IN VITRO IMMUNE SYNAPSE SYSTEM AND METHOD OF IN VITRO EVALUATING IMMUNE RESPONSE USING THE SAME
The present invention relates to an in vitro immune synapse system and a method of in vitro evaluating immune response using the same. The in vitro immune synapse system includes antigen-presenting cells (APCs) and at least one cell type of several specific T cell subtypes isolated from peripheral blood mononuclear cells (PBMCs), all of which is from a same individual of pigs. When a test sample is co-cultured in the in vitro immune synapse system for a given period, it can be determined that the test sample is immunogenic, immunostimulatory or not according to the immunization-related changes of these cells, thereby potentially replacing some kinds of animal experimentation.
1 . An in vitro immune synapse system, comprising:
an antigen-presenting cell; and
at least one cell type of a regulatory T cell, a killer cell and a helper T cell, and
wherein the antigen-presenting cell, the regulatory T cell, the killer cell and the helper T cell are primary cells derived from a same individual, and a number of the antigen-presenting cell is larger than a total number of the regulatory T cell, the killer cell and the helper T cell.
2 . The in vitro immune synapse system of claim 1 , wherein the same individual is a specific pathogen free (SPF) pig.
3 . The in vitro immune synapse system of claim 1 , wherein the antigen-presenting cell is an alveolar macrophage.
4 . The in vitro immune synapse system of claim 1 , wherein the number of the antigen-presenting cell is 2 to 10 times of the total number of the regulatory T cell, the killer cell and the helper T cell.
5 . The in vitro immune synapse system of claim 1 , further comprising a control cell, wherein the control cell is derived from the same individual, and the control cell has at least a CD4 − /CD25 − cell marker.
6 . The in vitro immune synapse system of claim 5 , wherein the control cell is a mononuclear cell.
7 . A method of in vitro evaluating immune response, comprising:
providing an in vitro immune synapse system, wherein the in vitro immune synapse system comprises:
an antigen-presenting cell;
at least one of a regulatory T cell, a killer cell and a helper T cell; and
a control cell, and
wherein the antigen-presenting cell, the regulatory T cell, the killer cell, the helper T cell and the control cell are primary cell derived from a same individual, and a number of the antigen-presenting cell is 2 to 10 times of a total number of the regulatory T cell, the killer cell and the helper T cell;
co-culturing a test sample in the immune synapse system for 24 hours; and
detecting the immune synapse system for an immunization-related change, wherein the immunization-related change comprises an level of toll-like receptor (TLR) gene expression and/or a cytokine concentration, and
wherein when any one of the regulatory T cell, the killer cell, the helper T cell has its immunization-related change significantly differ from the immunization-related change of the control cell, the test sample is determined to be immunogenic or immunostimulatory.
8 . The method of in vitro evaluating immune response of claim 7 , wherein the test sample comprises an antigen or an adjuvant.
9 . The method of in vitro evaluating immune response of claim 7 , wherein the same individual is a SPF pig.
10 . The method of in vitro evaluating immune response of claim 7 , wherein the antigen-presenting cell is an alveolar macrophage, the regulatory T cell has at least a CD4 + /CD25 + cell marker, the killer cell has at least a CD4 − /CD25 + cell marker, the helper T cell has at least a CD4 + /CD25 − cell marker, the control cell is a mononuclear cell, and the control cell has at least a CD4 − /CD25 − cell marker.
11 . The method of in vitro evaluating immune response of claim 7 , wherein the TLR gene expression level comprises the TLR3 gene expression level, the TLR7 gene expression level, and/or the TLR8 gene expression level.
12 . The method of in vitro evaluating immune response of claim 7 , wherein the cell cytokine concentration comprises an interleukin (IL) concentration.