TOLEROGENIC BONE MARROW-DERIVED DENDRITIC CELLS MODULATE ALLERGIC REACTIVITY

1
TOLEROGENIC BONE MARROW-DERIVED DENDRITIC CELLS
MODULATE ALLERGIC REACTIVITY OF LUNG CELLS FROM
MICE WITH SEVERE ALLERGIC LUNG DISEASE. Aarti
Nayyar, Xiaobei Zhang and John R. Gordon.
Immunology Research Group (Dept Vet
Microbiology), University of Saskatchewan,
Saskatoon.
HYPOTHESIS
TOLEROGENIC BONE MARROW-DERIVED DENDRITIC CELLS
CAN BE USED AS AN EFFECTIVE THERAPY FOR SEVERE
ALLERGIC LUNG DISEASE
OBJECTIVES

• To generate (through culture in
  IL-10-containing media) and characterize
  tolerogenic populations of mouse bone
  marrow-derived dendritic cells (DC)
• 2. To assess the impact of these DCs, as well as
  fully mature BMDC, on tolerance induction in a
  mouse model of allergic lung disease/asthma.

ALLERGIC LUNG DISEASE (ALD) MODEL
BALB/c mice were sensitized with OVA/alum (2
µg/mg, i.p.) on dy 0 14, exposed to 1 OVA
aerosols on days 28, 30, 32, then treated with
DCIL-10, DCGM-CSF or DCTNF on day 42 (Schneider
et al, 2001).
RESULTS
FIG 2. DCIL-10-TREATMENTS ABROGATE AHR IN MICE
WITH SEVERE ALD.
FIG. 1. FUNCTIONAL CHARACTERIZATION OF DCIL-10
IN VITRO
BALB/c mice with severe ALD (60 airway
eosinophils on airway allergen challenge) were
given 1x106 DCIL-10, DCGM-CSF, or DCTNF
transtracheally. Over the next 3 weeks they were
assess by head-out body plethysmo-graphy for AHR
to methacholine. This experiment is
representative of 8 others in which we have
found that, beginning at 15 -17 days
post-transplant, the AHR of DCIL-10-treated mice,
but not those treated with either DCGM-CSF or
DCTNF, disappears.
Bone marrow cells from BALB/c mice were cultured
in high (20 ng/ml) , then low (7.5 ng/ml) dose
GM-CSF IL-10 (50 ng/ml). After 15 dy, the
cells were analyzed (A) by FACS for multiple
markers (left panel), in vitro cytokine release
(right panel), (B) phagocytic capacity (left) and
chemokine receptor expression (right). These
DCIL-10 were compared in the FACS analysis with
immature DC GM-CSF and immunostimulatory, mature
DCTNF. DCIL-10 expressed slightly lower levels
of cell surface CD40, CD54 and MHC-II. They also
released significant amounts of IL-10 and TGFb
(A). They possessed functional phagocytosis and
strong chemotaxis to the inflammatory chemokine
MIP-1a (B).
DAY 21
DCIL-10
DCIL-10
DCIL-10
CYTOKINE SECRETION
A.
TGF-b
FACS analysis also confirmed that the DCIL-10
populations did not express neutrophil,
macrophage, B or T cell markers, and that they
did express low levels of CD11c, and DEC205, as
well as expected levels of CD11b, MHC-I CD45RB
DCGM-CSF and DCTNF populations did not express
appreciable levels of IL-10 or TGFb. DCTNF
expressed high levels of TNF and IL-12, while
DCGM-CSF expressed substantial amounts of IL-6,
but not the other cytokines.
B.
FITC-dextran PHAGOCYTOSIS
CHEMOKINE RECEPTORS
REFERENCES -Jonuleit H, Schmitt KE, Steinbrink K
and Enk AH(2001) Dendritic cells as a tool to
induce anergic and regulatory T cells. Trends
Immunol 22 394-400. -Steinbrink K, Matthias W,
Jonuleit H, Jurgen K and Enk AH (1997) Induction
of tolerance by IL-10 treated dendritic cells. J
Immunol 159 4772-80. -Schneider AM, Zhang X, Li
F, and Gordon JR. (2001) Differential induction
of allergen-specific IgA, AHR, or allergic airway
disease following sensitization with limiting
doses of ovalbumin-alum. Cell Immunol 212101-109
CONCLUSION DELIVERY OF DCIL-10, BUT NOT
DCGM-CSF OR DCTNF, INTO THE AIRWAYS OF MICE WITH
ALD ABROGATES AHR SUBSTANTIALLY AMELIORATES Th2
REACTIVITY
In MIP-3a chemotaxis (i.e., CCR7) assays,
DCGM-CSF and DCIL-10 were shown to express low,
but significant, levels of CCR7, while the DCTNF
responded very strongly via this receptor, as
expected (data not shown).
Both DCIL-10 and DCGM-CSF avidly phagocytosed
FITC-dextran, while the DCTNF did not.
SUPPORTED BY GRANTS FROM THE CANADIAN INSTITUTES
FOR HEALTH RESEARCH AND THE SASKATCHEWAN LUNG
ASSOCIATION