Objective

Introduction: We have developed two human in-vitro skin explant tests to assess the safety, efficacy and dose response to biopharmaceuticals from a hypersensitivity and immunogenicity perspective. The Skimune™ Mab assay tests biologics using human blood and non-artificial autologous human skin from a healthy donor to assess for histopathological damage. This can also be correlated to T cell proliferation and IFN-ƴ release screening assays which give a predictive read out for adverse immune reactions. This skin explant assay can be modified to also test immunomodulatory biologics, biosimilars, antibody-drug conjugates as well as to investigate biopharmaceutical mode of action. Methods: Briefly, Skimune™ Mab involves co-culture of peripheral blood mononuclear cells (PBMCs) in autologous human serum with autologous skin taken from healthy donors. Following co-culture the skin is fixed in formalin for 24 hours and Haematoxylin & Eosin stained. Skin sections are then graded according the Lerner classification system (grade I negative or grade II-IV positive) which correlates to immune damage. We have tested a wide range of therapeutic biologics in the Skimune™ Mab assay in different healthy volunteers (n=10), results have shown a statistically strong positive correlation with histopathological grading in Skimune™ Mab assay to expected clinical response (r=0.86, p=0.0001). Additionally, data from an analogue of TGN (TGN1412) showed a strong grade III response in the Skimune™ MAb assays, indicating that this assay could have predicted the serious life-threatening cytokine storm observed in the 2006 Northwick park trial disaster. We have also assessed the efficacy and dose response of immunomodulatory antibodies by setting up a Skimune™ Mab assay with alloreactive cells giving a Grade III response and adding different concentrations of a test immunomodulatory antibody 0.15 µg/ml and 1.5 µg/ml resulting in skin damage from a grade III to a grade I response, respectively. Conclusion: The novel Skimune™ skin models can provide vital information on the safety, efficacy and dose response to biopharmaceuticals including monoclonal antibodies, biosimilars and antibody drug conjugates, as well as providing information on adverse immune reactions from a hypersensitivity perspective. The novel Skimune™ skin models can provide vital information on the safety, efficacy and dose response to biopharmaceuticals including monoclonal antibodies, biosimilars and antibody drug conjugates, as well as providing information on adverse immune reactions from a hypersensitivity perspective.

Methods

Methods: Briefly, Skimune™ Mab involves co-culture of peripheral blood mononuclear cells (PBMCs) in autologous human serum with autologous skin taken from healthy donors. Following co-culture the skin is fixed in formalin for 24 hours and Haematoxylin & Eosin stained. Skin sections are then graded according the Lerner classification system (grade I negative or grade II-IV positive) which correlates to immune damage.

Results

We have tested a wide range of therapeutic biologics in the Skimune™ Mab assay in different healthy volunteers (n=10), results have shown a statistically strong positive correlation with histopathological grading in Skimune™ Mab assay to expected clinical response (r=0.86, p=0.0001). Additionally, data from an analogue of TGN (TGN1412) showed a strong grade III response in the Skimune™ MAb assays, indicating that this assay could have predicted the serious life-threatening cytokine storm observed in the 2006 Northwick park trial disaster. We have also assessed the efficacy and dose response of immunomodulatory antibodies by setting up a Skimune™ Mab assay with alloreactive cells giving a Grade III response and adding different concentrations of a test immunomodulatory antibody 0.15 µg/ml and 1.5 µg/ml resulting in skin damage from a grade III to a grade I response, respectively. Conclusion: The novel Skimune™ skin models can provide vital information on the safety, efficacy and dose response to biopharmaceuticals including monoclonal antibodies, biosimilars and antibody drug conjugates, as well as providing information on adverse immune reactions from a hypersensitivity perspective.

Conclusion

The novel Skimune™ skin models can provide vital information on the safety, efficacy and dose response to biopharmaceuticals including monoclonal antibodies, biosimilars and antibody drug conjugates, as well as providing information on adverse immune reactions from a hypersensitivity perspective.