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Immunology Group, International Center for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, India
We show in this study that incubation of freshly isolated bone
marrow cells with Mycobacterium tuberculosis (M.
tb) secretory Ag (MTSA), in the absence of any growth or
differentiation-inducing factor, differentiates them into dendritic
cell (DC)-like APCs. These DCs expressed moderate to high levels of
various markers typical of DCs. These included T cell costimulatory
molecules CD80, CD86, CD40, and CD54 and high levels of surface MHC
class I and II on CD11c+ cells. The levels and the kinetics
of up-regulation of these molecules were comparable with those of
GM-CSF-differentiated DCs. Furthermore, these DCs exhibited morphology
characteristics to DCs like the presence of dendritic processes. These
DCs were also potent stimulators of allogeneic T cells and
preferentially induced the secretion of IFN-
over IL-10 from the
interacting T cells. Interestingly, the differentiation of bone marrow
cells into DC-like APCs was obtained with many other M.
tb Ags, including whole cell extract of M. tb.
Further characterization of MTSA-differentiated DCs showed that they
were immature in nature, as stimulation of these DCs with TNF-
,
anti-CD40, or LPS further up-regulated the surface levels of
various molecules together with an increase in their T cell stimulatory
capacity. The Ag-specific T cell responses of MTSA-differentiated DCs
were mainly contributed by the CD4+ subset, indicating that
MTSA was largely MHC II restricted. Furthermore, stimulation of bone
marrow cells with MTSA induced the nuclear translocation of the
transcription factor NF-
B, thereby indicating its role during
MTSA-induced differentiation of DCs.
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