Table 1. Erbb Antibodies Used

Table 1. Erbb Antibodies Used

Table 1. erbB antibodies used.

Antibody / Source / Type / Clone / Reactant / Positive control / Dilution / Incubation time and temperature
NCL-EGFR-384 / Novocastra, Newcastle-upon-Tyne, UK / Monoclonal / EGFR.25 / EGFR (c-erbB1 protein) (internal domain) / Human skin / 1:100 / 60 min. at room temperature
NCL-EGFR / Novocastra / Monoclonal / EGFR.113 / EGFR (c-erbB1 protein) (external domain) / Human skin / 1:10 / 60 min. at room temperature
c-erbB-2 / Novocastra / Monoclonal / CB11 / c-erbB2 protein (internal domain) / SKBR-3 breast cancer cell line / 1:40 / 60 min. at room temperature
c-erbB-2 / Immunotech, Marseille, France / Monoclonal / 3B5 / c-erbB2 protein (internal domain) / SKBR-3 breast cancer cell line / Ready to use / 60 min. at room temperature
c-erbB-2 / Novocastra / Monoclonal / 5A2 / c-erbB2 protein (internal domain) / SKBR-3 breast cancer cell line / 1:50 / 60 min. at room temperature
NCL-c-erbB-3 / Novocastra / Monoclonal / RTJ1 / c-erbB-3 protein / Normal kidney / 1:10 / Over night at 4C
c-erbB-4/HER4 / NeoMarkers (Fremont, CA, USA) / Monoclonal / HFR-1 / c-erbB-4 protein / Breast carcinoma / 1:10 / Over night at 4C

Table 2. Patient characteristics.

Case no. / Age (years)/Sex / Prior brain tumor / KPS / Resection grade / Radiotherapy / Postoperative survival (months)
1 / 57/M / No / 80 / Partial / 1.8 Gy x 30 / 16.6
2 / 34/F / No / 80 / Partial / 1.8 Gy x 30 / 29.4
3 / 44/M / Grade II astrocytoma / 90 / Partial / No / 41.5
4 / 28/F / No / 90 / GTR / 1.8 Gy x 30 / 55.4
5 / 57/F / No / 70 / Partial / 1.8 Gy x 30 / 5.6
6 / 50/M / Grade II astrocytoma / 80 / Partial / 1.8 Gy x 30 / 23.7
7 / 67/F / No / 80 / Partial / 1.8 Gy x 30 / 12.1
8 / 39/F / No / 70 / GTR / N.D. / N.D.
9 / 53/M / No / 70 / Partial / 1.8 Gy x 30 / 24.6
10 / 78/F / No / 50 / Biopsy / 3.0 Gy x 13 / 9.8
11 / 34/F / No / 80 / Partial / 1.8 Gy x 30 / 95.8+
12 / 42/M / No / 70 / GTR / 1.8 Gy x 30 / 93.6+
13 / 78/M / No / 50 / Biopsy / No / 1.9
14 / 40/M / No / 70 / GTR / 1.8 Gy x 30 / 53.9
15 / 47/M / Grade II astrocytoma / 90 / GTR / No / 90+
16 / 49/M / No / 50 / Biopsy / No / 0.1
17 / 64/M / No / 70 / GTR / 1.8 Gy x 30 / 76.1+
18 / 48/F / No / 80 / GTR / 1.8 Gy x 30 / 8.3
19 / 57/F / No / 80 / Partial / 1.8 Gy x 30 / 63.8+
20 / 72/M / No / 60 / Biopsy / 1.8 Gy x 30 / 11.1
21 / 36/M / No / 70 / Partial / 3.0 Gy x 13 / 11.2
22 / 56/F / No / 70 / Partial / 2.0 Gy x 30 / 17.9
23 / 53/F / No / 80 / GTR / 2.0 Gy x 30 / 59.8+
24 / 44/F / No / 80 / GTR / No / 59.8+
25 / 37/M / No / 80 / GTR / No / 54.6+
26 / 58/M / No / 80 / Partial / 1.8 Gy x 30 / 28.6
27 / 32/F / No / 70 / Partial / 2.0 Gy x 30 / 16.2
28 / 38/M / No / 80 / Partial / 2.0 Gy x 30 / 35.3+
29 / 71/M / No / 50 / Biopsy / 3.0 Gy x 13 / 10.2
30 / 49/M / No / 70 / Partial / 2.0 Gy x 30 / 30.6+
31 / 44/M / Grade II astrocytoma / 80 / Partial / 1.8 Gy x 30 / 39.4+

M = Male, F = Female, KPS = Karnofsky Performance Status, GTR = Gross total resection, N.D. = No data. (+) denotes that the patient is still alive.

Table 3. Immunohistochemical and FISH data.

Case no. / c-erbB-1
(clone EGFR.25) (internal domain) / c-erbB-1
(clone EGFR.113) (external domain) / c-erbB-2
(clone CB11) (internal domain) / c-erbB-2
(clone 3B5) (internal domain) / c-erbB-2
(clone 5A2)
(internal domain) / c-erbB-3 / c-erbB-4 / EGFR gene amplification (FISH)
1 / 9 / 9 / 4 / 6 / 6 / 9 / 4 / Yes
2 / 9 / 1 / 0 / 9 / 3 / 4 / 2 / No
3 / 6 / 0 / 4 / 3 / 1 / 9 / 0 / No
4 / 6 / 1 / 1 / 9 / 0 / 0 / 0 / No
5 / 0 / 0 / 4 / 6 / 2 / 6 / 4 / No
6 / 1 / 9 / 1 / 4 / 0 / 4 / 0 / Yes
7 / 9 / 9 / 0 / 1 / 0 / 0 / 0 / No
8 / 6 / 2 / 0 / 6 / 4 / 6 / 4 / Yes
9 / 9 / 9 / 2 / 6 / 0 / 6 / 6 / No
10 / 9 / 9 / 0 / 9 / 1 / 9 / 4 / Yes
11 / 4 / 0 / 2 / 6 / 0 / 4 / 4 / Yes
12 / 9 / 4 / 0 / 6 / 2 / 9 / 2 / Yes
13 / 9 / 2 / 4 / 6 / 1 / 9 / 2 / No
14 / 4 / 0 / 0 / 1 / 0 / 0 / 0 / No
15 / 2 / 1 / 0 / 9 / 1 / 6 / 0 / No
16 / 6 / 0 / 0 / 4 / 0 / 6 / 1 / No
17 / 9 / 2 / 0 / 9 / 4 / 9 / 2 / No
18 / 4 / 1 / 6 / 9 / 1 / 4 / 4 / No
19 / 4 / 0 / 0 / 1 / 0 / 4 / 0 / Yes
20 / 6 / 1 / 1 / 6 / 0 / 6 / 1 / Yes
21 / 6 / 4 / 1 / 6 / 0 / 4 / 2 / No
22 / 9 / 6 / 2 / 6 / 6 / 4 / 2 / No
23 / 9 / 3 / 0 / 6 / 0 / 1 / 0 / No
24 / 6 / 0 / 0 / 6 / 3 / 6 / 2 / No
25 / 6 / 2 / 1 / 6 / 3 / 9 / 2 / No
26 / 6 / 4 / 6 / 6 / 2 / 4 / 1 / Yes
27 / 6 / 0 / 0 / 1 / 0 / 2 / 1 / No
28 / 6 / 1 / 0 / 4 / 0 / 4 / 1 / No
29 / 9 / 6 / 0 / 4 / 0 / 0 / 1 / No
30 / 3 / 0 / 0 / 4 / 0 / 2 / 2 / No
31 / 6 / 1 / 0 / 9 / 3 / 9 / 6 / No
Median Staining Index / 6 / 1 / 0 / 6 / 1 / 4 / 2
No. of positive cases/Total no. of cases / 30/31 / 22/31 / 14/31 / 31/31 / 16/31 / 27/31 / 23/31 / 9/31
Percentage positivity / 97 % / 71 % / 45 % / 100 % / 52 % / 97 % / 74 % / 29 %

Table 4.

Multiple Cox regression analyses were used to study the association between c-erbB1-4 expression and survival, adjusting for age at diagnosis, Karnofsky performance status scores, and extent of surgical resection. The association between EGFR amplification and survival was studied in the same manner.

Variable / Hazard ratio / 95% Confidence interval / P
c-erbB1 (clone EGFR.113) staining index / 1.076 / 0.92 to 1.26 / 0.37
Karnofsky performance status / 0.913 / 0.85 to 0.98 / 0.008
Gross total resection / 0.196 / 0.05 to 0.79 / 0.022
Age / 0.998 / 0.95 to 1.05 / 0.95
c-erbB1 (clone EGFR.25) staining index / 0.991 / 0.80 to 1.23 / 0.93
Karnofsky performance status / 0.912 / 0.85 to 0.98 / 0.009
Gross total resection / 0.199 / 0.05 to 0.82 / 0.026
Age / 1.007 / 0.96 to 1.06 / 0.79
c-erbB2 (clone CB11) staining index / 1.642 / 1.17 to 2.30 / 0.004
Karnofsky performance status / 0.854 / 0.78 to 0.94 / 0.001
Gross total resection / 0.324 / 0.08 to 1.38 / 0.13
Age / 0.973 / 0.92 to 1.03 / 0.36
c-erbB2 (clone 5A2) staining index / 1.036 / 0.79 to 1.35 / 0.80
Karnofsky performance status / 0.910 / 0.85 to 0.98 / 0.009
Gross total resection / 0.186 / 0.05 to 0.78 / 0.021
Age / 1.005 / 0.96 to 1.06 / 0.85
c-erbB2 (clone 3B5) staining index / 1.018 / 0.84 to 1.23 / 0.86
Karnofsky performance status / 0.912 / 0.85 to 0.98 / 0.009
Gross total resection / 0.190 / 0.05 to 0.77 / 0.020
Age / 1.006 / 0.96 to 1.06 / 0.81
c-erbB3 (clone RTJ1) staining index / 0.975 / 0.83 to 1.15 / 0.76
Karnofsky performance status / 0.914 / 0.85 to 0.98 / 0.010
Gross total resection / 0.204 / 0.05 to 0.81 / 0.024
Age / 1.008 / 0.96 to 1.06 / 0.74
c-erbB4 (clone HFR-1) staining index / 0.902 / 0.68 to 1.20 / 0.48
Karnofsky performance status / 0.907 / 0.85 to 0.97 / 0.005
Gross total resection / 0.178 / 0.05 to 0.72 / 0.015
Age / 1.003 / 0.96 to 1.05 / 0.90
EGFR gene amplification / 0.689 / 0.23 to 2.11 / 0.51
Karnofsky performance status / 0.919 / 0.86 to 0.99 / 0.021
Gross total resection / 0.175 / 0.04 to 0.73 / 0.017
Age / 1.013 / 0.96 to 1.07 / 0.63