Artificial lymphoid tissue equivalent
The present invention relates to methods of constructing an integrated artificial immune system that comprises appropriate in vitro cellular and tissue constructs or their equivalents to mimic the normal tissues that interact with vaccines in mammals. The artificial immune system can be used to test the efficacy of vaccine candidates in vitro and thus, is useful to accelerate vaccine development and testing drug and chemical interactions with the immune system.
1. A three-dimensional lymphoid tissue equivalent (LTE), comprising: a three-dimensional matrix and lymphoid cells from a single donor distributed therein, wherein said lymphoid cells consist of:
(i) a population of B cells and T cells, and
(ii) a population of dendritic cells, wherein said population of dendritic cells comprises both mature and immature dendritic cells.
2. The LTE of claim 1 , wherein said matrix is in the form of an inverse opal scaffold or a collagen matrix.
3. The LTE of claim 1 , wherein said matrix comprises a material selected from gelatin, collagen, synthetic ECM materials, PLGA, PGA, natural ECM materials, chitosan, protosan, and mixtures thereof.
4. The LTE of claim 1 , wherein the ratio of mature to immature dendritic cells is adjusted, prior to distribution of the dendritic cells in the three-dimensional matrix, so as to control the segregation and activation of the population of T cells and B cells.
5. The LTE of claim 4 , wherein the segregation of T cells is controlled by release of cytokines and/or chemokines by the immature dendritic cells.
6. The LTE of claim 4 , wherein the activation of T cells is controlled by the release of cytokines and/or chemokines by the mature dendritic cells.
7. The LTE of claim 5 , wherein the cytokines and/or chemokines is CXCL13.
8. The LTE of claim 6 , wherein the cytokines and/or chemokines is CXCL13.
9. The LTE of claim 1 , wherein said LTE further comprises stromal cells distributed in said matrix.
10. The LTE of claim 9 , wherein said stromal cells are BLS4 stromal cells.
11. The LTE of claim 9 , wherein said stromal cells are human stromal cells from the same donor as the lymphoid cells.
12. The LTE of claim 9 , wherein said stromal cells form a physical scaffold to support T cell and dendritic cell motility and interactions.
13. A method of evaluating the potential reaction of an animal to an agent, said method comprising:
administering an agent to the LTE of claim 1 ; and
evaluating the B cell and/or T cell responses to said agent.
14. The method of claim 13 , wherein said method further comprising administering one or more cytokines and/or chemokines to the LTE.
15. The method of claim 14 , wherein said one or more cytokines and/or chemokines comprises CCL21.
16. The method of claim 14 , wherein said chemokine is selected from the group consisting of CCL21 and CXCL13.
17. The method of claim 13 , wherein said agent is selected from the group consisting of a vaccine, an adjuvant, an immunotherapy candidate, a cosmetic, a drug, a biologic, and a chemical compound.
18. The LTE of claim 1 , wherein the dendritic cells are from a vaccination site culture.