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Biomedical subjects

T Kuratsuji

Publications and source records attributed to T Kuratsuji.

At least 19 recordsLinked to original sources
PubMed2004

Assessment of synthetic peptides of severe acute respiratory syndrome coronavirus recognized by long-lasting immunity.

In order to determine highly immunogenic severe acute respiratory syndrome coronavirus (SARS-CoV) epitope peptides capable of inducing long-lasting immunity, we first screened immunoglobulin-G (IgG) antibodies reactive to 197 different overlapping 15-mers from the SARS-CoV proteins in the sera of three infected patients. Forty-two peptides among them were reactive to the sera from all three patients. Consequently, we tested for the reactivity of these 42 peptides to patients' sera (n = 45) at 6-month post-infection. The significantly higher levels of IgG antibodies specific to three (S791, M207 and N161) of 42 peptides were detectable in the post-infection sera from 23 (51%), 27 (60%) and 19 (42%) of 45 patients, respectively. These three peptides, recognized by their long-lasting immunity, may provide a better understanding of the immunogenicity of SARS-CoV.

Enzyme-Linked Immunosorbent Assay↗
PubMed1997

[Improvement of respiratory burst by individual neutrophils from a patient with chronic granulomatous disease, type X91- under treatment by granulocyte colony-stimulating factor for multiple liver abscess].

A 20-year-old male with chronic granulomatous disease (CGD) was admitted with multiple liver abscesses. He had already been diagnosed as CGD, type X91-, when he was 10 years old. He was successfully treated with antibiotics and granulocyte colony-stimulating factor (G-CSF) combined with continuous drainage of abscess. Employing flow cytometry, respiratory burst by individual neutrophils was measured using 2', 7'-dichlorofluorescein. The fluorescence intensity in all individual neutrophils from the patient under G-CSF treatment was higher than the one without G-CSF. G-CSF can be one of effective therapies for infection in some patients with CGD such as X91-.

Adult↗
PubMed1996

Involvement of p40phox in activation of phagocyte NADPH oxidase through association of its carboxyl-terminal, but not its amino-terminal, with p67phox.

Phagocyte NADPH oxidase, dormant in resting cells, is activated upon cell stimulation to produce superoxide anion, a precursor of microbicidal oxidants. Active NADPH oxidase is found on the membrane as an enzyme complex, composed of membrane-integrated cytochrome b558 (gp91phox and p22phox subunits) and two cytosolic factors (p47phox and p67phox), each of the latter containing two src homology 3 (SH3) domains. Recently, we radioactively identified a third cytosolic factor, p40phox, as a molecule that associates with p67phox in human neutrophils. Although it has been found that this p40phox protein is defective in patients with chronic granulomatous disease (CGD) who lack p67phox, evidence to functionally relate it to the NADPH oxidase system has hitherto been lacking. In this study, we raised separate antibodies against both the COOH- and NH2-terminal polypeptides of p40phox as well as against the COOH-terminal polypeptide of p67phox to examine the mode of interaction between p40phox and p67phox in a complex. The antibody against the COOH terminus of p67phox was able to communoprecipitate p40phox in conjunction with p67phox itself as was expected. Very interestingly, however, the antibody against the COOH terminus of p40phox completely dissociated the p67phox molecule from the p40phox-p67phox complex unit without any detectable coimmunoprecipitation of p67phox, despite their tight association, whereas that against the NH2 terminus of p40phox had absolutely no dissociation effect. Similar results were found regarding their effects on the O2-generating ability of cytosol in a cell-free activation system, i.e., inhibition was noted with the COOH terminus antibody but not with that for the NH2 terminus of p40phox. However, this dissociation did not affect the translocation of the cytosolic components including p47phox to the membrane. Once the NADPH oxidase was activated, the antibody for the COOH terminus did not show any inhibitory effect on catalysis by the activated enzyme. The stimulators of NADPH oxidase, MA and SDS, did not dissociate the p40phox-p67phox complex. These results provide the first demonstration that p40phox is practically involved in the activation of NADPH oxidase through the association of its COOH-terminal, but not its NH2-terminal, with p67phox.

Cytosol↗
PubMed1994

A novel cytosolic component, p40phox, of respiratory burst oxidase associates with p67phox and is absent in patients with chronic granulomatous disease who lack p67phox.

Cytosolic components of human neutrophils, p47phox and p67phox, deficiencies of which lead to chronic granulomatous disease (CGD), potentiate respiratory burst oxidase translocating from cytosol to membrane upon cell stimulation. In this report we describe a novel cytosolic component, p40phox, which consistently behaves with p67phox through immunoprecipitation and column works, and is missing in patients with CGD who lack p67phox. Although actin has been reported to be involved in O2- generation, the p40phox profile did not correspond to that of actin. The tight association between p40phox and p67phox was not affected by treatment with a mixture of deoxycholate and Nonidet P-40, until subjected to SDS-PAGE. Addition of recombinant p67phox to cytosol did not produce any additional p40phox in the immunoprecipitate, unlike the additive increment in the band of p67phox. These results suggest that p40phox forms a complex with p67phox in a molar ratio of 1:1, without any free p40phox in the cytosol.

Cell Membrane↗
PubMed1994

Activation of respiratory burst oxidase is accompanied by desensitization of p47phox in nucleoside-triphosphate binding along with its translocation to cell membrane.

A cytosolic component of human neutrophils, p47phox, potentiates respiratory burst oxidase translocating from cytosol to membrane upon cell stimulation. In this study, the nucleotide-binding ability of p47phox was examined using [32P]GTP dialdehyde (oGTP), [32P]oATP, and [32P]oNADPH. p47phox showed affinities for both oGTP and oATP that were 14 times higher than that for oNADPH, suggesting that it is a nucleoside triphosphate (NTP)-binding protein rather than an NADPH-binding protein. Binding analysis of p47phox using either [32P]oGTP or [32P]oATP revealed an apparent binding constant for each individual NTP analogue and the same maximum binding value, which suggests that both NTPs share a common specific binding site. Stimulation of neutrophils with phorbol myristate acetate (PMA) resulted in enhancement of the oxidase activity to generate O2- anion and was accompanied by substantial translocation of p47phox to membrane. However, p47phox derived from the stimulated cell membrane had lost its NTP-binding ability, unlike that from the resting cytosol. These results suggest that the binding of NTP to p47phox may be involved in the process that activates the oxidase and is desensitized in translocated p47phox.

Adenosine Triphosphate↗