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PubMed2002

In vitro susceptibility of Cryptococcus neoformans serotypes to GM 237354 derivative of the sordarin class.

In vitro susceptibility to the sordarin derivative GM 237354 and amphotericin B were tested in a total of 190 Cryptococcus neoformans clinical isolates from different geographical areas of Spain and South American countries. Minimal inhibitory concentrations (MICs) were obtained using the NCCLS reference microbroth dilution method and analysed according the serotypes of Cr. neoformans. The MICs for amphotericin B were lower than 1.0 microg ml(-1) (MIC90% 0.5 microg ml(-1) , MIC50% 0.125 microg ml(-1)) but five isolates showed MICs of 2.0 microg ml(-1) to GM 237354 (MIC90% 1.0 microg ml(-1), MIC50% 0.5 microg ml(-1)). Cryptococcus neoformans var. gattii serotype B, was significantly less susceptible than A and AD serotypes (P = 0.047 and P = 0.022, respectively).

Amphotericin B↗

PubMed2002

Cryptococcus neoformans myositis without dissemination.

We report a case of isolated cryptococcal myositis involving the paraspinal muscle without evidence of disseminated disease in a patient with a large B-cell lymphoma diagnosed at the time of presentation. Biopsy of the muscle involved grew a pure culture of Cryptococcus neoformans and periodic acid-Schiff staining showed numerous budding yeast consistent with Cryptococcus spp. The patient responded to systemic antifungal therapy with complete resolution of his infection. We also present a review of 5 previously published cases of cryptococcal myositis.

Adult↗

PubMed2003

Cavernicolous habitats harbouring Cryptococcus neoformans: results of a speleological survey in Apulia, Italy, 1999-2000.

Twenty-five caves of speleological and palaeontological interest were investigated for the presence of Cryptococcus spp. within the Apulia region of Italy. Five hundred and forty-five specimens of soil, mud, animal faeces, water and decayed animal and plant remains were examined. Faecal specimens from bats, pigeons and foxes in three caves yielded Cryptococcus neoformans var. neoformans and C. laurentii, C. neoformans var. neoformans and C. albidus were isolated from two soil specimens. Only three caves were positive for the presence of C. neoformans, but the survey documents the finding of a possible natural reservoir in Apulia. It is the first record of occurrence of this yeast in association with cavernicolous habitats, and indicates the potential role of caves in exposing speleologists to life-threatening fungal infections.

Animals↗

PubMed2005

Follow-up study of dogs and cats with asymptomatic Cryptococcus gattii infection or nasal colonization.

The pathogenesis of Cryptococcus spp. infection following nasal colonization is unclear. This article reports follow-up data on a cohort of seven cats and five dogs identified in a previous study as sub-clinically infected with Cryptococcus spp. or colonized by C. gattii. Two cats progressed to clinical disease within four to six months of initial detection of antigenemia and nasal cavity colonization. The ten other animals remained asymptomatic but many were repeatedly positive on cryptococcal antigen testing or nasal fungal culture suggesting protracted infection or colonization. The results indicate that asymptomatically infected animals may clear the organism, remain sub-clinically infected or progress to clinical disease. Factors influencing the transition from exposure to disease require further investigation.

Animals↗

PubMed2002

Cryptococcus neoformans var. gattii meningitis in Singapore.

We describe here two cases of Cryptococcus neoformans var. gattii meningitis that occurred in Singapore. This variety of Cryptococcus neoformans differs from the more widely known C. n. var. neoformans in its ecology and epidemiology. Infections with this variety are still rare in Singapore.

Adult↗

PubMed2002

Fungemia in a cancer patient caused by fluconazole-resistant Cryptococcus laurentii.

We report the recent isolation of Cryptococcus laurentii from the blood of a patient given the diagnosis of ganglioneuroblastoma. The organism was identified using physiological and molecular characteristics, including morphology, carbohydrate and nitrate assimilation, urease activity, inability to form melanin on appropriate media, positive staining with diazonium blue B and sequence analysis of the D1/D2 domain of 26S ribosomal DNA. The isolate was resistant to fluconazole and 5-fluorocytosine using both the Etest and a broth microdilution assay. Repeated recovery of the organism from different blood cultures, and the patient's good response to treatment with amphotericin B support its etiological role. C. laurentii has rarely been implicated as a cause of clinically significant infections. The identity of reported isolates has not always been adequately documented, and some appear to have been isolated from lesions caused by Cryptococcus neoformans, emphasizing the true rarity of disease due to this fungus.

Adolescent↗

PubMed1991

Immunity to a pulmonary Cryptococcus neoformans infection requires both CD4+ and CD8+ T cells.

The role of CD4+ and CD8+ T cells in mediating pulmonary clearance of a cryptococcal infection was investigated. Intratracheal inoculation of BALB/c and C.B-17 mice with a moderately virulent strain of Cryptococcus neoformans (52D) resulted in a pulmonary infection, which was cleared by a T cell-dependent mechanism. During this clearance, there was a significant influx of both CD4+ and CD8+ T cells into the lungs. Depletion of CD4+ T cells by injections of CD4-specific monoclonal antibody (mAb) prevented pulmonary clearance and also resulted in significant colonization of the brain and spleen of infected mice. CD4 depletion did not prevent the influx of CD8+ T cells into the lungs. Surprisingly, depletion of CD8+ T cells by mAb also ablated pulmonary clearance. CD8-depleted mice also had a small but significant increase in brain and spleen colony-forming unit compared to control mice by the end of the study. CD4+ T cell pulmonary influx was independent of the presence of CD8+ T cells. The lungs of T cell-depleted mice were examined histologically. CD4+ and CD8+ T cells each mediated a degree of inflammatory influx seen in the lungs of infected mice and raised the possibility that CD4+ and CD8+ T cells may synergize to generate the inflammatory response in the lungs. Numerous phagocytized but intact cryptococci were seen in the inflammatory foci of CD8-depleted mice but not in control or CD4-depleted mice. We propose that CD4+ T cells may recruit and activate effector phagocytes while CD8+ T cells predominantly function to lyse cryptococcus-laden unactivated phagocytes similar to the function of CD8+ T cells during listeria and mycobacteria infections.

Animals↗

PubMed1992

CD4+ T cells cause multinucleated giant cells to form around Cryptococcus neoformans and confine the yeast within the primary site of infection in the respiratory tract.

The possible mechanisms by which CD4+ T cells prevent the dissemination of Cryptococcus neoformans from the primary site of infection in the respiratory tract were examined. It was found that even before fungicidal mechanisms are fully induced in the lungs, the host generates a CD4+ T cell-dependent inflammatory response that sequesters yeast within the pulmonary alveoli. This confinement is evident histopathologically and demonstrable objectively as a rapid decline in the ability to dislodge yeast from the lungs by bronchopulmonary lavage. One striking component of this response is the enclosure of cryptococci within multinucleated giant cells in granulomas. Studies in severe combined immunodeficient mice that were engrafted with selected lymphocyte subpopulations show that B cells, and hence anti-Cryptococcus antibodies, are not necessary for the CD4+ T cell-dependent responses that isolate and subsequently destroy this opportunistic pathogen in the lung parenchyma.

Animals↗

PubMed2001

Cryptococcus myelitis: atypical presentation of a common infection.

Cryptococcus neoformans is associated with as much as 45% of meningitis in patients admitted for hospital care in Zimbabwe, and it is an important opportunistic infection in patients infected with the human immunodeficiency virus. Cases of cryptococcosis presenting as a spinal cord syndrome have been reported from Zimbabwe and South Africa, but these were all cases of Cryptococcus vertebral osteomyelitis. We describe 3 unusual patients who presented with a myelitis-like syndrome without vertebral osteomyelitis.

AIDS-Related Opportunistic Infections↗

PubMed2005

Prevalence of clinical isolates of Cryptococcus gattii serotype C among patients with AIDS in Sub-Saharan Africa.

Cryptococcus gattii is a group of exogenous, neurotropic yeasts that possess the capsular serotype B or C. Isolates of serotype C are extremely rare and, until recently, were known to infect only immunocompetent individuals. We genotyped 176 isolates of Cryptococcus from patients in sub-Saharan Africa who had AIDS; 22 (13.7%) of 161 isolates from Botswana and 2 (13.3%) of 15 isolates from Malawi were C. gattii serotype C strains. All of these serotype C strains belong to the rare VGIV genotype, possess the MAT alpha mating-type allele, and exhibit little genetic diversity.

Acquired Immunodeficiency Syndrome↗

PubMed1994

Enhancement of HIV type 1 infectivity in vitro by capsular polysaccharide of Cryptococcus neoformans and Haemophilus influenzae.

High concentrations of the cryptococcal capsular polysaccharide (CCP) are present in the serum, cerebrospinal fluid or both in the majority of AIDS patients infected with Cryptococcus neoformans. Because the prognosis of AIDS patients infected with cryptococcus is poor, we investigated whether the presence of CCP enhanced HIV-1 infection. The presence of CCP markedly increased the infectivity of HIV-1-infected H9 cells and subsequent production of infectious HIV-1 and formation of syncytia. In addition to enhancing the infectivity of H9 cells infected with laboratory isolates of HIV-1, the presence of CCP also increased the infectivity of peripheral blood mononuclear cells (PBMCs) infected with primary field strains of HIV-1. The in vitro infectivity of PBMCs from 20 of 44 HIV-1-infected individuals was significantly increased when cultured with CCP. Furthermore, HIV-1 was isolated from the PBMCs of three of these individuals only when cultured in the presence of CCP. CCP increased the binding of HIV-1 and recombinant gp120 to H9 cells and recombinant CD4, respectively. Thus, it is possible that the enhancement of HIV-1 infectivity by CCP is due to its capacity to increase the adherence of HIV-1 to target cells. Whereas the capsular polysaccharide of Haemophilus influenzae also markedly enhanced the infectivity of HIV-1, the capsular polysaccharides of C. freundii or S. flexneri had minimal effects on the infectivity of HIV-1. This indicated that the capacity to enhance HIV-1 infectivity was a property of polysaccharides from some pathogens and not others.(ABSTRACT TRUNCATED AT 250 WORDS)

AIDS-Related Opportunistic Infections↗

PubMedDate not supplied

The ecology of Cryptococcus neoformans and the epidemiology of cryptococcosis.

The ecology of Cryptococcus neoformans and the epidemiology of cryptococcosis are reviewed. Two varieties of C. neoformans have been recognized. C. neoformans variety neoformans has been found in nature worldwide, primarily in association with bird droppings, although nonavian sources have also been found. Most cases of human cryptococcosis are caused by this variety. C. neoformans var. gattii has recently been isolated in nature in association with Eucalyptus trees. Infections caused by this variety occur mainly in tropical and subtropical regions. Because exposure to C. neoformans is probably common and clinically apparent cases of cryptococcosis in healthy hosts are rare, it is presumed that most people can mount adequate host defenses upon exposure to the organism. At least 5%-10% of patients with AIDS become infected with Cryptococcus; the epidemiology of this infection is different in many respects from that seen in patients without AIDS.

Acquired Immunodeficiency Syndrome↗

PubMed2000

Identification and characterization of a highly conserved calcineurin binding protein, CBP1/calcipressin, in Cryptococcus neoformans.

Calcineurin is the conserved target of the immunosuppressants cyclosporin A and FK506. Using the yeast two-hybrid system, we identified a novel calcineurin binding protein, CBP1, from the pathogenic fungus Cryptococcus neoformans. We show that CBP1 binds to calcineurin in vitro and in vivo, and FKBP12-FK506 inhibits CBP1 binding to calcineurin. Cryptococcus neoformans cbp1 mutant strains exhibit modest defects in growth under stress conditions and virulence, similar to but less severe than the phenotypes of calcineurin mutants. Saccharomyces cerevisiae mutants lacking the CBP1 homolog RCN1 are, like calcineurin mutants, sensitive to lithium cation stress. CBP1 shares a central peptide sequence motif, SPPxSPP, with related proteins in S.CEREVISIAE:, Schizosaccharomyces pombe, Drosophila melanogaster, Caenorhabditis elegans and humans, and peptides containing this motif altered calcineurin activity in vitro. Interestingly, the human CBP1 homolog DSCR1 is encoded by the Down's syndrome candidate region interval on chromosome 21, is highly expressed in the heart and central nervous system, and may play a role in calcineurin functions in heart development, neurite extension and memory.

Amino Acid Motifs↗

PubMed2001

Identification of virulence mutants of the fungal pathogen Cryptococcus neoformans using signature-tagged mutagenesis.

Cryptococcus neoformans var. neoformans is an important opportunistic fungal pathogen of patients whose immune system has been compromised due to viral infection, antineoplastic chemotherapy, or tissue transplantation. As many as 13% of all AIDS patients suffer a life-threatening cryptococcal infection at some time during the course of their HIV disease. To begin to understand the molecular basis for virulence in Cryptococcus neoformans var. neoformans serotype A, we have employed signature-tagged mutagenesis (STM) to identify mutants with altered virulence in a mouse model. The critical parameters of signature-tagged mutagenesis in C. neoformans are explored. Data are presented showing that at least 100 different strains can be mixed together in a single animal with each participating in the infection and that there is no apparent interaction between a virulent strain and an avirulent strain in our animal model. Using signature-tagged mutagenesis, we identified 39 mutants with significantly altered growth in a competitive assay. Molecular analyses of these mutants indicated that 19 (49%) contained an insertion in the actin promoter by homologous recombination from a single crossover event, creating a duplication of the actin promoter and the integration of single or multiple copies of the vector. Analysis of the chromosomal insertion sites of those mutants that did not have an integration event in the actin promoter revealed an approximately random distribution among the chromosomes. Individual challenge of the putative mutants in a mouse model revealed five hypovirulent mutants and one hypervirulent mutant.

Actins↗

PubMed1999

In-vitro activity of dicationic aromatic compounds and fluconazole against Cryptococcus neoformans and Candida spp.

We investigated the in-vitro activity of three selected dicationic aromatic compounds for nine clinical isolates of Cryptococcus neoformans and 93 clinical isolates of Candida spp., representing 12 different species, using a broth macrodilution method following NCCLS recommendations. All the clinical isolates were also tested for fluconazole susceptibility. The in-vitro data demonstrate that compounds 39 and 57 have excellent in-vitro activity for all tested strains (MIC 0.19-1.56 mg/L) except Candida pelliculosa. Moreover, compound 39 showed excellent in-vitro fungicidal activity against Candida krusei, Candida glabrata, Candida lusitaniae and Cryptococcus neoformans with MFCs in the range 0.39-6.25 mg/L. Both compounds 39 and 57 showed excellent in-vitro activity against fluconazole-resistant Candida albicans isolates, including a C. albicans strain that contains all known fluconazole-resistant mechanisms. Comparing MIC data from compounds 21, 39 and 57 with fluconazole, we found a statistically significant difference only with compound 39 (P = 0.043). However, comparing MFC data from compounds 21, 39 and 57 with fluconazole, we found statistically significant differences with all three compounds (P < 0.00001). These data indicate the potential antifungal breadth of two bis-benzimidazoles (compounds 39 and 57) as antifungal agents against yeasts. If it can be determined that compounds 39 and 57 are effective and non-toxic in vivo, the prospect of these compounds as clinically useful antifungal agents will be enhanced.

Antifungal Agents↗

PubMed2002

Cryptococcus nyarrowii sp. nov., a basidiomycetous yeast from Antarctica.

In December 1997, 196 soil and snow samples were collected from Vestvold Hills, Davis Base, Antarctica. Two isolates, CBS 8804T (pink colonies) and CBS 8805 (yellow colonies), were shown by proteome analysis and DNA sequencing to represent the same species. Results from the sequencing of the D1/D2 region of the large rDNA subunit placed this species in the hymenomycetous tree in a unique sister clade to the Trichosporonales and the Tremellales. The clade consists of Holtermannia corniformis CBS 6979 and CBS strains 8804T, 8805, 8016, 7712, 7713 and 7743. Morphological and physiological characteristics placed this species in the genus Cryptococcus, with characteristics including the assimilation of D-glucuronate and myo-inositol, no fermentation, positive Diazonium blue B and urease reactions, absence of sexual reproduction and production of starch-like compounds. Fatty acid analysis identified large proportions of polyunsaturated lipids, mainly linoleic (C18:2) and, to a lesser extent, linolenic (C18:3) acids. On the basis of the physiological and phylogenetic data, isolates CBS 8804T and CBS 8805 are described as Cryptococcus nyarrowii sp. nov.

Antarctic Regions↗

PubMed1988

Taxonomic relationships of Cryptococcus and Tremella based on fatty acid composition and other phenotypic characters.

The cellular long-chain fatty acids present in 33 strains, representing 15 species of Cryptococcus, and 4 species of Tremella, were determined by gas chromatography. According to the relative amounts of fatty acid methyl esters, the Cryptococcus species studied were divided into four main groups. Possible relationships between species representing the two genera are presented in a new model, where cellular long-chain fatty acid compositions and other phenotypic characteristics are included.

Chromatography, Gas↗

PubMed1994

Comparison of the electrophoretic karyotypes and chromosomal location of ten genes in the two varieties of Cryptococcus neoformans.

We compared multiple isolates of the two varieties of Cryptococcus neoformans, as well as previously characterized representative isolates, for their electrophoretic karyotypes using pulsed-field electrophoresis. The two varieties could be clearly distinguished based upon the size of the smallest chromosome. The smallest chromosome for isolates of the gattii variety (serotypes B and C) was found to be 400-700 kb in size. The smallest chromosome for isolates of the neoformans variety was consistently found to be larger, approximately 770 kb in size. Isolates of the gattii variety averaged 13 chromosomes while the neoformans variety averaged 12. The size of the Cryptococcus genome was found to be approximately 23 megabases. Isolates of C. neoformans var. neoformans tended to be more conserved than those of var. gattii with regard to gene position.

Chromosomes, Fungal↗