In vitro method for assessing cytokine storm responses
The present disclosure an in vitro method of assaying the stimulation of a cytokine storm response comprising the steps of: a. co-culturing PBMCs and matched differentiated endothelial cells to provide a system representative of human responses in vivo, and b. exposing the co-cultured cell system to a test agent, c. analyzing the system for the presence of one or more cytokines released after exposing the co-culture system to said test agent, and d. optionally evaluating the response to the test agent in comparison to a response to one or more control agents.
1. An in vitro method of assaying the stimulation of a cytokine storm response comprising:
(a) co-culturing peripheral blood mononuclear cells (PBMCs) and matched differentiated endothelial cells to provide a cell system representative of human responses in vivo;
(b) exposing the co-cultured cell system to a test agent;
(c) analyzing the system for presence of one or more cytokines released after exposing the co-culture system to the test agent; and
(d) optionally evaluating the response to the test agent in comparison to a response to one or more control agents;
wherein the endothelial cells are autologous to the PBMCs; and
wherein the endothelial cells are derived from blood outgrowth endothelial cells.
2. The method of claim 1 , wherein the endothelial cells are capable of being cultured to provide a phenotype; wherein the phenotype is characterized as elongated and aligned.
3. The method of claim 1 , wherein the response of the test agent is evaluated in comparison to one or more control agents.
4. The method of claim 3 , wherein the control agent is a TGN1412 anti-CD28 antibody or an anti-CD52 antibody.
5. The method of claim 4 , wherein the anti-CD52 antibody is alemtuzumab.
6. The method of claim 1 , wherein the analyzed cytokine is CXCL8.
7. The method of claim 1 , wherein exposing the co-cultured cell system to a test agent comprises exposing the co-cultured cell system to an agent which induces cytokine storm in the system, followed by the exposure of the system to a test inhibitor the induced response.
8. The method of claim 7 , further comprising identifying an inhibitor of cytokine storm in the in vitro co-cultured cell system.
9. The method of claim 4 , wherein the endothelial cells are from a biopsy.