bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Tumor Necrosis Factor-Inducing Potency of Lipopolysaccharide and Uronic Acid Polymers Is Increased when They Are Covalently Linked to Particles
Authors: Berntzen Gøril , Flo Trude H. , Medvedev Andrei , Kilaas Lars , Skjåk-Bræk Gudmund , Sundan Anders , Espevik Terje 
Published: December 2019

ABSTRACTLipopolysaccharide (LPS) and polymers of the uronic acid family stimulate monocytes to produce tumor necrosis factor (TNF). The TNF-inducing potency of these polysaccharides may depend on their supramolecular configuration. In this study detoxified LPS and uronic acid polymers have been covalently linked to particles which have been added to monocytes under serum-free conditions. Reducing the size of mannuronan from 350,000 to 5,500 Da (M-blocks) led to a 10- to 100-fold reduction in TNF-inducing potency. However, covalently linking the M-blocks to monodisperse suspensions of magnetic particles increased the TNF-inducing potency by up to 60,000-fold. Also, the TNF-inducing potency of glucuronic acid polymers was increased when they were linked to particles, but no potentiation was observed with guluronic acid blocks covalently attached to particles. Furthermore, O chains of LPS (detoxified LPS) became potent TNF inducers when they were presented to monocytes on a particle surface. No activation of the LPS-responsive SW480 adenocarcinoma cells was found with detoxified LPS or M-block particles, suggesting a preference for cells expressing CD14 and/or other membrane molecules. The potentiating effects were not restricted to polymers attached to aminated magnetic particles. Of particular interest, we found that short blocks of mannuronan induced TNF production also when covalently linked to biodegradable, bovine serum albumin particles.
Immunohistochemical Detection of Porcine Epidemic Diarrhea Virus Compared to Other Methods
Authors: Guscetti Franco , Bernasconi Curzio , Tobler Kurt , Van Reeth Kristien , Pospischil Andreas , Ackermann Mathias 
Published: December 2019

ABSTRACTAn immunohistochemistry method using formalin-fixed tissues, a direct immunofluorescence method using cryostat sections, an enzyme-linked immunosorbent assay (ELISA), and a PCR method were compared for diagnosis in a litter of weaned pigs that had been experimentally inoculated with wild-type porcine epidemic diarrhea virus (PEDV) and killed between 6 and 60 h after onset of diarrhea. The immunohistochemistry method proved to be as reliable as direct immunofluorescence for diagnosis of PEDV in tissues collected postmortem. The good reliability of ELISA for investigating clinical samples was confirmed, whereas the PCR method used was ineffective.
T Cells and T-Cell Cytokine Transcripts in the Synovial Membrane in Patients with Osteoarthritis
Authors: Sakkas Lazaros I. , Scanzello Carla , Johanson Norman , Burkholder Janet , Mitra Amitabha , Salgame Padmini , Katsetos Christos D. , Platsoucas Chris D. 
Published: December 2019

ABSTRACTThe synovial membrane in osteoarthritis (OA) often exhibits inflammatory infiltrates, but the role of T cells in these infiltrates is not known. T-cell activation antigens were analyzed by immunohistochemistry, and T-cell cytokine transcripts were measured by competitive PCR in synovial membranes from patients with OA and rheumatoid arthritis (RA). Lymphoid cell aggregates, containing primarily CD3+T lymphocytes, were found in 65% of patients with OA. Mononuclear cells expressing the activation antigens CD69, CD25, CD38, CD43, CD45RO, and HLA class II were present in both patient groups, although in higher numbers in patients with RA. Interleukin 2 (IL-2) transcripts were found in 10 of 18 patients with OA versus 12 of 13 patients with RA (P= 0.03). Gamma interferon (IFN-γ) transcripts were detected in 9 of 18 patients with OA versus 10 of 13 patients with RA (not significant), whereas IL-10 transcripts were found in nearly all patients. IL-4 and IL-5 were not detected in any patients. The levels of IFN-γ and IL-2 transcripts, normalized for T-cell number equivalents, were not statistically different between OA and RA, but the levels of IFN-γ, normalized for total cell number equivalents, were lower in OA than in RA (P= 0.01). Synovial membranes that expressed IL-2 and IFN-γ transcripts were more likely to have heavier infiltrations of T cells and cells bearing activation markers than synovial membranes that did not express these cytokines. The presence of activated T cells and TH1 cytokine transcripts in chronic joint lesions of patients with OA suggests that T cells contribute to chronic inflammation in a large proportion of these patients.
Intrathecal Synthesis of Immunoglobulins in Eosinophilic Meningoencephalitis Due to
Angiostrongylus cantonensis
Authors: Dorta-Contreras Alberto Juan , Reiber Hansotto 
Published: December 2019

ABSTRACT

Eosinophilic meningoencephalitis due to the nematode
Angiostrongylus cantonensis
, which is endemic to Cuba, occurs in children and is due to accidental contact with soil snails. The course is less often fatal than in adult patients in southeastern Asia. Cerebrospinal fluid (CSF) and serum samples from 24 pediatric patients were analyzed and evaluated in CSF/serum quotient diagrams (Reiber graphs) to characterize the neuroimmunological response and the blood-CSF barrier dysfunction that occur in the course of the disease. At the time of the first diagnostic lumbar puncture, together with eosinophilic pleocytosis (1,920 ± 400 cells/μl), intermediate blood-CSF barrier dysfunction (i.e., an increased CSF/serum albumin quotient) with no intrathecal immunoglobulin G (IgG), IgA, and IgM class response was observed in all cases. Seven days later, at the time of early clinical recovery, the blood-CSF barrier dysfunction was normalized in 75% of the patients, but meanwhile, intrathecal immunoglobulin synthesis emerged in all cases, as either a two-class response (IgG and IgA in 85% of the patients) or a three-class response (IgG, IgA, and IgM; 30%). The fraction of eosinophilic cells (40%) remained large despite a decreasing total cell count. The neuroimmunological pattern of this inflammatory response to the parasite and its toxins is discussed with regard to the CSF patterns of other infectious diseases caused by bacteria or viruses.
Human Immunodeficiency Virus Type 1 Disease Progression, CCR5 Genotype, and Specific Immune Responses
Authors: Visco-Comandini Ubaldo , Hultgren Catharina , Broström Christina , Birk Markus , Kim Soo , Sällberg Matti 
Published: December 2019

ABSTRACT

The correlation among the presence of a 32-bp deletion in the CC-chemokine receptor 5 (
CCR5
) gene, disease progression, and human immunodeficiency virus type 1 (HIV-1)-specific immune responses was analyzed for a cohort of 79 Caucasian HIV-1-infected patients. The
CCR5
genotype (
CCR5/CCR5
= wild type/wild type or Δ
32CCR5/CCR5
= 32-bp deletion/wild type) in peripheral blood mononuclear cells was determined by PCR, followed by sequencing of both wild-type and Δ
32CCR5
gene fragments. HIV-1-specific humoral responses to gp41 and V3
MN
peptides were determined by enzyme immunoassays. The prevalence of the Δ
32CCR5
allele was lower among 37 patients with rapid progression (progression to AIDS or to a CD4 cell count of <200 × 10
6
/liter in less than 9 years;
P
< 0.01) compared to that for 42 patients with slow progression (no AIDS and CD4 cell count of >200 × 10
6
/liter after at least 9 years from infection) or to that for 25 non-HIV-1-infected Swedish blood donors (
P
< 0.05). No differences were observed in the wild-type
CCR5
sequences between the different groups of patients. For three analyzed patients, the 32-bp Δ
32CCR5
gene deletions were identical. The antibody titers against gp41 and a V3
MN
peptide in patients with the Δ
32CCR5/CCR5
genotype were not significantly different from those in pair-matched
CCR5/CCR5
controls. However, in 13 analyzed patients, a stronger serum neutralizing activity was associated with the Δ
32CCR5/CCR5
genotype. Thus, a
CCR5/CCR5
genotype correlates with a shortened AIDS-free HIV-1 infection period and possibly with a worse neutralizing activity, without an evident influence on the antibody response to two major antigenic regions of HIV-1 envelope.
A Modified Enzyme-Linked Immunosorbent Assay for Measurement of Antibody Responses to Meningococcal C Polysaccharide That Correlate with Bactericidal Responses
Authors: Granoff Dan M. , Maslanka Susan E. , Carlone George M. , Plikaytis Brian D. , Santos George F. , Mokatrin Ahmad , Raff Howard V. 
Published: December 2019

ABSTRACTThe standardized enzyme-linked immunosorbent assay (ELISA) for measurement of serum immunoglobulin G (IgG) antibody responses to meningococcal C polysaccharide has been modified to employ assay conditions that ensure specificity and favor detection primarily of high-avidity antibodies. The modified and standard assays were used to measure IgG antibody concentrations in sera of toddlers vaccinated with meningococcal polysaccharide vaccine or a meningococcal C conjugate vaccine. The results were compared to the respective complement-mediated bactericidal antibody titers. In sera obtained after one or two doses of vaccine, the correlation coefficients,r, for the results of the standard assay and bactericidal antibody titers were 0.45 and 0.29, compared to 0.85 and 0.87, respectively, for the modified assay. With the standard assay, there were no significant differences between the geometric mean antibody responses of the two vaccine groups. In contrast, with the modified assay, 5- to 20-fold higher postvaccination antibody concentrations were measured in the conjugate than in the polysaccharide group. Importantly, the results of the modified assay, but not the standard ELISA, paralleled the respective geometric mean bactericidal antibody titers. Thus, by employing conditions that favor detection of higher-avidity IgG antibody, the modified ELISA provides results that correlate closely with measurements of antibody functional activity that are thought to be important in protection against meningococcal disease.
Concentrations of Circulating β-Chemokines Do Not Correlate with Viral Load in Human Immunodeficiency Virus-Infected Individuals
Authors: Kakkanaiah Vellalore N. , Ojo-Amaize Emmanuel A. , Peter James B. 
Published: December 2019

ABSTRACTThe CC or β-chemokines MIP-1α, MIP-1β, and RANTES are the primary components of human immunodeficiency virus type 1 (HIV-1)-suppressive soluble factors in vitro. We studied the relationship between the concentrations of MIP-1α, MIP-1β, and RANTES in plasma and HIV viral load in HIV-infected subjects. The HIV-positive patient group (n= 140) had significantly lower concentrations of all three β-chemokines (MIP-1α,P< 0.0005; MIP-1β,P< 0.005; RANTES,P< 0.0005) than the control group (n= 58 for MIP-1α,n= 27 for MIP-1β, andn= 59 for RANTES). In addition, we divided the patient group into three subgroups (high, moderate, and low) based on the number of HIV-1 RNA copies in the plasma (as measured by quantitative HIV RNA PCR). Again, all three subgroups had significantly lower concentrations of the β-chemokines than the HIV-negative control group. However, there was no significant difference in plasma β-chemokine concentrations among the three subgroups within the patient group (P< 0.3). Although our results demonstrate that HIV-infected individuals had significantly lower concentrations of circulating β-chemokines than healthy uninfected control subjects, we found no correlation between the concentrations of β-chemokines in plasma and HIV-1 viral load in HIV-infected individuals.
Oral Fluids as an Alternative to Serum for Measurement of Markers of Immune Activation
Authors: Nishanian Parunag , Aziz Najib , Chung Joanie , Detels Roger , Fahey John L. 
Published: December 2019

ABSTRACT
Oral fluids are convenient alternatives to blood sampling for evaluating significant metabolic components. Two forms of oral fluids, oral mucosal transudates (OMT) and saliva, were collected and compared for content of soluble products of immune activation. The data confirm that OMT and saliva represent distinct body fluids. The concentrations, outputs, and analyte/protein ratios of β-2-microglobulin (β2M), soluble tumor necrosis factor alpha receptor II (sTNFαRII), and neopterin were measured. Both the OMT and the saliva of most of the individuals in the control healthy populations had measurable levels of all three activation markers. When the immune system is activated, as in human immunodeficiency virus (HIV) infection, the levels of β2M and sTNFαRII are increased in both OMT and saliva compared to those in a healthy control population. OMT levels correlated better with levels in serum than did saliva and appear to reflect systemic immune activation in HIV infection. Because acquisition of oral fluids is noninvasive and easily repeatable, measurement of β2M and/or sTNFαRII content in OMT could be useful in the assessment of disease activity in patients with HIV infection or chronic inflammatory diseases.
Interpretations of Antibody Responses toSalmonella entericaSerotype Enteritidis gm Flagellin in Poultry Flocks Are Enhanced by a Kinetics-Based Enzyme-Linked Immunosorbent Assay
Authors: McDonough Patrick L. , Jacobson Richard H. , Timoney John F. , Mutalib Ahmed , Kradel David C. , Chang Yung-fu , Shin Sang J. , Lein Donald H. , Trock Susan , Wheeler Kaye 
Published: December 2019

ABSTRACTMany regulatory and diagnostic programs for the detection ofSalmonella entericaserotype Enteritidis infection in commercial poultry flocks have relied on rapid Pullorum agglutination tests to screen birds because of the shared antigens ofS. entericaEnteritidis andS. entericaPullorum and Gallinarum; however, the use of the enzyme-linked immunosorbent assay (ELISA) format affords better analytical sensitivity than crude agglutination tests. In this study, we adapted our earlier conventional indirect ELISA, using gm flagellin as the antigen, to a kinetics-based, computer-controlled ELISA (KELA). The KELA was used to screen for flagellin antibody from three commercial flocks: (i) a large flock involved in a U.S. Department of Agriculture trace back from a humanS. entericaEnteritidis foodborne outbreak (n= 3,209), (ii) a flock infected with the endemicS. entericaEnteritidis serotype but which also had multiple other salmonella serotypes (n= 65), and (iii) anS. entericaPullorum-infected flock (n= 12). The first flock (S. entericaEnteritidis prevalence of 2.45% based on culture) provided a field test of the KELA and allowed the calculation of diagnostic sensitivity (D-Sn) and diagnostic specificity (D-Sp). With a cutoff of 10 (used for screening flocks [i.e., high sensitivity]), the KELA has a D-Sn of 95.2% and a D-Sp of 18.5%; with a cutoff of 140 (used in confirmatory flock testing [i.e., high specificity]), the KELA has a D-Sn of 28.0% and a D-Sp of 99.1%. We found that with a cutoff of 60 (D-Sn = 63.1%; D-Sp = 91.6%), we could eliminate reactions in the KELA caused by other non-S. entericaEnteritidis salmonellae. The KELA was also compared to two commercial rapid Pullorum tests, the Solvay (D-Sn = 94.9%; D-Sp = 55.5%) and the Vineland (D-Sn = 62.0%; D-Sp = 75.3%).
Quantifying Serum Antiplague Antibody with a Fiber-Optic Biosensor
Authors: Anderson George P. , King Keeley D. , Cao Lynn K. , Jacoby Meagan , Ligler Frances S. , Ezzell John 
Published: December 2019

ABSTRACTThe fiber-optic biosensor, originally developed to detect hazardous biological agents such as protein toxins or bacterial cells, has been utilized to quantify the concentration of serum antiplague antibodies. This biosensor has been used to detect and quantify the plague fraction 1 antigen in serum, plasma, and whole-blood samples, but its ability to quantify serum antibodies has not been demonstrated. By using a competitive assay, the concentration of serum antiplague antibodies was ascertained in the range of 2 to 15 μg/ml. By making simple dilutions, concentrations for 11 serum samples whose antiplague antibody concentrations were unknown were determined and were found to be in good agreement with enzyme-linked immunosorbent assay results. The competitive assay method could be used to effectively determine the exposure to plague of animals or humans or could be applied to other diseases, such as hepatitis or AIDS, where the presence of antibodies is used to diagnose infection.
Analysis of Specific Immunoglobulin G Subclass Antibodies for Serological Diagnosis of Echinococcosis by a Standard Enzyme-Linked Immunosorbent Assay
Authors: Grimm Felix , Maly Friedrich E. , Lü Jian , Llano Roberto 
Published: December 2019

ABSTRACT
The potential roles of specific antibodies of the different immunoglobulin G (IgG) subclasses in the serological diagnosis of cystic echinococcosis (CE) and alveolar echinococcosis (AE) were investigated by an enzyme-linked immunosorbent assay based on hydatid fluid as antigen. Specific antibodies of subclass 1 were found to be of major importance. In sera collected at the time of diagnosis (i.e., before any therapeutic intervention was initiated) they could be demonstrated in 14 of 15 sera from patients with CE and in all 12 sera from patients with AE. The most discriminatory and the most specific antibodies found in this study belonged to IgG subclass 4. Only one false-positive reaction was observed with 253 sera from healthy volunteers, and no cross-reactions occurred in 80 sera from patients with different parasitic infections. Specific IgG4 antibodies could be demonstrated in 61.0 to 66.7% (CE) or 47.6 to 66.7% (AE) of the cases. Antibody levels of IgG subclass 2 were elevated only moderately, and subclass 3 antibodies were detected in a few cases only. In addition, nonspecific reactions in sera of healthy volunteers or patients with other parasitic infections could partially be attributed to antibodies of subclasses 2 and 3.
Enzyme Immunoassays for Serological Diagnosis of Bovine Brucellosis: A Trial in Latin America
Authors: Gall D. , Colling A. , Marino O. , Moreno E. , Nielsen K. , Perez B. , Samartino L. 
Published: December 2019

ABSTRACT

The results of a field trial conducted in Latin America with two indirect enzyme-linked immunosorbent assays (ELISAs) and two competitive ELISAs (CELISAs) for the detection of bovine antibody to
Brucella abortus
are reported. One of the CELISA formats performed most accurately. The percentage of positive reactions in the CELISA relative to the selected positive rose bengal agglutination test (RBT) and complement fixation test (CFT) results was 97.47%, the percentage of negatives relative to the selected negative RBT and CFT results for unexposed cattle was 98.32%, and the percentage of negatives in cattle vaccinated with
B. abortus
19 was 96.51%. The same assay format under Canadian conditions had an actual sensitivity of 100%, a specificity of 99.90% in nonvaccinates, and a specificity of 97.7% in a strain 19-vaccinated population. Overall, the CELISA performed as expected and the results were not dissimilar from the results obtained in the Canadian study. This provided further evidence that this CELISA can in many instances differentiate infected cattle from those that are vaccinated or infected with a cross-reacting organism while still giving very few false-positive or false-negative results.
Endothelial Cells Expressing an Inflammatory Phenotype Are Lysed by Superantigen-Targeted Cytotoxic T Cells
Authors: Riesbeck Kristian , Billström Anita , Tordsson Jesper , Brodin Thomas , Kristensson Karin , Dohlsten Mikael 
Published: December 2019

ABSTRACTThe objective of this study was to investigate whether the superantigen staphylococcal enterotoxin A (SEA), which binds to HLA class II and T-cell receptor Vβ chains, can direct cytotoxic T cells to lyse cytokine-stimulated endothelial cells (EC). In addition, we wanted to determine whether SEA-primed cytotoxic T cells could be targeted to EC surface molecules as a means of a novel cancer immunotherapy. Human umbilical vein EC (HUVEC), dermal microvascular EC (HMVEC), or the EC line EA.hy926 stimulated with gamma interferon (IFN-γ) or tumor necrosis factor alpha (TNF-α) displayed upregulated HLA class II and adhesion molecule (CD54 and CD106) expression, respectively. SEA-primed T cells induced a strong cytotoxicity against IFN-γ- and TNF-α-activated EA.hy926 which had been preincubated with SEA. Blocking of CD54 completely abrogated the T-cell attack. SEA-D227A, which has a mutated class II binding site, did not promote any cytotoxicity. A strong lysis was observed when a fusion protein consisting of protein A and SEA-D227A was added together with T cells to TNF-α-induced EA.hy926 and HUVEC precoated with monoclonal antibodies (MAb) directed against HLA class I, CD54, or CD106 molecules. Finally, an scFv antibody fragment reactive with an unknown EC antigen was fused with SEA-D227A. Both EA.hy926 and HMVEC were efficiently lysed by scFv-SEA-D227A-triggered cytotoxic T cells. Taken together, superantigen-activated T-cell-dependent EC killing was induced when EC expressed an inflammatory phenotype. Moreover, specific MAb targeting of the superantigen to surface antigens induced EC lysis. Our data suggest that directed T-cell-mediated lysis of unwanted proliferating EC, such as those in the tumor microvasculature, can be clinically useful.
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

ABSTRACT

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.