bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human Interleukin-15 Improves Engraftment of Human T Cells in NOD-SCID Mice
Authors: Sun Anyuan , Wei Haiming , Sun Rui , Xiao Weihua , Yang Yongguang , Tian Zhigang 
Published: February 2006

ABSTRACTHuman nonobese diabetic-severe combined immune deficiency (NOD-SCID) mouse chimeras have been widely used as an in vivo model to assess human immune function. However, only a small fraction of transferred human T lymphocytes can be detected in human peripheral blood lymphocyte (huPBL)-NOD-SCID chimeras. To improve the reconstitution of human T lymphocytes in NOD-SCID mice, the use of recombinant human interleukin-15 (rhIL-15) as a stimulator of human lymphocytes was explored. Administration of rhIL-15 after transplantation of huPBLs into NOD-SCID mice increased reconstitution of human T lymphocytes in a dose-dependent manner, with an optimal dosage of 1 μg/mouse. The number of human T lymphocytes (HLA-ABC+CD3+) in the lymphoid organs or tissue of rhIL-15-treated huPBL-NOD-SCID mice increased 11- to 80-fold, and phytohemagglutinin-induced T-lymphocyte proliferation and cytokine production were significantly enhanced. Additionally, although mature human cells have not been thought to enter the murine thymus, human T lymphocytes were detected in the huPBL-NOD-SCID thymus after rhIL-15 treatment. Thus, rhIL-15 can be used to optimize long-term peripheral T-cell engraftment in these human-mouse chimeras and may also be useful in clinical treatment of T-cell deficiencies.
Performance of the TruGene Human Immunodeficiency Virus Type 1 Genotyping Kit and OpenGene DNA Sequencing System on Clinical Samples Diluted to Approximately 100 Copies per Milliliter
Authors: Gale Howard B. , Kan Virginia L. , Shinol Rebecca C. 
Published: February 2006

ABSTRACT
The TruGene human immunodeficiency virus type 1 (HIV-1) genotyping kit/OpenGene DNA sequencing system (Bayer HealthCare, Tarrytown, NY) reliably produced clinically acceptable resistance profiles for reverse transcriptase and protease inhibitors on patient samples diluted to ∼100 copies/ml following extraction with the QIAamp viral RNA minikit (QIAGEN Inc., Valencia, CA). One modification of the standard protocol was made to guarantee PCR amplification: a centrifugation step to concentrate virus was added before RNA extraction. For genotypic antiretroviral resistance testing, no significant differences in the identification and sensitivity of detection for codon mutations, base mutations, and multibase sites were found between the original and diluted samples.
Modulation of Immunity-Related Gene Expression in Small Intestines of Mice by Oral Administration of Lactoferrin
Authors: Wakabayashi Hiroyuki , Takakura Natsuko , Yamauchi Koji , Tamura Yoshitaka 
Published: February 2006

ABSTRACTOral administration of lactoferrin (LF), an antimicrobial and immunomodulatory protein, shows a protective effect against infectious diseases, possibly via immunomodulation of the host. Initially, we confirmed an immunomodulatory effect of LF by observing changes in the number of cells in the leukocyte subsets in the peripheral blood and spleens of mice 1 day after oral administration of LF. Then we developed a quantitative reverse transcription-PCR method for 20 immunity-related genes of antimicrobial proteins, pattern recognition receptors, cytokines, and lymphocyte mobilization-related proteins, and we assessed the expression of these genes in the small intestines of mice 2 h after administration of water, bovine serum albumin (BSA), or LF. Expression of the LF gene was lower in mice administered LF than in mice administered water or BSA, implying a negative-feedback control. Expression of gamma interferon (IFN-γ) and interleukin-10 (IL-10) was lower in both BSA- and LF-administered mice than in water administered mice, suggesting a nonspecific effect of protein ingestion. Expression of NOD2, IFN-β, and IL-12p40 was higher with LF administration than with water or BSA administration. The expression levels of these three genes were correlated. This study indicated that oral administration of LF modulates the small intestinal expression of genes closely related to the host defense in a specific or a nonspecific manner.
Gamma Interferon Production in Response toMycobacterium bovisBCG andMycobacterium tuberculosisAntigens in Infants Born to Human Immunodeficiency Virus-Infected Mothers
Authors: Van Rie Annelies , Madhi Shabir A. , Heera Jayvant R. , Meddows-Taylor Stephen , Wendelboe Aaron M. , Anthony Fiona , Violari Avy , Tiemessen Caroline T. 
Published: February 2006

ABSTRACTIn utero sensitization to infectious pathogens can establish immunological memory and may influence the immune response to unrelated antigens. Little is known about the influence of intrauterine human immunodeficiency virus (HIV) exposure on the cellular immune response to mycobacterial antigens. Whole-blood culture gamma interferon (IFN-γ) production in response to mycobacterial antigens was measured at birth and 6 weeks of age to determine the characteristics of the IFN-γ response in HIV-exposed infants toMycobacterium bovisBCG and mycobacterial antigens. At birth, we observed an increased immune activation in response to phytohemagglutinin among HIV-exposed, uninfected infants. In a proportion of these infants, we also observed an increased immune activation in response to purified protein derivative, BCG, and early secreted antigen target 6. Increases in the IFN-γ response to the four antigens between birth and 6 weeks of age, observed in all HIV-unexposed infants, was absent in a substantial proportion of HIV-exposed, uninfected infants. The immunological differences persisted at 6 weeks of age, suggesting a sustained impact of in utero immune priming by HIV. Intrauterine exposure to HIV affects the infants' cellular immune response to mycobacterial antigens, either specifically or as a consequence of nonspecific, broadly reactive immune activation. Further studies will be important to help determine optimal vaccination and disease prevention strategies for this vulnerable population group.
Evaluation of a Tetraplex Microsphere Assay for
Bordetella pertussis
Antibodies
Authors: Prince Harry E. , Lapé-Nixon Mary , Matud Jose 
Published: February 2006

ABSTRACT

To increase testing efficiency, a microsphere-based multianalyte immune detection (MAID) system was developed to measure serum immunoglobulin G (IgG) and IgA recognizing two
Bordetella pertussis
antigens, pertussis toxin (PT) and filamentous hemagglutinin antigen (FHA). The assay was performed as two separate duplexes. One duplex measured IgG to PT and FHA, and the other measured IgA to PT and FHA. The two duplexes were then combined and analyzed as a tetraplex. The MAID system and an in-house enzyme-linked immunosorbent assay (ELISA) system were used to evaluate 100 sera from blood donors and 220 consecutive sera submitted for
B. pertussis
antibody testing. For both the MAID and ELISA systems, antibody levels were defined as increased if greater than the blood donor group 95th percentile value. The qualitative concordance rates between MAID and ELISA results for the 220 consecutively submitted sera were as follows: PT IgG, 99%; PT IgA, 94%; FHA IgG, 93%; FHA IgA, 94%. The overall concordance rate was 95% (836 of 880 result sets). For 29 of 44 (66%) discordant result sets, the discordant MAID result was supported by the MAID and ELISA results for other
B. pertussis
antibodies. The MAID and in-house ELISA systems were also used to evaluate 20 sera previously tested for pertussis antibodies at a pertussis vaccine research laboratory; MAID results for all four analytes did not significantly differ from results obtained by the research laboratory. These findings show that antibodies to
B. pertussis
antigens can be measured easily and accurately using a tetraplex microsphere system.
Kinetics and Diagnostic and Prognostic Potential of Quantitative Western Blot Analysis and Antigen-Specific Enzyme-Linked Immunosorbent Assay in Experimental Canine Leishmaniasis
Authors: Talmi-Frank D. , Strauss-Ayali D. , Jaffe C. L. , Baneth G. 
Published: February 2006

ABSTRACT

Quantitative computerized Western blot analysis of antibody responses during experimental canine
Leishmania infantum
infection distinguished between immunodominant and nonimmunodominant protein bands. Six infected beagles, positive by both PCR and parasite culture, were monitored over 75 weeks postinfection and during a 12-week allopurinol treatment course. All dogs were symptomatic at the time of treatment. Of 12 antigenic bands examined, the immunodominant bands (12, 14, 24, 29, 48, and 68 kDa) showed significantly increased intensities (
P
< 0.01) and higher frequencies of recognition than the nonimmunodominant bands at all time points. Detection of the former bands at 6 weeks postinfection preceded seroconversion by enzyme-linked immunosorbent assay (ELISA) both on crude
Leishmania
antigen or the recombinant proteins rK39 and HSP70. Reactivity with the 14-, 48-, and 68-kDa bands signified early infection, whereas increased reactivity with the 14-, 24-, and 29-kDa bands was associated with posttreatment parasite persistence and potential unfavorable prognosis. Total lane intensity (TLI) emerged as a sensitive marker for early infection and increased as early as 4 weeks postinfection. TLI had a significantly higher (
P
< 0.01) relative increase rate than crude
Leishmania
antigen or HSP70 or rK39 ELISA at all time points. These immunodominant antigens and TLI, as determined by quantitative Western blotting, will be valuable for early detection and treatment evaluation of canine leishmaniasis.
Detection of κ and λ Light Chain Monoclonal Proteins in Human Serum: Automated Immunoassay versus Immunofixation Electrophoresis
Authors: Jaskowski Troy D. , Litwin Christine M. , Hill Harry R. 
Published: February 2006

ABSTRACT
Recently, turbidimetric immunoassays for detecting and quantifying κ and λ free light chains (FLC) have become available and are promoted as being more sensitive than immunofixation electrophoresis (IFE) in detecting FLC monoclonal proteins. In this study, we assessed the ability of these turbidimetric assays to detect serum monoclonal proteins involving both free and heavy-chain-bound κ and λ light chains compared to standard immunofixation electrophoresis. Sera demonstrating a restricted band of protein migration (other than a definite M spike) by serum protein electrophoresis (SPE), which may represent early monoclonal proteins, were also examined. When compared to IFE, percent agreement, sensitivity, and specificity for the κ-FLC and λ-FLC were 94.6, 72.9, and 99.5% and 98.5, 91.4, and 99.7%, respectively, in detecting monoclonal proteins involving free and heavy-chain-bound light chains. The majority of sera (73.7%) demonstrating a restricted band of protein migration on SPE demonstrated abnormal IFE patterns suggestive of multiple myeloma or monoclonal gammopathy of unknown significance, but gave normal κ/λ FLC ratios using the turbidimetric immunoassays. In conclusion, the κ and λ FLC assays are significantly less sensitive (72.9 to 91.4%) than IFE, but specific in detecting serum monoclonal proteins. Moreover, the κ/λ ratio has little value in routine screening since the majority of sera with abnormal IFE patterns had normal κ/λ FLC ratios.
General Anesthesia Delays the Inflammatory Response and Increases Survival for Mice with Endotoxic Shock
Authors: Fuentes Joseph M. , Talamini Mark A. , Fulton William B. , Hanly Eric J. , Aurora Alexander R. , De Maio Antonio 
Published: February 2006

ABSTRACTAnesthesia is an indispensable component of any operative procedure. In this study, we demonstrate that continuous isoflurane anesthesia for 1 h after a lethal dose (20 mg/kg of body weight) ofEscherichia colilipopolysaccharide (LPS) results in a significant increase in survival of C57BL/6J (B6) mice in comparison with survival of nonanesthetized mice. Protection by anesthesia correlates with a delay in plasma LPS circulation, resulting in a delayed inflammatory response, particularly DNA binding activity of NF-κB and serum levels of tumor necrosis factor alpha, interleukin-6 (IL-6), and IL-10. Disparate classes of anesthetic agents produce the same effects on the inflammatory response, which is also independent of the inbred mouse strain used. These results suggest that anesthesia has an important impact on the outcome from endotoxemia. Moreover, the immunomodulatory effects of anesthetics should be considered when interpreting data from experimental animal models.
Anti-Interleukin-15 Prevents Arthritis inBorrelia-Vaccinated and -Infected Mice
Authors: Amlong Corey A. , Nardelli Dean T. , Peterson Sara Heil , Warner Thomas F. , Callister Steven M. , Schell Ronald F. 
Published: February 2006

ABSTRACTWe showed previously that interleukin-17 (IL-17) plays a significant role in the induction of arthritis associated withBorreliavaccination and challenge. Little information, however, is available about the chain of immunologic events that leads to the release of IL-17. The production of IL-17 has been linked to stimulation of memory cells by IL-15. Therefore, we hypothesized that IL-15 is involved in the induction of arthritis associated withBorreliavaccination and infection of mice. Here we present evidence that treatment ofBorrelia-vaccinated and -infected mice with anti-IL-15 antibody prevents swelling of the hind paws. More importantly, both anti-IL-15 antibody- and recombinant IL-15 receptor alpha-treatedBorrelia-vaccinated and -infected mice were free of major histopathologic indications of arthritis, including hyperplasia, hypertrophy, and vilus formation of the synovium. Similarly, the synovial space and perisynovium were free of inflammatory cells. By contrast, the synovium of nontreatedBorrelia-vaccinated and -infected mice had overt hyperplasia, hypertrophy, and vilus formation. Moreover, the synovial space and perisynovium were infiltrated with neutrophils, macrophages, and lymphocytes. Finally, we show that recombinant IL-15 stimulates the release of IL-17 from lymph node cells obtained near the arthritic site. These results suggest that IL-15 plays a major role in orchestrating IL-17 induction of arthritis associated withBorrelia-vaccinated and -infected mice.
Development of Specific Immunoglobulin Ga (IgGa) and IgGb Antibodies Correlates with Control of Parasitemia in
Babesia equi
Infection
Authors: Cunha Cristina W. , McGuire Travis C. , Kappmeyer Lowell S. , Hines Stephen A. , Lopez A. Marianela , Dellagostin Odir A. , Knowles Donald P. 
Published: February 2006

ABSTRACT

In this study, the kinetics of specific immunoglobulin G (IgG) isotypes were characterized in
Babesia equi
(
Theileria equi
)-infected horses. IgGa and IgGb developed during acute infection, whereas IgG(T) was detected only after resolution of acute parasitemia. The same IgG isotype profile induced during acute infection was obtained by equi merozoite antigen 1/saponin immunization.
Differential Expression of
Aspergillus nidulans
Ammonium Permease Genes Is Regulated by GATA Transcription Factor AreA
Authors: Monahan Brendon J. , Askin Marion C. , Hynes Michael J. , Davis Meryl A. 
Published: February 2006

ABSTRACT

The movement of ammonium across biological membranes is mediated in both prokaryotes and eukaryotes by ammonium transport proteins (AMT/MEP) that constitute a family of related sequences. We have previously identified two ammonium permeases in
Aspergillus nidulans
, encoded by the
meaA
and
mepA
genes. Here we show that
meaA
is expressed in the presence of ammonium, consistent with the function of MeaA as the main ammonium transporter required for optimal growth on ammonium as a nitrogen source. In contrast,
mepA
, which encodes a high-affinity ammonium permease, is expressed only under nitrogen-limiting or starvation conditions. We have identified two additional AMT/MEP-like genes in
A. nidulans
, namely,
mepB
, which encodes a second high-affinity ammonium transporter expressed only in response to complete nitrogen starvation, and
mepC
, which is expressed at low levels under all nitrogen conditions. The MepC gene product is more divergent than the other
A. nidulans
AMT/MEP proteins and is not thought to significantly contribute to ammonium uptake under normal conditions. Remarkably, the expression of each AMT/MEP gene under all nitrogen conditions is regulated by the global nitrogen regulatory GATA factor AreA. Therefore, AreA is also active under nitrogen-sufficient conditions, along with its established role as a transcriptional activator in response to nitrogen limitation.
MNN5
Encodes an Iron-Regulated α-1,2-Mannosyltransferase Important for Protein Glycosylation, Cell Wall Integrity, Morphogenesis, and Virulence in
Candida albicans
Authors: Bai Chen , Xu Xiao-Li , Chan Fong-Yee , Lee Raymond Teck Ho , Wang Yue 
Published: February 2006

ABSTRACT

The cell walls of microbial pathogens mediate physical interactions with host cells and hence play a key role in infection. Mannosyltransferases have been shown to determine the cell wall properties and virulence of the pathogenic fungus
Candida albicans
. We previously identified a
C. albicans
α-1,2-mannosyltransferase, Mnn5, for its novel ability to enhance iron usage in
Saccharomyces cerevisiae
. Here we have studied the enzymatic properties of purified Mnn5 and characterized its function in its natural host. Mnn5 catalyzes the transfer of mannose to both α-1,2- and α-1,6-mannobiose, and this activity requires Mn
2+
as a cofactor and is regulated by the Fe
2+
concentration. An
mnn5Δ
mutant showed a lowered ability to extend O-linked, and possibly also N-linked, mannans, hypersensitivity to cell wall-damaging agents, and a reduction of cell wall mannosylphosphate content, phenotypes typical of many fungal mannosyltransferase mutants. The
mnn5Δ
mutant also exhibited some unique defects, such as impaired hyphal growth on solid media and attenuated virulence in mice. An unanticipated phenotype was the
mnn5Δ
mutant's resistance to killing by the iron-chelating protein lactoferrin, rendering it the first protein found that mediates lactoferrin killing of
C. albicans
. In summary,
MNN5
deletion impairs a wide range of cellular events, most likely due to its broad substrate specificity. Of particular interest was the observed role of iron in regulating the enzymatic activity, suggesting an underlying relationship between Mnn5 activity and cellular iron homeostasis.
Heterologous Expression of Membrane and Soluble Proteins Derepresses
GCN4
mRNA Translation in the Yeast
Saccharomyces cerevisiae
Authors: Steffensen Lotte , Pedersen Per Amstrup 
Published: February 2006

ABSTRACT

This paper describes the first physiological response at the translational level towards heterologous protein production in
Saccharomyces cerevisiae
. In yeast, the phosphorylation of eukaryotic initiation factor 2α (eIF-2α) by Gcn2p protein kinase mediates derepression of
GCN4
mRNA translation. Gcn4p is a transcription factor initially found to be required for transcriptional induction of genes responsible for amino acid or purine biosynthesis. Using various
GCN4-lacZ
fusions, knockout yeast strains, and anti-eIF-2α-P/anti-eIF-2α antibodies, we observed that heterologous expression of the membrane-bound α
1
β
1
Na,K-ATPase from pig kidney, the rat pituitary adenylate cyclase seven-transmembrane-domain receptor, or a 401-residue soluble part of the Na,K-ATPase α
1
subunit derepressed
GCN4
mRNA translation up to 70-fold.
GCN4
translation was very sensitive to the presence of heterologous protein, as a density of 1‰ of heterologous membrane protein derepressed translation maximally. Translational derepression of
GCN4
was not triggered by misfolding in the endoplasmic reticulum, as expression of the wild type or temperature-sensitive folding mutants of the Na,K-ATPase increased
GCN4
translation to the same extent. In situ activity of the heterologously expressed protein was not required for derepression of
GCN4
mRNA translation, as illustrated by the expression of an enzymatically inactive Na,K-ATPase. Two- to threefold overexpression of the highly abundant and plasma membrane-located endogenous H-ATPase also induced
GCN4
translation. Derepression of
GCN4
translation required phosphorylation of eIF-2α, the tRNA binding domain of Gcn2p, and the ribosome-associated proteins Gcn1p and Gcn20p. The increase in Gcn4p density in response to heterologous expression did not induce transcription from the
HIS4
promoter, a traditional Gcn4p target.
Role of N-Terminal Hydrophobic Region in Modulating the Subcellular Localization and Enzyme Activity of the Bisphosphate Nucleotidase from
Debaryomyces hansenii
Authors: Aggarwal Monika , Mondal Alok K. 
Published: February 2006

ABSTRACT

3′, 5′-Bisphosphate nucleotidase is a ubiquitous enzyme that converts 3′-phosphoadenosine-5′-phosphate to adenosine-5′-phosphate and inorganic phosphate. These enzymes are highly sensitive to sodium and lithium and, thus, perform a crucial rate-limiting metabolic step during salt stress in yeast. Recently, we have identified a bisphosphate nucleotidase gene (
DHAL2
) from the halotolerant yeast
Debaryomyces hansenii
. One of the unique features of Dhal2p is that it contains an N-terminal 54-amino-acid-residue hydrophobic extension. In this study, we have shown that Dhal2p exists as a cytosolic as well as a membrane-bound form and that salt stress markedly influences the accumulation of the latter form in the cell. We have demonstrated that the N-terminal hydrophobic region was necessary for the synthesis of the membrane-bound isoform. It appeared that an alternative translation initiation was the major mechanism for the synthesis of these two forms. Moreover, the two forms exhibit significant differences in their substrate specificity. Unlike the cytosolic form, the membrane-bound form showed very high activity against inositol-1,4-bisphosphate. Thus, the present study for the first time reports the existence of multiple forms of a bisphosphate nucleotidase in any organism.
Retention and Loss of Amino Acid Biosynthetic Pathways Based on Analysis of Whole-Genome Sequences
Authors: Payne Samuel H. , Loomis William F. 
Published: February 2006

ABSTRACT

Plants and fungi can synthesize each of the 20 amino acids by using biosynthetic pathways inherited from their bacterial ancestors. However, the ability to synthesize nine amino acids (Phe, Trp, Ile, Leu, Val, Lys, His, Thr, and Met) was lost in a wide variety of eukaryotes that evolved the ability to feed on other organisms. Since the biosynthetic pathways and their respective enzymes are well characterized, orthologs can be recognized in whole genomes to understand when in evolution pathways were lost. The pattern of pathway loss and retention was analyzed in the complete genomes of three early-diverging protist parasites, the amoeba
Dictyostelium
, and six animals. The nine pathways were lost independently in animals,
Dictyostelium
,
Leishmania
,
Plasmodium
, and
Cryptosporidium
. Seven additional pathways appear to have been lost in one or another parasite, demonstrating that they are dispensable in a nutrition-rich environment. Our predictions of pathways retained and pathways lost based on computational analyses of whole genomes are validated by minimal-medium studies with mammals, fish, worms, and
Dictyostelium
. The apparent selective advantages of retaining biosynthetic capabilities for amino acids available in the diet are considered.