Populations

(Antibodies used)

Untreated rats (%)

LF15-0195
treated rats (%)

CD4+ T cells
(R7.3 + W3.25)

41.6 2.8

39.5 1.6

p= 0.52

CD8+ T cells
(R7.3 + Ox 8)

23.6 0.8

23.5 0.9

p= 0.6

B cells
(Ox33 + Ox12)

14.9 0.8

12.2 2

p= 0.14

NK cells
(3.2.3 + Ox 8)

4.5 0.2

4.2 0.5

p= 0.6

1.2 0.1

1.5 0.3

p= 0.85

Monocytes
-Macrophages
(ED3 + Ox 42)

Table I : FACS analysis of LF15-0195-treated Buff/Mna (at the end of the treatment) versus
untreated age-matched Buff/Mna rats. Results are expressed as the percentage of positive
cells for the two noted antibodies.

Rat mRNA

Forward primer

Reverse primer

HPRT

CCTTGGTCAAGCAGTACAGCC

TTCGCTGATGACACAAACATGA

CD25

GCTCCAGGCACTGACCCTAG

CTGGGCTGACAATGAGGACTC

cPLA2

GCAGCAGAGGAAAGAGAACATAA

GTAGGTCGCACAATCCAAAATC

CTLA4

GGCAGACAAATGACCAAGTGAC

TCTGAATCTGGGCATGGTTCT

FoxP3

CCCAGGAAAGACAGCAACCTT

CTGCTTGGCAGTGCTTGAGAA

GITR

GCAGACTTTGGACCAACTGTTC

AGCGGCTGGGTATTGACCT

IDO

GCTGCCTCCCATTCTGTCTT

TGCGATTTCCACCATTAGAGAG

IKB

CAGCGTCTGAAGATTTTCCA

AGTAACCTACCAGGGCTACTCC

IL10

TGCTATGTTGCCTGCTCTTACTG

TCAAATGCTCCTTGATTTCTGG

IL13

TATGGAGCGTGGACCTGACA

GCGGAAAAGTTGCTTGGAGTA

iNOS

GACCAAACTGTGTGCCTGGA

TACTCTGAGGGCTGACACAAGG

NF-KB

GCATCCACAGCTTCCAGAAC

GCACGGTTATCAAAAATCGG

TNF-

CTTATCTACTCCCAGGTTCTCTTCAA

GAGACTCCTCCCAGGTACATGG

Table II: List of primers used in this article

Legends
Figure a-c: Effect of CS (a), CsA (b), and the CS-CsA combination (c) on proteinuria in
Buff/Mna rats. Each treatment was administered for 60 days. Each group consisted of 5
animals. Proteinuria is expressed as the ratio of the urinary proteins (g/l) to urinary creatinine
concentration (mmol/l). All values are expressed as mean SEM (scale bar). * indicates a
significant decrease in proteinuria (vs Day 0).
During these treatments, serum creatinine remained stable and within the normal range (CS:
26.2 3.5 mol at Day 0 vs. 29 2.9 mol at Day 60; CsA: 29.4 4 mol vs. 29 3.3 mol
at Day 60; p=ns). Histological examination of kidneys from CS and/or CsA-treated rats failed
to reveal any ultrastructural changes compared with vehicle-treated rats
Figure d: Effect of anti-T lymphocyte treatment using R7.3 Ab on proteinuria in Buff/Mna
rats. R7.3 Ab (IP; 50 g/d) and CsA (per os ; 10 mg/kg/d; to slow down the anti-R7.3
immunization) was injected simultaneously for 20 days. The isotypic control Ab (3G8)
associated with olive oil (CsA vehicle) was administered as a control group. Each group
consisted of 7 animals. Proteinuria is expressed as the ratio of the urinary proteins (g/l) to
urinary creatinine concentration (mmol/l). All values are expressed as mean SEM (scale
bar). * indicates a significant decrease in proteinuria (vs Day 0).
The administration of R7.3 Ab rapidly decreased both T cells (90.5 3.6% of WBC at Day 0
vs. 22.3 3.3 % at day 28, a 4.5 fold decrease) and proteinuria (65% of reduction), but this
decrease was transient and did not reach normalization.
Figure e: Effect of LF15-0195 on proteinuria of Buff/Mna rats with different initial proteinuria
levels. 59 animals were treated. We grouped together them according to their initial
proteinuria range. Proteinuria is expressed as the ratio of the urinary proteins (g/l) to urinary
creatinine concentration (mmol/l). All values are expressed as mean SEM (scale bar).
Figure f: Effect of PDTC on Buff/Mna proteinuria. Treatment was administered for 30 days.
The group consisted of 5 animals. Proteinuria is expressed as the ratio of the urinary proteins
(g/l) to urinary creatinine concentration (mmol/l). All values are expressed as mean SEM
(scale bar).
Figure g: Effect of irradiation (4 gray) on proteinuria of Buff/Mna rats. The group consisted of
5 animals. Proteinuria is expressed as the ratio of the urinary proteins (g/l) to urinary
creatinine concentration (mmol/l). All values are expressed as mean SEM (scale bar).