Primary culture Schwann cell IgG & IgM ICC are performed to test paraneoplastic neuropathy patients’ staining pattern (our unit’s two patients have been included in the experiment)
PATIENTS
The following patients were analyzed by IgG:
- A1) 95-43
- A2)94-292
- A3) 97-543
- A4) 93-40
- A5) 92-147
- A6) 94-55
- B1) 93-38
- B2) 92-133
- B3) 01-848
- B4) 00-1289
- B5) 98-14
- B6) 03-1167
- C1) 04-2
- C2) 13-1127
- C3) 15-2420
- C4) 15-1131
- C5) 11-1167
- C6) 00-307
- D1) 15-2373
- D2) 08-530
- D3) 00-1248
- D4) 90-92
- D5) 04-726
- D6) 92-38
- E1) 98-164
- E2) 03-838
- E3) 05-409
- E4) 04-1323
- E5) 16-3512
- E6) 01-203
- F1) 93-94
- F2) 97-506
- F3) 02-616
- F4) IRB 15-467
- F5) AMC 15-107
Only three patients were tested for IgM staining:
- F1) 93-94
- F4) IRB 15-467
- F5) AMC 15-107
Materials
- Rat primary culture of Schwann grown in coverslips. Fixed with PFA 4% and blocked with GOAT 5% in PBS.
- PBS
- Blocking: GOAT 5% in PBS
- Patient’s sera diluted 1/100 (for IgG) OR 1/40 (for IgM) with GOAT 5% in PBS
- Secondary Abs: GAH 488 IgG (1/1000) (box 25) or GAH 488 IgM (box 21)
- Vectashield mounting medium
RAT PRIMARY CULTURE SCHWANN CELL ICC
- Primary culture Schwann cells grown in coverslips were fixed and blocked with GOAT 5% in PBS and frozen at -80ºC.
- Thaw these cells with a little GOAT 5% in PBS
- Perform ICC:
- Add patient’s sera 1/100 1h at RT.
- Wash 3 times with PBS.
- Add secondary Ab GAH 488 IgG (1/1000) or GAH 488 IgM (1/1000) and incubate 1h at RT.
- Wash 3 times with PBS.
- Mount with Vectashield mounting medium.
RESULTS
All patients are negative except otherwise stated below:
IgG staining:
- A6) 94-55 +
- B4) 00-1289 +
- C4) 15-1131 +
- C6) 00-307 +
- D1) 15-2373 +
- D4) 90-92 +
- E3) 05-409 +
- E4) 04-1323 +
IgM staining:
- F1) 93-94 +++ (IP already performed)
- F4) IRB 15-467 ++ (not as intense as to be an IP candidate)
- F5) AMC 15-107 ++ (not as intense as to be an IP candidate)
