Objective. Dendritic cells (DCs) are derived from bone marrow precursor cells and
migrate to lymphoid and non-lymphoid organs via blood. DCs have been shown to play
an important role in inducing immunity and/or tolerance. Several DC subgroups with
different biological roles have been identified. Myeloid DCs usually induce immune
response and tolerance is usually induced by lymphoid DCs. Differences in frequency
and ratio of DC subtypes appears to be one of the reasons for the difference in immune
response in various lymphoid tissues. Considering the recent applications of different
DC subgroups for the induction of immune response or tolerance induction and their
use as vaccine for the immunotherapy of tumors or autoimmune diseases.
Materials and Methods. In this study, it was tried to investigate the function and
phenotype of Splenic DCs, these cells were separated by mechanical and enzymatic
methods from mouse spleen and their surface markers using tlowcytometry. To study
the ability of separated DCs in immune response induction, their functional capability
was investigated by lymphocyte transformation test.
Result. Following collagenase digestion of Splenic tissue, using density gradient
centrifugation (13% nycodenz) and adherance characteristic of DCs to tissue culture
dish, more than 7-9 X 105 DCs were separated from each spleen with purity of 90-95
percent. The phenotypic analysis of these cells showed that at least three DC
subpopulations: CD11c+, CD 8α, CD11b+: CD11c+, CD8α+, CD11b-: CD11c+, CD8α+, CD11b+ could be found in spleen. The ratio of lymphoid (CD11c+, CD&i+, CD11b-)
and myeloid (CD11c+, CD8α-, CD11b+) DCs subgroups was almost equal in spleen. Lymphocyte transformation test showed that lymphocytes proliferation in response to ova albumin is higher in ova albumin pulsed DCs injected mice compared to control groups (P<0.004).
Conclusion. Considering the obtained results it can be said that Splenic DCs can
process and present the antigen in vivo and induce antigen specific response in vitro.
Similar ratio of lymphoid and myeloid DCs in spleen may be implicated in regulation of
and homeostasis of immune response by this organ.