scholarly journals Cryptococcosis Serotypes Impact Outcome and Provide Evidence of Cryptococcus neoformans Speciation

mBio ◽  
2015 ◽  
Vol 6 (3) ◽  
Author(s):  
Marie Desnos-Ollivier ◽  
Sweta Patel ◽  
Dorothée Raoux-Barbot ◽  
Joseph Heitman ◽  
Françoise Dromer ◽  
...  
Keyword(s):  
Cryptococcus Neoformans ◽  
Clinical Data ◽  
Mating Type ◽  
Severe Disease ◽  
Nationwide Survey ◽  
High Serum ◽  
Content Type ◽  
Serotype A ◽  
Population Structures ◽  
Ad Hybrids

ABSTRACTCryptococcus neoformansis a human opportunistic fungal pathogen causing severe disseminated meningoencephalitis, mostly in patients with cellular immune defects. This species is divided into three serotypes: A, D, and the AD hybrid. Our objectives were to compare population structures of serotype A and D clinical isolates and to assess whether infections with AD hybrids differ from infections with the other serotypes. For this purpose, we analyzed 483 isolates and the corresponding clinical data from 234 patients enrolled during the CryptoA/D study or the nationwide survey on cryptococcosis in France. Isolates were characterized in terms of ploidy, serotype, mating type, and genotype, utilizing flow cytometry, serotype- and mating type-specific PCR amplifications, and multilocus sequence typing (MLST) methods. Our results suggest thatC. neoformansserotypes A and D have different routes of multiplication (primarily clonal expansion versus recombination events for serotype A and serotype D, respectively) and important genomic differences. Cryptococcosis includes a high proportion of proven or probable infections (21.5%) due to a mixture of genotypes, serotypes, and/or ploidies. Multivariate analysis showed that parameters independently associated with failure to achieve cerebrospinal fluid (CSF) sterilization by week 2 were a high serum antigen titer, the lack of flucytosine during induction therapy, and the occurrence of mixed infection, while infections caused by AD hybrids were more likely to be associated with CSF sterilization. Our study provides additional evidence for the possible speciation ofC. neoformansvar.neoformansandgrubiiand highlights the importance of careful characterization of causative isolates.IMPORTANCECryptococcus neoformansis an environmental fungus causing severe disease, estimated to be responsible for 600,000 deaths per year worldwide. This species is divided into serotypes A and D and an AD hybrid, and these could be considered two different species and an interspecies hybrid. The objectives of our study were to compare population structures of serotype A and serotype D and to assess whether infections with AD hybrids differ from infections with serotype A or D isolates in terms of clinical presentation and outcome. For this purpose, we used clinical data and strains from patients diagnosed with cryptococcosis in France. Our results suggest that, according to the serotype, isolates have different routes of multiplication and high genomic differences, confirming the possible speciation of serotypes A and D. Furthermore, we observed a better prognosis for infections caused by AD hybrid than those caused by serotype A or D, at least for those diagnosed in France.

scholarly journals Serotype, mating type and ploidy of Cryptococcus neoformans strains isolated from patients in Brazil

2002 ◽  
Vol 44 (6) ◽  
pp. 299-302 ◽  
Author(s):  
Misako OHKUSU ◽  
Naomi TANGONAN ◽  
Kanji TAKEO ◽  
Eriko KISHIDA ◽  
Masami OHKUBO ◽  
...  
Keyword(s):  
Cryptococcus Neoformans ◽  
Mating Type ◽  
Sao Paulo ◽  
São Paulo ◽  
Aids Patients ◽  
Serotype A ◽  
Tertiary Teaching ◽  
Tertiary Teaching Hospital ◽  
Serotype B ◽  
Alpha Type

Serotype, mating type and ploidy of 84 strains of Cryptococcus neoformans isolated from 61 AIDS and 23 non-AIDS patients admitted in a tertiary teaching hospital in São Paulo, Brazil were examined. Among 61 strains isolated from AIDS patients, 60 strains were var. grubii (serotype A). Only one strain was var. gattii (serotype B). No var. neoformans (serotype D) was found. Among 23 strains isolated from non-AIDS patients, 15 were var. grubii (serotype A) and the remaining 8 were var. gattii, all of which were serotype B. Seventy-three of the 75 serotype A strains were the heterothallic alpha type (MATalpha) and the remaining 2 were untypable (asexual). Most of the MATalpha strains (69/73) were haploid and the remaining 4 strains were diploid. Similarly, both of the 2 asexual strains among the 75 serotype A strains were haploid. There were no alpha-mating type (MATalpha) strains among the 84 isolates. All of the 8 var. gattii strains were serotype B and haploid. Among a total of 84 strains tested, neither serotype AD nor serotype D were found. Neither triploid nor tetraploid were found. These results suggest that the serological, sexual and ploidy characteristics in C. neoformans strains isolated from AIDS patients in São Paulo were rather simple, whereas strains isolated from non-AIDS patients presented serotype A and B with predominance of serotype A.

scholarly journals Mating-Type-Specific and Nonspecific PAK Kinases Play Shared and Divergent Roles in Cryptococcus neoformans

2002 ◽  
Vol 1 (2) ◽  
pp. 257-272 ◽  
Author(s):  
Ping Wang ◽  
Connie B. Nichols ◽  
Klaus B. Lengeler ◽  
Maria E. Cardenas ◽  
Gary M. Cox ◽  
...  
Keyword(s):  
Cryptococcus Neoformans ◽  
Mating Type ◽  
Fungal Pathogen ◽  
Sexual Cycle ◽  
Type Locus ◽  
Serotype A ◽  
A Cells ◽  
Genes Encoding ◽  
Opportunistic Fungal Pathogen ◽  
Synthetically Lethal

ABSTRACT Cryptococcus neoformans is an opportunistic fungal pathogen with a defined sexual cycle involving fusion of haploid MATα and MATa cells. Virulence has been linked to the mating type, and MATα cells are more virulent than congenic MATa cells. To study the link between the mating type and virulence, we functionally analyzed three genes encoding homologs of the p21-activated protein kinase family: STE20α, STE20a, and PAK1. In contrast to the STE20 genes that were previously shown to be in the mating-type locus, the PAK1 gene is unlinked to the mating type. The STE20α, STE20a, and PAK1 genes were disrupted in serotype A and D strains of C. neoformans, revealing central but distinct roles in mating, differentiation, cytokinesis, and virulence. ste20α pak1 and ste20a pak1 double mutants were synthetically lethal, indicating that these related kinases share an essential function. In summary, our studies identify an association between the STE20α gene, the MATα locus, and virulence in a serotype A clinical isolate and provide evidence that PAK kinases function in a MAP kinase signaling cascade controlling the mating, differentiation, and virulence of this fungal pathogen.

scholarly journals Impact of Mating Type, Serotype, and Ploidy on the Virulence of Cryptococcus neoformans

2008 ◽  
Vol 76 (7) ◽  
pp. 2923-2938 ◽  
Author(s):  
Xiaorong Lin ◽  
Kirsten Nielsen ◽  
Sweta Patel ◽  
Joseph Heitman
Keyword(s):  
Cryptococcus Neoformans ◽  
Mating Type ◽  
Pathogenic Fungi ◽  
Mating Types ◽  
Type Locus ◽  
Virulence Potential ◽  
The Impact ◽  
Ad Hybrids

ABSTRACT Hybridization with polyploidization is a significant biological force driving evolution. The effect of combining two distinct genomes in one organism on the virulence potential of pathogenic fungi is not clear. Cryptococcus neoformans, the most common cause of fungal infection of the central nervous system, has a bipolar mating system with a and α mating types and occurs as A (haploid), D (haploid), and AD hybrid (mostly diploid) serotypes. Diploid AD hybrids are derived either from a-α mating or from unisexual mating between haploid cells. The precise contributions of increased ploidy, the effect of hybridization between serotypes A and D, and the combination of mating types to the virulence potential of AD hybrids have remained elusive. By using in vitro and in vivo characterization of laboratory-constructed isogenic diploids and AD hybrids with all possible mating type combinations in defined genetic backgrounds, we found that higher ploidy has a minor negative effect on virulence in a murine inhalation model of cryptococcosis. The presence of both mating types a and α in AD hybrids did not affect the virulence potential, irrespective of the serotype origin. Interestingly, AD hybrids with only one mating type behaved differently, with the virulence of αADα strains similar to that of other hybrids, while aADa hybrids displayed significantly lower virulence due to negative epistatic interactions between the Aa and Da alleles of the mating type locus. This study provides insights into the impact of ploidy, mating type, and serotype on virulence and the impact of hybridization on the fitness and virulence of a eukaryotic microbial pathogen.

scholarly journals Diploids in the Cryptococcus neoformans Serotype A Population Homozygous for the α Mating Type Originate via Unisexual Mating

2009 ◽  
Vol 5 (1) ◽  
pp. e1000283 ◽  
Author(s):  
Xiaorong Lin ◽  
Sweta Patel ◽  
Anastasia P. Litvintseva ◽  
Anna Floyd ◽  
Thomas G. Mitchell ◽  
...  
Keyword(s):  
Cryptococcus Neoformans ◽  
Mating Type ◽  
Serotype A

scholarly journals Galactoxylomannans from Cryptococcus neoformans Varieties neoformans and grubii Are Structurally and Antigenically Variable

2010 ◽  
Vol 9 (7) ◽  
pp. 1018-1028 ◽  
Author(s):  
Magdia De Jesus ◽  
Siu-Kei Chow ◽  
Radames J. B. Cordero ◽  
Susana Frases ◽  
Arturo Casadevall
Keyword(s):  
Light Scattering ◽  
Cryptococcus Neoformans ◽  
Capsular Polysaccharide ◽  
Growth Conditions ◽  
Radius Of Gyration ◽  
Effective Diameter ◽  
Content Type ◽  
Serotype A ◽  
Multiangle Laser Light Scattering ◽  
And Diffusion

ABSTRACT Prior studies have established that the Cryptococcus neoformans capsular polysaccharide component galactoxylomannan (GalXM) manifests serotype-related structural differences that translate into antigenic differences. We analyzed GalXM from acapsular serotype A and D strains by carbohydrate analysis and static and dynamic light scattering to determine mass, effective diameter, polydispersity, and diffusion coefficients. Multiangle laser light scattering showed that GalXM from C. neoformans var. grubii strain cap59 (serotype A) had larger molecular mass (4.21 × 106 ± 0.95 × 106 g/mol) and radius of gyration (207 ± 27 nm) than GalXM from C. neoformans var. neoformans cap67 (serotype D). cap67 GalXM had corresponding values of 0.70 × 106 ± 0.05 × 106 g/mol and 120 ± 22 nm, respectively. The effective diameter for GalXM and polydispersity from the two strains varied depending on temperature and medium growth conditions, indicating that GalXM structure can vary within a strain, depending on its environment. Zeta potential determinations were negative for GalXM from both strains under all conditions, consistent with the recently reported presence of glucuronic acid. These results imply that C. neoformans GalXM, like glucuronoxylomannan, can manifest variety- and growth condition-related variations. Analysis of 16 C. neoformans and 7 Cryptococcus gattii strains with polyclonal antibody to a GalXM strain revealed antigenic similarities among the C. neoformans variety neoformans and grubii strains and no reactivity with C. gattii. As a result of the deleterious effects of GalXM on immune function, structural and antigenic variability between serotypes may translate into differences in immunomodulatory effects.

scholarly journals Laccase Affects the Rate of Cryptococcus neoformans Nonlytic Exocytosis from Macrophages

mBio ◽  
2020 ◽  
Vol 11 (5) ◽  
Author(s):  
Stefânia de Oliveira Frazão ◽  
Herdson Renney de Sousa ◽  
Lenise Gonçalves da Silva ◽  
Jéssica dos Santos Folha ◽  
Kaio César de Melo Gorgonha ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cryptococcus Neoformans ◽  
Laccase Activity ◽  
Severe Disease ◽  
Cell Types ◽  
Great Variability ◽  
Melanin Production ◽  
Content Type ◽  
Fungal Melanin ◽  
Laccase Enzyme

ABSTRACT Nonlytic exocytosis is a process in which previously ingested microbes are expelled from host phagocytes with the concomitant survival of both cell types. This process has been observed in the interaction of Cryptococcus spp. and other fungal cells with phagocytes as distant as mammalian, bird, and fish macrophages and ameboid predators. Despite a great amount of research dedicated to unraveling this process, there are still many questions about its regulation and its final benefits for host or fungal cells. During a study to characterize the virulence attributes of Brazilian clinical isolates of C. neoformans, we observed great variability in their rates of nonlytic exocytosis and noted a correlation between this process and fungal melanin production/laccase activity. Flow cytometry experiments using melanized cells, nonmelanized cells, and lac1Δ mutants revealed that laccase has a role in the process of nonlytic exocytosis that seems to be independent of melanin production. These results identify a role for laccase in virulence, independent of its role in pigment production, that represents a new variable in the regulation of nonlytic exocytosis. IMPORTANCE Cryptococcus neoformans is a yeast that causes severe disease, primarily in immunosuppressed people. It has many attributes that allow it to survive and cause disease, such as a polysaccharide capsule and the dark pigment melanin produced by the laccase enzyme. Upon infection, the yeast is ingested by cells called macrophages, whose function is to kill them. Instead, these fungal cells can exit from macrophages in a process called nonlytic exocytosis. We know that this process is controlled by both host and fungal factors, only some of which are known. As part of an ongoing study, we observed that C. neoformans isolates that produce melanin faster are more-frequent targets of nonlytic exocytosis. Further experiments showed that this is probably due to higher production of laccase, because fungi lacking this enzyme are nonlytically exocytosed less often. This shows that laccase is an important signal/regulator of nonlytic exocytosis of C. neoformans from macrophages.

scholarly journals Sexual Cycle of Cryptococcus neoformans var. grubii and Virulence of Congenic a and α Isolates

2003 ◽  
Vol 71 (9) ◽  
pp. 4831-4841 ◽  
Author(s):  
Kirsten Nielsen ◽  
Gary M. Cox ◽  
Ping Wang ◽  
Dena L. Toffaletti ◽  
John R. Perfect ◽  
...  
Keyword(s):  
Cryptococcus Neoformans ◽  
Mating Type ◽  
Pathogenic Fungus ◽  
Sexual Cycle ◽  
Protein Signaling ◽  
Serotype A ◽  
Human Fungal Pathogen ◽  
The Central Nervous System ◽  
Human Pathogenic Fungus ◽  
Unusual Strain

ABSTRACT Cryptococcus neoformans is a human-pathogenic fungus that has evolved into three distinct varieties that infect most prominently the central nervous system. A sexual cycle involving haploid cells of a and α mating types has been reported for two varieties (C. neoformans var. neoformans, serotype D, and C. neoformans var. gattii, serotypes B and C), yet the vast majority of infections involve a distinct variety (C. neoformans var. grubii, serotype A) that has been thought to be clonal and restricted to the α mating type. We recently identified the first serotype A isolate of the a mating type which had been thought to be extinct (strain 125.91). Here we report that this unusual strain can mate with a subset of pathogenic serotype A strains to produce a filamentous dikaryon with fused clamp connections, basidia, and viable recombinant basidiospores. One meiotic segregant mated poorly with the serotype A reference strain H99 but robustly with a crg1 mutant that lacks a regulator of G protein signaling and is hyperresponsive to mating pheromone. This meiotic segregant was used to create congenic a and α mating type serotype A strains. Virulence tests with rabbit and murine models of cryptococcal meningitis showed that the serotype A congenic a and α mating type strains had equivalent virulence in animal models, in contrast to previous studies linking the α mating type to increased virulence in congenic serotype D strains. Our studies highlight a role for sexual recombination in the evolution of a human fungal pathogen and provide a robust genetic platform to establish the molecular determinants of virulence.

scholarly journals Serotype AD Strains of Cryptococcus neoformans Are Diploid or Aneuploid and Are Heterozygous at the Mating-Type Locus

2001 ◽  
Vol 69 (1) ◽  
pp. 115-122 ◽  
Author(s):  
Klaus B. Lengeler ◽  
Gary M. Cox ◽  
Joseph Heitman
Keyword(s):  
Cryptococcus Neoformans ◽  
Mating Type ◽  
Yeast Cells ◽  
Sexual Cycle ◽  
Pcr Analysis ◽  
Environmental Isolates ◽  
Type Locus ◽  
Serotype A ◽  
Mating Type Locus ◽  
Normal Meiosis

ABSTRACT Cryptococcus neoformans is a pathogenic basidiomycete with a defined sexual cycle involving mating between haploid yeast cells with a transient diploid state. C. neoformans occurs in four predominant serotypes (A, B, C, and D), which represent different varieties or species. Rare clinical and environmental isolates with an unusual AD serotype have been reported and suggested to be diploid. We found by fluorescence-activated cell sorter analysis that serotype AD strains are aneuploid or diploid. PCR analysis with primers specific for serotype A or D alleles of theCNA1, CLA4, and GPA1 genes revealed that both alleles are often present in serotype AD strains. PCR analysis with primers specific for genes in the MATa orMATα mating-type loci revealed that serotype AD strains are heterozygous for the mating-type locus. Interestingly, in several serotype AD strains, the MATα locus was derived from the serotype D parent and the MATa locus was inherited from a serotype A parent that has been thought to be extinct. Basidiospores from a self-fertile serotype AD strain bearing the putative serotype AMATa locus showed a very low viability (∼5%), and no fertile serotype A MATa strain could be recovered. Serotype AD strains were virulent in a murine model. Hybrid AD strains could readily be isolated following a laboratory cross between a serotype A strain and a serotype D strain. In summary, serotype AD strains ofC. neoformans are unusual aneuploid or diploid strains that result from matings between serotype A and D strains. Self-fertile isolates fail to undergo normal meiosis because of genetic divergence. Our findings further suggest that serotype A MATa strains may exist in nature.

Sign in / Sign up

Export Citation Format

Share Document