Follicular dendritic cells mediated maintenance of primary lymphocyte cultures for long-term analysis of a functional in vitro immune system.
Primary lymphocyte cultures are important for analysis of cellular and molecular events occurring during immune responses. However, the lymphoid cells (especially B cells) typically only survive for a few days in vitro which limits studies. Establishment of long-term primary lymphocyte cultures where a functioning humoral immune responses can be maintained and regulated is still a challenge. Follicular dendritic cells (FDC) are immune accessory cells that reside in the follicles of secondary lymphoid organs and are known to protect lymphocytes from apoptosis. We hypothesized that addition of FDC to primary lymphocyte cultures may help maintain humoral immune responses in vitro as they do in vivo. To test the hypothesis, freshly isolated lymphocytes were cultured with or without FDC. The B cells in cultures were labeled using B220 and apoptotic cells were labeled using the TUNEL assay. Antibody production was monitored in supernatant fluids using ELISA. The results showed that FDC reduced apoptosis and helped sustain primary lymphocyte cultures and antibody production was maintained throughout the entire period (e.g., 8 weeks). This FDC dependent system should be useful for analysis of cellular and molecular events over extended periods in vitro.
Department of Microbiology and Immunology, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298-0678, USA.
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Qin
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Wu
G F
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Burton
A K
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Szakal
J G
JG
Tew
D000818:Animals; D017209:Apoptosis; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D003713:Dendritic Cells / Q000166:cytology / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007107:Immune System / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D008214:Lymphocytes / Q000166:cytology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010047:Ovalbumin / Q000276:immunology; D013997:Time Factors
D007074:Immunoglobulin G; D010047:Ovalbumin