Animals ; Leukocyte Count ; Leukocytes ; drug effects ; Male ; Mice ; Nerve Growth Factor ; pharmacology ; Skin ; pathology ; radiation effects ; Survival Rate ; Whole-Body Irradiation ; adverse effects ; mortality ; Wound Healing ; drug effects

Long-term survivors of hematopoietic stem cell transplantation (HSCT) during childhood and adolescence are at risk of developing endocrine complications. The purpose of this study was to evaluate the long-term endocrine complications and their associated risk factors among such patients. We reviewed the data from 111 patients (59 males and 52 females) who underwent HSCT at the mean age of 8.3+/-4.1 yr. Thirty patients (27.0%) had growth impairment, and seven (21.2%) out of 33 patients who attained final height reached final height below 2 standard deviation (SD). The final height SD score of the patients conditioned with total body irradiation (TBI) was significantly lower than that of the patients conditioned without TBI (-1.18+/-1.14 vs. -0.19+/-0.78, P=0.011). Thirteen patients (11.7%) developed hypothyroidism (11 subclinical, 2 central) 3.8+/-1.8 (range 1.6-6.2) yr after HSCT. Nineteen (65.5%) out of 29 females had evidence of gonadal dysfunction, and 18 (64.3%) out of 28 males had evidence of gonadal dysfunction. The risk for gonadal dysfunction was significantly higher in females conditioned with busulfan/cyclophosphamide (P=0.003). These results suggest that the majority of patients treated with HSCT during childhood and adolescence have one or more endocrine complications. Therefore, multiple endocrine functions should be monitored periodically after HSCT until they reach adult age.
Adolescent ; Adult ; Body Height ; Child ; Child, Preschool ; Endocrine System Diseases/*etiology/physiopathology ; Female ; Gonadal Disorders/etiology ; Growth Disorders/etiology ; Hematopoietic Stem Cell Transplantation/*adverse effects ; Humans ; Infant ; Male ; Thyroid Diseases/etiology ; Transplantation Conditioning/adverse effects ; Whole-Body Irradiation/adverse effects

The objective of this study was to investigate the function and mechanism of hyperbaric oxygen (HBO) in antagonizing acute graft-versus-host disease (aGVHD) and improving the rate of survival. The lethally irradiated C57BL/6 recipients were injected with bone marrow and lymphocyte of spleen from BALB/c donors and were treated with HBO, cyclosporine A (CsA) and methotrexate (MTX). T lymphocytes and subsets, adhesion molecules and cytokines were detected by flow cytometry, ELISA and RT-PCR respectively. The results showed that the survival rate in HBO group was much higher than that in allogenetic bone marrow transplantation (allo-BMT) group and CsA + MTX group; the numbers of CD3(+), CD4(+), CD8(+), CD4(+)CD11a(+), CD4(+)CD18(+), CD8(+)CD11a(+), CD8(+)CD18(+) lymphocytes in spleen were decreased markedly by HBO and CsA + MTX (p < 0.05); the levels of IL-2 and TNFalpha mRNA and their serum concentrations in HBO group were much lower than those in allo-BMT group but were higher than those in CsA + MTX group; the levels of IL-4 and IL-10 mRNA in HBO group were much higher than those in allo-BMT group and CsA + MTX group. It is concluded that HBO has more remarkable advantage in improving the rate of survival than CsA + MTX, its mechanism of anti-aGVHD is tightly correlated with the transform of T cell and its subsets and the expression of adhesion molecules and cytokines.
Acute Disease ; Animals ; Bone Marrow Transplantation ; adverse effects ; Cytokines ; biosynthesis ; Female ; Graft vs Host Disease ; etiology ; therapy ; Hyperbaric Oxygenation ; Lymphocyte Transfusion ; adverse effects ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; T-Lymphocytes ; immunology ; Whole-Body Irradiation

OBJECTIVETo study the radioprotective effect of catechines against radiation injury in mice.

METHODSCatechines were administered in mice intragastrically at the daily dose of 200 mg/kg for 10 consecutive days before whole body irradiation with 6 Gy X-rays. The body weight changes, survival time, 30-day survival rate, and counts of peripheral white blood cells were recorded.

RESULTSThe mice with catechine pre-treatment before X-ray exposure suffered less body weight loss than those without the treatment before exposure. Catechines markedly increased the survival time of the irradiated mice, and raised the 30-d survival rate of the irradiated mice to 53.33% as compared with the rate of 13.33% in the radiated mice without catechine pre-treatment. Catechines significantly promoted recovery of peripheral white blood cells.

CONCLUSIONCatechines have definite radioprotective effect against radiation injury in mice.

Animals ; Catechin ; pharmacology ; Female ; Leukocyte Count ; Mice ; Radiation Injuries, Experimental ; blood ; mortality ; prevention & control ; Radiation-Protective Agents ; pharmacology ; Random Allocation ; Survival Analysis ; Survival Rate ; Time Factors ; Whole-Body Irradiation ; adverse effects

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

OBJECTIVETo test whether vaccination performed during irradiation or chemotherapeutics-induced lymphopenia-driven T cell proliferation could augment the antitumor immunity.

METHODSThe study composed of two parts, investigating the anti-tumor efficacy of performing tumor vaccination during early immune reconstitution period following sublethal total body irradiation and cyclophosphamide (Cy)-induced lymphopenia, respectively. Mice were subsequently reconstituted with naïve splenocytes from syngeneic mice and were named RLM (Reconstituted lymphopenic mice). Immunization/vaccination (F10) and adoptive immunotherapy (D5-G6) were used to explore anti-tumor immune responses in vaccinated irradiation/RLM and vaccinated Cy/RLM, respectively. Both normal C57BL/6 mice and RLM were vaccinated with irradiated, weakly immunogenic F10 melanoma cells and subsequently challenged with F10 cells. In addition, to determine the role of CD4(+) and CD8(+) T cells in the protective anti-tumor immune response, irradiation/RLM were depleted of these subpopulations by administration of the appropriate mAb around challenge. In the second part, adoptive immunotherapy was used to evaluate the anti-tumor immune responses under chemotherapeutics-induced lymphopenic condition. Both normal mice and RLM (Cy-treated) were vaccinated with GM-CSF-modified D5 melanoma cells (D5-G6) and tumor vaccine draining lymph nodes (TVDLN) were harvested 9-10 days later. Effector T cells were generated in vitro from TVDLN cells and adoptively transferred to mice bearing 3-day pre-established pulmonary metastases (D5). Recipient mice were sacrificed 2 weeks later after tumor inoculation and pulmonary metastases were enumerated.

RESULTSSignificantly greater protection was induced in vaccinated irradiation/RLM, compared to vaccinated normal mice (63.2% vs 16.7%). Protective immunity in RLM depended on CD8(+) T cells. Increase in the interval between reconstitution and vaccination significantly decrease the protection. Effector T cells generated from vaccinated Cy-treated RLM demonstrated significantly higher in vivo anti-tumor efficacy over those of vaccinated normal mice.

CONCLUSIONThis study suggests that vaccination of RLM could elicit augmented antitumor immunity compared to normal hosts, highlighting the potential clinical benefit of performing tumor vaccination during irradiation or chemotherapeutics-induced lymphopenia in cancer patients.

Animals ; CD8-Positive T-Lymphocytes ; immunology ; Cancer Vaccines ; therapeutic use ; Cyclophosphamide ; adverse effects ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; immunology ; Immunotherapy, Adoptive ; methods ; Lymphopenia ; etiology ; therapy ; Melanoma, Experimental ; drug therapy ; immunology ; radiotherapy ; Mice ; Mice, Inbred C57BL ; Whole-Body Irradiation

The purpose of the study was to investigate the influencing factors of mortality rate in the bone marrow transplantation in mice. The recipient mice receiving whole-body irradiation of gamma-ray were infused with the same strain of bone marrow cells or the mixture of the bone marrow cells and splenocytes respectively. Experiments were carried out in four batches, with different strains of mice used, respectively. The manifestations and the appearance of graft-versus-host disease (GVHD) were observed, as well as the mortality rate within 35 d of the transplantation in the recipient mice. The mortality rate of the first group of recipient mice was the lowest, the mortality rate of the second group of recipient mice was the highest and the obvious GVHD performance was observed before death. In the third group, the mortality rate declined and there was statistical significance compared to that of the second group. The mortality rate of the fourth group of mice was higher than that of the third group, but still lower than that of the second group of mice and there is a statistical significance. This evidence suggested that mouse genetic purity, splenocytes, the ratio of the bone marrow cells and splenocytes and the week-old of the mouse could be the important influencing factors of the mortality rate in mouse bone marrow transplantation.
Animals ; Bone Marrow Transplantation ; adverse effects ; methods ; mortality ; Cell Transplantation ; Female ; Graft vs Host Disease ; immunology ; mortality ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred ICR ; Mice, Transgenic ; Spleen ; cytology ; Survival Rate ; Whole-Body Irradiation

BACKGROUNDRadiation-induced injury after accidental or therapeutic total body exposure to ionizing radiation has serious pathophysiological consequences, and currently no effective therapy exists. This study was designed to investigate whether transplantation of allogeneic murine compact bone derived-mesenchymal stem cells (CB-MSCs) could improve the survival of mice exposed to lethal dosage total body irradiation (TBI), and to explore the potential immunoprotective role of MSCs.

METHODSBALB/c mice were treated with 8 Gy TBI, and then some were administered CB-MSCs isolated from C57BL/6 mice. Survival rates and body weight were analyzed for 14 days post-irradiation. At three days post-irradiation, we evaluated IFN-γ and IL-4 concentrations; CD4(+)CD25(+)Foxp3(+) regulatory T cell (Treg) percentage; CXCR3, CCR5, and CCR7 expressions on CD3(+) T cells; and splenocyte T-bet and GATA-3 mRNA levels. CB-MSC effects on bone marrow hemopoiesis were assessed via colony-forming unit granulocyte/macrophage (CFU-GM) assay.

RESULTSAfter lethal TBI, compared to non-transplanted mice, CB-MSC-transplanted mice exhibited significantly increased survival, body weight, and CFU-GM counts of bone marrow cells (P < 0.05), as well as higher Treg percentages, reduced IFN-γ, CXCR3 and CCR5 down-regulation, and CCR7 up-regulation. CB-MSC transplantation suppressed Th1 immunity. Irradiated splenocytes directly suppressed CFU-GM formation from bone marrow cells, and CB-MSC co-culture reversed this inhibition.

CONCLUSIONAllogeneic CB-MSC transplantation attenuated radiation-induced hematopoietic toxicity, and provided immunoprotection by alleviating lymphocyte-mediated CFU-GM inhibition, expanding Tregs, regulating T cell chemokine receptor expressions, and skewing the Th1/Th2 balance toward anti-inflammatory Th2 polarization.

Animals ; Bone and Bones ; cytology ; Cytokines ; metabolism ; Female ; Granulocyte-Macrophage Progenitor Cells ; cytology ; Male ; Mesenchymal Stem Cell Transplantation ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Whole-Body Irradiation ; adverse effects

To explore the effect of bone marrow camouflaged with methoxy polyethylene glycol (mPEG) on allogeneic bone marrow transplantation, 60 BALB/c(H-2d) mice were randomly divided into 3 groups after irradiation by 8.0 Gy of (60)Co gamma ray. A group was given RPMI 1640 0.5 ml in tail vein. B group was infused with the bone marrow cells (1 x 10(7)) mixed with the spleen cells (1 x 10(7)) of donor 615(H-2k) mice. C group was transplanted with same dose cells, which were camouflaged with mPEG before infusion. Severity GVHD was determined by total manifestation of mice, survival rate, survival time and histo-pathological microscopy, and engraftment of allogeneic bone marrow was evaluated by chromosome examination. The results showed that 75% mice in B group had severe adverse manifestations, such as hunched posture, diarrhea and loss of hair. Occurrence of the same adverse manifestations in C group was 35% and significantly lower than that in B group (P Animals ; Bone Marrow Transplantation ; adverse effects ; methods ; Female ; Graft vs Host Disease ; etiology ; mortality ; prevention & control ; Leukocyte Count ; Male ; Mice ; Mice, Inbred BALB C ; Polyethylene Glycols ; therapeutic use ; Random Allocation ; Survival Analysis ; Survival Rate ; Transplantation, Homologous ; Whole-Body Irradiation

To explore whether the complete donor chimerism could be achieved and graft-versus-host disease could be alleviated by donor lymphocyte infusion which was sensitized by the skin of the recipient, female C57BL/6 mice (H-2(b), B6) as recipients received total body irradiation (TBI) of 5.5 Gy ((60)Co gamma-ray) on day 0 followed by allogeneic hematopoietic stem cell transplantation (allo-HSCT). The allo-grafts consisted of 2 x 10(7) peripheral hematopoietic stem cells from mobilized male BALB/c (H-2(d)) donor mice with the granulocyte colony-stimulating factor (G-CSF). Day 2 after allo-HSCT, the recipient mice were given 200 mg/kg cyclophosphamide intraperitoneally. Afterwards these recipient mice were infused 2 x 10(6) sensitized or unsensitized-donor lymphocytes at the 28 days after transplantation. The results showed that the mice receiving sensitized-donor lymphocyte infusion did not suffer from GVHD and the phenotypic character of the recipient mice (black color) converted to that of the donor mice (white color), and to become full-donor chimerism. It was found that the ratio of CD4(+)/CD8(+) T lymphocytes of them decreased at the earlier period and increased after half month, but which were also lower than that of the normal value. While various grades of acute GVHD was observed in that of the control group and the mixed-chimeras were maintained, though it increased a little, and the ratio of CD4(+)/CD8(+) T lymphocytes increased at first, then decreased to the normal level half month later. It is concluded that sensitized DLI converted mixed to complete donor chimerism without GVHD, and the rate of CD4(+)/CD8(+) has close relation to the incidence of GVHD.
Animals ; CD4-CD8 Ratio ; Chimerism ; Female ; Graft vs Host Disease ; prevention & control ; Graft vs Leukemia Effect ; Lymphocyte Transfusion ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Stem Cell Transplantation ; adverse effects ; methods ; Transplantation Conditioning ; methods ; Whole-Body Irradiation