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Lei Peng-Cheng Takashi Yoshiike Hitoshi Yaguchi Hideoki Ogawa MD PhD 《Mycopathologia》1993,122(2):89-93
Defense mechanisms againstSporothrix schenckii were studied using mouse models. After an intracutaneous injection of the yeast form ofS. schenckii to the dorsal skin of the congenitally athymic nude and normal heterozygote littermate mice, nodules were formed. They regressed and disappeared in 10 weeks in the case of normal mice. On the other hand, nodules and then ulceration developed progressively in nude mice until all animals expired by dissemination of microorganisms at the 11th week of inoculation. Histopathologically the migrated cells were similar in both the normal and the nude mice, particularly during the early phase (within 24 h), with infiltration by PMNs being predominant. Fragmentation ofS. schenckii commenced early during the 12–24 h stage of inoculation in the normal mice, while such fragmentation was scarce in nude mice even though numerous PMNs accumulated. Microscopic observations in the early stages (within 24 h of inoculation) suggested that the lack of killing activity by PMNs in nude mice contributes more to the impaired defense than the lack of macrophage activation by T-cells. 相似文献
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Takashi Yoshiike Peng-Cheng Lei Hisano Komatsuzaki Hideoki Ogawa MD PhD 《Mycopathologia》1993,123(2):69-73
Sporothrix schenckii produces two extracellular proteinases, namely proteinase I and II. Proteinase I is a serine proteinase, inhibited by chymostatin, while proteinase II is an aspartic proteinase, inhibited by pepstatin. Studies on substrate specificity and the effect of proteinase inhibitors on cell growth suggest an important role for these proteinases in terms of fungal invasion and growth. There has, however, been no evidence presented demonstrating thatS. schenckii produces 2 extracellular proteinases in vivo. In order to substantiate the in vivo production of proteinases and to attempt a preliminary serodiagnosis of sporotrichosis, serum antibodies against 2 proteinases were assayed usingS. schenckii inoculated hairless mice. Subsequent to an intracutaneous injection ofS. schenckii to the mouse skin, nodules spontaneously formed and disappeared for a period of 4 weeks. Histopathological examination results were in accordance with the microscopic observations. Micro-organisms disappeared during the fourth week. Serum antibody titers against purified proteinases I and II were measured weekly, using enzyme-linked immunosorbent assay (EIA). As a result, the time course of the antibody titers to both proteinases I and II were parallel to that of macroscopic and microscopic observations in an experimental mouse sporotrichosis model. These results suggest thatS. schenckii produces both proteinases I and II in vivo. Moreover, the detection of antibodies against these proteinases can contribute to a serodiagnosis of sporotrichosis. 相似文献
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Karyotyping by PFGE of clinical isolates of Sporothrix schenckii 总被引:3,自引:0,他引:3
Takeshi Tateishi Somay Yamagata Murayama Fujio Otsuka Hideyo Yamaguchi 《FEMS immunology and medical microbiology》1996,13(2):147-154
Abstract From October 1991 to December 1992 we had eight patients with sporotrichosis at Tsukuba University Hospital in Japan. With 8 strains isolated from these patients, PFGE (pulsed-field gel electrophoresis) analyses were carried out to examine whether the karyotype of S. schenckii is distinguished by our method and whether this molecular approach is a useful means of biotyping of S. schenckii strains. Chromosomes were separated by contour-clamped homogeneous electric field (CHEF) gel electrophoresis. The strains had six to eight chromosomes and a total genome size was approx. 28 Mbp. Although these karyotypes of all the isolates looked closely similar to each other, they were grouped into three types. 相似文献
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In an attempt to elucidate the effects of Sporothrix schenckii infection on the immune response, our laboratory has developed a murine model of disseminated sporotrichosis. Helper T cells
can be further subdivided into Th1 and Th2 phenotypes. The differentiation of two subsets of T lymphocytes is driven by IL-12
and IL-4 cytokines, respectively. Th1 cells produce IFN-γ that activate macrophages and promote cell-mediated immunity. In
addition, we found low levels of iNOS and NO production in the initial (1st and 2nd weeks) and final (9th and 10th weeks)
periods of the infection, in contrast with the period of week 4 to 7 of elevated values. The determination of IFN-γ and IL-12
are in agreement with NO/iNOS detection, showing the presence of cellular immune response throughout the infectious process.
However, the production of IL-4 shows an increase in levels after the 5th and 6th weeks suggesting a participation of Th2
response in this period as well. Regarding these results, the study demonstrated that in experimental sporotrichosis infection
the cellular immune response participated throughout the period analyzed as a nitric oxide dependent mechanism. In contrast,
the presence of Th2 response began in the 5th week, suggesting the participation of humoral immune response in advanced stages
of sporotrichosis. 相似文献
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Mitochondrial DNA(mtDNA) diversity was investigated in 257 clinical isolants of Sporothrix schenckii obtained from 4 districts in Japan. S. schenckii was classified into 10 types based on Hae III restriction profiles. Phylogeny of types constructed by the method of Fitch and Margoliash [1] on the estimated sequence divergence within mtDNA using the methods of Nei and Li [2], showed that S. schenckii are grouped into 2 clusters, one group consisting of types 1, 2 and 3, and the other group consisting of the other seven types. In addition, types 1, 2, and 5 were correlated with their geographic origin, whereas type 4 was present throughout Japan. 相似文献
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Protein kinase C (PKC) plays an important role in the control of proliferation and differentiation of a wide range of cell
types, and fungi are no exception. Previous results reported by us on the effects of the phorbol ester, 12-myristate-13-acetate
phorbol (PMA) and other PKC effector molecules, on dimorphism in Sporothrix schenckii suggested the presence of this enzyme
in the fungus and its involvement in the control of morphogenetic transitions. The work summarized here confirms the presence
of PKC in yeast and mycelium extracts of S. schenckii. Different isoforms of this enzyme were found to be present in the yeast
and mycelium forms of the fungus and were identified by Western blot analysis using affinity purified anti-PKC isoforms specific
antibodies: the γ and ζ isoforms were detected in both the yeast and mycelium forms of the fungus, while the β isoform was
only detected in the yeast form. The presence of PKC was confirmed biochemically by measuring total enzyme activity in both
forms of the fungus. No significant differences were observed for the PKC activity level recorded for both the mycelium and
yeast forms of the fungus (p ≤ 0.05). These data confirm the presence of PKC activity in Sporothrix schenckii and constitutes
the first evidence concerning the differential expression of PKC isoforms in the mycelium and yeast forms of a dimorphic fungus,
supporting the possible involvement of this important signal transduction enzyme in the control of morphogenesis in this fungus.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
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A dextranase (EC 3.2.1.11) was purified and characterized from the IP-29 strain of Sporothrix schenckii, a dimorphic pathogenic fungus. Growing cells secreted the enzyme into a standard culture medium (20 °C) that supports the
mycelial phase. Soluble bacterial dextrans substituted for glucose as substrate with a small decrease in cellular yield but
a tenfold increase in the production of dextranase. This enzyme is a monomeric protein with a molecular mass of 79 kDa, a
pH optimum of 5.0, and an action pattern against a soluble 170-kDa bacterial dextran that leads to a final mixture of glucose
(38%), isomaltose (38%), and branched oligosaccharides (24%). In the presence of 200 mM sodium acetate buffer (pH 5.0), the
K
m for soluble dextran was 0.067 ± 0.003% (w/v). Salts of Hg2+, (UO2)2+, Pb2+, Cu2+, and Zn2+ inhibited by affecting both V
max and K
m. The enzyme was most stable between pH values of 4.50 and 4.75, where the half-life at 55 °C was 18 min and the energy of
activation for heat denaturation was 99 kcal/mol. S. schenckii dextranase catalyzed the degradation of cross-linked dextran chains in Sephadex G-50 to G-200, and the latter was a good
substrate for cell growth at 20 °C. Highly cross-linked grades (i.e., G-10 and G-25) were refractory to hydrolysis. Most strains
of S. schenckii from Europe and North America tested positive for dextranase when grown at 20 °C. All of these isolates grew on glucose at
35 °C, a condition that is typically associated with the yeast phase, but they did not express dextranase and were incapable
of using dextran as a carbon source at the higher temperature.
Received: 29 December 1997 / Accepted: 4 March 1998 相似文献