May 23, 2008

Simultaneous gene expression profiling in human macrophages infected with Leishmania major parasites using SAGE

BMC Genomics
Fatma Z GuerfaliKoussay Dellagi

Abstract

Leishmania (L) are intracellular protozoan parasites that are able to survive and replicate within the harsh and potentially hostile phagolysosomal environment of mammalian mononuclear phagocytes. A complex interplay then takes place between the macrophage (MPhi) striving to eliminate the pathogen and the parasite struggling for its own survival. To investigate this host-parasite conflict at the transcriptional level, in the context of monocyte-derived human MPhis (MDM) infection by L. major metacyclic promastigotes, the quantitative technique of serial analysis of gene expression (SAGE) was used. After extracting mRNA from resting human MPhis, Leishmania-infected human MPhis and L. major parasites, three SAGE libraries were constructed and sequenced generating up to 28,173; 57,514 and 33,906 tags respectively (corresponding to 12,946; 23,442 and 9,530 unique tags). Using computational data analysis and direct comparison to 357,888 publicly available experimental human tags, the parasite and the host cell transcriptomes were then simultaneously characterized from the mixed cellular extract, confidently discriminating host from parasite transcripts. This procedure led us to reliably assign 3,814 tags to MPhis' and 3,666 tags to ...Continue Reading

  • References50
  • Citations31

References

Mentioned in this Paper

Glutaredoxin
Infectious Process
Real-Time Polymerase Chain Reaction
RPL13A gene
Loxogramme mexicana
Immune Response
FCGR3B protein, human
Pathogenic Aspects
Biochemical Pathway
Microarray Analysis

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