Method to identify antigen-specific immune cells
This disclosure is directed to methods for retrieving and using at least one lymphocyte. Additionally, cell receptor sequences identified with this strategy could be used for antibody development, TCR discovery, or appropriate therapeutics development or evaluation.
1. A method comprising:
collecting a sample suspected of containing at least one immune cell;
re-stimulating the at least one immune cell with an aliquot of an antigen, the at least one immune cell having been stimulated with the antigen prior to at least the re-stimulating step; and
obtaining an image of the at least one immune cell that was re-stimulated during the re-stimulating step.
2. The method of claim 1 , further comprising:
stimulating the at least one immune cell with a separate aliquot of the antigen; and
washing away any excess antigen of the separate aliquot,
wherein the stimulating and washing steps are performed before the re-stimulating step.
3. The method of claim 2 , further comprising the step of incubating the at least one immune cell with the separate aliquot of the antigen.
4. The method of claim 3 , wherein the incubation step is performed after the washing step.
5. The method of claim 1 , further comprising:
stimulating the at least one immune cell with a separate aliquot of the antigen; and
resting the at least one immune cell,
wherein the stimulating step is performed in vivo and before the collecting step,
wherein the resting step is performed after the collecting step, and
wherein the re-stimulating is performed in vitro or ex vivo.
6. The method of claim 1 , further comprising the step of labeling the at least one immune cell with at least one fluorescent probe comprising a fluorescent molecule bound to a ligand which is bound to a biomarker on the at least one immune cell.
7. The method of claim 6 , wherein the labeling step occurs after the re-stimulating step.
8. The method of claim 6 , wherein the obtaining step occurs after the labeling step and before the re-stimulating step.
9. The method of claim 6 , wherein the obtaining step determines that the at least one immune cell was re-stimulated in response to the separate aliquot of the antigen.
10. The method of claim 9 , wherein the obtaining step occurs after the labeling and re-stimulating steps.
11. The method of claim 1 , further comprising the step of generating an assay based on the at least one immune cell that was re-stimulated during the re-stimulating step.
12. The method of claim 1 , wherein the image is a picture, a dot plot, a scatter plot, or a histogram.
13. The method of claim 1 , further comprising the step of enumerating the at least one immune cell present on or within the image.
14. The method of claim 1 , wherein the immune cell is B cell or a T cell.
15. The method of claim 1 , further comprising retrieving the at least one immune cell.
16. The method of claim 15 , wherein the retrieving step is performed after the obtaining step.
17. The method of claim 16 , further comprising the step of analyzing the at least one immune cell that was re-stimulated during the re-stimulating step with an assay.
18. The method of claim 1 , further comprising sequencing a receptor of the at least one immune cell.
19. The method of claim 18 , further comprising generating a receptor-dependent substance based on the sequence of the receptor.
20. The method of claim 19 , wherein the receptor-dependent substance is at least one of a monoclonal antibody, a modified cell expressing the same or substantially the same receptor sequence, or a soluble T Cell Receptor.