Based on the co-inoculated model , LIDE can provide in vitro induction using monocytes isolated from the same donor with huPBMC into indicated type of Mφ (e.g.> M2 type), and co-inoculation using activated huPBMC, induced Mφ, as well as cancer cells for immunotherapy that could mediate Mφ (de-)polarization and/or phagocytosis.
Flowchart of huPBMC+Mφ+cancer cells co-inoculating model
|M2 type of Mφ was induced in vitro using monocytes isolated from huPBMC.
|MDA-MB-231 human breast cancer cells were seeded in cultural dish, while human PBMC from a healthy volunteer was isolated and co-cultured with MDA-MB-231. Monocytes isolated from PBMC of the same donor were induced in vitro to form M2 type of macrophages. Then the activated huPBMC, induced M2 type of macrophages, and MDA-MB-231 cells were co-inoculated into the right flank in NCG mice. Mice were randomized right after tumor inoculation and dosing were initiated accordingly. Test ab was administrated at 30 mg/kg via i.p., whereas anti-CD47 ab was dosed at 20 μg/dose through i.v. All dosing were performed every other day for a total of 18 injections, while tumor volume and body weight were measured twice a week.
Application of “wildtype” mice for macrophage-involved in vivo efficacy evaluation
As cross reaction between humanized ab (e.g.> anti-CD47 ab versus murine macrophage, or Fc versus murine NK cells) and murine immune cells, specific strain of mice (e.g.> NCG mice with functional murine macrophage, despite the deficiency of T, B, and NK cells, or SCID mice with both NK cells and macrophage, etc.) can be utilized for in vivo evaluation of anti-tumor effect exerted by antibody.
|NALM-6 human B lymphoblastic leukemia cells were inoculated into the right flank in NCG mice. Mice were randomized when average tumor volume grown up to ~120 mm3 and dosing were initiated accordingly. Anti-CD47 ab was dosed at 5 mg/kg through i.v. Dosing were performed bi-weekly for a total of 8 injections, while tumor volume and body weight were measured twice a week.
|#LD1-0029-361915 human multiple myeloma PDX model was inoculated into the right flank in CB17.SCID mice. Mice were randomized when average tumor volume grown up to ~150 mm3 and dosing were initiated accordingly. Anti-CD47 ab and anti-CD38 ab, either in monotherapies or in combination, were dosed at 1 mg/kg through i.p. Dosing were performed bi-weekly for a total of 5 injections, while tumor volume and body weight were measured twice a week.
Macrophage-involved binding and ADCP in vitro using fresh tissues directly from patients
For macrophage-involved in vitro assay, LIDE provides unique platforms for binding and ADCP assays using fresh tissues generated directly from patients.