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Clinical and Vaccine Immunology, May 2006, p. 568-574, Vol. 13, No. 5
1071-412X/06/$08.00+0     doi:10.1128/CVI.13.5.568-574.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Evaluation of Various Methods of Maternal Placental Blood Collection for Immunology Studies

Kenya Medical Research Institute, Kisumu, Kenya,¹ Nyanza Provincial General Hospital, Kisumu, Kenya,² Roll Back Malaria, World Health Organization, Geneva, Switzerland,³ Department of Infectious Diseases, College of Veterinary Medicine, and Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, Georgia,⁴ Division of Parasitic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia⁵

Received 2 December 2005/ Returned for modification 9 January 2006/ Accepted 16 March 2006

The collection of maternal placental intervillous blood (IVB), without contamination of fetal blood and with an accurate mononuclear cell profile, is essential for immunological studies of placental malaria and other infectious diseases of the placenta. We have compared five documented methods of IVB collection: perfusion, incision, biopsy, tissue grinding, and puncture (prick) for fetal blood contamination and mononuclear cell profiles using flow cytometry. Twenty-five placentas were obtained from Plasmodium falciparum and human immunodeficiency virus-negative primigravid and secundigravid women delivering at Nyanza Provincial Hospital in Kisumu, western Kenya. Each of the five methods was performed on the same placenta. Fetal red blood cell contamination was significantly lower for the prick and perfusion methods (4.1% and 8.3%, respectively) than for incision (59.5%), biopsy (42.6%), and tissue grinding (19.9%). Significant variation was noted among the five methods in the percentages of monocytes, total T cells, CD4⁺ and CD8⁺ T cells, B cells, and NK cells. Further, a pairwise comparison of prick and perfusion, the two methods with low fetal blood contamination, showed that they were not different for fetal red blood cell contamination levels; however, prick yielded significantly higher percentages of CD4 T cells and CD4 memory T cells than perfusion. Collection by prick was determined to be the best method of intervillous blood collection for immunology studies, and perfusion represented the next best method of choice due to high sample volume yield. Overall, in considering the advantages/disadvantages of the two methods with low fetal cell contamination, we conclude that a combination of prick and perfusion is most suitable for immunology studies.

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