bluuebunny/crossref_metadata_2025_split huggingface dataset:
Reproducibility of Immunological Tests Used To Assess Multiple Chemical Sensitivity Syndrome
Authors: Hoover Donald R. , Donnay Albert , Mitchell Clifford S. , Ziem Grace , Rose Noel R. , Sabath Daniel E. , Yurkow Edward J. , Nakamura Robert , Vogt Robert F. , Waxdal Myron , Margolick Joseph B. 
Published: November 2003

ABSTRACTWhether persons with multiple chemical sensitivity syndrome (MCS) have immunological abnormalities is unknown. To assess the reliability of selected immunological tests that have been hypothesized to be associated with MCS, replicate blood samples from 19 healthy volunteers, 15 persons diagnosed with MCS, and 11 persons diagnosed with autoimmune disease were analyzed in five laboratories for expression of four T-cell surface activation markers (CD25, CD26, CD38, and HLA-DR) and in four laboratories for autoantibodies (to smooth muscle, thyroid antigens, and myelin). For T-cell activation markers, the intralaboratory reproducibility was very good, with 90% of the replicates analyzed in the same laboratory differing by ≤3%. Interlaboratory differences were statistically significant for all T-cell subsets except CD4+cells, ranging from minor to eightfold for CD25+subsets. Within laboratories, the date of analysis was significantly associated with the values for all cellular activation markers. Although reproducibility of autoantibodies could not be precisely assessed due to the rarity of abnormal results, there were inconsistencies across laboratories. The effect of shipping on all measurements, while sometimes statistically significant, was very small. These results support the reliability of fresh and shipped samples for detecting large (but perhaps not small) differences between groups of donors in the T-cell subsets tested. When comparing markers that are not well standardized, it may be important to distribute samples from different study groups evenly over time.
Genetic Variation at the Mitochondrial Large-Subunit rRNA Locus of
Pneumocystis
Isolates from Simian Immunodeficiency Virus-Infected Rhesus Macaques
Authors: Norris Karen A. , Wildschutte Hans , Franko Jennifer , Board Kathryn F. 
Published: November 2003

ABSTRACT

The nucleotide sequences of a segment of the
Pneumocystis
mitochondrial large-subunit (mt LSU) rRNA gene from rhesus macaques coinfected with simian immunodeficiency virus (SIV) and
Pneumocystis carinii
were examined. Of 12 isolates examined, 3 were found to be identical and the others showed substantial sequence variation, with up to 13% divergence among variants. We identified two general sequence types that differed at several sites, including a conserved 26-nucleotide insertion. Four monkeys had evidence of two
Pneumocystis
variants present simultaneously, indicative of a mixed infection. There was a high degree of variance between the rhesus macaque-derived
Pneumocystis
mt LSU rRNA gene sequence and the cognate sequences in
Pneumocystis
organisms derived from other hosts. Analysis of the mt LSU rRNA genes of
Pneumocystis
organisms derived from rhesus macaques and several other mammalian hosts supports the observation that rhesus macaque-derived
Pneumocystis
is most closely related to human-derived
Pneumocystis
. In addition, the data identify the mt LSU rRNA gene as an informative locus for transmission and epidemiological studies of the SIV-rhesus macaque model of
Pneumocystis
infection.
Detection of Antibodies toAnaplasma phagocytophilumandEhrlichia chaffeensisAntigens in Sera of Korean Patients by Western Immunoblotting and Indirect Immunofluorescence Assays
Authors: Park Jin-ho , Heo Eun-jeong , Choi Kyoung-seong , Dumler J. Stephen , Chae Joon-seok 
Published: November 2003

ABSTRACTTwo hundred seventy one serum samples from South Korean patients were tested to detect antibodies againstAnaplasma phagocytophilum(the human granulocytic ehrlichiosis agent) andEhrlichia chaffeensis(the human monocytic ehrlichiosis agent) by indirect fluorescent-antibody assay (IFA) and the Western blot assay. These sera were collected from patients with symptoms of high fever. The rate of seropositivity forOrientia tsutsugamushiwas 50.9% by IFA at the Public Health & Environmental Research Institute and National Institute of Health in South Korea. By IFA, 30 (11.1%) and 39 (14.4%) of the serum samples reacted withA. phagocytophilumandE. chaffeensisantigens, respectively. By the Western blot assays, 24 (8.9%) and 29 (10.7%) of the serum samples reacted with purifiedA. phagocytophilumandE. chaffeensisprotein antigens, respectively. This report strengthens other evidence regarding the presence ofA. phagocytophilumandE. chaffeensisinfections in humans in South Korea.
CD56+-T-Cell Responses to Bacterial Superantigens and Immune Recognition of Attenuated Vaccines
Authors: Saikh Kamal U. , Dyas Beverly , Kissner Teri , Ulrich Robert G. 
Published: November 2003

ABSTRACTNatural killer T (NKT) cells, coexpressing natural killer (NK) and T-cell receptors (TCR), are associated with immunity to viruses, tumors, and parasites. A well-characterized subclass of these NKT cells expresses biased TCR and recognizes glycolipids such as α-galactoceramide, which is found naturally only in marine sponges and presented by the cell surface glycoprotein CD1d. However, a larger number of T cells present in human blood coexpress the NK marker CD56 and unbiased TCR and do not appear to require CD1 for antigen presentation. Observing high frequencies of CD4 and CD8 coreceptor expression in human CD56+T cells, we examined the potential role of major histocompatibility complex (MHC) class II molecules in the activation of these cells. Activation of mononuclear cells with bacterial superantigens presented by MHC class II molecules resulted in increased frequency of CD56+T cells. Primarily, CD4+cells within the CD56+-T-cell population responded to the bacterial superantigens, and cytokine expression profiles were Th1-like. Further, increased levels of T cells expressing CD56 were observed in mononuclear cell cultures responding to aStaphylococcus aureusvaccine or tetanus toxoid. Collectively, our data suggest that a significant number of CD56+T cells recognize pathogen-associated ligands in association with MHC class II molecules.
Diagnosis of Dengue Virus Infection by the Visual and Simple AuBioDOT Immunoglobulin M Capture System
Authors: Vázquez Susana , Lemos Gilda , Pupo Maritza , Ganzón Oscar , Palenzuela Daniel , Indart Adriana , Guzmán María G. 
Published: November 2003

ABSTRACTThe Dengue IgM Capture ELISA (MAC-ELISA) is the immunoenzymatic system recommended by the Pan American Health Organization and the World Health Organization for the serological diagnosis of dengue virus infection due to its high sensitivity, ease of performance, and use of a single acute-phase serum sample. However, tests with this enzyme-linked immunosorbent assay (ELISA) system are time-consuming and require equipment for washing, incubation, and reading of the results. AuBioDOT is a multistep visual diagnostic immunoassay that uses technology based on the immunoglobulin M (IgM) capture ELISA principle. This system uses white polyethylene opaque plates as the solid phase, colloidal gold as the marker, and silver ion amplification. It does not require special equipment, it is totally manually operated, and it can be performed in less than 1 h. The sensitivity and specificity of AuBioDOT for the detection of anti-dengue virus IgM antibodies were studied with a panel of 336 serum samples (150 serum samples from patients with suspected or serologically confirmed dengue virus infection, 186 serum samples from healthy blood donors and patients without dengue virus infection). The results were compared with those obtained by the MAC-ELISA. A sensitivity of 97.7% and a specificity of 97.1% were obtained. The concordance of the two tests was 97.3%, with a kappa index of 0.94. The application of AuBioDOT for the detection of anti-dengue virus IgM antibodies is recommended as an alternative method for the diagnosis of dengue virus infection, both for clinical diagnosis and for seroepidemiological surveillance. The system is useful under field conditions and in laboratories and requires little equipment.
Mimotopes of the Vi Antigen ofSalmonella entericaSerovar Typhi Identified from Phage Display Peptide Library
Authors: Tang Swee-Seong , Tan Wen-Siang , Devi Shamala , Wang Lin-Fa , Pang Tikki , Thong Kwai-Lin 
Published: November 2003

ABSTRACTThe capsular polysaccharide Vi antigen (ViCPS) is an essential virulence factor and also a protective antigen ofSalmonella entericaserovar Typhi. A random 12-mer phage-displayed peptide library was used to identify mimotopes (epitope analogues) of this antigen by panning against a ViCPS-specific monoclonal antibody (MAb) ATVi. Approximately 75% of the phage clones selected in the fourth round carried the peptide sequence TSHHDSHGLHRV, and the rest of the clones harbored ENHSPVNIAHKL and other related sequences. These two sequences were also obtained in a similar panning process by using pooled sera from patients with a confirmed diagnosis of typhoid fever, suggesting they mimic immunodominant epitopes of ViCPS antigens. Binding of MAb ATVi to the mimotopes was specifically blocked by ViCPS, indicating that they interact with the same binding site (paratope) of the MAb. Data and reagents generated in this study have important implications for the development of peptide-base diagnostic tests and peptide vaccines and may also provide a better understanding of the pathogenesis of typhoid fever.
Monoclonal Antibodies That Distinguish Antigenic Variants of
Canine Parvovirus
Authors: Nakamura Masato , Nakamura Kazuya , Miyazawa Takayuki , Tohya Yukinobu , Mochizuki Masami , Akashi Hiroomi 
Published: November 2003

ABSTRACT

Canine parvovirus
(CPV) is classified as a member of the feline parvovirus (FPV) subgroup. CPV isolates are divided into three antigenic types: CPV type 2 (CPV-2), CPV-2a, and CPV-2b. Recently, new antigenic types of CPV were isolated from Vietnamese leopard cats and designated CPV-2c(a) or CPV-2c(b). CPV-2c viruses were distinguished from the other antigenic types of the FPV subgroup by the absence of reactivity with several monoclonal antibodies (MAbs). To characterize the antigenicity of CPV-2c, a panel of MAbs against CPV-2c was generated and epitopes recognized by these MAbs were examined by selection of escape mutants. Four MAbs were established and classified into three groups on the basis of their reactivities: MAbs which recognize CPV-2a, CPV-2b, and CPV-2c (MAbs 2G5 and 20G4); an MAb which reacts with only CPV-2b and CPV-2c(b) (MAb 21C3); and an MAb which recognizes all types of the FPV subgroup viruses (MAb 19D7). The reactivity of MAb 20G4 with CPV-2c was higher than its reactivities with CPV-2a and CPV-2b. These types of specificities of MAbs have not been reported previously. A mapping study by analysis of neutralization-resistant mutants showed that epitopes recognized by MAbs 21C3 and 19D7 belonged to antigenic site A. Substitution of the residues in site B and the other antigenic site influenced the epitope recognized by MAb 2G5. It was suggested that the epitope recognized by MAb 20G4 was related to antigenic site B. These MAbs are expected to be useful for the detection and classification of FPV subgroup isolates.
Novel Microtechnique for Assessment of Postnatal Maturation of the Phagocytic Function of Neutrophils and Monocytes
Authors: Muniz-Junqueira Maria Imaculada , Peçanha Lídia Maria Figueira , da Silva-Filho Valeriano Luiz , de Almeida Cardoso Maria Cecília , Tosta Carlos Eduardo 
Published: November 2003

ABSTRACTWe describe a simple test for the evaluation of phagocytosis and provide a chart of reference values to evaluate normal phagocytosis by age. We assessed the postnatal maturation of phagocytic function of neutrophils and monocytes. Phagocytosis was evaluated in newborn children delivered vaginally or by cesarean section, infants, preschool children, schoolchildren, and adult subjects. Two drops of blood were placed on a microscope slide and incubated withSaccharomyces cerevisiaeyeasts, and phagocytosis was evaluated by microscopy. Our technique showed results comparable to or better than those obtained by other usual techniques. The neutrophils of newborn children delivered by cesarean section showed a phagocytic capacity 45% higher than those of neonates delivered vaginally, whereas neutrophils from children in the latter group showed the lowest phagocytic capacity of all age groups. Phagocytosis by neutrophils reached the levels seen in adults at about the first year of life, while there were no important variations in phagocytosis by monocytes in the different age groups. The technique described is reliable and fast, uses only a few drops of blood, and allows better preservation of cell function due to the minimal manipulation to which the cells are submitted. The delayed maturation of the phagocytic function by neutrophils may account for the high levels of susceptibility of newborn and infant children to bacterial infections. This practical method of assessment of phagocytosis may allow the diagnosis of primary or secondary phagocytic deficiencies to be made more easily and may allow better monitoring and treatment of those with dysfunctions of these cells.
Evolution of Two Amino Acid Positions Governing Broad Neutralization Resistance in a Strain of Feline Immunodeficiency Virus over 7 Years of Persistence in Cats
Authors: Pistello Mauro , Matteucci Donatella , Giannecchini Simone , Bonci Francesca , Sichi Olimpia , Presciuttini Silvano , Bendinelli Mauro 
Published: November 2003

ABSTRACT
Fresh isolates of lentiviruses are characterized by an outstanding resistance to antibody-mediated neutralization. By investigating the changes that occurred in a neutralization-sensitive tissue culture-adapted strain of feline immunodeficiency virus after it was reinoculated into cats, a previous study had identified two amino acid positions of the surface glycoprotein (residues 481 and 557) which govern broad neutralization resistance (BNR) in this virus. By extending the follow-up of six independently evolving in vivo variants of such virus for up to 92 months, we now show that the changes at the two BNR-governing positions not only were remarkably stereotyped but also became fixed in an ordered sequential fashion with the duration of in vivo infection. In one variant, the two positions were also seen to slowly alternate at determining BNR. Evidence that evolution at the BNR-governing positions was accompanied, and possibly driven, by changes in the antigenic makeup of the viral surface brought about by the mutations at such positions is also presented.
Anti-HBs Cellular Immune Response in Kidney Recipients before and 4 Months after Transplantation
Authors: Aguilar Patricia , Renoult Edith , Jarrosson Loraine , Kolopp-Sarda Marie Nathalie , Mathieu Christine Prin , Faure Gilbert C. , Kessler Michěle , Bene Marie C. , Kohler Chantal , Kennel De March Anne 
Published: November 2003

ABSTRACTPatients with renal failure represent a population at risk for hepatitis B, since only 50 to 60% of them develop protective humoral responses after vaccination. As this could be due to an altered regulation of cellular immune responses, the objectives of the present study were to evaluate the proliferative abilities of lymphocytes from patients with chronic renal failure after stimulation in vitro with a mitogen (pokeweed mitogen [PWM]) or HBsAg. In order to differentiate between the immunodeficiency associated with renal failure and that due to immunosuppression posttransplantation, the same subjects were tested before and 4 months after kidney transplantation. The lymphoproliferation assay used was performed by flow cytometry, which is based on sequential analysis of the cell cycle and which allows analysis of cytokine production. Serologically, the group of 36 patients tested comprised 22% nonresponders, 30% poor responders, and 48% responders. Lymphocyte growth was observed for all patients after stimulation with PWM, indicating that these cells had the capacity to proliferate in vitro. The level of lymphoproliferation in response to PWM was significantly reduced after transplantation, yet both before and after transplantation, all serologic nonresponders developed cellular responses to at least two vaccines. No correlation between humoral and cellular responses was shown. Proliferating cells were lymphocytes, which mostly secreted interleukin 4 (IL-4) and IL-10 for the three serologic groups. This study suggests that even when repeated vaccination fails to induce significant antibody levels in patients with renal failure, specific HBs cellular responses develop, and these may prove to be efficient in protecting these patients against hepatitis B.
Quantification of CCR5 mRNA in Human Lymphocytes and Macrophages by Real-Time Reverse Transcriptase PCR Assay
Authors: Lai Jian-Ping , Yang Ji-Hong , Douglas Steven D. , Wang Xu , Riedel Eric , Ho Wen-Zhe 
Published: November 2003

ABSTRACT

CCR5, a β-chemokine receptor, plays an important role in human immunodeficiency virus (HIV) infection of human immune cells, as it is a primary coreceptor for HIV entry into macrophages. We have applied a newly developed real-time reverse transcriptase PCR (RT-PCR) assay for the quantification of CCR5 mRNA in human blood immune cells. The CCR5 real-time RT-PCR assay has a sensitivity of 100 mRNA copies, with a dynamic range of detection between 10
2
and 10
6
copies of the CCR5 mRNA transcripts per reaction. The assay is highly reproducible, with an intra-assay coefficient of variation of the threshold cycle of less than 2.07%. When used for quantification of CCR5 mRNA in human monocyte-derived macrophages (MDM) and peripheral blood lymphocytes (PBL), the assay has precision and reproducibility. MDM expressed higher levels of CCR5 mRNA than did PBL. Thus, this assay has the potential and a wide application for the investigation of the role of CCR5 in inflammatory diseases and viral infections, including HIV disease.
Effect of Antigen Coating Conditions on Enzyme-Linked Immunosorbent Assay for Detection of Immunoglobulin G Antibody to
Neisseria meningitidis
Serogroup Y and W135 Capsular Polysaccharide Antigens in Serum
Authors: Giardina Peter C. , Evans Renee E. , Sikkema Daniel J. , Madore Dace , Hildreth Stephen W. 
Published: November 2003

ABSTRACT
Human sera collected from 28 consenting adult volunteers were used to define assay conditions for meningococcal vaccine clinical trial serology. Immunoassay parameters were optimized with these test sera and the standard reference serum, CDC1992. Coating conditions for serogroup Y and W135 polysaccharide antigens were found to influence the predicted serum immunoglobulin G (IgG) antibody concentrations. Sera that displayed IgG antibody binding profiles most unlike that of CDC1992 were influenced the most by coating conditions. Our results suggest that presentation of specific epitopes is influenced by antigen-coating concentrations for serogroup Y and W135 polysaccharides.
ImmunochromatographicBrucella-Specific Immunoglobulin M and G Lateral Flow Assays for Rapid Serodiagnosis of Human Brucellosis
Authors: Smits Henk L. , Abdoel Theresia H. , Solera Javier , Clavijo Encarnacion , Diaz Ramon 
Published: November 2003

ABSTRACTTo fulfill the need for a simple and rapid diagnostic test for human brucellosis, we used the immunochromatographic lateral flow assay format to develop two assays, one for the detection ofBrucella-specific immunoglobulin M (IgM) antibodies and one for the detection ofBrucella-specific IgG antibodies. The diagnostic values of these tests were examined. The tests are shown to detect acute, persistent, and relapsing disease and can be used to monitor treatment. The sensitivity ofBrucellaIgM and IgG flow assays calculated for the combined assay results is 96%, and specificity amounts to 99%. The flow assay requires neither specialized training nor equipment, the assay is very easy to perform and to read, and the components are stable without a requirement for refrigeration and well standardized. Together these characteristics indicate that theBrucellaIgM and IgG flow assays are ideal for use in clinical settings in rural and suburban areas in which brucellosis is endemic.
Cell-Mediated Immune Responses of Healthy Laboratory Volunteers to Sonicate Antigens Prepared from the Most Prevalent Strains ofMycobacterium tuberculosisfrom South India Harboring a Single Copy of IS6110
Authors: Rajavelu Priya , Das Sulochana D. 
Published: November 2003

ABSTRACTOur restriction fragment length polymorphism (RFLP) studies have shown that the most prevalent (40%) strains ofMycobacterium tuberculosisfrom South India contain a single copy of the IS6110insertion sequence and are of importance in studying virulence and immunity. Sonicate antigens from seven such strains were used to study in vitro T-cell proliferation and gamma interferon (IFN-γ) and interleukin-12 (IL-12) secretion as markers of protective immunity in 25 healthy subjects positive for purified protein derivative (PPD). The standard PPD and heat-killed H37Rv antigens induced the maximum levels of T-cell proliferation and IFN-γ secretion but low levels of IL-12. All sonicate antigens induced T-cell proliferation and IFN-γ secretion with strong positive correlation. Our results suggest that sonicate antigens from the most prevalent and recent strains ofM.tuberculosisfrom clinical isolates have the potential to induce T-cell activation and may allow newer and specific antigens to be further characterized for diagnosis and vaccine development.
Prokaryotic Expression and Antigenic Characterization of Three RecombinantLeishmaniaAntigens for Serological Diagnosis of Canine Leishmaniasis
Authors: Rosati S. , Ortoffi M. , Profiti M. , Mannelli A. , Mignone W. , Bollo E. , Gradoni L. 
Published: November 2003

ABSTRACTThree recombinant antigens ofLeishmania chagasi(=L. infantum) were expressed in prokaryotic systems and evaluated (using a panel of dog sera characterized by parasitological and serological immunofluorescent antibody test [IFAT] techniques) as diagnostic markers of infection. The whole open reading frame encoding K9, the gene fragment encoding the repetitive sequence of K26, and the 3′-terminal gene fragment encoding a single 39-amino-acid subunit of the kinesin-related protein K39 (K39sub) were amplified fromL. infantumDNA and cloned into a pGEX-2T expression vector in frame with glutathioneS-transferase (GST). The sensitivity and specificity of enzyme-linked immunosorbent assays (ELISAs) using K26 as an antigen (evaluated with sera from 20 parasitologically positive and 20 parasitologically negative dogs) were both 100% (95% confidence interval [CI] = 83.2 to 100). When K9 and K39sub were used, sensitivity was 95% (95% CI = 75.1 to 99.9) and specificity was 100% (95% CI = 83.2 to 100). Using 182 field sera, a good agreement was found between the recombinant K26 ELISA and IFAT (K= 0.92; 95% CI = 0.86 to 0.98) results and between the K9 and K39sub ELISA (used in parallel) and IFAT (K= 0.87; 95% CI = 0.80 to 0.95) results. The results demonstrate that each antigen carries immunodominant epitopes and that their combination may further increase the sensitivity of currently available serological tests.