Ex-vivo treatment of peripheral plasmacytoid dendritic cells with IFN-lambda
The present invention provides an ex vivo method of treating plasmacytoid dendritic cells (pDC) in Th2- or Th17-associated diseases by modulating the cytokine expression or secretion using interferon lambda (IFN-λ). For the Th-2 or Th17-associated diseases, pDC cells from a patient having the disease are exposed ex vivo with IFN-λ in an effective amount to inhibit cytokine releases. The IFN-λ exposed pDC are administered back into the patient. The present invention also provides a method of ex vivo IFN-λ treatment of pDC, in conjunction with co-administration of a composition comprising IFN-λ.
1. A method of ex vivo treatment of plasmacytoid dendritic cells (pDC), comprising the steps of:
(a) obtaining peripheral blood from a human subject;
(b) isolating mononuclear cells from said peripheral blood;
(c) isolating pDC from said isolated mononuclear cells using a negative cell selection;
(d) exposing said isolated pDC to interferon-lambda (IFN-λ); and
(e) administering said exposed pDC to said human subject,
wherein said IFN-λ is present in an amount sufficient to inhibit pDC-supported cytokine release in a mixed lymphocyte reaction assay, and
wherein said pDC-supported cytokine is selected from the group consisting of interferon-gamma (IFN-γ), interleukin-13 (IL-13), interleukin-10 (IL-10) and interleukin-17 (IL-17).
2. The method of claim 1 , wherein said negative cell selection is performed by (i) exposing said isolated mononuclear cells to a magnetic bead that is coupled with a monoclonal antibody against a cell surface molecule that is present on said mononuclear cells but absent on pDC, and (ii) removing said mononuclear cells that are bound with said magnetic bead.
3. The method of claim 2 , wherein said cell surface molecule is selected from the group consisting of CD1a, CD3, CD11c, CD14, CD16, CD19, CD20, CD56, BDCA1, BDCA3, and glycophorin A.
4. The method of claim 1 , wherein said IFN-λ is one compound selected from the group consisting of IFN-λ1, IFN-λ2, and IFN-λ3.
5. The method of claim 1 , wherein said IFN-λ amount is in the range about 1 ng/mL to about 200 μg/mL.
6. The method of claim 1 , wherein said IFN-λ amount is in the range of about 100 ng/mL to about 10 μg/mL.
7. The method of claim 1 , wherein said exposing step is performed for about 1 hour to about 24 hours.
8. The method of claim 1 , wherein said exposing step is performed for about 6 hours to about 12 hours.
9. The method of claim 1 , wherein said IL-17 is IL-17A or IL-17F.
10. The method of claim 1 , wherein said human subject is suffering from asthma.
11. The method of claim 10 , wherein said asthma is virus-induced or allergen-induced.
12. The method of claim 1 , wherein said isolated pDC has a cell purity of greater than 95%.
13. The method of claim 1 , wherein said isolated pDC has a cell purity of greater than 97%.
14. The method of claim 1 , wherein said exposed pDC are administered to a human subject in the range of about 5×10 4 cells to about 5×10 6 cells.
15. The method of claim 1 , wherein said exposed pDC are administered to a human subject in the range of about 1×10 6 cells.
16. The method of claim 1 , further comprising the step of: (f) administering a composition comprising IFN-λ to said human subject.
17. The method of claim 16 , wherein said IFN-λ composition is administered nasally, intravaneously, or orally.
18. The method of claim 16 , wherein said step (f) is performed simultaneously or sequentially with step (e).