%0 Journal Article
%T Var transcription profiling of Plasmodium falciparum 3D7: assignment of cytoadherent phenotypes to dominant transcripts
%A Uta GŁżlnitz
%A Letusa Albrecht
%A Gerhard Wunderlich
%J Malaria Journal
%D 2008
%I BioMed Central
%R 10.1186/1475-2875-7-14
%X P. falciparum 3D7 parasites were panned on several transfected CHO-cell lines and their var transcripts analysed by i) reverse transcription/PCR/cloning/sequencing using a universal DBL¦Á specific oligonucleotide pair and ii) by reverse transcription followed by quantitative PCR using 57 different oligonucleotide pairs.Each cytoadherence selected parasite line also adhered to untransfected CHO-745 cells and upregulation of the var gene PFD995/PFD1000c was consistently associated with cytoadherence to all but one CHO cell line. In addition, parasites panned on different CHO cell lines revealed candidate var genes which reproducibly associated to the respective cytoadherent phenotype. The transcription profile obtained by RT-PCR/cloning/sequencing differed significantly from that of RT-quantitative PCR.Transfected CHO cell lines are of limited use for the creation of monophenotypic cytoadherent parasite lines. Nevertheless, 3D7 parasites can be reproducibly selected for the transcription of different determined var genes without genetic manipulation. Most importantly, var transcription analysis by RT-PCR/cloning/sequencing may lead to erroneous interpretation of var transcription profiles.Plasmodium falciparum the causative agent of tropical malaria still kills over one million people per year, mostly children under five years in sub-Saharan Africa [1]. One major virulence factor of this parasite is the highly variant Plasmodium falciparum erythrocyte membrane protein-1 (PfEMP-1) family [2]. Members of this family appear at the surface infected red blood cell, during the trophozoite stage. Their large, highly variable ectodomains, consist of several subdomains (termed Duffy binding-like, DBL, and cystein rich interdomain regions, CIDR), which interact with a number of cellular host receptors such as CD36, ICAM-1, CSA, PECAM, E-Selectin, VCAM and others (reviewed by [3]). This interaction causes the retention of mature blood stage forms in the tissue which express the c
%U http://www.malariajournal.com/content/7/1/14