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Analysis of the early cellular and humoral responses of Galleria mellonella larvae to infection by Candida albicans

ABSTRACT: Galleria mellonella larvae were administered an inoculum of Candida albicans and the cellular and proteomic response to infection over the first 24 hours was monitored. The yeast cell density in infected larvae declined initially but replication commenced six hours post-infection. The hemocyte density decreased from 5.2 × 106/ml to 2.5 × 106/ml at 2 hours but increased to 4.2 × 106 at 6 hours but decreased subsequently. Administration of β - glucan to larvae also resulted in an increase in hemocyte density (5.1 ± 0.22 × 106/ml to 6.25 ± 0.25 × 106/ml, p < 0.05) and the population showed an increase in the density of small, granular type cells at 24 hours (p < 0.05). Hemocytes from larvae inoculated with β - glucan showed faster killing of C. albicans cells (53 ± 4.1% (p < 0.01), 64 ± 3.7%, (p < 0.01), respectively) than hemocytes from control larvae (24 ± 11%) at 60 min. Proteomic analysis indicated increased abundance of immune related proteins cecropin-A (5 fold) and prophenoloxidase-activating proteinase-1 (5 fold) 6 hours post infection but by 24 hours there was elevated abundance of muscle (tropomyosin 2 (141 fold), calponin (66 fold), troponin I (62 fold), troponin T (61 fold)) and proteins indicative of cellular stress (glutathione-S-transferase-like protein (114 fold)), fungal dissemination (muscle protein 20-like protein (174 fold)) and tissue breakdown (mitochondrial cytochrome c (10 fold)). Proteins decreased in abundance at 24 hour included β - 1,3 - glucan recognition protein precursor (29 fold), prophenoloxidase subunit 2 (25 fold) and lysozyme-like protein 1 (5 fold). By characterizing the early responses to infection, G. mellonella larvae may be used to model the initial stages of infection processes in mammals.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Galleria Mellonella

TISSUE(S): Blood

SUBMITTER: Gerard Sheehan

LAB HEAD: Prof. Kevin Kavanagh

PROVIDER: PXD006879 | Pride | 2017-09-06

REPOSITORIES: Pride

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Analysis of the early cellular and humoral responses of Galleria mellonella larvae to infection by Candida albicans.

Sheehan Gerard G Kavanagh Kevin K

Virulence 20170921 1

Galleria mellonella larvae were administered an inoculum of Candida albicans and the response to infection over 24 hours was monitored. The yeast cell density in infected larvae declined initially but replication commenced six hours post-infection. The hemocyte density decreased from 5.2 × 10<sup>6</sup>/ml to 2.5 × 10<sup>6</sup>/ml at 2 hours but increased to 4.2 × 106 at 6 hours and decreased subsequently. Administration of β - glucan to larvae also caused a fluctuation in hemocyte density (5 ...[more]

PMID: 28872999

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Project description:Background All animals must be able to adapt to changes in nutrient quality and supply. One striking aspect of this adaptive response is the extension of lifespan when caloric intake is reduced. These animals are also more resistant to various stresses, including starvation. We have characterized the response of adult Drosophila males and females to starvation and sugar conditions using high density microarrays. Results We have organized the regulated genes into specific metabolic and physiologic pathways. Most of the highly regulated genes have a strong sex bias. In addition to expected sex specific changes, such as egg production in females, we found unexpected biases in sensory systems, amino acid and purine metabolism, and signaling pathways. Comparison of starvation to caloric restriction revealed numerous overlaps. Conclusions Our results support the notion that starvation response shares similarities to caloric restriction. They have relevance for sex specific differences in longevity and in response to nutrient conditions which alter lifespan. Nutrient Conditions and Fly handling Larvae were kept on apple juice agar plates with yeast, and the larvae were raised at 25°C. For the developmental control larvae were collected 39-41 h AEL, for the developmental time-points at 51-53 hours and every 12 hours until 99-101 hours. Adult flies were kept at 25°C in 50 ml PS-tubes flat bottom (Greiner bio-one, Frickenhausen, Germany) on standard fly food (16.5 g yeast, 81,5 g cornmeal, 8 g agar, 100 ml sugar beet sirup and 200 ml 10% Nipagin in EtOH). Flies were collected that had hatched in 24 hours and kept in groups of ~200 flies/tube. After 5 days we separated males and females and put them in groups of 80/28 ml PS-tube. We allowed for one day CO2 recovery on standard fly food before flies were moved to 1) tubes with yeast paste on PBS soaked filter paper (control) 2) tubes with only PBS soaked filter paper (starvation) or 3) tubes with filter paper soaked in PBS with 20% Sucrose (sugar). After 24 or 48 hours flies were shock frozen in liquid nitrogen and stored at - 80°C for further handling. Males and females were assayed separately. For females, we performed 24 and 48 starvation, and 24 and 48 hours sugar. For the males only 24 hour starvation was done, since most died upon 48 hour starvation; and 24 and 48 hour sugar. Briefly, flies were collected within a 24 hour period, allowed to stay together for 4 days, separated into males and females, and allowed one day recovery in normal fly food. Afterwards, the flies were placed into three conditions: fresh yeast plus PBS, PBS or PBS plus 20 % sucrose. These experiments essentially parallel the experimental set up of previous larval experiments (7). The starvation and PBS were then compared with yeast (control). Keywords: ordered

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Analysis of the early cellular and humoral responses of Galleria mellonella larvae to infection by Candida albicans

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Analysis of the early cellular and humoral responses of Galleria mellonella larvae to infection by Candida albicans.

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