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Infection and Immunity, August 2006, p. 4875-4883, Vol. 74, No. 8
0019-9567/06/$08.00+0     doi:10.1128/IAI.01978-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Transcribed var Genes Associated with Placental Malaria in Malawian Women{dagger}

Michael F. Duffy,1,{ddagger}* Aphrodite Caragounis,1,{ddagger} Rintis Noviyanti,2 Helen M. Kyriacou,3 Ee Ken Choong,1 Katja Boysen,1 Julie Healer,1,§ J. Alexandra Rowe,3 Malcolm E. Molyneux,4,5 Graham V. Brown,1 and Stephen J. Rogerson1

Department of Medicine (RMH), University of Melbourne, Melbourne, Australia,1 The Eijkman Institute for Molecular Biology, Eijkman Building, Jl. Diponegoro 69, Jakarta, Indonesia 10430,2 Institute of Immunology and Infection Research, School of Biological Sciences, University of Edinburgh, King's Buildings, West Mains Rd., Edinburgh, United Kingdom,3 Malawi-Liverpool-Wellcome Trust Clinical Research Programme, College of Medicine, University of Malawi, Blantyre, Malawi,4 School of Tropical Medicine, University of Liverpool, Liverpool, United Kingdom5

Received 6 December 2005/ Returned for modification 24 January 2006/ Accepted 13 May 2006

Determining the diversity of PfEMP1 sequences expressed by Plasmodium falciparum-infected erythrocytes isolated from placentas is important for attempts to develop a pregnancy-specific malaria vaccine. The DBL{gamma} and var2csa DBL3x domains of PfEMP1 molecules are believed to mediate placental sequestration of infected erythrocytes, so the sequences encoding these domains were amplified from the cDNAs of placental parasites by using degenerate oligonucleotides. The levels of specific var cDNAs were then determined by quantitative reverse transcription-PCR. Homologues of var2csa DBL3x were the predominant sequences amplified from the cDNAs of most placental but not most children's parasites. There was 56% identity between all placental var2csa sequences. Many different DBL{gamma} domains were amplified from the cDNAs of placental and children's isolates. var2csa transcripts were the most abundant var transcripts of those tested in 11 of 12 placental isolates and 1 of 6 children's isolates. Gravidity did not affect the levels of var2csa transcripts. We concluded that placental malaria is frequently associated with transcription of var2csa but that other var genes are also expressed, and parasites expressing high levels of var2csa are not restricted to pregnant women. The diversity of var2csa sequences may be important for understanding immunity and for the development of vaccines for malaria during pregnancy.

* Corresponding author. Mailing address: Department of Medicine (RMH), University of Melbourne, Post Office, Royal Melbourne Hospital, Victoria 3050, Australia. Phone: 61 3 8344 3267. Fax: 61 3 9347 1863. E-mail: mduffy{at}unimelb.edu.au.

Editor: W. A. Petri, Jr.

{dagger} Supplemental material for this article may be found at http://iai.asm.org/.

{ddagger} M.F.D. and A.C. contributed equally to this work.

§ Present address: The Walter and Eliza Hall Institute of Medical Research, 19 Royal Parade, Parkville, Victoria, Australia 3050.

Infection and Immunity, August 2006, p. 4875-4883, Vol. 74, No. 8
0019-9567/06/$08.00+0     doi:10.1128/IAI.01978-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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