bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Whole-Blood Assay for Qualitative and Semiquantitative Measurements of CD69 Surface Expression on CD4 and CD8 T Lymphocytes Using Flow Cytometry
Authors: Lim Lony C. L. , Fiordalisi Michelle N. , Mantell Janet L. , Schmitz John L. , Folds James D. 
Published: December 2019

ABSTRACTA whole-blood flow cytometry-based assay was utilized to assess CD4 and CD8 T-lymphocyte activation in response to phytohemagglutinin (PHA) stimulation. T-lymphocyte activation was assessed by qualitative (percent CD69) and semiquantitative (anti-CD69 antibody binding capacity) measurements of CD69 surface expression. Whole-blood samples from 21 healthy and 21 human immunodeficiency virus (HIV)-infected (<500 absolute CD4 counts per mm3) individuals were stimulated with 20 μg of PHA per ml for 18 to 24 h. The proportions of activated CD4 and CD8 T lymphocytes expressing CD69 (percent CD69) and the levels of CD69 expression on each T-lymphocyte subset (anti-CD69 antibody binding capacity) were measured. By using this assay system, T-lymphocyte activation was impaired in both CD4 and CD8 T-lymphocyte subsets of HIV-infected individuals. The proportions of CD69-positive CD4 and CD8 T lymphocytes were 43 and 27% lower, respectively, in samples from HIV-infected individuals compared to samples from healthy individuals. Similarly, the levels of CD69 expression on each activated CD4 and CD8 T-lymphocyte subset were 48 and 51% lower, respectively. These results suggest that both qualitative and semiquantitative measurements of CD69 surface expression by flow cytometry can be used to assess T-lymphocyte activation.
Adenosine Deaminase Deficiency and Purine Nucleoside Phosphorylase Deficiency in Common Variable Immunodeficiency
Authors: Fleischman Amy , Hershfield Michael S. , Toutain Stephan , Lederman Howard M. , Sullivan Kathleen E. , Fasano Mary Beth , Greene Jeff , Winkelstein Jerry A. 
Published: December 2019

ABSTRACTThe clinical presentations of adenosine deaminase deficiency and purine nucleoside phosphorylase deficiency are widely variable and include clinical and immunologic findings compatible with common variable immunodeficiency. The screening of 44 patients with common variable immunodeficiency failed to identify any individuals with deficiencies of these enzymes.
Serum Cytokines in Patients withLegionellaPneumonia: Relative Predominance of Th1-Type Cytokines
Authors: Tateda Kazuhiro , Matsumoto Tetsuya , Ishii Yoshikazu , Furuya Nobuhiko , Ohno Akira , Miyazaki Shuichi , Yamaguchi Keizo 
Published: December 2019

ABSTRACTSerum samples from 14 patients withLegionellapneumonia were examined for the presence of cytokines. In spite of high levels of serum C-reactive protein in all patients during the acute phase in only four cases (one involving interleukin-1β [IL-1β], three involving IL-6, and none involving tumor necrosis factor alpha) was the concentration of cytokines more than 100 pg/ml. Th2 cytokines IL-4 and IL-10 were detected in only one patient each. In contrast, significant increases of serum gamma interferon (IFN-γ) and IL-12 levels were observed during the acute phase in 6 and 11 cases, respectively. Interestingly, although serum IFN-γ levels diminished thereafter, in seven cases IL-12 levels remained high or increased further during the convalescent phase. In an additional 22 cases clinically suspected to be but not diagnosed asLegionellapneumonia, increases of serum IL-12 levels were observed in 16 cases, whereas the remaining 6 cases showed no detectable IL-12. Our results demonstrate the relative predominance of Th1 cytokine production inLegionellapneumonia. Although the role and significance of prolonged increases in IL-12 levels inLegionelladisease are unknown, our results should prompt further investigation of the host immune response in terms of Th1 and Th2 balance in legionellosis.
Primary Epitopes of Chicken Egg Yolk Antibodies to Peptidophosphogalactomannan
Authors: Tuekam Brigitte A. , Bonetti Sandra J. , Gander John E. 
Published: December 2019

ABSTRACT

Egg yolks from hens immunized with peptidophosphogalactomannan (pPGalMan
ii
), which contains 10 phosphocholine diester residues and is secreted by
Penicillium fellutanum
, contain antibodies against 5-
O
-β-
d
-galactofuranosyl epitopes. These epitopes were the only significant determinants in pPGalMan
ii
. Approximately 60-fold less pPGalMan
ii
(1.6 μM galactofuran chains) was required for 50% inhibition than galactofurano-oligosaccharides or pPGalMan containing two galactofuranosyl residues per chain.
Seroprevalence of Antibodies to
Bartonella henselae
in Patients with Cat Scratch Disease and in Healthy Controls: Evaluation and Comparison of Two Commercial Serological Tests
Authors: Sander Anna , Posselt Miriam , Oberle Karin , Bredt Wolfgang 
Published: December 2019

ABSTRACT

Serologic testing for the presence of antibodies to
Bartonella henselae
is a widely accepted diagnostic procedure for laboratory confirmation of the diagnosis of cat scratch disease (CSD). In this study a commercially available indirect immunofluorescence assay (IFA) based on
B. henselae
-infected human larynx carcinoma cells (test A) was evaluated. Sera from 42 patients with CSD (20 confirmed by PCR) and 270 sera from healthy controls (consisting of 63 cat owners, 65 individuals whose last close contact with cats was >6 months previously, and 142 persons who had never been exposed to cats) were investigated for antibodies to
B. henselae
. All patients with CSD had titers of immunoglobulin G (IgG) to
B. henselae
of 128 or higher (test A; sensitivity, 100%). Of the 270 controls 189 (70%) were seronegative (titer, <64), 38 (14.1%) had titers of 64, 30 (11.1%) had titers of 128, 9 (3.3%) had titers of 256, and 4 (1.5%) had high titers, 512 (test A; specificity, 70%). Of the cat owners and individuals who had never had close contact with cats, 71.4 and 71.12%, respectively, were seronegative, and titers of 64, 128, 256, and 512 were found in 14.3 and 16.2%, 1.6 and 10.5%, 9.5 and 0.7%, and 3.2 and 1.4%, respectively. The sera from the patients and from the first 100 healthy adults without a history of close contact with cats were additionally tested with a second commercially available IFA, based on Vero cells infected with
B. henselae
and
Bartonella quintana
(test B). The sensitivity and specificity of test B were 93 and 73%, respectively. For patients with CSD the cross-reactivity between
B. henselae
and
B. quintana
in this test was 95%. Both systems are highly sensitive but less specific for detection of IgG antibodies to
B. henselae
in samples from patients with clinically apparent CSD. For detection of IgM antibodies, test A seems to be more sensitive (88%) and more specific (95%) than test B (sensitivity and specificity of 64 and 86%, respectively). The data show that the seroprevalence of antibodies to
B. henselae
in German individuals is high (30%). Low antibody levels are not sufficient evidence of active or prior infection.
Expression and Refolding of Truncated Recombinant Major Outer Membrane Protein Antigen (r56) of
Orientia tsutsugamushi
and Its Use in Enzyme-Linked Immunosorbent Assays
Authors: Ching W.-M. , Wang H. , Eamsila C. , Kelly D. J. , Dasch G. A. 
Published: December 2019

ABSTRACT

The variable 56-kDa major outer membrane protein of
Orientia tsutsugamushi
is the immunodominant antigen in human scrub typhus infections. The gene encoding this protein from Karp strain was cloned into the expression vector pET11a. The recombinant protein (r56) was expressed as a truncated nonfusion protein (amino acids 80 to 456 of the open reading frame) which formed an inclusion body when expressed in
Escherichia coli
BL21. Refolded r56 was purified and compared to purified whole-cell lysate of the Karp strain of
O. tsutsugamushi
by immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) for reactivity with rabbit sera prepared against eight antigenic prototypes of
O. tsutsugamushi
as well as several other species of
Rickettsiales
and nonrickettsial antigens. Refolded r56 exhibited broad reactivity with the rabbit antisera against the
Orientia
prototypes, and the ELISA reactions with the r56 and Karp whole-cell lysate antigens correlated well (
r
= 0.81,
n
= 22, sensitivity compared to that of standard ELISA of 91%). Refolded r56 did not react with most antisera against other rickettsial species or control antigens (specificity = 92%,
n
= 13) using a positive cutoff value determined with eight uninfected rabbit sera. Refolded r56 was evaluated further by ELISA, using 128 sera obtained from patients with suspected scrub typhus from Korat, Thailand, and 74 serum specimens from healthy Thai soldiers. By using the indirect immunoperoxidase assay as the reference assay, the recombinant antigen exhibited a sensitivity and specificity of 93% or greater for detection of both IgG and IgM in the ELISA at 1:400 serum dilution. These results strongly suggest that purified r56 is a suitable candidate for replacing the density gradient-purified, rickettsia-derived, whole-cell antigen currently used in the commercial dipstick assay available in the United States.
Lack of Maternal Antibodies to P Serotypes May Predispose Neonates to Infections with Unusual Rotavirus Strains
Authors: Ramachandran Madhumati , Vij Aarti , Kumar Ramesh , Das Bimal K. , Gentsch Jon R. , Bhan M. K. , Glass Roger I. 
Published: December 2019

ABSTRACT
Rotavirus (RV) strains infecting newborns often have unique neutralization antigens (P serotypes) on their outer capsids that are distinct from those found on RV strains that cause diarrhea in older children. We examined the hypothesis that unusual RV strains preferentially infect newborns because the newborns lack maternal neutralizing antibodies to these strains. To test this hypothesis, sera and saliva samples collected from neonates infected with 116E-like (P[11]G9) strains in the maternity ward of the All India Institute of Medical Sciences (AIIMS) hospital in New Delhi were tested for neutralizing antibodies against common RV strains and those infecting newborns and these titers were compared with those of newborns who did not become infected (controls). The infected neonates had significantly lower levels of cord blood neutralizing antibodies to 116E than the controls, suggesting that immunity to neonatal RV infection is acquired transplacentally through maternal antibodies. Further, this study confirmed the immunogenicity of the AIIMS neonatal strain 116E, a vaccine candidate, in its ability to evoke a potent RV-specific immunoglobulin A and neutralizing antibody response in serum and saliva among the infected babies. Our findings have important implications for the development of an effective RV vaccine. In India, where G9 strains are common in the community, the use of 116E as a vaccine, together with the rhesus tetravalent vaccine, may provide a broader protection against all the circulating RV serotypes, including serotype G9, which is not represented in the current rhesus RV tetravalent vaccine (G1-G4).
Neutrophil Chemiluminescence during Phagocytosis Is Inhibited by Abnormally Elevated Levels of Acetoacetate: Implications for Diabetic Susceptibility to Infections
Authors: Saeed Fawzia A. , Castle Georgina E. 
Published: December 2019

ABSTRACT
Human neutrophils by a chemiluminescence assay exhibit diminished phagocytic activity in the presence of abnormally high levels of the serum metabolite acetoacetate. These findings, along with our previous evidence demonstrating myeloperoxidase inhibition by acetoacetate, implicate metabolic ketosis in the inhibition of neutrophil microbicidal activity and thus in increased susceptibility to infections.
Antigenic Structure of the Nucleocapsid Protein of Porcine Reproductive and Respiratory Syndrome Virus
Authors: Wootton Sarah K. , Nelson Eric A. , Yoo Dongwan 
Published: December 2019

ABSTRACT

A collection of 12 monoclonal antibodies (MAbs) raised against porcine reproductive and respiratory syndrome (PRRS) virus was used to study the antigenic structure of the virus nucleocapsid protein (N). The full-length N gene, encoded by open reading frame 7, was cloned from the Canadian PRRS virus, PA-8. Deletions were introduced into the N gene to produce a series of nine overlapping protein fragments ranging in length from 25 to 112 amino acids. The individual truncated genes were cloned as glutathione
S
-transferase fusions into a eukaryotic expression vector downstream of the T7 RNA polymerase promoter. HeLa cells infected with recombinant vaccinia virus expressing T7 RNA polymerase were transfected with plasmid DNA encoding the N protein fragments, and the antigenicity of the synthesized proteins was analyzed by immunoprecipitation. Based on the immunoreactivities of the N protein deletion mutants with the panel of N-specific MAbs, five domains of antigenic importance were identified. MAbs SDOW17, SR30, and 5H2.3B12.1C9 each identified independent domains defined by amino acids 30 to 52, 69 to 123, and 37 to 52, respectively. Seven of the MAbs tested specifically recognized the local protein conformation formed in part by the amino acid residues 52 to 69. Furthermore, deletion of 11 amino acids from the carboxy terminus of the nucleocapsid protein disrupted the epitope configuration recognized by all of the conformation-dependent MAbs, suggesting that the carboxy-terminal region plays an important role in maintaining local protein conformation.
Effect of Ethanol on Monocytic Function in Human Immunodeficiency Virus Type 1 Infection
Authors: Chen Houchu , George Italas , Sperber Kirk 
Published: December 2019

ABSTRACTWe have developed a novel system to study monocytic function after human immunodeficiency virus type 1 (HIV-1) infection by infecting a series of human macrophage hybridoma cell lines with HIV-1. Since ethanol has detrimental effects on immune function, we investigated the effect of ethanol and its metabolites acetaldehyde and acetate on monocytic function by utilizing one human macrophage hybridoma cell line, clone 43, as well as primary monocytes. Pretreatment of clone 43 and primary monocytes with ethanol and its metabolites resulted in diminished accessory cell function for mitogen-, anti-CD3-, and antigen-induced T-cell proliferation. The decreased accessory cell function was associated with reduced interleukin 1α (IL-1α), IL-1β, and tumor necrosis factor alpha production with loss of intracellular cytokine and mRNA production and the induction of transforming growth factor β. In ethanol-, acetaldehyde-, and acetate-treated HIV-1-infected clone 43 cells (43HIV), there was a more rapid loss (3 days after infection) of accessory cell function at a lower infecting dose of HIV-1 than that in untreated 43HIVcells. We also observed a more rapid loss of surface class II antigen expression in the ethanol-, acetaldehyde-, and acetate-treated 43HIVcells, but no change in surface expression of CD80 or CD86. Ethanol-induced impairment of monocytic function may compound the immunologic defects of AIDS, making the infected individual more susceptible to the complications of the disease.
Improved Immunodiagnosis of Human Candidiasis by an Enzyme-Linked Immunosorbent Assay Using a
Candida albicans
52-Kilodalton Metallopeptidase
Authors: El Moudni Brahim , Rodier Marie-Helene , Daniault Gyslaine , Jacquemin Jean Louis 
Published: December 2019

ABSTRACT

An immunogenic aminopeptidase of
Candida albicans
was purified by high-performance liquid chromatography. It was then used for the development of an enzyme-linked immunosorbent assay to detect antibodies directed against this antigen in sera from patients with candidiasis. This enzyme specifically cleaves the
l
-Arg–7-amino-4-methyl-coumarin substrate at pH 7.4 and was detected in the crude extract of different
C. albicans
isolates. Sera used for this study were obtained from healthy blood donors or from patients with one of the following: systemic candidiasis, aspergillosis, cryptococcosis, toxoplasmosis, or malaria. The statistical analysis demonstrates significant differences between absorbency values obtained with sera from patients with candidiasis and with sera from the other groups (
P
= 0.000001). Diagnostic parameters show high diagnostic specificity of 97% and a sensitivity of 83% at a cutoff value of 0.425 and suggest the usefulness of this aminopeptidase for the diagnosis of systemic candidiasis.
Antineuronal Antibodies in Rheumatic Chorea
Authors: Kotby A. A. , El Badawy N. , El Sokkary S. , Moawad H. , El Shawarby M. 
Published: December 2019

ABSTRACT

The objectives of our study were to examine the sera of rheumatic chorea (RhCh) patients (those with acute or chronic RhCh or with a past history of RhCh) for the presence of antineuronal antibodies (ANeurA) and to correlate the results with disease activity, chronicity, and the number and durations of choreic attacks. Subjects were inpatients of the Pediatric Hospital, Ain Shams University, and outpatients of the Outpatient Pediatric Cardiology Clinic (both in Cairo, Egypt). Forty children with RhCh (mean age, 10.9 years) and 40 healthy controls were tested. An indirect-immunofluorescence technique was used for the detection of ANeurA. ANeurA were present in the sera of 100, 93, and 44% of the patients with acute, chronic, and past histories of RhCh, respectively. A definition of chronic chorea is presented for the first time. None of the control subjects had ANeurA in their sera. The presence of ANeurA correlated with disease activity. A statistically significant increase (
P
< 0.01) in the prevalence of ANeurA was found for patients with active chorea (acute and chronic) compared with the prevalence in patients with past histories of RhCh (controlled chorea). ANeurA were present in the sera of both patients with acute RhCh and patients with chronic RhCh, yet patients with acute RhCh showed more brightness and cell staining than chronic patients. The severity, number, and duration of each attack were not related to the presence of ANeurA. These results strengthen further the concept of autoimmunity being the basis for the pathogenesis of RhCh. The presence of ANeurA correlated with the activity of RhCh but not with the severity, number, or duration of attacks. Humoral immunity definitely plays a role in RhCh; thus, routine administration of corticosteroids to patients with acute RhCh is suggested to prevent neuron damage and chronicity. The chronicity of chorea is not due to a further increase in ANeurA but is probably due to sensitivity to these antibodies.
Assessment of Markers of the Cell-Mediated Immune Response after Influenza Virus Infection in Frail Older Adults
Authors: McElhaney Janet E. , Gravenstein Stefan , Krause Peggy , Hooton Jonathan W. , Upshaw Craig M. , Drinka Paul 
Published: December 2019

ABSTRACTThe purpose of this study was to determine whether measures of the cell-mediated immune response to influenza virus could be used as markers of influenza virus infection. We studied 23 subjects who developed upper respiratory, lower respiratory, or systemic symptoms during a small outbreak of influenza in a nursing home population. Influenza virus culture from nasopharyngeal swabs yielded influenza virus isolates from 7 of the 23 subjects. Only three of the subjects had a fourfold rise in antibody titer to the influenza virus antigen positivity after the infection. Granzyme B and cytokine levels were measured in peripheral blood mononuclear cells (PBMC) obtained from all subjects and stimulated with live influenza virus. Elevated granzyme B levels in virus-stimulated PBMC in combination with lower respiratory tract or systemic symptoms in study subjects was a significant predictor of culture-confirmed influenza virus infection compared to those from whom influenza virus could not be identified. Cytokine levels did not distinguish between the two groups in a similar type of analysis. Granzyme B in combination with the clinical profile of symptoms may be a useful retrospective marker for influenza virus infection.
Longitudinal Studies of Neutralizing Antibody Responses to Rotavirus in Stools and Sera of Children following Severe Rotavirus Gastroenteritis
Authors: Coulson Barbara S. 
Published: December 2019

ABSTRACT
Rotavirus-neutralizing antibody responses in sera and stools of children hospitalized with rotavirus gastroenteritis and then monitored longitudinally were optimally detected by using local rotavirus strains. Stool responses were highest on days 5 to 8 after the onset of diarrhea. Longitudinal monitoring suggested that serum neutralizing antibody responses were a more useful measure of severely symptomatic rotavirus infection than stool responses but that stool antibody responses may be a useful measure of rotavirus immunity.
Ceramidase Activity in Bacterial Skin Flora as a Possible Cause of Ceramide Deficiency in Atopic Dermatitis
Authors: Ohnishi Yoshinori , Okino Nozomu , Ito Makoto , Imayama Shuhei 
Published: December 2019

ABSTRACT

A marked decrease in the content of ceramide has been reported in the horny layer of the epidermis in atopic dermatitis (AD). This decrease impairs the permeability barrier of the epidermis, resulting in the characteristic dry and easily antigen-permeable skin of AD, since ceramide serves as the major water-holding molecule in the extracellular space of the horny layer. On the other hand, the skin of such patients is frequently colonized by bacteria, most typically by
Staphylococcus aureus
, possessing genes such as those for sphingomyelinase, which are related to sphingolipid metabolism. We therefore tried to identify a possible correlation between the ceramide content and the bacterial flora obtained from the skin of 25 patients with AD versus that of 24 healthy subjects, using a thin-layer chromatographic assay of the sphingomyelin-associated enzyme activities secreted from the bacteria. The findings of the assay demonstrated that ceramidase, which breaks ceramide down into sphingosine and fatty acid, was secreted significantly more from the bacterial flora obtained from both the lesional and the nonlesional skin of patients with AD than from the skin of healthy subjects; sphingomyelinase, which breaks sphingomyelin down into ceramide and phosphorylcholine, was secreted from the bacterial flora obtained from all types of skin at similar levels for the patients with AD and the healthy controls. The finding that the skin of patients with AD is colonized by ceramidase-secreting bacteria thus suggests that microorganisms are related to the deficiency of ceramide in the horny layer of the epidermis, which increases the hypersensitivity of skin in AD patients by impairing the permeability barrier.