European consensus study 20062007

1
European consensus study 2006/2007

• Sera were sent to 43 participants, 41 reported
  results
• Images were sent to 43 participants, 39 reported
  results
• Data entry sheet was used (kindly provided by the
  lab of Prof. Seelig at Karlsruhe)
• All participants returned results electronically
• Data were reported on a scale between 0 and 4
• Data were analyzed using an excel program

2
European consensus study 2006/2007

• Preliminary summary of the results
• Consensus is defined as more than 60 of results
• No consensus results were reported between
  negative and positive without clear distribution
• Results are shown if 3 or more labs reported data
• IIFT patterns for serum 4 and 5 are not correct
  in the handouts!

3
European consensus study 2006/2007Summary of
results
4
European consensus study 2006/2007

• Special emphasis on vasculitis associated
  antibodies 6 sera
• Specificities that will not be mentioned were
  negative

5
Serum 1

• No clinical data

6
Serum 1
ANA (IIFT) n40
Pattern n47
b2 homogenous/neg nucleoli b4 coarse speckled b5
fine speckled c1 homogenous nucleolar c2 clumpy
nucleolar c3 punctate nucleolar
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
7
Serum 1
SS-A/Ro n36 positive
SS-B/La n40 positive
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
8
Serum 1
SS-A/Ro 52 n20 positive
SS-A/Ro 60 n20 positive
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
9
Serum 1
SL/Ki n5 No consensus
1 x CIE, 1 x IB
3 x CIE
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
10
Summary serum 1

• Consensus on SS-A/Ro (Ro52 Ro60) and SS-B/La
  strongly positive
• SL/Ki no consensus
• IgM and IgG rheumatoid factor negative
• IgA rheumatoid factor majority negative but 3 out
  of 11 positive
• Clinical diagnosis?
• Primary Sjögrens syndrome
• Ki/SL positive in 2-8 of primary SS, also seen
  in SLE

11
Serum 2

• No clinical diagnosis

12
Serum 2
ANA (IIFT) n40
Pattern n54
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
b5 fine speckled b9 multiple nuclear dots
b10 coiled body c1 homogenous nucleolar
13
Serum 2
SS-A/Ro n34 positive
SS-B/La n40 low positive
60
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
14
Serum 2
SS-A/Ro 52 n20 positive
SS-A/Ro 60 n20 low positive
70
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
15
Serum 2
Sp100 n6 No consensus
ELISA (IMTEC or INOVA recombinant ag, both
ELISAs are comparable Two labs IB lineblot
from IMTEC, purified antigen from HeLa cells, one
positive, one borderline result One lab IB with
human full length recombinant protein, positive
0 Negative, 1 Borderline, 2 low positive, 3
positive
16
Serum 2
Multiple nuclear dots or coiled body?
Sp100 low positive (3 out of 6)
Coilin p80 negative n2
17
Serum 2

• Consensus on ANA pos
• Pattern fine speckled, multiple nuclear dots,
  coiled body?
• Coilin p80 negative (n2, both IB)
• No consensus on Sp100
• Recombinant versus natural ag?
• SS-A (Ro52 and Ro60) positive, SS-B low positive
• Clinical suggestions
• Sjögren Syndrome
• Primary biliary cirrhosis

18
Serum 3

• No clinical data

19
Serum 3
ANA (IIFT) n40
Pattern n60
b1 homogenous/pos nucleoli b2 homogenous/neg
nucleoli b3 large speckled b4 coarse
speckled b5 fine speckled e1 fine speckled
cytoplasmic e2 diffuse cytoplasmic
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
20
Serum 3
Jo-1 n39 Positive
95
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
21
Serum 3
ds-DNA n40 Very diverse
ss-DNA n6 Majority positive
67
0 Negative 1 Borderline 2 low positive 3 positive
4 high positive
22
Serum 3
Method details a-ds-DNA
CLIFT n21

• ELISA and CLIFT perform similar
• Unicap majority borderline
• Farr majority low positive

0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
23
Serum 3
a-C1q n1 Positive
Nucleosomes n9 No consensus
Histones n10 Majority negative
50
0 Negative 1 Borderline 2 low positive 3 positive
4 high positive
24
Serum 3
MPO n38 Majority negative
p-ANCA n32 30 positive
n21
n32
n9
n4
n1
n1
cathepsin G, lactoferrin, BPI, elastase and
lysozyme negative
25
Summary serum 3

• ANA positive, homogenous / fine speckled
• Jo-1 positive
• ds-DNA diverse, ss-DNA majority positive
• Nucleosomes and histones - majority negative
• C1q positive?
• P-ANCA pattern, no a-MPO
• Clinical suggestions?
• Polymyositis
• SLE

26
Serum 4

• Clinical data scleroderma

27
Serum 4
ANA (IIFT) n40 Low positive
Pattern n39
75
b1 homogenous/pos nucleoli b2 homogenous/neg
nucleoli b5 fine speckled b6 grainy-SCL70
like
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
28
Serum 4
SCL-70 final result, n39 Low positive?
46
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
29
Method details SCL-70
Serum 4

• CIE all negative
• ELISA 38 negative
• FPIA 50 positive
• ALBIA and ID 67 positive
• IB majority positive

76
38
50
67
67
30
Summary serum 4

• ANA low positive
• Pattern diverse, homogenous/pos nucleoli,
  homogenous/ neg nucleoli, fine speckled, grainy
  SCL-70 like
• SCL-70 low positive, IB most sensitive technique!
  
• ELISA, FPIA, ID and ALBIA intermediate
  sensitivity
• CIE low sensitivity, not suitable?
• Usually, anti-SCL-70 present in high titers in
  scleroderma patients
• Patient might have longstanding disease

31
Serum 5

• No clinical data

32
Serum 5
ANA (IIFT) n40
Pattern n37
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
b1 homogenous/pos nucleoli b2 homogenous/neg
nucleoli b5 fine speckled e3 mitochondrial like
33
Serum 5
MPO n41 Strongly positive
P-ANCA n33 High levels
0 Negative, 1 Borderline, 2 low positive, 3
positive, 4 high positive
34
Summary serum 5

• ANA positive
• Pattern homogenous / fine speckled
• P-ANCA, MPO highly positive
• Clinical suggestion
• Systemic vasculitis
• Microscopic polyangiitis

35
Serum 6

• No clinical data

36
Serum 6
ANA (IIFT) n39
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
37
Serum 6
PR-3 n39 Strongly positive
C-ANCA n34 High levels
0 Negative, 1 Borderline, 2 low positive, 3
positive, 4 high positive
38
Summary serum 6

• ANA negative
• C-ANCA, PR-3 highly positive
• Clinical suggestion
• Wegener granulomatosis

39
Serum 7

• No clinical data

40
Serum 7
GBM n27 Highly positive
ANA (IIFT) n40
0 Negative, 1 Borderline, 2 low positive, 3
positive, 4 high positive
41
Summary serum 7

• ANA negative
• GBM high levels (all labs)
• Clinical suggestion
• Anti-GBM disease

42
Serum 8

• Clinical data
• 72 years old woman with acute renal failure
• Antecedents of hypertension and hypothyroidism
• Diagnosis Probable microscopic PAN
• Sudden death during dialysis due to heart attack
• Pathological study heart rupture and massive
  hemopericardial effusion producing death

43
Serum 8
ANA (IIFT) n39
Pattern n36
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
b1 homogenous/neg nucleoli b8 centromere
44
Serum 8
Centromere (IIFT) n38 Positive
CENP-B n28 Positive
0 Negative, 1 Borderline, 2 low positive, 3
positive, 4 high positive
CENP-A n3 positive, CENP-C n1 positive
45
Serum 8
p-ANCA n34 Intermediate levels
MPO n39 Intermediate positive
0 Negative, 1 Borderline, 2 low positive, 3
positive, 4 high positive
46
Serum 8
GBM n41 Highly positive
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
47
Summary serum 8

• ANA positive, centromere pattern
• CENP-A, B and C positive
• p-ANCA and MPO positive
• GBM positive
• Clinical suggestions given by participants
• anti-GBM disease
• systemic vasculitis
• limited scleroderma

48
Serum 9

• Clinical diagnosis Wegeners granumolatosis

49
Serum 9
ANA (IIFT) n39
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
50
Serum 9
PR-3 n40 Intermediate positive
C-ANCA n34 Intermediate levels
0 Negative, 1 Borderline, 2 low positive, 3
positive, 4 high positive
51
Summary serum 9

• Consensus on C-ANCA, PR-3 positive
• Fits with diagnosis

52
Serum 10

• Clinical data renal failure

53
Serum 10
ANA (IIFT) n40
0 Negative 1 Borderline 2 low positive 3
positive 4 high positive
54
Serum 10
Atypical ANCA n27 Negative
C-ANCA n34 Majority negative
P-ANCA n34 Majority negative
0 Negative 1 Borderline 2 low positive 3 positive
4 high positive
C-ANCA P-ANCA atypical ANCA gt1 n25 Thus
majority ANCA IIFT positive, but low level
55
Serum 10
MPO n40 Majority positive
PR-3 n40 Negative
75
56
Summary serum 10

• MPO positive
• ANCA IIFT positive but pattern difficult to
  interpret
• C-ANCA or atypical pattern can be seen with a-MPO
• Clinical suggestion
• Systemic vasculitis

57
Summary vasculitis associated ab

• 1 serum (number 7) GBM positive, very good
  consensus
• 2 sera (number 5 and 8) p-ANCA and MPO positive
• One strong and one intermediate positive
• Good consensus
• 2 sera (number 6 and 9) C-ANCA and PR-3 positive
• One strong and one intermediate positive
• Good consensus
• 1 serum (number 10) ANCA IIFT positive but
  pattern difficult (low ab level), MPO positive
  (75 of participants)

58
Thanks to all participants!