Objectives
To test some of the most positive neuron-staining Zika-GBS patients for IgG and IgM positivity towards human AXL and GAS6 in HEK 293 transfected cells.
Patients
- ZN004A (lab id # 5)
- ZN0018A (lab id # 13)
- ZN008A-5 (LAB ID #18)
- ZN007A (lab id # 21)
- ZN052A (lab id # 22)
- N009A-4 (lab id #26)
AXL ICC
- Human AXL transfected HEKs 293 grown in coverslips (plates). Fixed with PFA 4%, permeabilized and blocked with 5% goat serum in PBS.
- PBS
- Blocking: 5% goat serum in PBS
- Patient’s sera diluted 1/100 (IgG) or 1/40 (IgM) with 5% goat serum in PBS.
- Primary Ab: Mouse myc (1/200)
- Secondary Abs: GAM 488 (1/500)+ GAH 594 (1/500) IgG or IgM
- Vectashield mounting medium
Note: due to restrictions in the number of transfected plates, patient N009A-4 (lab id #26) could not be tested for IgM.
GAS6 ICC
- Human GAS6 transfected HEKs 293 grown in coverslips ( plates). Fixed with PFA 4%, permeabilized and blocked with 5% goat serum in PBS.
- PBS
- Blocking: 5% goat serum in PBS
- Patient’s sera diluted 1/100 (IgG) or 1/40 (IgM) with5% goat serum in PBS
- Primary Ab: Mouse myc (1/200)
- Secondary Abs: GAM 488 (1/500)+ GAH 594 (1/500) IgG or IgM
- Vectashield mounting medium
Note: due to restrictions in the number of transfected plates, patient N009A-4 (lab id #26) could not be tested for IgM.
RESULTS
All samples are negative. In future experiments the remaining Zika-GBS patients will be tested.
