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Serum Removal from Culture Induces Growth Arrest, Ploidy Alteration, Decrease in Infectivity and Differential
Pedro J Alcolea1, Ana Alonso1, Miguel A Moreno-Izquierdo1
1Department of Molecular Microbiology and Biology of Infections, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas (CIB-CSIC), Madrid. Spain.
Abstract:
Leishmania infantum is one of the species responsible for visceral leishmaniasis. This species is distributed basically in the Mediterranean basin. A recent outbreak in humans has been reported in Spain. Axenic cultures are performed for most procedures with Leishmania spp. promastigotes. This model is stable and reproducible and mimics the conditions of the gut of the sand fly host, which is the natural environment of promastigote development. Culture media are undefined because they contain mammalian serum, which is a rich source of complex lipids and proteins. Serum deprivation slows down the growth kinetics and therefore, yield in biomass. In fact, we have confirmed that the growth rate decreases, as well as infectivity. Ploidy is also affected. Regarding the transcriptome, a high-throughput approach has revealed a low differential expression rate but important differentially regulated genes. The most remarkable profiles are: up-regulation of the GINS Psf3, the fatty acyl-CoA synthase (FAS1), the glyoxylase I (GLO1), the hydrophilic surface protein B (HASPB), the methylmalonyl-CoA epimerase (MMCE) and an amastin gene; and down-regulation of the gPEPCK and the arginase. Implications for metabolic adaptations, differentiation and infectivity are discussed herein.
Insights
Serum deprivation impacts Leishmania infantum growth, infectivity, and ploidy. Key genes involved in metabolism and differentiation are altered, offering insights into parasite adaptation.
Area of Science:
- Parasitology
- Molecular Biology
- Infectious Diseases
Background:
- Leishmania infantum causes visceral leishmaniasis, prevalent in the Mediterranean.
- Axenic cultures of promastigotes are standard, mimicking the sand fly gut environment.
- Serum-containing media are complex, but serum deprivation affects parasite biology.
Purpose of the Study:
- To investigate the effects of serum deprivation on Leishmania infantum growth, infectivity, and ploidy.
- To analyze transcriptomic changes in response to serum deprivation.
- To understand metabolic adaptations and differentiation.
Main Methods:
- Axenic culture of Leishmania infantum promastigotes under serum-deprived conditions.
- Assessment of growth kinetics, infectivity, and ploidy.
- High-throughput transcriptomic analysis.
Main Results:
- Serum deprivation reduced growth rate, biomass yield, and infectivity.
- Ploidy was affected by serum deprivation.
- Transcriptome analysis revealed significant differential expression of genes related to metabolism (e.g., FAS1, GLO1, MMCE), differentiation (e.g., HASPB, amastin), and virulence.
Conclusions:
- Serum deprivation significantly impacts Leishmania infantum physiology and infectivity.
- Altered gene expression highlights adaptive metabolic and differentiation pathways.
- Findings provide insights into parasite survival strategies and potential therapeutic targets.
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