This study was aimed to investigate a new method of avoiding graft-vs-host disease (GVHD) through selective elimination of alloreactive donor lymphocytes by using total body irradiation (TBI) and cyclophosphamide (Cy). Female (BALB/c x C57BL/6) F1 mice (H-2(d/b)) as recipients received (60)Co gamma-ray sublethal TBI of 4 Gy on day 0 followed by being inoculated with P388D1 leukemia cell line on day 1, injection of allogeneic splenocytes from C57BL/6 male mice (H-2(b)) was carried out for induction of graft-vs-leukemia (GVL) effect prior to stem cell transplantation (SCT), intraperitoneally injection of cyclophosphamide (Cy) (200 mg/kg) and TBI (9 Gy) was given on day 6. One day later, treated mice were rescued with bone marrow hematopoietic stem cells from (BALB/c x C57BL/6) F1 male mice (H-2(d/b)). The results showed that recipients had no occurrence of leukemia and GVHD through selective elimination of alloreactive donor lymphocytes by Cy and TBI, survived more than 210 days, the complete-donor chimerism occurred on day 21 after transplantation. The ratio of chimerism descended subsequently, but still displayed mixed-chimerism at 90 days. Control mice died of GVHD, leukemia or other death-related-transplantation within 20 to 36 days (P<0.01). It is concluded that to induce GVL effects by MHC mismatched splenocytes given before syngeneic bone marrow transplantation followed by selective elimination of alloreactive donor lymphocytes through TBI and Cy, graft-vs-host disease was thus avoided.
Animals ; Cyclophosphamide ; therapeutic use ; Female ; Graft vs Host Disease ; prevention & control ; Graft vs Tumor Effect ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Leukemia P388 ; therapy ; Lymphocyte Depletion ; Lymphocytes ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Whole-Body Irradiation

To investigate the effect of bone marrow mesenchymal stem cells (BMMSC) on acute graft versus host disease (aGVHD) and graft versus leukemia (GVL) after allogeneic bone marrow transplantation (allo-BMT), both bone marrow cells and BMMSC obtained after three to four weeks of culture from donor mice were transplanted into the recipient mice injected with acute lymphocytic leukemia cells 5 days before, the control group was injected with bone marrow cells alone. The survial time after allo-BMT was recorded; the general manifestation and pathological changes of aGVHD in recipient mice were observed; the effects of BMMSC on the quatity of CD4(+) and CD8(+) T cell in vivo after allo-BMT were evaluated by flow cytometry; chimerism was detected by sex chromosome. The results showed BMMSC could increase obviously the survival time, and delay onset of aGVHD, BMMSC could decrease the amount of CD4(+) T cell and increase CD8(+) T cell in vivo. It is concluded that cotransplantation of bone marrow cells with BMMSC from the same donor mice has GVL effect. BMMSC can alleviate aGVHD and maintain GVL effect after allo-BMT.
Animals ; Bone Marrow Transplantation ; adverse effects ; methods ; CD4-Positive T-Lymphocytes ; cytology ; immunology ; CD8-Positive T-Lymphocytes ; cytology ; immunology ; Cell Line, Tumor ; Female ; Flow Cytometry ; Graft vs Host Disease ; etiology ; immunology ; prevention & control ; Graft vs Leukemia Effect ; immunology ; Leukemia P388 ; immunology ; pathology ; surgery ; Male ; Mesenchymal Stem Cell Transplantation ; adverse effects ; methods ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Transplantation, Homologous

OBJECTIVETo investigate the effect of immature dendritic cells (inDC) genetically modified to express sTNFR I on acute graft-versus-host disease (aGVHD) and the graft-versus-leukemia (GVL) effect ofter allogeneic bone marrow transplantation (allo-BMT) in leukemic mice and its mechanism.

METHODSAn EL4 leukemia allo-BMT model was established with the BALB/c (H-2d) donor mice (DM)and C57BL/6 (H-2b) recipient mice (RM). The RM received DM bone marrow (BM) cells at a 1:1 ratio with spleen cells intravenously via tail vein at 4 h after TBI. Fifty DM were separated randomly into five groups: (1) Group A: total body irradiation (TBI) group, (2) Group B: lymphoma cell leukemia group, (3) Group C: allo-BMT group, (4) Group D: pXZ9-DC group, (5) Group E: sTNFR I-DC group. Acute GVHD scores, incidence of leukemic cell infiltration, histopathological analysis, survival rate, and survival rate of the recipients were estimated after allo-BMT. Enzyme-linked immunosorbent assay (ELISA) method was used to detect cytokines (INF-gamma and IL-4 ) production. Flow cytometry (FCM) analysis was used to detect allogeneic chimerism.

RESULTS(1) The mice in group A and group B all died of the BM failure and lymphoma cell leukemia, respectively. The mice in group C developed typical clinical signs of a GVHD after BMT with an average survival time(AST) of (11.50 +/- 3.50) d. The signs of aGVHD were less evident in the group D and E, and their AST (21.70 +/- 5.80 and 25.80 +/- 5.20 days, respectively) were all longer than that in group C (P < 0.05). AST of group E was the longest (P < 0.05). The mice in group B all died of leukemia within 18 days after engraftment of EL4 cells. There was was no significant difference in groups C, D and E in the incidence of leukemia (P > 0.05). (2) Serum IFN-gamma level reached peak value. At + 12 d, then decreased gradually in group C, D, and E, and then reached the nadir at +18 d post-BMT, with the lowest in group E (P < 0.05), and the level was significantly lower in group D than in group C (P < 0.05). After BMT, serum IL-4 level slightly decreased in group C, but gradually elevated in group D and E and reached their peak at +12 d, and even more significantly increased in group E (P < 0.05). There was no statistical significance in the pair wise comparison among three group (P < 0.05). (3) The average proportion of H-2d positive cells in RM was 95%-100% on day 30 post-BMT, with complete donor-type implantation.

CONCLUSIONImmature DC can induce immuno tolerance. Immature DC genetically modified to express sTNFR I has been shown to prevent acute GVHD in lethally irradiated mice reconstituted with allogeneic bone marrow grafts while maintaining the GVL response.

Animals ; Bone Marrow Transplantation ; adverse effects ; methods ; Dendritic Cells ; immunology ; Female ; Graft vs Host Disease ; prevention & control ; Graft vs Leukemia Effect ; Immune Tolerance ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Receptors, Tumor Necrosis Factor, Type I ; genetics ; Transplantation, Homologous

BACKGROUND: It is apparent that more stem cells can be harvested by mobilization treatment with recombinant human G-CSF and/or GM-CSF from normal healthy donors in allogeneic peripheral blood stem cell transplantation (allo-PBSCT) compared to allogeneic bone marrow transplantation (allo-BMT). It is also known to be more effective in inducing the graft-vs-tumor effects than allogeneic BMT because of higher T cell content. METHODS: A variety of clinical applications with allo-PBSCT was done for patients with he matological malignancies with a high risk of relapse in single transplantation center. We reported the preliminary results on trial of allo-PBSCT followed by planned prophylactic G- and/or GM-CSF primed donor lymphocyte infusion additionally reserved at harvest in hematological malignancies with a high risk of relapse and also presented the successful trial of non-myeloablative transplantation for old aged AML patient in 4th complete remission and cases with 2nd transplantation with allo-PBSCs. RESULTS: Seventeen patients with hematological malignancies with a high risk of relapse were enrolled in trial of allo-PBSCT followed by prophylactic donor lymphocyte infusion. All patients received allogeneic PBSCT from HLA- matched sibling donors. Seven out of 17 patients received additional PBSCs with a median number of CD3+ cells of 5.0x107/kg (range, 3.0 to 9.9x107/kg), between day 41 and day 120. Four surviving patients (4/7) were free of disease when last assessed (median follow-up duration, 538 days), but were suffering from chronic GVHD (1 limited and 3 extensive). A 56 year old acute myeloid leukemia patient in the 4th complete remission was successfully treated with allo-PBSCT with non-myeloablative conditioning regimen. One of 2 patients who received second transplantation with allo-PBSCT has shown a long term disease free survival. CONCLUSION: A merit of allo-PBSCT would allow us to design a variety of clinical applications. Allo-PBSCT might be preferable in special clinical setting such as non-myeloablative transplantation, second transplantation, or the situation in need of the strong GVL effect. And also CSF-primed PBSCs can be used for the purpose of donor lymphocyte infusion.
Bone Marrow Transplantation ; Disease-Free Survival ; Follow-Up Studies ; Graft vs Tumor Effect ; Granulocyte Colony-Stimulating Factor ; Granulocyte-Macrophage Colony-Stimulating Factor ; Hematologic Neoplasms ; Humans ; Leukemia, Myeloid, Acute ; Lymphocytes ; Middle Aged ; Peripheral Blood Stem Cell Transplantation* ; Recurrence ; Siblings ; Stem Cells ; Tissue Donors

Graft vs Host Disease ; metabolism ; surgery ; Graft vs Tumor Effect ; HLA Antigens ; genetics ; Hematopoietic Stem Cell Transplantation ; Humans ; Receptors, KIR ; genetics ; metabolism

Animals ; Antigens, Neoplasm ; immunology ; Cancer Vaccines ; immunology ; Graft vs Tumor Effect ; Humans ; Immunotherapy ; methods ; Neoplasms ; immunology ; therapy ; Th1 Cells ; cytology ; immunology ; transplantation ; Transplantation, Homologous ; immunology

OBJECTIVETo investigate whether whole tumor cell vaccination strategies in combination with bone marrow transplantation (BMT) can stimulate graft-versus-tumor effect (GVT).

METHODSTwenty-six BALB/c mice were randomly divided into 3 groups: BMT group (group A, n = 10), BMT + vaccination group (group B, n = 10), control group (group C, n = 6). (BALB/c × C57BL/6) F1 mice [CB6F1, H-2K(b/d)] were used as donors. BALB/c mice of group C were only inoculated with Renca cell (2.6 × 10(6)). Mice of group A and B were conditioned with 8 Gy irradiation, followed by infusion by bone marrow cell of CB6F1 mice on day 1, then inoculated with Renca cell (2.6 × 10(6)) on day 8. All mice of group B were immunized subcutaneous on the back with 5 × 10(5) irradiated Renca tumor cells on day 9 and day 16. All mice of group C were inoculated with Renca cell (2.6 × 10(6)) on day 8. In group A and B, all mice were analyzed by fluorescence activated cell sorter (FACS) on day 14, and 28 day after BMT. Mice were killed on day 32 after inoculation with tumor cell and collected blood sample. All tumors were taken out to be weighed and then fixed in 10% buffered formalin, embedded in paraffin, and cut into 5 µm slices. The slices were stained with HE and examined by TdT mediated-dUTP nick end labeling (TUNEL). Liver, skin, intestine, and spleen were biopsied for histopathological examination.

RESULTSThe results of chimera showed that engraftments of group A, B were full donor chimerism, and the chimerism of those remained above 90% and preserved even after 28 days. The tumor weight, tumor volume increment in the group B was lower than group A and C (P < 0.05). The tumor suppressing rates of the group A and B were 54%, 60% respectively. The area ratio of tumor necrosis and apoptosis index (AI) of the tumor in the group B were higher than group A and C (P < 0.05). Graft-versus-host disease was not observed in each group.

CONCLUSIONThe mechanism of GVT after haploidentical allogeneic bone marrow transplantation with tumor vaccination may be the promotion of tumor necrosis and apoptosis.

Animals ; Bone Marrow Transplantation ; immunology ; Cancer Vaccines ; immunology ; Carcinoma, Renal Cell ; immunology ; therapy ; Cells, Cultured ; Disease Models, Animal ; Graft vs Tumor Effect ; immunology ; Kidney Neoplasms ; immunology ; therapy ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Transplantation Chimera ; immunology

In treating of leukemia and controlling of minimal-residual disease (MRD), graft versus leukemia effect (GVL) plays a critical role, and complicated mechanisms are involved in this immunology process. When graft cells are infused into recipients, the evoked GVL effect must be inevitably influenced by many factors derived from allogeneic effect between donor and receptor. To utilize GVL more efficiently in future clinical practice and to improve the curative effect of allo-HSCT, it is necessary to recognize these factors. Some potential factors influencing GVL such as chimerism patterns, autocytotoxic cells, dynamics of immune cells in patients, the cytokines and so on are reviewed in this article.
Graft vs Leukemia Effect ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia ; immunology ; surgery

Graft vs Host Disease ; prevention & control ; Graft vs Leukemia Effect ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Transplantation, Homologous ; methods

OBJECTIVETo explore the dissociation of graft-versus-leukemia (GVL) effects from graft-versus-host disease (GVHD) in the patients who experienced GVHD during leukemia relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSThe primary disease, disease status, GVHD, response to donor lymphocyte infusion (DLI) and prognosis were analysed in 11 leukemia patients who relapsed with GVHD after allo-HSCT.

RESULTSOf the 11 relapsed, 5 were acute lymphoblastic leukemia and 6 acute myeloid leukemia. Five received DLI before relapse and all developed post-DLI GVHD, including 2 grade II acute GVHD (aGVHD), 1 limited chronic GVHD (cGVHD) plus grade II aGVHD, and 2 extensive cGVHD. After relapse of the 5 patients, 2 received Chemo-DLI, one achieved CR with extensive cGVHD and then relapsed again, the other didn't achieved CR. The other 6 patients didn't received DLI before relapse and also developed post-HSCT GVHD while relapsing, including 3 extensive cGVHD, 1 grade I aGVHD and 2 grade II-IV aGVHD. After relapse, these 6 patients received Chemo-DLI, 2 achieved CR and then relapsed again, 4 didn't achieved CR.

CONCLUSIONThe elicited GVHD after allo-HSCT may not accompany effective GVL effects inhibiting leukemic relapse.

Graft vs Host Disease ; Graft vs Leukemia Effect ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myeloid, Acute ; Precursor Cell Lymphoblastic Leukemia-Lymphoma