An improved enrichment method for functionally competent, highly purified peripheral blood dendritic cells and its application to HIV-infected blood samples.
Dendritic cells (DC) were purified from human peripheral blood using a rapid and simple method based on magnetic depletion of phagocytes with carbonyl iron, followed by centrifugation of nonphagocytic cells on a Percoll density gradient and depletion of lymphocytes and macrophages/monocytes with a panel of MoAbs and immunomagnetic beads. Enriched DC were obtained with > 99% purity as judged by non-specific esterase (NSE) staining. After isolation, these cells, representing 0.4% of the starting mononuclear cell population, still function as potent antigen-presenting cells for purified T lymphocytes. The present results confirm the ability of human peripheral blood DC to present soluble antigens to T cells including microbial antigens and show, further, that DC are more potent soluble antigen-presenting cells than monocytes. The method was successfully applied to the purification of DC from the blood of HIV-infected individuals. We could not detect decreased numbers of DC in four individuals with early HIV infection and no replicating HIV was detected by in situ hybridization in the DC.
Karhumäki
E
E
Department of Pathology, University of Tampere, Finland.
Viljanen
M E
ME
Cottler-Fox
M
M
Ranki
A
A
Fox
C H
CH
Krohn
K J
KJ
D000938:Antigen-Presenting Cells / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D003713:Dendritic Cells / Q000276:immunology* / Q000648:ultrastructure*; D015658:HIV Infections / Q000097:blood* / Q000276:immunology; D015497:HIV-1 / Q000235:genetics / Q000276:immunology; D006801:Humans; D008213:Lymphocyte Activation / Q000276:immunology; D009000:Monocytes / Q000276:immunology; D011205:Povidone; D012367:RNA, Viral / Q000235:genetics; D012822:Silicon Dioxide; D013601:T-Lymphocytes / Q000276:immunology
D012367:RNA, Viral; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone