG2, and IgG3 antibodies in plasma. In feces, the response to homologous LPS occurred in both groups of patients with the IgA1 and IgA2 subclasses, with 55 to 67% of patients showing a positive response.V. choleraeO1- and O139-infected patients showed CT-specific ASC responses of the different IgG and IgA subclasses in the circulation, and the pattern followed the order IgG1 > IgA1 > IgG2 > IgA2, with low levels of IgG3 and IgG4 ASC. Plasma anti-CT antibody responses in all subclasses were seen by day 11 after onset of disease. Although there were no increases in CT-specific ASC of the IgG3 (NS) and IgG4 (NS) subtypes, there were significant increases of these two subclasses in plasma (P≤ 0.001). The response to CT in the fecal extracts was contributed to by both IgA1 and IgA2 isotypes, with 67 to 75% of the patients responding. Thus, the mucosa-derived ASC and fecal antibodies to LPS and CT were of both the IgA1 and IgA2 subclasses; in plasma, the contribution from IgA2 was lower. Very little difference in the B-cell responses to LPS and CT in the different subclasses was seen in the two groups of cholera patients. Vaccines against O1 and O139 cholera ideally should stimulate antibody subclasses that are likely to offer protection.
Changes in Endotoxin-Binding Proteins during Major Elective Surgery: Important Role for Soluble CD14 in Regulation of Biological Activity of Systemic Endotoxin
Authors: Hiki Naoki , Berger Dieter , Dentener Mieke A. , Mimura Yoshikazu , Buurman Wim A. , Prigl Claus , Seidelmann Manuela , Tsuji Eiichi , Kaminishi Michio , Beger Hans G. 
Published: December 2019

ABSTRACTAssessment of circulating endotoxin during the perioperative period, which is only demonstrated by theLimulusamebocyte lysate (LAL) test, may be modulated by several endotoxin-binding proteins. Endotoxin-neutralizing capacity (ENC) and the plasma levels of soluble CD14 (sCD14), lipopolysaccharide-binding protein, and bactericidal/permeability-increasing protein (BPI) were determined in 40 patients 6 h prior to skin incision for major abdominal surgery. The bioactivity of plasma endotoxin was tested by the polymyxin B-inhibited stimulatory activity of the plasma samples on healthy monocytes as measured by the release of tumor necrosis factor alpha. Plasma endotoxin levels in almost all patients increased from 0.05 ± 0.01 to 0.23 ± 0.03 experimental units (EU) per ml (P< 0.001); more specifically, 17 of 40 samples showed endotoxin levels of greater than 0.2 EU per ml and corresponding reductions in ENC. Soluble CD14 plasma levels were decreased from 5.6 ± 0.3 to 4.6 ± 0.3 μg per ml (P< 0.05). ENC was strongly correlated with the sCD14 plasma concentration throughout the period of observation. The addition of sCD14-neutralizing monoclonal anti-sCD14 antibodies reduced ENC both pre- and postoperatively. No correlation could be established between ENC and the plasma levels of BPI, high-density lipoproteins, or low-density lipoproteins determined by measuring the concentrations of apoprotein A and apoprotein B. Biologically active endotoxin was found in only 6 of 17 samples with endotoxin levels greater than 0.2 EU per ml in the LAL test. These samples could be characterized by their perioperative loss of at least 35% of their sCD14. No change in sCD14 was detected in the remaining 11 samples. The perioperative loss of ENC is partly caused by the loss of sCD14 resulting from its consumption by endotoxin reaching the bloodstream. This study demonstrated the role of sCD14 on the bioactivity of circulating endotoxin in a human model of endotoxemia after major abdominal surgery.
Identification of a Subset of Common Variable Immunodeficiency Patients with Impaired B-Cell Protein Tyrosine Phosphorylation
Authors: Schwartz Rivka , Porat Yael Ben-Anat , Handzel Zeev , Sthoeger Zeev , Garty Ben-Zion , Confino-Cohen Ronit , Levy Jacov , Zan-Bar Israel 
Published: December 2019

ABSTRACTThe mechanisms responsible for common variable immunodeficiency syndrome (CVID) are as yet unknown. In the present study, we show that the B-cell dysfunction in a subset of CVID patients is caused by defective protein tyrosine phosphorylation (PTP). We demonstrated that the PTP level and immunoglobulin (Ig) secretion malfunctions can be successfully repaired when normal plasma membrane components are implanted into these patients’ B cells. Stimulation of CVID patients’ peripheral blood mononucleated cells with anti-Ig antibody revealed that 7 of 11 patients had lower PTP levels than those found in the normal donor cells. Plasma membrane implantation to the cells of these patients resulted in elevated PTP levels which reached normal levels upon stimulation with anti-human Ig antibody. The results revealed two distinct groups of CVID patients. The first group included patients whose B cells expressed low PTP levels after Ig stimulation. In these patients the plasma membrane implantation restored the normal PTP level as well as the ability to secrete IgM and/or IgG after B-cell stimulation. In the second group, patients whose B cells expressed a normal PTP level after Ig stimulation, with no restoration of their ability to secrete Ig upon plasma membrane implantation and lipopolysaccharide stimulation. We conclude that the first group has an early signal transduction defect located in the B-cell plasma membrane, while in the second group the defect is located elsewhere.
Sterols of
Pneumocystis carinii hominis
Organisms Isolated from Human Lungs
Authors: Kaneshiro Edna S. , Amit Zunika , Chandra Jyotsna , Baughman Robert P. , Contini Carlo , Lundgren Bettina 
Published: December 2019

ABSTRACT

The opportunistic pathogen
Pneumocystis carinii
causes pneumonia (
P. carinii
pneumonia, or PCP) in immunocompromised individuals such as AIDS patients. Rat-derived
P. carinii carinii
organisms have distinct sterols which are not synthesized by mammals and not found in other microbes infecting mammalian lungs. The dominant sterol present in the organism is cholesterol (which is believed to be scavenged from the host), but other sterols in
P. carinii carinii
have an alkyl group at C-24 of the sterol side chain (C
28
and C
29
24-alkylsterols) and a double bond at C-7 of the nucleus. Recently, pneumocysterol (C
32
), which is essentially lanosterol with a C-24 ethylidene group, was detected in lipids extracted from a formalin-fixed human
P. carinii
-infected lung, and its structures were elucidated by gas-liquid chromatography, mass spectrometry, and nuclear magnetic resonance spectrometry in conjunction with analyses of chemically synthesized authentic standards. The sterol composition of isolated
P. carinii hominis
organisms has yet to be reported. If
P. carinii
from animal models is to be used for identifying potential drug targets and for developing chemotherapeutic approaches to clear human infections, it is important to determine whether the 24-alkylsterols of organisms found in rats are also present in organisms in humans. In the present study, sterol analyses of
P. carinii hominis
organisms isolated from cryopreserved human
P. carinii
-infected lungs and from bronchoalveolar lavage fluid were performed. Several of the same distinct sterols (e.g., fungisterol and methylcholest-7-ene-3β-ol) previously identified in
P. carinii carinii
were also present in organisms isolated from human specimens. Pneumocysterol was detected in only some of the samples.
The Glutamate-Rich Protein (GLURP) ofPlasmodium falciparumIs a Target for Antibody-Dependent Monocyte-Mediated Inhibition of Parasite Growth In Vitro
Authors: Theisen Michael , Soe Soe , Oeuvray Claude , Thomas Alan W. , Vuust Jens , Danielsen Steffen , Jepsen Søren , Druilhe Pierre 
Published: December 2019

ABSTRACTMonocyte-dependent as well as direct inhibitory effects of antimalarial antibodies point toward antigens accessible at the time of merozoite release as targets for biologically active antibodies capable of mediating protection againstPlasmodium falciparum. The glutamate-rich protein (GLURP), being an antigen associated with mature schizont-infected erythrocytes, was therefore the object of the present investigation, in which we analyzed whether anti-GLURP antibodies can either interfere directly with merozoite invasion or act indirectly by promoting a monocyte-dependent growth inhibition, antibody-dependent cellular inhibition. GLURP-specific human immunoglobulin G (IgG) antibodies, from pooled IgG of healthy Liberian adults who were clinically immune to malaria, were purified by affinity chromatography on columns containing R0 (N-terminal nonrepetitive region of GLURP) or R2 (C-terminal repetitive region of GLURP) recombinant protein or synthetic peptides as ligands. Analysis of the pattern of reactivity of highly purified anti-GLURP antibodies led to the definition of at least four B-cell epitopes. One epitope was specific for R0, two were specific for R2, and the fourth displayed cross-reactivity between R0 and R2. None of the purified IgG antibodies had direct invasion-inhibitory effects, even at high concentrations. In contrast, when allowed to cooperate with monocytes, all anti-GLURP IgG preparations mediated a strong monocyte-dependent parasite growth inhibition in a dose-dependent manner.
Binding of Salivary Glycoprotein-Secretory Immunoglobulin A Complex to the Surface Protein Antigen of
Streptococcus mutans
Authors: Oho Takahiko , Yu Hao , Yamashita Yoshihisa , Koga Toshihiko 
Published: December 2019

ABSTRACT

The interaction between a surface protein antigen (PAc) of
Streptococcus mutans
and human salivary agglutinin was analyzed with a surface plasmon resonance biosensor. The major component sugars of the salivary agglutinin were galactose, fucose, mannose,
N
-acetylglucosamine,
N
-acetylgalactosamine, and
N
-acetylneuraminic acid. Binding of salivary agglutinin to PAc was calcium dependent and heat labile and required a pH greater than 5. Binding was significantly inhibited by
N
-acetylneuraminic acid and α2,6-linked sialic acid-specific lectin derived from
Sambucus sieboldiana
in a dose-dependent manner. Pretreatment of the salivary agglutinin with sialidase reduced the binding activity of the agglutinin to the PAc molecule. The agglutinin was dissociated into high-molecular-mass glycoprotein and secretory immunoglobulin A (sIgA) components by electrophoretic fractionation in the presence of 1% sodium dodecyl sulfate and 1% 2-mercaptoethanol. Neither of the components separated by electrophoretic fractionation, high-molecular-mass glycoprotein or sIgA, bound to the PAc molecule. Furthermore, the high-molecular-mass glycoprotein strongly inhibited the binding of the native salivary complex to PAc. These results suggest that the complex formed by the high-molecular-mass salivary glycoprotein and sIgA is essential for the binding reaction and that the sialic acid residues of the complex play an important role in the interaction between the agglutinin and PAc of
S. mutans.