P72
IL-17-/- MICE ARE PROTECTED FROM NEPHROTOXIC NEPHRITIS AND ARE
RESISTANT TO DISEASE INDUCTION BY ADOPTIVE CELL TRANSFER
Hamour, S1, Cook, T1, Iwakura, Y2, Pusey, C1, Salama, A1
1
Imperial College Kidney and Transplant Institute, London, 2Centre for Experimental
Medicine, University of Tokyo
INTRODUCTION: Accelerated nephrotoxic nephritis (NTN) is a well-established animal model of
immune-complex mediated glomerulonephritis. Th17 cells are a distinct lineage of CD4+ T-helper
cells characterised by the production of IL-17, implicated in autoimmunity. This group and others
have previously shown that IL-17-/- mice are protected from NTN. Here we show that IL-17-/- mice
remain resistant to disease even following adoptive transfer with wild-type (WT) splenocytes or T
cells.
METHODS: Standard protocol for induction of accelerated NTN was followed. From WT or IL-17-/animals we isolated and transferred 8, 12 or 18 million splenocytes; 3 or 5 million purified T cells or 8
million non-T cells into WT and IL-17-/- animals undergoing induction of NTN. Cells were transferred
on the same day as pre-immunisation with sheep IgG/CFA or following isolation from pre-immunised
animals, two days prior to NTS injection. Purity of cell fractions was confirmed by FACS. Cell
fractions were labelled with PKH26 red fluorescent dye for in vivo tracking.
RESULTS: IL-17-/- mice were significantly protected from NTN as demonstrated by preserved renal
function, minimal proteinuria(Figure) and diminished glomerular thrombosis. Following adoptive
transfer of WT cells, PKH26-labelled cells were demonstrable in the kidney of IL-17-/- mice 2 weeks
after cell transfer confirming that cells were viable and had trafficked to the kidney. Despite high
numbers of cells transferred, IL-17-/- animals remained significantly resistant to NTN with preserved
renal function, less proteinuria (Figure) and less glomerular thrombosis than WT animals receiving
similar WT cell transfer. Moreover, transfer of 3x106 IL-17-/- T cells into WT animals conferred no
protection from disease, suggesting that a regulatory T cell population was not responsible.
45
40
35
30
25
20
15
10
5
0
10.0
serum urea
mmol/L
7.5
5.0
2.5
***
0.0
WT
IL-17-/-
n=7
n=9
*
*
6
5
4
3
2
1
20
10
/ls
to
to
IL
-1
7-
IL
-1
7/-
IL
-1
7/to
ce
l
no
nT
T
T
ce
lls
W
T
T
W
W
en
oc
yt
es
sp
le
T
W
W
T
W
no
n
T
T
T
ce
no
cy
t
lls
es
to
to
IL
17
W
T
-/-
-/IL
17
to
ce
l
te
s
oc
y
sp
le
n
T
W
ls
to
te
s
to
IL
17
W
-/-
T
0
oc
y
sp
le
n
T
IL17-/-
n=10
30
0
W
WT
n=8
40
serum urea mmol/24h
proteinuria mg/24h
7
**
sp
l
Proteinuria mg/24hrs
12.5
CONCLUSIONS: IL-17-/- mice are resistant to induction of NTN and remain protected despite
adoptive transfer of WT splenocytes, purified WT T cells or non-T cell fractions. These data suggest
that there may be other intrinsic mechanisms of disease protection, unrelated to Th17 cell induction.
Further investigation is underway to define the mechanisms by which this protection is mediated.