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Timecourse of mAbs 18B7 or MOPC-21 (control) binding to Cryptococcus neoformans serotype D strains JEC21 or 24067 (52D) after 1, 2 or 4 hours at 37 C.

ABSTRACT: Changes in gene expression of serotype D strains JEC21 or 24067 (52D) were examined after incubation with mAb 18B7 or the control MOPC-21 for 1, 2 or 4 hours at 37 C.

ORGANISM(S): Cryptococcus neoformans var. neoformans Cryptococcus neoformans

PROVIDER: GSE35241 | GEO | 2012/02/23

SECONDARY ACCESSION(S): PRJNA151545

REPOSITORIES: GEO

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Timecourse of mAbs 18B7 or MOPC-21 (control) binding to Cryptococcus neoformans serotype D strains JEC21 or 24067 (52D) after 1, 2 or 4 hours at 37 C.

Project description:Changes in gene expression of serotype D strains JEC21 or 24067 (52D) were examined after incubation with mAb 18B7 or the control MOPC-21 for 1, 2 or 4 hours at 37 C. Analysis used incubation with a non-binding IgG1 mAb as a control for incubation with a protective IgG1 mAb 18B7 at different timepoints.

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Project description:Cryptococcus neoformans and Cryptococcus gattii cause life-threatening meningoencephalitis or lung diseases in immunocompetent individuals or immunocompromised ones. C. neoformans and C. gattii are subdivided into five serotypes based on their capsular glucuronoxylomannan (GXM). C. neoformans consists of serotypes A, D, and AD hybrid, and C. gattii consists of serotypes B and C. Given structural differences of GXM between C. neoformans and C. gattii, it remains unclear that how innate immune system recognizes GXM. Here, we report that C-type lectin receptor Dectin-3 (MCL encoded by Clec4d) is a direct receptor for GXMs from C. neoformans serotype AD (C.n-AD) and C. gattii serotype B (C.g-B). GXMs from C.n-AD and C.g-B activated NF-κB and ERK pathways to induce pro-inflammatory cytokine production, whereas it was completely abolished due to deficiency of Dectin-3 or caspase recruitment domain family member 9 (CARD9). Upon pulmonary C.n-AD and C.g-B infection, Dectin-3- and CARD9-deficient mice were highly susceptible and showed augmented lung injury due to impairment of alveolar macrophage accumulation and killing activities. Our study provides the first biological and genetic evidence demonstrating that Dectin-3 recognizes GXM of C.n-AD and C.g-B to initiate host defense against cryptococcosis.

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Project description:Cryptococcus neoformans interactions with murine macrophages are critical for disease. In this project we analyzed fungal proteins which were co-purified with murine host proteins after interaction. H99 C. neoformans was opsonized with mAb 18B7 and addedd to murine macrophages. Then murine cells were lysed and cell extracts submitted to proteomics.

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Expression Profiling of the Response to Short-Term Exposure to Fluconazole in Cryptococcus neoformans Serotype A

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Project description:Cryptococcus neoformans is an opportunistic human fungal pathogen and serves as a model organism for studies of eukaryotic microbiology and microbial pathogenesis. C. neoformans species complex is classified into serotype A, serotype D, and AD hybrids, which are currently considered different subspecies. Different serotype strains display varied phenotypes, virulence, and gene regulation. Genetic investigation of important pathways is often performed in both serotype A and D reference strains in order to identify diversification or conservation of the interrogated signaling network. Many genetic tools have been developed for C. neoformans serotype A reference strain H99, including the gene free "safe haven" (SH) regions for DNA integration identified based on genomic features. However, no such a genomic safe haven region has been identified in serotype D strains. Here, capitalizing on the available genomic, transcriptomic, and chromatin data, we identified an intergenic region named as SH3 for the serotype D reference strains JEC21 and XL280. We also designed a sgRNA and a vector facilitating any alien gene integration into SH3 through a CRISPR-Cas9 system. We found that gene inserted in this region complemented the corresponding gene deletion mutant. Fluorescent reporter gene inserted in SH3 can also be expressed efficiently. Insertion in SH3 itself did not alter the expression of adjacent genes and did not affect the growth or mating of C. neoformans. Thus, SH3 provides a resource for genetic manipulations in serotype D strains and will facilitate comparative analyses of gene functions in this species complex. In addition, the incorporation of the multi-omic data in our selection of the safe haven region could help similar studies in other organisms.

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