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1.
Cryptococcus neoformans was instilled intranasally into mice which were periodically sacrificed to determine the course of infection. Cryptococci persisted within the nasal passages throughout the 90 day study. Extranasal dissemination began 14–28 days after instillation and was still demonstrable 90 days post-exposure. Ten percent mortality was observed in mice receiving 106 cryptococci, while no mortality was observed in mice exposed to 103 or 104 cryptococci. Our research suggests that nasal colonization with C. neoformans can precede pulmonary and systemic cryptococcosis by weeks or months. 相似文献
2.
The histopathology of murine cryptococcosis was observed until the 55th day and particular attention was paid to whether or not cysts, which had been formed in the brain, could change to granulomas. Cryptococcus neoformans RIB-12M was used in this experiment. As experimental animals, five-week-old male BALB/c mice, weighing 20–22 g, were used. An infective inoculum was prepared by adjusting the number of cryptococci to 106 or 5 × 106/0.2 ml. Each mouse was inoculated intravenously with 0.2 ml of the cell suspension, and the colony forming unit of the brain and liver, and the histopathological findings in various visceral organs were investigated.40 × 104 colonies grew from 100 mg of the brain tissue of the eighth day. Thereafter, the number increased gradually. It reached 500 × 104 on the 20th day. The colony forming unit from the liver reached a peak on the 12th day (250 × 104) and thereafter the number decreased gradually.Histopathologically, the brain and liver were severely affected with the fungus. In the brain cysts with cryptococci continued to increase until the end of the experiment. On the other hand, in the liver several purulent foci appeared on the second day. On the eighth day numerous mononuclear cells accumulated at the foci and their lesions changed to granulomatous ones with cryptococci. The number of granulomatous lesions reached a peak on the 16th day in the mice inoculated with 5 × 106 cryptococci, and thereafter showed a tendency to decrease gradually. 相似文献
3.
Cryptococcus laurentii is one of the non-neoformans cryptococci that have rarely been isolated from humans. We report a case of repeated colonization of the oropharynx by Cr. laurentii in a patient with erythroleukaemia. The isolate was identified by phenotypic and genotypic tests and showed resistance to
fluconazole.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
4.
Phagocytosis by cells of the innate immune system, such as macrophages, and the subsequent successful maturation of the phagosome, is key for the clearance of pathogens. The fungal pathogen Cryptococcus neoformans is known to overcome killing by host phagocytes and both replicate within these cells and also escape via a non‐lytic process termed vomocytosis. Here we demonstrate that, during intracellular growth, cryptococci modify phagolysosome maturation. Live cryptococci, but not heat‐killed pathogens or inert targets, induce the premature removal of the early phagosome markers Rab5 and Rab11. In addition, significant acidification of the phagosome, calcium flux and protease activity is hindered, thus rendering the phagosome permissive for cryptococcal proliferation. Interestingly, several attenuated cryptococcal mutants retain this ability to subvert phagosomal maturation, suggesting that hitherto unidentified pathogen mechanisms regulate this process. 相似文献
5.
Brendan J. McMullan Desmarini Desmarini Julianne T. Djordjevic Sharon C-A. Chen Michael Roper Tania C. Sorrell 《PloS one》2015,10(1)
Background
Cryptococcus neoformans is the commonest cause of fungal meningitis, with a substantial mortality despite appropriate therapy. Quantitative culture of cryptococci in cerebrospinal fluid (CSF) during antifungal therapy is of prognostic value and has therapeutic implications, but is slow and not practicable in many resource-poor countries.Methods
We piloted two rapid techniques for quantifying viable cryptococci using mixtures of live and heat-killed cryptococci cultured in vitro: (i) quantitative microscopy with exclusion staining using trypan blue dye, and (ii) flow cytometry, using the fluorescent dye 2′-7′-Bis-(2-carboxyethyl)-5-(6)-carboxyfluorescein, acetoxymethyl ester (BCECF-AM). Results were compared with standard quantitative cryptococcal cultures. Quantitative microscopy was also performed on cerebrospinal fluid (CSF) samples.Results
Both microscopy and flow cytometry distinguished between viable and non-viable cryptococci. Cell counting (on log scale) by microscopy and by quantitative culture were significantly linearly associated (p<0.0001) and Bland-Altman analysis showed a high level of agreement. Proportions of viable cells (on logit scale), as detected by flow cytometry were significantly linearly associated with proportions detected by microscopy (p<0.0001) and Bland-Altman analysis showed a high level of agreement.Conclusions
Direct microscopic examination of trypan blue-stained cryptococci and flow-cytometric assessment of BCECF-AM-stained cryptococci were in good agreement with quantitative cultures. These are promising strategies for rapid determination of the viability of cryptococci, and should be investigated in clinical practice. 相似文献6.
Our previous reports have shown that animals inoculated for 8 weeks with 1×105 pseudohyphal, avirulentC. neoformans exhibited prolonged survival upon challenge with virulent cryptococci. This paper described a transient phase of immunosuppression which occurs during the initial weeks of the immunization protocol. Animals injected with pseudohyphal organisms had depressed responses to T and B-cell mitogens. In addition, they had lowered responses to immunization with sheep erythrocytes. Both humoral and cell mediated responses were affected. 相似文献
7.
The ultrastructure of Cryptococcus neoformans in the cerebrospinal fluid, which was obtained from a patient suffering from systemic lupus erythematosus and cryptococcal meningitis, before treatment and on the 10th and 20th day after the start of treatment was studied. Numerous cryptococci were detected in the cerebrospinal fluid before treatment. However, in most of them their biological activity was low and their organelles were not clear. A few yeasts preserved well their organelles. Such cells showed a tendency to develop vesicular membrane structures (lomasomes) touching the internal part of the cell wall. In the cerebrospinal fluid on the 20th day all of the yeasts were dead, even though numerous yeasts were observed by an India-ink method. At this time no colonies were recovered from the fluid. 相似文献
8.
Previous reports have demonstrated that natural killer (NK) cells are capable of inhibiting the growth of Cryptococcus neoformans in vitro, and recent studies indicate that adoptively transferred NK cell-enriched spleen cell populations enhance clearance of cryptococci from the tissues of cyclophosphamide-pretreated recipients. The primary objective of these studies was to confirm that NK cells participate in early clearance of C. neoformans in vivo. Secondarily, the anti-cryptococcal activities of polymorphonuclear leukocytes and macrophages were examined. Seven-week-old C57BL/6 bg/+ mice, which have normal levels of NK cell activity, were compared with their bg/bg littermates, which have impaired NK cell function. One and 3 days after injecting both groups of mice i.v. with 2 X 10(4) cryptococci, we assessed the NK cell activities in spleens, lungs, and livers and clearance of the organism from corresponding tissues as determined by the mean log10 numbers of cryptococcal colony-forming units (CFU) per organ. Three days postinfection, the mean numbers of cryptococcal CFU in lungs and spleens of bg/+ mice were significantly lower than in the corresponding organs of bg/bg mice. NK cell activities in spleens and lungs of bg/+ mice were significantly higher than were the NK cell activities in similar cell populations from bg/bg mice. In contrast, the mean numbers of cryptococcal CFU in livers of the two groups of animals were nearly equivalent, a situation not unexpected, since liver NK cell activities were extremely low and similar in both groups of animals. Although these data indicated a correlation between early clearance of cryptococci from tissues and levels of NK cell activities in the corresponding tissues, it was also possible that differences in phagocytic cell function between the bg/+ and bg/bg animals could account for the observed differences in clearance of cryptococci from the tissues. Therefore, phagocytic cells from the two groups of animals were compared with respect to their abilities to phagocytize and inhibit the growth of cryptococci and to their abilities to respond to chemotactic stimuli in vivo. Peritoneal PMNL from bg/+ and bg/bg mice were similar in their abilities to phagocytize and inhibit the growth of cryptococci, as well as in their chemotactic responses to viable cryptococci or sodium caseinate. In addition, there were no differences in splenic macrophage functions between the two groups of mice.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
9.
Macrophage-mediated fungistasis in vitro: requirements for intracellular and extracellular cytotoxicity 总被引:25,自引:0,他引:25
D L Granger J R Perfect D T Durack 《Journal of immunology (Baltimore, Md. : 1950)》1986,136(2):672-680
Macrophage cytotoxicity for Cryptococcus neoformans was investigated by culturing mouse peritoneal macrophages with a thin-capsuled clone of cryptococcus under conditions permitting efficient phagocytosis. Yeast replication was quantitated by electronic particle counting after detergent lysis of macrophages, and viability was determined by quantitative plate counts. Under appropriate conditions, reproduction was completely inhibited; stasis began at 2 hr after addition of yeasts and lasted for 30 hr. During this time organisms in medium alone proliferated rapidly, doubling their number every 2.5 hr. After removal from macrophages, 60 to 100% of macrophage-inhibited cryptococci formed colonies, indicating that the cytotoxic effect was primarily fungistatic. When yeast cells were removed from macrophages, replication recommenced within 5 hr. Supernatant medium from fungistatic co-cultures was not inhibitory for fresh yeast cells. Conditions required for complete fungistasis were 1) peritoneal macrophages induced by peptone from BCG-infected mice, 2) endotoxin in nanogram per milliliter range added to serum-containing cell culture medium, 3) confluent macrophage monolayers, and 4) macrophage:cryptococci ratios of 20 to 100:1. Fungistasis occurred without phagocytosis but was more efficient when cryptococci were engulfed. For efficient fungistasis, macrophages must differentiate to and be maintained in the activated state. These results with yeast cells agree with the known requirements for macrophage effector function against neoplastic target cells. 相似文献
10.
Takaki Fukuizumi Hiromasa Inoue Yuichi Anzai Toshiyuki Tsujisawa Choji Uchiyama 《Microbiology and immunology》1995,39(5):351-359
We have shown that the palatine tonsil effectively incorporates exogenous foreign substances instilled at its surface. It is not clear whether antigen-specific IgA can be induced by the instillation. Sheep red blood cells (SRBC) were instilled at the palatine tonsil every three days as the antigen, and the agglutination titer of specific IgA in saliva was examined. Nasal or intragastric administration, which have been shown to induce specific antibody in saliva, were done as control experiments. Anti-SRBC antibody in saliva from the tonsillar instillation group was detected in the second week, and the agglutination titer reached a maximum in the 6th week after the instillation. The maximum titers in the tonsillar instillation group and nasal administration group were 16 (P<0.01, n=7) and 4 times (P<0.01, n = 7) higher, respectively, than that in the intragastric administration group. In the tonsillar instillation group, the number of specific antibody-producing cells per 105 lymphocytes was the highest in the parotid glands compared with the lymphoid tissues such as the retropharyngeal lymph nodes, nasal mucosa, mesenteric lymph nodes, Peyer's patches, cervical lymph nodes, palatine tonsil and spleen. In the nasal administration group, the number of lymphocytes was the highest in the nasal mucosa. The results indicate that tonsillar instillation was more effective than nasal administration in inducing specific IgA in saliva. 相似文献
11.
O. V. Isaeva A. M. Glushakova S. A. Garbuz A. V. Kachalkin I. Yu. Chernov 《Biology Bulletin》2010,37(1):26-34
It was found that plant storage tissues (fleshy sugar-containing fruits, subsurface metamorphically altered plant organs (storage
roots, tubers, etc.), and starch-containing seed lobes) nearly always contain yeasts that are able to actively reproduce in
these tissues causing no visible damage. Within storage tissues, yeast cells were detected both in the intercellular space
and inside plant cells. In the tissues of fleshy fruits, endophytic yeasts are represented by the same species as epiphytic
ones; cryptococci of the order Filobasidiales and ascomycetes belonging to the genera Hanseniaspora and Metschnikowia are predominant. In subsurface plant organs, red pigmented basidiomycetous yeasts of the genus Rhodotorula prevail. Selective growth of representatives of one species, Candida railenensis, is typical of starch-containing storage tissues of seeds. The results obtained change the established notion of the distributional
patterns of yeast fungi in natural habitats and suggest that internal storage tissues of plants can be considered as a new
interesting model for studies of coevolving plant-microbial associations. 相似文献
12.
Background
The aim of the present study was to assess possible health effects of airway exposures to Bacillus thuringiensis (Bt) based biopesticides in mice. Endpoints were lung inflammation evaluated by presence of inflammatory cells in bronchoalveolar lavage fluid (BALF), clearance of bacteria from the lung lumen and histological alterations of the lungs. Hazard identifications of the biopesticides were carried out using intratracheal (i.t.) instillation, followed by an inhalation study. The two commercial biopesticides used were based on the Bt. subspecies kurstaki and israelensis, respectively. Groups of BALB/c mice were i.t instilled with one bolus (3.5 × 105 or 3.4 × 106 colony forming units (CFU) per mouse) of either biopesticide. Control mice were instilled with sterile water. BALFs were collected and the inflammatory cells were counted and differentiated. The BALFs were also subjected to CFU counts. 相似文献13.
Marie Louise Grndahl Martin Andreas Unmack Helga Berglind Ragnarsdttir Mark Berner Hansen John Elmerdahl Olsen Erik Skadhauge 《Comparative biochemistry and physiology. Part A, Molecular & integrative physiology》2005,141(4):476
The effects of nitric oxide (NO) in the secretory response to the endogenous secretagogue 5-hydroxytryptamine (5-HT), the enterotoxins heat-labile enterotoxigenic Escherichia coli (ETEC) toxin (LT) and cholera toxin (CT), and various cultures of ETEC and Salmonella serotype Typhimurium in the porcine small intestine (Sus scrofa) were investigated. In anaesthetized pigs, jejunal tied-off loops were instilled with 5-HT, LT, CT, various cultures of ETEC or S. Typhimurium. Pigs were given intravenously isotonic saline or isotonic saline containing the NO synthase inhibitor, Nω-nitro-l-arginine methyl ester (L-NAME). L-NAME significantly induced an increased fluid accumulation in loops induced by 5-HT, ETEC and stn-mutated S. Typhimurium. Fluid accumulation in loops instilled with wild-type S. Typhimurium was increased by L-NAME, although not significantly, while there was no effect on fluid accumulation induced by an invH-mutated isogenic strain. No significant effect of L-NAME was observed on the fluid accumulation induced by the purified enterotoxins LT and CT. The results also demonstrated a relatively large difference in the ability to induce fluid accumulation between the bacteria strains. Diastolic, systolic and mean blood pressures were significantly increased and the body temperature was significantly decreased in groups of pigs treated with L-NAME. In conclusion, the results suggest that NO has a proabsorptive effect in the intact porcine jejunum and is involved in the systemic vascular tone. 相似文献
14.
Germination,Viability and Clearance of Stachybotrys chartarum in the Lungs of Infant Rats 总被引:1,自引:0,他引:1
Observing that the conidia of Stachybotrys chartarum can germinate in the lung of infant rats, it became important to ascertain whether an infection can ensue. Viable conidia of S. chartarum were instilled into the lungs of 4 and 14 day-old rat pups. Germination was observed frequently in the lungs of 4 day-old but rarely in the 14 day-old pups. In the 4 day-old pups, pulmonary inflammation with hemorrhagic exudates was observed and resulted in about 15% mortality rate compared to 0% for the controls instilled with phosphate buffered saline. Acute neutrophilic inflammation and intense interstitial pneumonia with poorly formed granulomas observed three days following exposure were associated with fungal hyphae and conidia. The surviving experimental pups showed significantly slower weight gain for seven days. Dilution plating and quantitative PCR analysis were used to follow total fungal load in the rat pups lung homogenates. In the 4 day-old rat pups viable fungi decreased rapidly and were less than 1% by day seven. Similarly, fungal DNA decreased exponentially and was only 0.03% by fourteen days after exposure. However, 14 day-old rat pups showed neither the lethal effects of exposures to viable conidia of S. chartarum nor the slower weight gain, and the fungal load decreased even more rapidly. We conclude that S. chartarum conidia can initially germinate and form hyphae but even in the immature rat pups do not establish an effective infection, although a very limited persistence cannot be excluded.This revised version was published online in October 2005 with corrections to the Cover Date. 相似文献
15.
J. S. Watkins M. J. Campbell D. Gardner-Medwin H. R. Ingham I. G. Murray 《BMJ (Clinical research ed.)》1969,3(5661):29-31
Cryptococcal meningitis occurred in two patients in the North of England. One, an elderly woman who was ill for 12 months with an obscure indolent meningitis until use of steroid drugs resulted in an acute exacerbation during which cryptococci were isolated, was treated successfully with amphotericin B. The other, a student from New Guinea with subacute meningitis, could not tolerate amphotericin B but responded to two courses of oral treatment with 5-fluorocytosine 100–200 mg./kg./ day without any appreciable side-effects. Further trials of this drug in the treatment of cryptococcosis are recommended. 相似文献
16.
Susceptibility of congenitally immunodeficient mice to a nonencapsulated strain of Cryptococcus neoformans. 总被引:2,自引:0,他引:2
The susceptibility of congenitally immunodeficient mice to a nonencapsulated strain of Cryptococcus neoformans (strain M7) was evaluated. Gnotobiotic mice with defined congenital defects in innate immunity (beige) or cell-mediated immunity (athymic) or with combined defects in innate and cellular immunity (beige athymic) were i.v. challenged with C. neoformans M7. The nonencapsulated strain of C. neoformans produced a persistent low-grade infection in the brains of all immunodeficient and immunocompetent mice used in this study. Immunocompetent mice (nu/+;bg/+) and immunodeficient bg/bg mice readily cleared nonencapsulated cryptococci from their kidneys, liver, lungs, and spleen. In contrast to nu/+ mice, nu/nu mice had a reduced capacity to clear nonencapsulated cryptococci from their kidneys and liver after i.v. challenge. Both bg/bg-nu/nu and bg/bg-nu/+ mice developed a low-grade infection in their kidneys, liver, lungs, and spleen, which was maintained throughout the 21-day study. Persistent infections were not due to reversion to an encapsulated state. These data indicate that a capsule may not always be necessary for C. neoformans to survive, in vivo, in tissues of immunodeficient and immunocompetent mice. 相似文献
17.
Our previous reports have shown that animals inoculated for 8 weeks with 1 × 105 pseudohyphal, avirulentC. neoformans exhibited prolonged survival upon challenge with virulent cryptococci. This paper described a transient phase of immunosuppression which occurs during the initial weeks of the immunization protocol. Animals injected with pseudohyphal organisms had depressed responses to T and B-cell mitogens. In addition, they had lowered responses to immunization with sheep erythrocytes. Both humoral and cell mediated responses were affected. 相似文献
18.
Stachylysin is a proteinaceous hemolytic agent that is produced by Stachybotrys chartarum. Stachylysin was found, using immunohistochemical and immunocytochemical methods, to be localized in S. chartarum spores/mycelia primarily in the inner wall suggesting that it is constitutively produced. Spores instilled in mouse or rat lung tissues resulted in granuloma formation, which showed the highest stachylysin concentration in the inner wall of the spore and near the spore, with less at distance indicating that it had diffused out from the spore. The in vitro high stachylysin producing strain (58-06) was also highest in vivo, based on immunohistochemistical staining. More stachylysin was observed in the mouse lung tissue at 72 h than at 24 h indicating that production/release is a relatively slow process. The localization of stachylysin in macrophage phagolysosomes suggests that these cells may be involved with hemolysin inactivation. This would be consistent with what is known about asp-hemolysin produced by Aspergillus fumigatus.This revised version was published online in October 2005 with corrections to the Cover Date. 相似文献
19.
A 24-year-old woman was found to have cryptococcal meningitis and Cushing''s syndrome due to an adrenal adenoma. Her meningitis was successfully arrested with fluorouracil. Treatment with metyrapone decreased her cortisol production and produced clinical remission of Cushing''s syndrome. On admission her peripheral T lymphocytes were few and hyporeactive. When the overproduction of cortisol ceased the numbers of T lymphocytes and their reactivity returned to normal and she developed in-vitro lymphocyte responsiveness to the cryptococci. 相似文献
20.
A central nervous system isolate from an acquired immunodeficiency syndrome (AIDS) patient of 103
Cryptococcus neoformans cells was instilled intranasally into guinea pigs. These were killed to evaluate the fate of the organisms and the response of the olfactory mucosa. Olfactory epithelium prevented the penetration ofCryptococcus neoformans and showed focal hyperplastic responses. The organisms, which manifested an affinity for the olfactory portion of the nasal cavities, were cleared from the olfactory space with no intervention from the immune system cells. By the end of the fifth week almost no organisms could be found and there was no histological evidence of dissemination. In contrast, destruction of the olfactory epithelium with zinc sulfate allowed the invasion of the subepithelial tissues, demonstrating the role of the olfactory mucosa in preventing infection withCryptococcus neoformans through the nasal route. The results and the model described in this report may be useful in clarifying the pathogenic mechanisms of cryptococcosis and the non immune mediated host responses toCryptococcus neoformans. 相似文献