bluuebunny/crossref_metadata_2025 huggingface dataset:
A Bereavement Support Group Intervention Is Longitudinally Associated with Salutary Effects on the CD4 Cell Count and Number of Physician Visits
Authors: Goodkin Karl , Feaster Daniel J. , Asthana Deshratn , Blaney Nancy T. , Kumar Mahendra , Baldewicz Teri , Tuttle Raymond S. , Maher Kevin J. , Baum Marianna K. , Shapshak Paul , Fletcher Mary Ann
Published: December 2019

A randomized, controlled, clinical trial was conducted to examine the impact of a semistructured, 10-week, once weekly, 90-min/session bereavement support group intervention on immunological, neuroendocrine, and clinical health status in human immunodeficiency virus type 1-seropositive (HIV-1+) and HIV-1-seronegative (HIV-1−) homosexual men, compared to a standard of care control condition. A total of 119 homosexual men (74 HIV-1+and 45 HIV-1−) were assessed at baseline, 10 weeks, and 6 months follow-up. At the 6-month follow-up assessment, the intervention groups exhibited significant beneficial effects compared to controls on changes in CD4 cell, total T-lymphocyte, and total lymphocyte counts, when baseline levels, antiretroviral medication use, CDC stage of disease, and other potentially confounding factors were accounted for. There was no statistically significant effect on the CD4/CD8 ratio or on the CD8 cell count. The effect on CD4 cell count was associated with group attendance and with changes in plasma cortisol level. Plasma cortisol levels decreased significantly among intervention subjects, compared to controls. A significantly reduced number of health care visits over the 6-month follow-up period among the intervention subjects supported the clinical relevance of the immunological changes observed for both HIV-1+and HIV-1−individuals. These results indicate that behavioral interventions may have salutary immunological and clinical health effects following bereavement among HIV-1-infected individuals. The effect in HIV-1−individuals suggests that this bereavement support group intervention might have similar salutary effects in the general population. Potential effects of such interventions on clinical HIV disease progression are of interest and should be studied.
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

A 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

The 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

Serum 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

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

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

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

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

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

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

We 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

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

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

The 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

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

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.
Evaluation of a Western Blot Test in an Outbreak of Acute Pulmonary Histoplasmosis
Authors: Pizzini Claudia V. , Zancopé-Oliveira Rosely M. , Reiss Errol , Hajjeh Rana , Kaufman Leo , Peralta José Mauro
Published: December 2019

A western blot (WB) test was evaluated for detection of antibodies against native glycosylated and chemically deglycosylated M and H antigens of
Histoplasma capsulatum
in serum obtained from patients during the acute phase of pulmonary histoplasmosis that occurred during an outbreak. Of 275 serum samples tested by immunodiffusion and complement fixation (CF) samples from 40 patients affected during this outbreak and from 37 negative controls were tested by WB test. A group of patients whose sera were negative for CF antibodies and precipitins early in the acute stage of histoplasmosis but who all seroconverted during convalescence 6 weeks later were tested with the WB test. Antibodies against untreated H and M antigens were detected at a 1:100 dilution by WB test in 45% of the 20 acute-phase serum samples and in all 20 of the convalescent-phase specimens. The WB test’s sensitivity for acute-phase specimens increased to 90% (18 of 20 specimens) when H and M antigens were treated by periodate oxidation to inactivate susceptible carbohydrate epitopes. When native glycosylated antigens were used in the WB test, positive reactions were observed in negative control serum specimens (3 of 37 specimens; 8%) and in serum specimens obtained from asymptomatic persons screened as part of the outbreak investigation (13 of 20 specimens; 65%). These positive reactions were also attributed to glycosidic epitopes since the specificity of the WB test increased from 78 to 100% when periodate-treated H and M antigens were used. WB test with deglycosylated H and M antigens of histoplasmin provides a rapid, sensitive, and specific test to diagnose acute pulmonary histoplasmosis before precipitins can be detected.
Humoral Response in Toscana Virus Acute Neurologic Disease Investigated by Viral-Protein-Specific Immunoassays
Authors: Magurano Fabio , Nicoletti Loredana
Published: December 2019

The Toscana virus (family
Bunyaviridae
, genus
Phlebovirus
) is the only sandfly-transmitted virus that demonstrates neurotropic activity. Clinical cases ranging from aseptic meningitis to meningoencephalitis caused by Toscana virus are yearly observed in central Italy during the summer, and several cases have been reported among tourists returning from zones of endemicity (Italy, Portugal, Spain, and Cyprus). In Toscana virus patients, immunoglobulin M (IgM) antibodies, usually present at the onset of symptoms, can reveal elevated titers by enzyme-linked immunosorbent assay and can persist for at least 1 year. IgG antibodies can be absent at the onset of symptoms: titers rise in convalescent sera and persist for many years. At least five proteins have been identified in Toscana virus-infected cells: nucleoprotein N, glycoproteins G1 and G2, a large protein (L) assumed to be a component of the polymerase, and two nonstructural proteins, NSm and NSs. We report results of a study on the antibody response to individual viral proteins in patients with Toscana virus-associated acute neurologic disease. Immunoblotting and semiquantitative radioimmunoprecipitation assay (RIPA) allow identification of nucleoprotein N as the major antigen responsible for both IgM and IgG responses. Antibodies to proteins other than nucleoprotein N are detected only by RIPA. Antibodies to glycoproteins are detected in about one-third of patients, and whereas their presence always predicts neutralization, some serum samples with neutralizing activity have undetectable levels of antibodies to G1-G2. Antibodies to nonstructural proteins NSm and NSs are also identified. The results obtained raise some questions about antigenic variability and relevant neutralization epitopes of Toscana virus.
Validation of the Indirect MAP1-B Enzyme-Linked Immunosorbent Assay for Diagnosis of Experimental
Cowdria ruminantium
Infection in Small Ruminants
Authors: Mboloi Martin M. , Bekker Cornelis P. J. , Kruitwagen Cas , Greiner Matthias , Jongejan Frans
Published: December 2019

The major antigenic protein 1 fragment B (MAP1-B) enzyme-linked immunosorbent assay (ELISA) for the diagnosis of
Cowdria ruminantium
infections was validated to determine cutoff values and evaluate its diagnostic performance with sheep and goat sera.
Cowdria
-infected populations consisted of 48 sheep and 44 goats, while the noninfected populations consisted of 64 sheep and 107 goats. Cutoff values were determined by two-graph receiver-operating characteristic (TG-ROC) curves. The cutoff value was set at 31 and 26.6% of the positive control reference samples for sheep and goat sera, respectively. The test’s diagnostic performance was evaluated with measurements of the area under the concentration-time curve (AUC) of the ROC curves and by the valid range proportion (VRP). The AUCs were 0.978 for sheep sera and 0.989 for goat sera. The VRP for both sheep and goat sera was approximately 1.0. The intermediate range (IR), which defines results that are neither positive nor negative, was 0 for goat sera and 2.81 for sheep sera. In an ideal test, the AUC and VRP would be 1.0 and the IR would be 0. In this study these parameters were close to those of an ideal test. It is concluded that the MAP1-B ELISA is a useful test for the diagnosis of
C. ruminantium
infection in small ruminants.
Immunodeficiency Due to a Unique Protracted Developmental Delay in the B-Cell Lineage
Authors: Goldman Armond S. , Miles Stephen E. , Rudloff Helen E. , Palkowetz Kimberly H. , Schmalstieg Frank C.
Published: December 2019

A unique immune deficiency in a 24-month-old male characterized by a transient but protracted developmental delay in the B-cell lineage is reported. Significant deficiencies in the number of B cells in the blood, the concentrations of immunoglobulins in the serum, and the titers of antibodies to T-dependent and T-independent antigens resolved spontaneously by the age of 39 months in a sequence that duplicated the normal development of the B-cell lineage: blood B cells followed by immunoglobulin M (IgM), IgG, IgA, and specific IgG antibodies to T-independent antigens (pneumococcal polysaccharides). Because of the sequence of recovery, the disorder could have been confused with other defects in humoral immunity, depending on when in the course of disease immunologic studies were conducted. Investigations of X-chromosome polymorphisms suggested that the disorder was not X linked in that the mother appeared to have identical X chromosomes. An autosomal recessive disorder involving a gene that controls B-cell development and maturation seems more likely. In summary, this case appears to be a novel protracted delay in the development of the B-cell lineage, possibly due to an autosomal recessive genetic defect.
Development of a Western Blot Assay for Detection of Bovine Immunodeficiency-Like Virus Using Capsid and Transmembrane Envelope Proteins Expressed from Recombinant Baculovirus
Authors: Abed Y. , St-Laurent G. , Zhang H. , Jacobs R. M. , Archambault D.
Published: December 2019

A 120-amino-acid polypeptide selected from the transmembrane protein region (tTM) and the major capsid protein p26 of bovine immunodeficiency-like virus (BIV) were expressed as fusion proteins from recombinant baculoviruses. The antigenic reactivity of both recombinant fusion proteins was confirmed by Western blot with bovine and rabbit antisera to BIV. BIV-negative bovine sera and animal sera positive for bovine syncytial virus and bovine leukemia virus failed to recognize the recombinant fusion proteins, thereby showing the specificity of the BIV Western blot. One hundred and five bovine serum samples were tested for the presence of anti-BIV antibodies by the recombinant protein-based Western blot and a reference Western blot assay using cell culture-derived virions as test antigens. There was a 100% concordance when the p26 fusion protein was used in the Western blot. However, the Western blot using the tTM fusion protein as its test antigen identified four BIV-positive bovine sera which had tested negative in both the p26 recombinant-protein-based and the reference Western blot assays. This resulted in the lower concordance of 96.2% between the tTM-protein-based and reference Western blot assays. The results of this study showed that the recombinant p26 and tTM proteins can be used as test antigens for the serodetection of BIV-infection in animals.
Predominant Immunoglobulin A Response to Phase II Antigen of
Coxiella burnetii
in Acute Q Fever
Authors: Fournier Pierre-Edouard , Raoult Didier
Published: December 2019

Diagnosis of acute Q fever is usually confirmed by serology, on the basis of anti-phase II antigen immunoglobulin M (IgM) titers of ≥1:50 and IgG titers of ≥1:200. Phase I antibodies, especially IgG and IgA, are predominant in chronic forms of the disease. However, between January 1982 and June 1998, we observed anti-phase II antigen IgA titers of ≥1:200 as the sole or main antibody response in 10 of 1,034 (0.96%) patients with acute Q fever for whom information was available. In order to determine whether specific epidemiological or clinical factors were associated with these serological profiles, we conducted a retrospective case-control study that included completion of a standardized questionnaire, which was given to 40 matched controls who also suffered from acute Q fever. The mean age of patients with elevated phase II IgA titers was significantly higher than that usually observed for patients with acute Q fever (
P
= 0.026); the patients were also more likely than controls to live in rural areas (
P
= 0.026) and to have increased levels of transaminase in blood (
P
= 0.03). Elevated IgA titers are usually associated with chronic Q fever and are directed mainly at phase I antigens. Although the significance of our findings is unexplained, we herein emphasize the fact that IgA antibodies are not specific for chronic forms of Q fever and that they may occasionally be observed in patients with acute disease. Moreover, as such antibody profiles may not be determined by most laboratories, which test only for total antibody titers to phase I and II antigens, the three isotype-specific Ig titers should be determined as the first step in diagnosing Q fever.
Timing of Development of Measles-Specific Immunoglobulin M and G after Primary Measles Vaccination
Authors: Helfand Rita F. , Kebede Senait , Gary Howard E. , Beyene Hagos , Bellini William J.
Published: December 2019

A standard method for diagnosing measles is to detect measles-specific immunoglobulin M (IgM) in the serum of infected persons. Interpreting a positive IgM result from a person with suspected measles can be difficult if the person has recently received a measles vaccine. We have previously demonstrated that measles-specific IgM may persist for at least 8 weeks after primary vaccination, but it is unknown how quickly IgM appears. This study determined the timing of the rise of measles-specific IgM and IgG after primary measles vaccination with Schwartz vaccine. Two hundred eighty 9-month-old children from Ethiopia presenting for routine measles vaccination were enrolled. Sera were collected before and either 1, 2, 3, or 4 weeks after vaccination and tested for measles-specific antibodies by an IgM capture enzyme immunoassay (EIA) and by an indirect IgG EIA. A total of 209 of the 224 children who returned for the second visit had prevaccination sera that were both IgM and IgG negative. The postvaccination IgM positivity rates for these 209 children were 2% at 1 week, 61% at 2 weeks, 79% at 3 weeks, and 60% at 4 weeks. The postvaccination IgG positivity rates were 0% at 1 week, 14% at 2 weeks, 81% at 3 weeks, and 85% at 4 weeks. We conclude that an IgM-positive result obtained by this antibody capture EIA is difficult to interpret if serum is collected between 8 days and 8 weeks after vaccination; in this situation, the diagnosis of measles should be based on an epidemiologic linkage to a confirmed case or on the detection of wild-type measles virus.
Increased Expression of Regeneration and Tolerance Factor in Individuals with Human Immunodeficiency Virus Infection
Authors: DuChateau Brian K. , Lee Gerald W. , Westerman Maxwell P. , Beaman Kenneth D.
Published: December 2019

Regeneration and tolerance factor (RTF) plays a pivotal role in successful pregnancy outcome and has potent immunomodulating properties. During pregnancy, it is abundantly expressed in the placenta and on peripheral B lymphocytes. Several lines of evidence suggest that both successful pregnancy outcome and progression from human immunodeficiency virus (HIV) infection to AIDS are associated with a Th2-type response. As a result, we hypothesized that the cellular expression of RTF may also be increased during infection with HIV. Using flow cytometric analysis, we showed a significantly (P< 0.01) increased expression of RTF on CD3+cells obtained from individuals with HIV over that for individuals without HIV. On average, 32.1% of the CD3+cells from individuals with HIV expressed high levels of RTF. In contrast, an average of only 6.7% of the CD3+cells from individuals without HIV expressed high levels of RTF. Similar results were obtained when CD19+cells from individuals with (mean, 44.1%) and without (mean, 25.8%) HIV were evaluated. Linear regression analysis suggested that high levels of RTF expression by CD3+cells correlated better with viral load (rvalue, 0.46) than with absolute CD4 count (rvalue, 0.09). While additional experiments are necessary to delineate the precise immunologic role of RTF, our current data suggest that RTF expression during HIV infection may be a useful marker of immune activation.
Immunoglobulin Subclass Distribution and Diagnostic Value ofLeishmania donovaniAntigen-Specific Immunoglobulin G3 in Indian Kala-Azar Patients
Authors: Anam Khairul , Afrin Farhat , Banerjee Dwijadas , Pramanik Netai , Guha Subhasis K. , Goswami Rama P. , Gupta Pratap N. , Saha Shiben K. , Ali Nahid
Published: December 2019

Visceral leishmaniasis, or kala-azar, a fatal tropical disease, remains problematic, as early diagnosis is difficult and treatment often results in drug resistance and relapse. We have developed a sensitive enzyme-linked immunosorbent assay (ELISA), using leishmanial membrane antigenic extracts (LAg) to detect specific antibody responses in 25 untreated Indian visceral leishmaniasis patients. To investigate the pathogenetic significance of isotype markers in kala-azar, relative levels of specific immunoglobulin G (IgG), IgM, IgA, IgE, and IgG subclasses were analyzed under clinically established diseased conditions. Since LAg showed higher sensitivity for specific IgG than lysate, the immunoglobulin isotype responses were evaluated, with LAg as antigen. Compared to 60 controls, which included patients with malaria, tuberculosis, leprosy, and typhoid and healthy subjects, visceral leishmaniasis patients showed significantly higher IgG (100% sensitivity, 85% specificity), IgM (48% sensitivity, 100% specificity), and IgE (44% sensitivity, 98.3% specificity) responses. Low levels of IgA in visceral leishmaniasis patients contrasted with a 13-fold-higher reactivity in sera from patients with leprosy. Among IgG subclasses, IgG1, -3, and -4 responses were significantly higher in visceral leishmaniasis patients than in the controls. IgG2 response, however, was significantly higher (twofold) in leprosy than even visceral leishmaniasis patients. The rank orders for sensitivity (IgG = IgG1 = IgG3 = IgG4 > IgG2 > IgM > IgE > IgA) and specificity (IgM = IgG3 > IgE > IgG4 > IgG2 > IgG > IgG1 > IgA) for LAg-specific antibody responses suggest the potentiality of IgG3 as a diagnostic marker for visceral leishmaniasis.
Cytokines and Inflammatory Mediators Do Not Indicate Acute Infection in Cystic Fibrosis
Authors: Wolter Joanne M. , Rodwell Robyn L. , Bowler Simon D. , McCormack Joseph G.
Published: December 2019

Various treatment regimens and difficulties with research design are encountered with cystic fibrosis (CF) because no standard diagnostic criteria exist for defining acute respiratory exacerbations. This study evaluated the role of serial monitoring of concentrations of selected cytokines and inflammatory mediators in serum and sputum as predictors of respiratory exacerbation, as useful outcome measures for CF, and to guide therapy. Interleukin-8 (IL-8), tumor necrosis factor alpha (TNF-α), neutrophil elastase-α-1-protease inhibitor complex (NE complex), protein, and α-1-protease inhibitor (α-1-PI) were measured in serum and sputum collected from CF patients during respiratory exacerbations and periods of well-being. Levels of NE complex, protein, and α-1-PI in sputum rose during respiratory exacerbations and fell after institution of antibiotic therapy (
P
= 0.078, 0.001, and 0.002, respectively). Mean (± standard error of the mean) levels of IL-8 and TNF-α were extremely high in sputum (13,780 ± 916 and 249.4 ± 23.5 ng/liter, respectively) but did not change significantly with clinical deterioration of the patient (
P
> 0.23). IL-8 and TNF-α were generally undetectable in serum, and therefore these measures were unhelpful. Drop in forced expiratory volume in 1 s was the only clinical or laboratory parameter that was close to being a determinant of respiratory exacerbation (
P
= 0.055). This study provides evidence of intense immunological activity occurring continually within the lungs of adult CF patients. Measurement of cytokines and inflammatory mediators in CF sputum is not helpful for identifying acute respiratory exacerbations.
Marked Suppression of T Cells by a Benzothiophene Derivative in Patients with Human T-Lymphotropic Virus Type I-Associated Myelopathy/Tropical Spastic Paraparesis
Authors: Makino Masahiko , Azuma Miyuki , Wakamatsu Shin-Ichi , Suruga Yukio , Izumo Shuji , Yokoyama Mitchel M. , Baba Masanori
Published: December 2019

In a search for new anti-autoimmune agents that selectively suppress activation of autoreactive T cells, one such agent, 5-methyl-3-(1-methylethoxy)benzo[b]thiophene-2-carboxamide (CI-959-A), was found to be effective. This compound, which is known to suppress tumor necrosis factor alpha (TNF-α)-induced CD54 expression, inhibited the primary proliferative response of the T cell to antigen (Ag)-presenting cells (APCs) including allogenic dendritic cells (DCs), autologous Epstein-Barr virus-infected B cells, and human T lymphotropic virus type I (HTLV-I)-infected T cells. Autoreactive T cells from patients with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) spontaneously proliferate in vitro, and their activation is reported to be associated with CD54 expression. The spontaneous proliferation of T cells from patients with HAM/TSP was entirely blocked by CI-959-A. However, in this study, the T-cell proliferation in 15 patients with HAM/TSP was found to depend more extensively on major histocompatibility complex (MHC) class II and CD86 than on CD54 Ags. Since most important APCs for the development of HAM/TSP are DCs and HTLV-I-infected T cells, the effect of CI-959-A on DC generation and on the expression of surface molecules on activated T cells is examined. CI-959-A suppressed recombinant granulocyte-macrophage colony stimulating factor (GM-CSF)- and recombinant interleukin-4-dependent differentiation of DCs from monocytes and inhibited the expression of CD54 and, more extensively, MHC class II and CD86 Ags. CI-959-A showed little toxicity toward lymphoma or HTLV-I-infected T-cell lines or toward monocytes and cultured DCs. These results suggest that CI-959-A might be a potent anti-HAM/TSP agent.
Immunoglobulin G (IgG) Subclass Distribution and IgG1 Avidity of Antibodies in Human Immunodeficiency Virus-Infected Individuals after Revaccination with Tetanus Toxoid
Authors: Kroon F. P. , van Tol M. J. D. , Jol-van der Zijde C. M. , van Furth R. , van Dissel J. T.
Published: December 2019

In human immunodeficiency virus (HIV)-infected individuals the amount of antibodies formed after vaccination with T-cell-dependent recall antigens such as tetanus toxoid is proportional to the peripheral blood CD4+T-lymphocyte counts. To investigate whether the immunoglobulin G (IgG) subclass distribution and avidity of the antibodies produced after vaccination are affected as well, we gave 13 HIV-infected adults with low CD4+T-lymphocyte counts (<200 × 106/liter; group I), 11 HIV-infected adults with intermediate CD4+T-lymphocyte counts (≥200 × 106/liter; group II), and 5 healthy controls booster immunizations with tetanus toxoid. The prevaccination antibody concentrations against tetanus toxoid were similar in the HIV-infected and healthy adults. After vaccination the total IgG and the IgG1 anti-tetanus toxoid antibody concentrations were significantly lower in group I than in group II and the controls. The avidity of the IgG1 anti-tetanus toxoid antibodies formed by HIV-infected adults was within the range for healthy controls, irrespective of their CD4+T-lymphocyte counts.
Significance of Fatty Acids in Pregnancy-Induced Immunosuppression
Authors: Crocker Ian , Lawson Nigel , Daniels Ian , Baker Philip , Fletcher John
Published: December 2019

Pregnancy can exert suppressive effects on chronic inflammatory conditions. We have previously demonstrated a depression in polymorphonuclear leukocyte (PMN) respiratory burst during pregnancy which could explain this amelioration. To elucidate the biochemical mechanism, we have examined PMN phospholipase A2(PLA2) activity and its relationship to cellular and circulating fatty acids in pregnant women (30 to 34 weeks) and nonpregnant controls. PMN PLA2activity was determined by arachidonic acid (AA) and leukotriene B4(LTB4) release, respiratory burst activity was determined by lucigenin-enhanced chemiluminescence, and total serum and PMN fatty acid levels were determined by gas-liquid chromatography. AA release was significantly reduced for pregnancy PMNs in response toN-formyl-met-leu-phe (fMLP) under unprimed and tumor necrosis factor alpha (TNF-α)- or interleukin 8-primed conditions. Similarly, LTB4liberation was significantly reduced in response to fMLP and phorbol myristate acetate in unprimed and TNF-α-primed pregnancy PMNs. All major fatty acid classes were altered in the pregnant state. Of these differences in PMNs, oleic acid and α-linolenic acid showed a significant increase (13 and 26%, respectively) and stearic acid and AA showed a significant decrease (8 and 30%, respectively). The stearic acid, oleic acid, and AA compositions of all cells analyzed correlated with their corresponding changes in serum fatty acid levels. Crossover serum incubations modified both fatty acid profiles and the PMN respiratory burst accordingly, while individual fatty acid incorporation studies highlighted the importance of polyunsaturated fatty acids for NADPH oxidase efficiency. These findings indicate that the attenuation of PMN function in pregnancy may originate from a reduction in the available pool of cellular fatty acids. Furthermore, this reduction arises as a direct result of a pregnancy-induced shift in circulating fatty acids from polyunsaturated to monounsaturated forms.