GVHD after chemotherapy conditioning in allogeneic transplanted mice

Abstract

GVHD is a major complication in allogeneic SCT. Available GVHD models are mainly based on radiotherapy-conditioning and/or immune deficient mice. GVHD models based on chemotherapy-based regimens remain poorly studied, despite 50% of all transplantations being chemotherapy based. Our aim was to develop a GVHD model using chemotherapy as conditioning. Female BALB/c (H-2Kd) were conditioned with BU-CY and transplanted with 2 Â 10 7 BM and 3 Â 10 7 spleen cells from either C57BL/6 (H-2 Kb) mice (allogeneic setting) or from male BALB/c to serve as a control group for regimen-related toxicity and engraftment. GVHD manifestations and histopathological changes were evaluated. Chimerism and donor T cells presence in skin, intestine and liver were studied using FACS-, FISH analysis and immunohistochemistry. Allogeneic transplanted mice developed lethal GVHD starting from day þ 7 with both histological and clinical signs. Donor T cells accumulated in recipient skin and intestine with GVHD progression. BM-failure, apoptosis and T-lymphocyte infiltration into target organs were significantly higher in allogeneic when compared with the syngeneic group. No toxicity or GVHD signs were observed in the syngeneic setting. We report a mouse model of GVHD using BU-CY conditioning that represents the most common myeloablativeconditioning regimen in clinical SCT. This model can be utilized to study the role of conditioning on mechanisms underlying GVHD.

Early-phase GVHD gene expression profile in target versus non-target tissues: kidney, a possible target?

“…17 Briefly, recipient mice received BU (80 mg/kg) followed by CY (200 mg/kg). At day 0, recipient mice were injected via the lateral tail vein with 2 Â 10 7 and 3 Â 10 7 viable cells of bone marrow cell (BMC) and splenocyte (SP) from C57BL/6 (allogeneic) or BALB/c (syngeneic) as donor, respectively.…”

Omega-3 from fish oil augments GVHD through the enhancement of chemotherapy conditioning regimen and selective FoxP3 depletion

“…To elucidate the effects of these compounds on the host immune system during conditioning chemotherapy and BMT, we used our previously established chemotherapy-based GVHD mouse model. 35 Several novel findings of potential clinical relevance emerged.…”

“…On day 14, 15 mice from each group received a myeloablative BU-CY conditioning regimen as previously described. 35 Briefly, animals were administered BU (80 mg/kg) (Sigma-Aldrich, Stockholm, Sweden) for 4 days followed by CY (200 mg/kg) (SigmaAldrich, Stockholm, Sweden) for 2 days. Thereafter, the animals were rested for 1 day before BMT.…”

“…Thereafter, all the transplanted animals were fed a standard diet and monitored for the development of GVHD as described previously. 35 For the 2-month exposure period, FO-, CO-and standard-fed groups were conditioned with BU-CY on day 49 and killed on day 56.…”

“…After transplantation, all groups were monitored for the development of GVHD as described previously. 35 Preparation of cell suspensions from the BM and spleen BM cells were flushed out of the femurs using sterile phosphate-buffered saline containing 2% fetal calf serum and 0.01 mM EDTA by means of a syringe connected to a 27-gauge needle and subsequently washed twice with phosphate-buffered saline. The spleens were cut into smaller pieces and gently passed through a 70-mm strainer (BD Falcon, BD, Franklin Lakes, NJ, USA) using a syringe plunger.…”