Combination Of Chemotherapeutics Of Busulfan And Cyclophsphamide To Establish Model Of GVHD In Allogeneic Mice

GVHD is a severe clinical complication in bone marrow transplantation, which iscritically related to the immunesuppression of host. In vivo, it has been successfullyestablished that a model of immunesuppression mouse caused by Total BodyIrradiation (TBI). And this model has been widely used in varied researches aboutorgan transplantation of mouse.However, chemotherapeutics capable of immune suppression are increasinglyused to acquire the immunesuppression instead of TBI in the clinical practice.Therefore, there is an essential need for a mouse model of GVHD based onchemotherapeutics that mimics the clinical settings and reduces the difference fromdifferent conditioning regime.In the present research, female BALB/c mice have been used for the host,received IP BU-CY to produce the immunesuppression. And male C57BL/6miceused for the donor, amount of BMC and spleen cells have been extracted andtransplanted to the host to mimic the clinical state of BMT. After engraftment, GVHDmanifestations and histopathological changes were observed and evaluated. Byusing immunochemistry (IHC) and flow cytometry (FCM)，further details of damage on organs including skin, large intestine, and liver can be observed. Another controlgroup was set with tail vein injection of anti-HMGB1in consideration of GVHDbeing a prospective inflammation.The results showed that the host mice conditioned with BU-CY and allogeneictransplanted exhibited features of GVHD including hunched posture, hair loss, ruffledfur and weight loss, the average survival is12.23±2.41days. Morever, IHC and FCMhave provided further more pathological manifestations, which completely fulfilledthe criteria for GVHD. The present model is reliable and can be developed afterchemotherapy conditioning in allogeneic transplanted mice and to evaluateinvestigation therapies.