bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of Immunoglobulin A Antibodies to
Helicobacter pylori
in Serum and Gastric Juice in Relation to Mucosal Inflammation
Authors: Hayashi Shunji , Sugiyama Toshiro , Yokota Kenji , Isogai Hiroshi , Isogai Emiko , Oguma Keiji , Asaka Masahiro , Fujii Nobuhiro , Hirai Yoshikazu
Published: December 2019

Helicobacter pylori
is a major etiologic agent in gastroduodenal disorders. In this study, immunoglobulin A (IgA) antibodies to
H. pylori
antigens were evaluated in serum and gastric juice specimens obtained from patients with gastritis or peptic ulcers by utilizing antibody capture enzyme-linked immunosorbent assays (ACELISAs). Urease α subunit (UA), urease β subunit (UB), the 66-kDa heat shock protein (HSP), and the 25-kDa protein (25K) were used as antigens for the ACELISAs. The antibody titers of the ACELISAs reflect the ratio of
H. pylori
-specific IgA to total IgA. The ratio is stable, although the antibody concentration fluctuates in gastric juice. By using ACELISAs it was possible to evaluate quantitatively not only serum IgA antibodies but also gastric juice secretory IgA (S-IgA) antibodies. In both serum IgA and gastric juice S-IgA ACELISAs, the titers of antibody to HSP and 25K were remarkably correlated with the histologic grade of gastritis, whereas those to UA and UB were not strongly correlated with histologic grade. Thus, it is useful for estimating the histologic grade of gastritis to quantify serum IgA and gastric juice S-IgA antibodies to HSP and 25K.
Age-Related Changes in Blood Lymphocyte Subsets of Saudi Arabian Healthy Children
Authors: Shahabuddin S. , Al-Ayed I. , El-Rab M. O. Gad , Qureshi M. I.
Published: December 2019

The age-related changes in absolute and percentage values of lymphocyte subsets in the peripheral blood of healthy children of different ages (1 month to 13 years) were studied by flow cytometry. The absolute and percentage values for most lymphocyte subpopulations differed substantially with age. Comparisons among age groups from infants through adults revealed progressive declines in the absolute numbers of leukocytes, total lymphocytes, and T, B, and natural killer (NK) cells. The percentages of T cells increased with age. Within the T-lymphocyte population, the CD8+subset increased but the CD4+subset decreased, resulting in a declining CD4+/CD8+ratio. The percentage of B cells declined, but that of NK cells remained unchanged. The percentage of HLA-DR+T cells increased over time, but their number changed inconsistently. Our findings confirm and extend earlier reports on age-related changes in lymphocyte subpopulations. These data should be useful in the interpretation of disease-related changes, as well as therapy-dependent alterations, in lymphocyte subsets in children of different age groups.
Serodiagnosis of Neosporosis in Individual Cows and Dairy Herds: A Comparative Study of Three Enzyme-Linked Immunosorbent Assays
Authors: Wouda W. , Brinkhof J. , van Maanen C. , de Gee A. L. W. , Moen A. R.
Published: December 2019

The performance of three enzyme-linked immunosorbent assays (ELISA) for detection of antibodies toNeospora caninumin bovine sera was evaluated by using various categories of sera. Two commercial ELISA methods, one based on chemically fixed intact tachyzoites and one based on a sonicate lysate of whole tachyzoites, were compared with an in-house ELISA based on a detergent lysate of whole tachyzoites. A brief description of the development of the latter ELISA is also given. There was good agreement among all three tests with regard to postabortion sera. By using acute-phase abortion sera from cows with confirmedN. caninum-induced and non-N. caninum-induced abortions, satisfactory levels of sensitivity and specificity were calculated for all tests. In addition, similar test results were obtained with postpartum samples from dams and calves. However, considerable differences were found between test results of sequential samples and cross-sectional and total-herd samples. Apparently, these discrepancies were due to different sensitivities of the tests for detection of low antibody levels in chronically infected animals. It is suggested that these differences were primarily due to the use of different antigens and different test sample dilutions. It is concluded that all tests are applicable as an additional diagnostic tool in cases of abortion in cattle and for monitoring of congenitally infected calves. For herd screening, the lysate-based ELISAs appear to be more adequate because of their higher sensitivities.
rK39 Enzyme-Linked Immunosorbent Assay for Diagnosis of
Leishmania donovani
Infection
Authors: Zijlstra E. E. , Daifalla N. S. , Kager P. A. , Khalil E. A. G. , El-Hassan A. M. , Reed S. G. , Ghalib H. W.
Published: December 2019

The rK39 enzyme-linked immunosorbent assay (ELISA) was compared with the direct agglutination test (DAT) for
Leishmania donovani
infection in the Sudan. rK39 ELISA proved more sensitive than DAT in diagnosis of kala-azar (93 and 80%, respectively); both tests may remain positive up to 24 months after treatment. For patients with post-kala-azar dermal leishmaniasis and individuals with subclinical infection, rK39 ELISA performed as well as DAT but could detect infection 6 months earlier in ∼40% of patients. Conversion in DAT and rK39 ELISA also occurred in leishmanin skin test (LST)-positive individuals, suggesting active parasite replication (rK39 is an amastigote antigen) in these presumably immune individuals. In contrast to DAT, rK39 ELISA also detected infection in randomly selected LST-positive individuals (in four of six) and endemicity (LST-negative) controls (in one of five). rK39 ELISA appears more sensitive than DAT and may prove an important tool in epidemiological studies.
Antigenic Variation in
Bacteroides forsythus
Detected by a Checkerboard Enzyme-Linked Immunosorbent Assay
Authors: Sims Tom J. , Mancl Lloyd A. , Braham Pamela H. , Page Roy C.
Published: December 2019

Evidence indicating that multiple serotypes of
Bacteroides forsythus
participate in rapidly progressing periodontal infections has not been reported previously. Our aim was to develop an assay for detecting subsets of
B. forsythus
clinical isolates which differ in serogroup membership and subsets of patients with immunoglobulin G (IgG) responses which differ in serogroup recognition. A checkerboard enzyme-linked immunosorbent assay (ELISA) was used to assess variation in the IgG binding profiles of 22 clinical isolates in sera from 28 patients with early-onset rapidly progressive periodontitis. To accommodate the maximum number of isolates and sera in a given assay run, a multiplate assay grid with standard 96-well microtest plates was established. Single dilutions of individual sera were placed in rows crossing columns of isolate-coated wells, and antigen-specific IgG immobilized in the wells was measured as ELISA absorbance. Pooled sera and isolates were assayed in parallel to serve as negative controls for variation in IgG binding profiles. Correlation and hierarchical cluster analysis of the absorbance data matrix showed that the isolates could be sorted into at least four clusters based on variations in their IgG binding profiles across different sera. Furthermore, at least two patient clusters were defined by variations in their serum IgG antigen recognition profiles across different isolates. We conclude that multiple serogroups of
B. forsythus
exist and that different serogroups are dominant in the antibody response of different patients. The method applied here could be used to serologically classify clinical isolates of other species which evoke a serum antibody response in patients.
Levels of Cytokines and Immune Activation Markers in Plasma in Human Immunodeficiency Virus Infection: Quality Control Procedures
Authors: Aziz Najib , Nishanian Parunag , Fahey John L.
Published: December 2019

Procedures for quality control (QC) in a laboratory that concentrates on cytokine and soluble marker measurements in biological fluids are outlined. Intra-assay, interassay, and interlaboratory experiences are presented. Plasma and serum β2-microglobulin (β2M) and neopterin test data are presented in greatest detail, along with substantial tumor necrosis factor alpha (TNF-α), gamma interferon, soluble interleukin-2 receptor-α (sIL-2Rα), sTNF-RII, IL-4, and IL-6 data. Recommended QC procedures for cytokine and soluble-marker testing include replicate testing of two or more reference samples provided by the kit manufacturer, replicate testing of in-house frozen reference QC samples that represent normal and abnormal analyte contents, retesting 15 to 20% of randomly selected samples, and comparing normal reference ranges each year. Also, eight cytokines and soluble markers were evaluated in human immunodeficiency virus (HIV)-seronegative and HIV-seropositive individuals stratified on the basis of CD4 T-cell numbers. Levels of some but not all cytokines in serum increased in HIV infection. There was a tendency for cytokines to increase with more advanced disease, defined by reduced CD4 T-cell numbers. Cytokine changes did not relate closely to CD4 level, indicating that separate information was provided by the measurements of TNF-α, sTNF-RII, sIL-2Rα, β2M, and neopterin. Serum IL-4 and TNF-α levels were not increased. The quality of laboratory data can impact on clinical relevance. Interlaboratory comparisons revealed substantial differences at some sites and documented the need for external proficiency-testing quality assurance programs.
Development of a Fluorescent Focus Identification Assay Using Serotype-Specific Monoclonal Antibodies for Detection and Quantitation of Rotaviruses in a Tetravalent Rotavirus Vaccine
Authors: Yang D. P. , Goldberg K. M. , Ma Xiang-Dong , Magargle W. , Rappaport R.
Published: December 2019

A fluorescent focus identification assay (FFIDA) was developed for use in experimental studies and for quantitation of the components in a tetravalent live oral rotavirus vaccine. The assay utilizes four serotype-specific neutralizing monoclonal antibodies (MAb) to detect and quantify individual rotaviruses by immunofluorescence staining of fixed virus-infected monkey kidney cells. In mixed virus infections, all four MAb, W1 (serotype 1), 1C10 (serotype 2), R1 (serotype 3), and S4 (serotype 4), specifically stain the relevant homologous serotype without exhibiting any cross-reactivity against the other serotypes. Furthermore, the test is sensitive enough to differentiate at least twofold (0.3 log) differences in virus titer. The results of testing four individual experimental vaccine lots three or more consecutive times showed that all four lots contained similar proportions of the four vaccine strains as detected by the classical plaque neutralization identification test. The rapidity and efficiency of the FFIDA are desirable attributes that make it suitable for use in studies requiring identification and quantitation of one or more of the four major rotavirus serotypes.
Accumulation of Acid-Fast Lipochrome Bodies in Glial Cells of the Midbrain Nigral Lesion in Parkinson’s Disease
Authors: Kohbata Shunro , Tamura Tomokazu , Hayashi Ryoichi
Published: December 2019

To confirm or refute the proposed link between nocardiae and Parkinson’s disease (PD), we investigated the presence of acid-fast spherical structures similar to filterable nocardiae at the midbrain nigral lesions of three patients with PD. Many clusters of acid-fast lipochrome bodies were dense around blood vessels in the two patients with Hoehn and Yahr stage II and III PD. These clusters were present in the vicinity of melanin-pigmented neurons in the three PD patients studied. Examination of adjacent hematoxylin-and-eosin-stained sections indicated that they consisted of yellow-green granules, bodies, and aggregates in ballooned glial cells. On the other hand, no clusters of acid-fast lipochrome bodies were observed at the compacta region of three control patients. Our results suggest that the immunological and genetic relationship between the acid-fast lipochrome bodies and filterable nocardiae should be investigated.
Angiocentric CD3+T-Cell Infiltrates in Human Immunodeficiency Virus Type 1-Associated Central Nervous System Disease in Children
Authors: Katsetos Christos D. , Fincke John E. , Legido Agustin , Lischner Harold W. , de Chadarevian Jean-Pierre , Kaye Edward M. , Platsoucas Chris D. , Oleszak Emilia L.
Published: December 2019

A significant proportion of brain tissue specimens from children with AIDS show evidence of vascular inflammation in the form of transmural and/or perivascular mononuclear-cell infiltrates at autopsy. Previous studies have shown that in contrast to inflammatory lesions observed in human immunodeficiency virus type 1 (HIV-1) encephalitis, in which monocytes/macrophages are the prevailing mononuclear cells, these infiltrates consist mostly of lymphocytes. Perivascular mononuclear-cell infiltrates were found in brain tissue specimens collected at autopsy from five of six children with AIDS and consisted of CD3+T cells and equal or greater proportions of CD68+monocytes/macrophages. Transmural (including endothelial) mononuclear-cell infiltrates were evident in one patient and comprised predominantly CD3+T cells and small or, in certain vessels, approximately equal proportions of CD68+monocytes/macrophages. There was a clear preponderance of CD3+CD8+T cells on the endothelial side of transmural infiltrates. In active lesions of transmural vasculitis, CD3+T-cell infiltrates exhibited a distinctive zonal distribution. The majority of CD3+cells were also CD8+and CD45RO+. Scattered perivascular monocytes/macrophages in foci of florid vasculitis were immunoreactive for the p24 core protein. In contrast to the perivascular space, the intervening brain neuropil was dominated by monocytes/macrophages, microglia, and reactive astrocytes, containing only scant CD3+CD8+cells. Five of six patients showed evidence of calcific vasculopathy, but only two exhibited HIV-1 encephalitis. One patient had multiple subacute cerebral and brainstem infarcts associated with a widespread, fulminant mononuclear-cell vasculitis. A second patient had an old brain infarct associated with fibrointimal thickening of large leptomeningeal vessels. These infiltrating CD3+T cells may be responsible for HIV-1-associated CNS vasculitis and vasculopathy and for endothelial-cell injury and the opening of the blood-brain barrier in children with AIDS.
Anticytomegalovirus (Anti-CMV) Immunoglobulin G Avidity in Identification of Pregnant Women at Risk of Transmitting Congenital CMV Infection
Authors: Lazzarotto Tiziana , Spezzacatena Patrizia , Varani Stefania , Gabrielli Liliana , Pradelli Paola , Guerra Brunella , Landini Maria Paola
Published: December 2019

In this work, we show that the determination of the anticytomegalovirus antibody avidity carried out before week 18 of gestation is a helpful tool to identify women for enrollment in prenatal diagnosis. This procedure can identify all pregnant women who will give birth to an infected newborn.
Dysregulated Synthesis of Intracellular Type 1 and Type 2 Cytokines by T Cells of Patients with Cutaneous T-Cell Lymphoma
Authors: Lee Bang-Ning , Duvic Madeleine , Tang Chih-Kwang , Bueso-Ramos Carlos , Estrov Zeev , Reuben James M.
Published: December 2019

Mycosis fungoides (MF) and Sezary syndrome (SS) are the two main clinical entities of cutaneous T-cell lymphoma (CTCL). As the disease progresses from MF to SS, a switch from a type 1 (interleukin [IL]-2 and gamma interferon [IFN-γ]) to a type 2 (IL-4) cytokine production profile occurs. Although roles for type 1 and type 2 cytokines in the pathogenesis of CTCL have been proposed, the cellular origins of these cytokines are unclear. Using flow cytometry to identify individual T-cell subsets, we studied cytokine synthesis by the T cells of 13 patients with SS and 12 with MF and 9 hematologically healthy donors. Upon activation with phorbol 12-myristate 13-acetate (PMA), the numbers of T cells synthesizing IL-2 were similar for all study groups. Whereas the predominant T-cell producing IL-2 in healthy donors and in those with MF was CD7+, in patients with SS, it was CD7−. Although the number of IL-4+CD4+T cells was low for all study groups, there was a significantly higher number of IL-4+CD8+T cells in patients with MF than in those with SS or healthy donors. There was a decline in the number of IFN-γ-producing T cells in CTCL donors compared to that in healthy donors. More importantly, there was a significant decrease in the number of IFN-γ-producing T cells with disease progression from MF to SS. The inability of these T cells to synthesize IFN-γ may have prognostic value in CTCL, since it may be responsible for the progression of the disease from MF to SS.
Protection against Influenza Virus Infection of Mice Fed
Bifidobacterium breve
YIT4064
Authors: Yasui Hisako , Kiyoshima Junko , Hori Tetuji , Shida Kan
Published: December 2019

Mice fed
Bifidobacterium breve
YIT4064 and immunized orally with influenza virus were more strongly protected against influenza virus infection of the lower respiratory tract than ones immunized with influenza virus only. The number of mice with enhanced anti-influenza virus immunoglobulin G (IgG) in serum upon oral administration of
B. breve
YIT4064 and oral immunization with influenza virus was significantly greater than that upon oral immunization with influenza virus only. These findings demonstrated that the oral administration of
B. breve
YIT4064 increased anti-influenza virus IgG antibodies in serum and protected against influenza virus infection. The oral administration of
B. breve
YIT4064 may enhance antigen-specific IgG against various pathogenic antigens taken orally and induce protection against various virus infections.
A Powerful DNA Extraction Method and PCR for Detection of Microsporidia in Clinical Stool Specimens
Authors: Müller Andreas , Stellermann Kerstin , Hartmann Pia , Schrappe Matthias , Fätkenheuer Gerd , Salzberger Bernd , Diehl Volker , Franzen Caspar
Published: December 2019

The diagnosis of intestinal microsporidiosis has traditionally depended on direct visualization of the parasite in stool specimens or intestinal biopsy samples by light and/or electron microscopy. Limited information about the specificity and sensitivity of PCR for the detection microsporidia in clinical stool specimens is available. To establish a sensitive and specific method for the detection of microsporidia in clinical samples, we studied clinical stool specimens of 104 randomly selected human immunodeficiency virus-infected patients with diarrhea to compare light microscopy and PCR. Fluorochrome Uvitex 2B staining was used for light microscopy. To raise the sensitivity of PCR, we used a powerful and fast DNA extraction method including stool sedimentation, glass bead disruption, and proteinase K and chitinase digestion. PCR was performed with primer pairs V1-PMP2, V1-EB450, and V1-SI500, and the nature of the PCR products was confirmed by Southern blot hybridization. Microsporidiosis was diagnosed by light microscopy in eight patients. Ten patients tested positive for microsporidiosis by PCR.
Enterocytozoon bieneusi
was found in seven cases, and
Encephalitozoon intestinalis
was found in four cases. In one case a double infection with
E. bieneusi
and
E. intestinalis
was diagnosed by PCR, whereas light microscopy showed only
E. bieneusi
infection. PCR testing of stool specimens is useful for diagnosis and species differentiation of intestinal microsporidiosis in HIV patients.
Alloreactivity and Association of Human Natural Killer Cells with the Major Histocompatibility Complex
Authors: Mavoungou Elie , Sall Aicha , Poaty-Mavoungou Virginie , Toure Fousseyni S. , Yaba Philippe , Delicat Andre , Lansoud-Soukate Joseph
Published: December 2019

All NK cells potentially lytic for autologous cells but not expressing self-major histocompatibility complex (MHC)-reactive receptors could be eliminated by a negative selection mechanism during ontogeny. This idea is based on the existence of a NK cell subset expressing a specific inhibitory receptor for allogeneic MHC alleles. As ancestral haplotypes of the MHC appear to define identical MHC haplotypes in unrelated individuals, unrelated individuals having the same ancestral haplotype should also have the same NK-defined allospecificities that have been shown to map to the human MHC. To test this prediction, multiple cell lines from unrelated individuals having the same ancestral haplotypes were tested for the NK-defined allospecificities. It was found that cells having the same ancestral haplotypes do have the same NK-defined specificities. Furthermore, the NK-defined phenotype of cells that possess two different ancestral haplotypes can be predicted from the NK-defined phenotypes of unrelated cells that are homozygous for the ancestral haplotypes concerned. Although the group 1 and 2 NK-defined allospecificities can be explained to some extent by HLA-C alleles, evidence is presented that additional genes may modify the phenotype conferred by HLA-C.
Variants of aCryptococcus neoformansStrain Elicit Different Inflammatory Responses in Mice
Authors: Chen Lin-Chi , Casadevall Arturo
Published: December 2019

The virulence ofCryptococcus neoformansisolates with high and low extracellular proteolytic activity was investigated in mice. No consistent relationship between proteolytic activity and virulence was observed, but isolates derived from one strain were shown to elicit different inflammatory responses.
A Murine Model of Renal Abscess Formation
Authors: Mussalli George M. , Brunnert Steven R. , Hirsch Emmet
Published: December 2019

We developed a murine model of kidney abscess by direct renal injection of either
Escherichia coli
(1 × 10
6
to 7 × 10
6
organisms) or sterile medium. Bacterial infection produced renal abscesses, bacteremia, and late-onset leukocytosis in all animals. Controls were unaffected. This model may be useful for the study of various sequelae of kidney infection.
Recombinant Human Gamma Interferon in Human Immunodeficiency Virus-Infected Children: Safety, CD4+-Lymphocyte Count, Viral Load, and Neutrophil Function (AIDS Clinical Trials Group Protocol 211)
Authors: Shearer William T. , Kline Mark W. , Abramson Stuart L. , Fenton Terence , Starr Stuart E. , Douglas Steven D.
Published: December 2019

Nineteen children with human immunodeficiency virus (HIV) infection were treated with recombinant human gamma interferon (rIFN-γ) (50 μg/m2subcutaneously three times each week during weeks 1 through 12 and 100 μg/m2subcutaneously three times each week during weeks 13 through 24) in a phase I/II clinical trial. All children continued to receive previously prescribed therapy with oral zidovudine or didanosine. Children were assessed clinically and with laboratory studies during 24 weeks of study treatment and for 12 weeks after completion of rIFN-γ therapy. In general, rIFN-γ therapy was well tolerated. There were two clinical or laboratory adverse events thought to be possibly or probably study drug associated. One child developed acute pancreatitis; another child developed granulocytopenia. Median CD4+-lymphocyte counts and plasma HIV RNA concentrations did not change significantly during therapy. In vitro neutrophil bactericidal activity againstStaphylococcus aureusand superoxide production were not significantly affected by rIFN-γ therapy. We conclude that rIFN-γ therapy in HIV-infected children receiving single-agent antiretroviral therapy is safe and does not produce consistent changes in CD4+-lymphocyte count, plasma HIV RNA concentration, or in vitro neutrophil function.
RNAs in the Sera of Persian Gulf War Veterans Have Segments Homologous to Chromosome 22q11.2
Authors: Urnovitz Howard B. , Tuite James J. , Higashida Jean M. , Murphy William H.
Published: December 2019

Reverse transcriptase PCR (RT-PCR) was used for polyribonucleotide assays with sera from deployed Persian Gulf War veterans with the Gulf War Syndrome and a cohort of nonmilitary controls. Sera from veterans contained polyribonucleotides (amplicons) that were obtained by RT-PCR and that ranged in size from 200 to ca. 2,000 bp. Sera from controls did not contain amplicons larger than 450 bp. DNA sequences were derived from two amplicons unique to veterans. These amplicons, which were 414 and 759 nucleotides, were unrelated to each other or to any sequence in gene bank databases. The amplicons contained short segments that were homologous to regions of chromosome 22q11.2, an antigen-responsive hot spot for genetic rearrangements. Many of these short amplicon segments occurred near, between, or in chromosome 22q11.2 Alu sequences. These results suggest that genetic alterations in the 22q11.2 region, possibly induced by exposures to environmental genotoxins during the Persian Gulf War, may have played a role in the pathogenesis of the Gulf War Syndrome. However, the data did not exclude the possibility that other chromosomes also may have been involved. Nonetheless, the detection of polyribonucleotides such as those reported here may have application to the laboratory diagnosis of chronic diseases that have a multifactorial etiology.
Immunization with a Peptide Corresponding to Chlamydial Heat Shock Protein 60 Increases the Humoral Immune Response in C3H Mice to a Peptide Representing Variable Domain 4 of the Major Outer Membrane Protein ofChlamydia trachomatis
Authors: Motin Vladimir L. , de la Maza Luis M. , Peterson Ellena M.
Published: December 2019

C3H (H-2k) mice are susceptible to a vaginal challenge with human strains ofChlamydia trachomatisand thus are a useful strain for testing potentialChlamydiavaccine candidates. However, C3H mice are fairly poor responders in terms of the level of antibody resulting from immunization with potential protective peptides representing variable domains (VDs) of the major outer membrane protein (MOMP). C57BL/6 (H-2b) mice, on the other hand, are moderately resistant to a vaginal challenge but are good responders to the chlamydial MOMP VDs. Peptides representing universal T-cell helper epitopes were employed to determine whether the antibody response to a peptide representing VD4 of the MOMP, which has been shown to contain neutralizing epitopes, could be enhanced in C3H and C57 mice. Universal T-cell helper peptides from tetanus toxin, the pre-S2 region of hepatitis B virus, and the mouse heat shock protein 60, as well as the corresponding segment of theChlamydiaheat shock protein 60 (hspct), were coadministered with the VD4 peptide. Peptides were coencapsulated in liposomes containing the adjuvant monophosphoryl lipid A and administered by using a combination of mucosal and intramuscular injection. The only T-cell helper peptide that improved the immune response as judged by antibody level, in vitro neutralization assays, and T-cell proliferation was hspct. The response in the C57BL/6 strain was not significantly enhanced with hspctover levels achieved with VD4 alone; however, in C3H mice the levels of serum antibody toC. trachomatisincreased to that seen in C57 mice. However, the molecular specificity and immunoglobulin subclass distribution differed from those of the C57 response, and the neutralizing titers and T-cell proliferation responses were lower. In both strains of mice, titers of vaginal antibody toC. trachomatiswere low. In summary, of the T-helper peptides used, only hspctsignificantly enhanced the immune response of C3H mice to the VD4 peptide, but it had only a modest effect on the immune response of C57 mice.
Use of Protein AG in an Enzyme-Linked Immunosorbent Assay for Screening for Antibodies against Parapoxvirus in Wild Animals in Japan
Authors: Inoshima Yasuo , Shimizu Shinya , Minamoto Nobuyuki , Hirai Katsuya , Sentsui Hiroshi
Published: December 2019

Using protein AG in an enzyme-linked immunosorbent assay (ELISA), we tried to detect antibodies against parapoxvirus in 9 species of wild animals in Japan: the Japanese badger (Meles meles anakuma), Japanese black bear (Ursus thibetanus japonicus), Japanese deer (Cervus nippon centralis), Japanese monkey (Macaca fuscata), Japanese raccoon dog (Nyctereutes procyonoides viverrinus), Japanese serow (Capricornis crispus), Japanese wild boar (Sus scrofa leucomystax), masked palm civet (Paguma larvata), and nutria (Myocastor coypus). A total of 272 serum samples were collected over the period from 1984 to 1995 and were tested by the protein AG-ELISA, the agar gel immunodiffusion test, and an indirect immunofluorescence assay. The protein AG-ELISA was effective in a serological survey for parapoxvirus in wild animals, and antibodies were detected only in Japanese serows. A total of 24 of 66 (36.4%) Japanese serows reacted positively, and they were found in almost all prefectures in all years tested. These results suggest that epizootic cycles of parapoxvirus exist widely in Japanese serows and that they could be reservoirs for the virus in the field in Japan. Moreover, it is probable that they might carry the virus to domestic animals such as cattle, sheep, and goats.
Production, Characterization, and Epitope Mapping of a Monoclonal Antibody against Aspartic Proteinase of
Candida albicans
Authors: Na Byoung-Kuk , Chung Gyung-Tae , Song Chul-Yong
Published: December 2019

A monoclonal antibody (MAb; MAb CAP1) that was reactive with extracellular aspartic proteinase of
Candida albicans
(CAP) was produced. The MAb showed strong sensitivity and reactivity to CAP but not to the aspartic proteinases of
Candida parapsilosis
,
Candida tropicalis
, and
Aspergillus fumigatus
or to human cathepsin D or porcine pepsin. The epitope of the CAP recognized by the MAb was the proteinaseous part of CAP and the putative epitope of the MAb was located in the Asp
77
to Gly
103
sequence. This antibody could be useful for the characterization of CAP and would be a valuable probe for the detection of CAP antigen in the sera of patients with invasive candidiasis.
Inhibition of the Activities of Matrix Metalloproteinases 2, 8, and 9 by Chlorhexidine
Authors: Gendron Renée , Grenier Daniel , Sorsa Timo , Mayrand Denis
Published: December 2019

Matrix metalloproteinases (MMPs) are a host cell-derived proteolytic enzyme family which plays a major role in tissue-destructive inflammatory diseases such as periodontitis. The aim of the present study was to evaluate the inhibitory effect of chlorhexidine (CHX) on MMP-2 (gelatinase A), MMP-9 (gelatinase B), and MMP-8 (collagenase 2) activity. Heat-denatured type I collagen (gelatin) was incubated with pure human MMP-2 or -9 activated with
p
-aminophenylmercuric acetate (APMA), and the proteolytic degradation of gelatin was monitored by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Coomassie blue staining. The effect of CHX on MMP-8 activity was also studied with a cellular model addressing the ability of phorbol myristate acetate (PMA)-triggered human peripheral blood neutrophils (polymorphonuclear leukocytes [PMNs]) to degrade native type I collagen. CHX inhibited the activities of both gelatinases (A and B), but MMP-2 appeared to be more sensitive than MMP-9. Adding calcium chloride to the assay mixtures almost completely prevented the inhibition of MMP-9 activity by CHX, while the inhibition of MMP-2 activity could be reversed only when CHX was used at a low concentration. This observation suggests that CHX may act via a cation-chelating mechanism. CHX dose-dependently inhibited collagenolytic activity of MMP-8 released by PMA-triggered PMNs. MMP-8 without APMA activation was inhibited clearly more efficiently than APMA-activated MMP-8. Our study suggests that the direct inhibition of the MMPs’ activities by CHX may represent a new valuable effect of this antimicrobial agent and explains, at least in part, the beneficial effects of CHX in the treatment of periodontitis.
Cell Surface Display of Human Immunodeficiency Virus Type 1 gp120 on
Escherichia coli
by Using Ice Nucleation Protein
Authors: Kwak Young-Don , Yoo Seung-Ku , Kim Eui-Joong
Published: December 2019

A new system designed for cell surface display of recombinant proteins on
Escherichia coli
has been evaluated for expression of eukaryotic viral proteins. Human immunodeficiency virus type 1 (HIV-1) gp120 was fused to the C terminus of ice nucleation protein (INP), an outer membrane protein of
Pseudomonas syringae
. Western blotting, immunofluorescence microscopy, fluorescence-activated cell-sorting analysis, whole-cell enzyme-linked immunosorbent assay, and ice nucleation activity assay confirmed the successful expression of HIV-1 gp120 on the surface of
Escherichia coli
. This study shows that the INP system can be used for the expression of eukaryotic viral proteins. There is also a possibility that the INP system can be used as an AIDS diagnostic system, an oral vaccine delivery system, and an expression system for various heterologous higher-molecular-weight proteins.
Correlation of Cytokine Elaboration with Mononuclear Cell Adhesion to Platelet Storage Bag Plastic Polymers: a Pilot Study
Authors: ElKattan Ikbal , Anderson James , Yun J. K. , Colton E. , Yomtovian Roslyn
Published: December 2019

The basis for many febrile nonhemolytic transfusion reactions associated with platelet transfusion therapy is cytokine elaboration and accumulation in the storage bag, which correlate with the leukocyte content and the length of platelet storage. We propose that a possible additional variable in the elaboration and accumulation of cytokines is the differential adhesion of mononuclear cells to the plastic substrate of the platelet storage bag. We hypothesize that mononuclear cell adhesion-induced cytokine release is greater in random-donor platelet bags composed of the polyolefin polymer compared to the single-donor apheresis platelet bags composed of the polyvinyl chloride polymer with the tri-(2-ethylhexyl) trimellitate (TEHTM) plasticizer. For four blood donors, we demonstrate preferential mononuclear cell adhesion, in vitro, to discs of polyolefin polymer versus discs of polyvinyl chloride polymer with the TEHTM plasticizer. Scanning electron microscopy corroborates this. In addition, proinflammatory cytokine (interleukin 1β [IL-1β] and tumor necrosis factor alpha [TNF-α]) levels are greater in culture wells containing discs of polyolefin polymer than in those containing discs of polyvinyl chloride polymer with the TEHTM plasticizer, and even more so in storage bags containing polyolefin polymer versus polyvinyl chloride polymer with the TEHTM plasticizer (IL-1β, TNF-α, IL-6, and IL-8). This study suggests, for the first time, that differential plastic substrate mononuclear cell adhesion may contribute to cytokine release during platelet storage. This may represent an additional variable in the pathophysiology of febrile nonhemolytic transfusion reactions in patients receiving stored platelet units.
Immunoglobulin G Avidity in Diagnosis of Toxoplasmic Lymphadenopathy and Ocular Toxoplasmosis
Authors: Paul Malgorzata
Published: December 2019

Traditional serological techniques have some limitations in evaluating the duration of
Toxoplasma gondii
infection in pregnant women, patients with lymphadenopathy, and older children suspected of having congenital toxoplasmosis. In these three groups of patients, two variants of
T. gondii
immunoglobulin G (IgG) avidity tests were used: an EIA Kit (Labsystems) and a noncommercial enzyme-linked immunosorbent assay specially elaborated in the laboratory. The avidity of specific IgG in sera from 23 patients with a known recently acquired infection (mainly pregnant women) was low (less than 30%), whereas that in sera from 19 patients with toxoplasmic lymphadenopathy of 3 weeks to 6 months in duration (mean, 8.3 weeks) covered a large range (between 0.2 and 57.8%; mean, 25.7%); high avidity results were observed for 10 of 19 patients (52.6%). The large range of IgG avidity in patients with toxoplasmic lymphadenopathy suggests various durations of infection in these patients, with a tendency for a chronic phase of toxoplasmosis. According to the avidity marker, five patients with lymphadenopathy for less than 3 months did not have a recent
Toxoplasma
infection. In 6 of 19 patients with lymphadenopathy (31.6%), low IgG avidity values persisted until 5 months after the first serological examination. In all four patients with a documented chronic course of
Toxoplasma
infection (6 months to 8 years after the first positive serology), high IgG avidity values were observed. Among sera from 10 children and young immunocompetent adults suspected of having ocular reactivation of congenital toxoplasmosis, all had high IgG avidity values (over 40%), suggesting congenitally acquired ocular infection rather than noncongenital infection. In conclusion, the avidity of IgG is a valuable marker of recent toxoplasmosis in pregnant women, suggests the duration of invasion in patients with lymphadenopathy, and may be helpful for differentiation between reactivation of congenital infection and recently acquired ocular toxoplasmosis in immunocompetent patients. A low IgG avidity does not always identify a recent case of toxoplasmosis, but a high IgG avidity can exclude primary infections of less than 5 months’ duration.
A Reverse-Sandwich Enzyme-Linked Immunosorbent Assay for Verocytotoxin 1 and 2 Antibodies in Human and Bovine Sera
Authors: Miyazawa H. , Bannai H. , Yanase T. , Morita C. , Satoh S. , Sugiyama J. , Taniguchi S. , Inouye S.
Published: December 2019

A reverse-sandwich enzyme-linked immunosorbent assay (ELISA), in which an antibody is sandwiched by antigens, was established for the titration of antibodies to verocytotoxins (VT) in human and animal sera. This assay has two advantages over a conventional indirect ELISA: (i) higher specificity and sensitivity and (ii) the ability to comparably titrate antibodies from different species. The VT1 (Shiga-like toxin 1) antibody-positive rates were 5% in 202 normal adult humans and 99% in 93 normal cattle at a dairy farm. This ELISA is most suitable for seroepidemiologic studies of infections with VT-producing
Escherichia coli
in humans and various animal species.
Identification of an IS711Element Interrupting thewboAGene ofBrucella abortusVaccine Strain RB51 and a PCR Assay To Distinguish Strain RB51 from OtherBrucellaSpecies and Strains
Authors: Vemulapalli Ramesh , McQuiston John R. , Schurig Gerhardt G. , Sriranganathan Nammalwar , Halling Shirley M. , Boyle Stephen M.
Published: December 2019

Brucella abortusvaccine strain RB51 is a natural stable attenuated rough mutant derived from the virulent strain 2308. The genetic mutations that are responsible for the roughness and the attenuation of strain RB51 have not been identified until now. Also, except for an assay based on pulsed-field gel electrophoresis, no other simple method to differentiate strain RB51 from its parent strain 2308 is available. In the present study, we demonstrate that thewboAgene encoding a glycosyltransferase, an enzyme essential for the synthesis of O antigen, is disrupted by an IS711element inB. abortusvaccine strain RB51. Exploiting this feature, we developed a PCR assay that distinguishes strain RB51 from all otherBrucellaspecies and strains tested.
Complement Fixation Test To Assess Humoral Immunity in Cattle and Sheep Vaccinated withBrucella abortusRB51
Authors: Adone Rosanna , Ciuchini Franco
Published: December 2019

The live attenuatedBrucella abortusstrain RB51 is a rifampin-resistant, lipopolysaccharide (LPS) O-chain-deficient mutant of virulentB. abortus2308. The reduced O-chain content in RB51 prevents this bacterium from inducing antibodies detectable by the conventional serologic tests for bovine brucellosis diagnosis that mainly identify antibodies to LPS. The absence of available serologic tests for RB51 also complicates the diagnosis of possible RB51 infections in humans exposed to this strain. The purpose of this study was to evaluate the suitability of a complement fixation (CF) test performed with the rough strainB. abortusRB51, previously deprived of anticomplementary activity, in detecting anti-B. abortusRB51 antibodies in cattle and sheep experimentally vaccinated with this strain. The results of this study showed that a CF test with RB51 as the antigen is able to specifically detect antibodies following RB51 vaccination in cattle and sheep. In addition, this method could be a useful tool for detectingB. abortusRB51 infection in humans.
Exogenous Cyclic AMP, Cholera Toxin, and Endotoxin Induce Expression of the Lipopolysaccharide Receptor CD14 in Murine Bone Marrow Cells: Role of Purinoreceptors
Authors: Pedron Thierry , Girard Robert , Chaby Richard
Published: December 2019

Little is known about the mechanisms of lipopolysaccharide (LPS) signaling in immature cells that do not express the LPS receptor CD14 yet. Bone marrow granulocytes do not constitutively express CD14 but can be stimulated by low doses of LPS in the absence of serum and then express an inducible form of LPS receptor (iLpsR). We show that in addition to LPS, cholera toxin (CT) and various cyclic AMP (cAMP) analogs can also induce the expression of iLpsR, which was identified as CD14. Induction was independent of intracellular cAMP. The hypothesis that cAMP analogs act via a cell surface receptor was suggested by the unresponsiveness of trypsin-treated cells to these inducers and by the specific binding of [3H]cAMP to the cells. This binding was not inhibited by LPS or CT but was inhibited by various purine derivatives. However, the receptor involved is not a conventional purinoreceptor since both an agonist and an antagonist of such receptors were able to induce iLpsR expression. The results suggest that cAMP analogs and other purine derivatives induce iLpsR after interaction with an unconventional, trypsin-sensitive, purinoreceptor distinct from LPS and CT receptors.
Use of Monoclonal Antibody in Diagnosis of Candidiasis Caused by
Candida albicans
: Detection of Circulating Aspartyl Proteinase Antigen
Authors: Na Byoung-Kuk , Song Chul-Yong
Published: December 2019

To develop a serological diagnosis of invasive candidiasis based on detection of circulating secreted aspartyl proteinase (SAP) antigen of
Candida albicans
, three different enzyme-linked immunosorbent assays (ELISAs) were compared. The first was a standard ELISA to detect anti-SAP antibodies, and the others were an antigen capture ELISA and an inhibition ELISA to detect circulating SAP antigen with monoclonal antibody (MAb) CAP1, which is highly specific for SAP. These tests were applied to 33 serum samples retrospectively selected from 33 patients with mycologically and/or serologically proven invasive candidiasis caused by
C. albicans
. Serum samples from 12 patients with aspergillosis and serum samples from 13 healthy individuals were also included. The sensitivities and specificities were 69.7 and 76.0% for the standard ELISA and 93.9 and 92.0% for the antigen capture ELISA, respectively. However, these values reached 93.9 and 96.0%, respectively, for the inhibition ELISA. Serum samples from 31 of 33 patients had detectable SAP antigen, with concentrations ranging from 6.3 to 19.0 ng/ml. These results indicate that the inhibition ELISA with MAb CAP1 is effective in detection of circulating SAP antigen and that this assay may be useful for diagnosis and treatment monitoring of invasive candidiasis.
Increased Levels of ɛ and γ Isoforms of 14-3-3 Proteins in Cerebrospinal Fluid in Patients with Creutzfeldt-Jakob Disease
Authors: Takahashi Hidehiro , Iwata Toshinari , Kitagawa Yoshinori , Takahashi Reisuke H. , Sato Yuko , Wakabayashi Hideki , Takashima Miwa , Kido Hiroshi , Nagashima Kazuo , Kenney Kimbra , Gibbs Clarence J. , Kurata Takeshi
Published: December 2019

We established four hybridoma cell lines producing monoclonal antibodies (MAbs) against 14-3-3 proteins. Immunoblot analysis revealed that ɛ and γ isoforms were specifically increased in premortem cerebrospinal fluid samples from patients with sporadic Creutzfeldt-Jakob disease. Furthermore, dot immunoblot analysis showed that MAbs were more specific for native antigen than polyclonal antibodies were.
Temporal Variation of the Merozoite Surface Protein-2 Gene ofPlasmodium falciparum
Authors: Eisen Damon , Billman-Jacobe Helen , Marshall Vikki F. , Fryauff Dave , Coppel Ross L.
Published: December 2019

Extensive polymorphism of key parasite antigens is likely to hamper the effectiveness of subunit vaccines againstPlasmodium falciparuminfection. However, little is known about the extent of the antigenic repertoire of naturally circulating strains in different areas where malaria is endemic. To address this question, we conducted a study in which blood samples were collected from parasitemic individuals living within a small hamlet in Western Irian Jaya and subjected to PCR amplification using primers that would allow amplification of the gene encoding merozoite surface protein-2 (MSP2). We determined the nucleotide sequence of the amplified product and compared the deduced amino acid sequences to sequences obtained from samples collected in the same hamlet 29 months previously.MSP2genes belonging to both major allelic families were observed at both time points. In the case of the FC27MSP2family, we observed that the majority of individuals were infected by parasites expressing the same form of MSP2. Infections with parasites expressing 3D7MSP2family alleles were more heterogeneous. NoMSP2alleles observed at the earlier time point were detectable at the later time point, either for the population as a whole or for individuals who were assayed at both time points. We examined a subset of the infected patients by using blood samples taken between the two major surveys. In no patients could we detect reinfection by a parasite expressing a previously encountered form of MSP2. Our results are consistent with the possibility that infection induces a form of strain-specific immune response against the MSP2 antigen that biases against reinfection by parasites bearing identical forms of MSP2.
Differential Responses of Human Mononuclear Phagocytes to Mycobacterial Lipoarabinomannans: Role of CD14 and the Mannose Receptor
Authors: Bernardo John , Billingslea Andrea M. , Blumenthal Robin L. , Seetoo Kurt F. , Simons Elizabeth R. , Fenton Matthew J.
Published: December 2019

CD14 is a signaling receptor for both gram-negative bacterial lipopolysaccharide (LPS) and mycobacterial lipoarabinomannan (LAM) that lacks terminal mannosyl units (AraLAM). In contrast, terminally mannosylated LAM (ManLAM) binds the macrophage mannose receptor (MMRc), although the ability of the MMRc to serve as a signaling receptor has not been previously reported. We compared the abilities of AraLAM and ManLAM to induce distinct responses in two monocytic cell populations, freshly isolated human peripheral blood monocytes (PBM) and monocyte-derived macrophages (MDM). The responses examined were chemotaxis and transient changes in free cytosolic calcium ([Ca2+]in). We found that AraLAM but not ManLAM was chemotactic for both PBM and MDM. Migration of these cells in vitro to AraLAM was specifically blocked by an anti-CD14 monoclonal antibody, suggesting that CD14 mediates the chemotactic response to AraLAM. Subsequently, we found that AraLAM induced a transient rise in [Ca2+]inlevels within a subpopulation of PBM but not MDM. This response was blocked by anti-CD14 antibodies. In contrast, ManLAM induced a transient rise in [Ca2+]inlevels within a subpopulation of MDM but not PBM. This response was blocked by either anti-CD14 or anti-MMRc antibodies. These data suggest that the MMRc can serve as a signaling receptor and that coligation of both CD14 and the MMRc is required to elicit a specific response. Thus, one response to LAM (chemotaxis) can be elicited solely by engaging CD14, whereas a different response (changes in [Ca2+]inlevels) depends on both the differentiation state of the cells and concomitant engagement of CD14 and the MMRc.