Ž . Journal of Immunological Methods 209 1997 93–104 A simple perfusion technique for isolation of maternal intervillous blood mononuclear cells from human placentae J.M. Moore a , B. Nahlen a,b , A.V.O. Ofulla c , J. Caba d , J. Ayisi b , A. Oloo b , A. Misore e , A.J. Nahmias d , A.A. Lal a , V. Udhayakumar a, ) a Immunology Branch, DiÕision of Parasitic Diseases, National Center for Infectious Diseases, Centers for Disease Control and PreÕention, Public Health SerÕice, U.S. Department of Health and Human SerÕices, Atlanta, GA 30333, USA b Vector Biology and Control Research Center, Kenya Medical Research Institute, Kisumu, Kenya c Zoology Department, Maseno UniÕersity College, Maseno, Kenya d Department of Pediatrics, Emory UniÕersity School of Medicine, Atlanta, GA 30303, USA e New Nyanza ProÕincial General Hospital, Kisumu, Kenya Received 9 May 1997; revised 19 September 1997; accepted 23 September 1997 Abstract Ž . A noninvasive perfusion method for the recovery of maternal placental intervillous blood for use in immunologic assays Ž . is described. 60% of the perfused blood samples tested for fetal red blood cell RBC contamination were found to be pure maternal blood; in the remainder, fetal RBC contamination, with a single exception, was less than 6%. The intervillous Ž . mononuclear cells IVBMC isolated from this blood were of predominantly maternal origin as demonstrated by a polymerase chain reaction-based DNA typing technique. The number of IVBMC obtained was within the range of 9 to 55 =10 6 cells. Phenotypic analysis of IVBMC surface antigens revealed that 61% of the cells were CD3 q T-cells and 18% were CD19 q B-cells. The CD4 q and CD8 q T-lymphocyte subsets accounted for 28 and 26% of the IVBMC, respectively. The IVBMC were functionally competent as evidenced by in vitro lymphoproliferation and cytokine production in response to mitogen and PPD stimulation. This technique allows for rapid and safe isolation of large numbers of IVBMC which are functionally active up to 12 h post-delivery, thus representing a significant improvement over previously described methods. It should facilitate more vigorous research in the study of uteroplacental immunity and infectious disease research, particularly in field settings where sample collection and laboratory facilities are distant. q 1997 Elsevier Science B.V. Keywords: Pregnancy; Immunology; Placenta; Mononuclear cells; Lymphoproliferation; Cytokines Abbreviations: IVBMC, intervillous blood mononuclear cells; HIV, human immunodeficiency virus; PBS, phosphate-buffered saline; EDTA, ethylenediaminetetraacetic acid; UVC, umbilical vessel catheter; PCR, polymerase chain reaction; PHA, phytohemagglutinin; mAbs, monoclonal antibodies; PPD, purified protein derivative; ELISA, enzyme-linked immunosorbent assay; SI, stimulation index; cpm, counts per minute; IFN-g , g-interferon; IL, interleukin; PBMC, peripheral blood mononuclear cells ) Corresponding author. DPD, CDC, Mail Stop F-12, 4770 Buford Highway, Chamblee, GA 30341, USA. Tel.: q1-770-4884030; fax: q1-770-4884454; e-mail: vxu0@cdc.gov 0022-1759r97r$17.00 q 1997 Elsevier Science B.V. All rights reserved. Ž . PII S0022-1759 97 00162-2