|
| Method | Protocol | Readout | Reference |
|
| Neutrophil activation in vitro | Incubation of isolated neutrophils with immune complexes of anti-COL7 antibodies and COL7 | (i) ROS release
(ii) Elastase release | [61] |
|
| Neutrophil activation ex vivo | Incubation of cryosections of human/mouse skin with antibodies against COL7 (serum, IgG, affinity-purified IgG from patients, monoclonal anti-COL7 antibodies, or rabbit anti-COL7 IgG) | (i) Deposition of formazan (ROS detection)
(ii) Extent of dermal-epidermal separation | [60, 62] |
|
|
Anti-COL7 IgG transfer into mice | (a) Repetitive injection of rabbit anti-mouse COL7 IgG into C57Bl/6, BALB/c, or BALB/cnude mice | (i) Body surface area affected by skin lesions
(ii) Several secondary endpoint measurements | [44] |
| (b) Repetitive injection of rabbit anti-human COL7 IgG into SKH1 mice | Same as above | [63] |
| (c) Repetitive injection of human anti-human COL7 IgG into SKH1 mice | Same as above | [64] |
| (d) Repetitive injection of human affinity-purified (CMP) anti-human COL7 IgG into SKH1 mice | Same as above | [57] |
| (e) Repetitive injection of human affinity-purified (Fn3-like domain) anti-human COL7 IgG into SKH1 mice | Same as above | [65] |
| (f) Repetitive injection of rabbit anti-mouse COL7 IgG (vWFA2-like specific) into several in- or outbred mice | Same as above | [66] |
| (g) Repetitive injection of rabbit anti-human COL7 IgG into COL7-humanized mice | Same as above | [67] |
|
|
Immunization of mice with COL7 | (a) Repetitive immunization of SJL/J, BALB/c, and Fc gamma receptor IIB-deficient mice (antigen located within the Fn3-like repeats) | (i) Autoantibody production
(ii) Body surface area affected by skin lesions
(iii) Several secondary endpoint measurements | [45] |
| (b) Single immunization of SJL/J, B6.SJL-H2s, C57Bl/10.s, and MRK/MpJ mice (antigen located within the Fn3-like repeats) | Same as above | [68, 69] |
| (c) Single immunization of SJL/J, B6.SJL-H2s, mice (antigen: vWFA2-like domain) | Same as above | [66] |
|
