T and B cell responses to cytomegalovirus antigens in healthy blood donors and bone marrow transplant recipients

Abstract We measured the production of interferon-gamma (IFN-γ) from single T cells and the T cell proliferative response to different cytomegalovirus (CMV) antigens in healthy blood donors and bone marrow transplant recipients. The antigens consisted of a CMV nuclear antigen (CMV na) containing the pp65-kDa matrix protein and the immediate early antigens but lacking CMV glycoproteins, and an antigen comprising native CMV glycoproteins (CMV gp). We also measured the IgG antibodies to CMV na and CMV gp. The T cells reacted to CMV na in CMV seropositive blood donors both with the production of IFN-γ and with proliferation, while bone marrow transplant recipients had a deficient T cell response. After stimulation with CMV gp, no T cell response could be observed in CMV seropositive subjects. IgG antibodies to CMV na coexisted in plasma with similar levels of antibodies to CMV gp.     

Analyzing the cellular contribution of bone marrow to fracture healing using bone marrow transplantation in mice

The bone marrow is believed to play important roles during fracture healing such as providing progenitor cells for inflammation, matrix remodeling, and cartilage and bone formation. Given the complex nature of bone repair, it remains difficult to distinguish the contributions of various cell types. Here we describe a mouse model based on bone marrow transplantation and genetic labeling to track cells originating from bone marrow during fracture healing. Following lethal irradiation and engraftment of bone marrow expressing the LacZ transgene constitutively, wild type mice underwent tibial fracture. Donor bone marrow-derived cells, which originated from the hematopoietic compartment, did not participate in the chondrogenic and osteogenic lineages during fracture healing. Instead, the donor bone marrow contributed to inflammatory and bone resorbing cells. This model can be exploited in the future to investigate the role of inflammation and matrix remodeling during bone repair, independent from osteogenesis and chondrogenesis.     

Effect of repeated allogeneic bone marrow mononuclear cell transplantation on brain injury following transient focal cerebral ischemia in rats

Aims

Transplantation of bone marrow mononuclear cells (BMMCs) exerts neuroprotection against cerebral ischemia. We examined the therapeutic timepoint of allogeneic BMMC transplantation in a rat model of focal cerebral ischemia, and determined the effects of repeated transplantation outside the therapeutic window.

Main methods

Male Sprague–Dawley rats were subjected to 90 minute focal cerebral ischemia, followed by intravenous administration of 1 × 10⁷ allogeneic BMMCs or vehicle at 0, 3 or 6 h after reperfusion or 2 × 10⁷ BMMCs 6 h after reperfusion. Other rats administered 1 × 10⁷ BMMCs at 6 h after reperfusion received additional BMMC transplantation or vehicle 9 h after reperfusion. Infarct volumes, neurological deficit scores and immunohistochemistry were evaluated 24 or 72 h after reperfusion.

Key findings

Infarct volumes at 24 h were significantly decreased in transplantation rats at 0 and 3 h, but not at 6 h, after reperfusion, compared to vehicle-treatment. Even high dose BMMC transplantation at 6 h after reperfusion was ineffective. Repeated BMMC transplantation at 6 and 9 h after reperfusion reduced infarct volumes and significantly improved neurological deficit scores at 24 and 72 h. Immunohistochemistry showed repeated BMMC transplantation reduced ionized calcium-binding adapter molecule 1, 4-hydroxy-2-nonenal and 8-hydroxydeoxyguanosine expression at 24 and 72 h after reperfusion.

Significance

Intravenous allogeneic BMMCs were neuroprotective following transient focal cerebral ischemia, and the therapeutic time window of BMMC transplantation was > 3 h and < 6 h after reperfusion in this model. Repeated transplantation at 6 and 9 h after reperfusion suppressed inflammation and oxidative stress in ischemic brains, resulting in improved neuroprotection.     

移植途径对大鼠骨髓间充质干细胞归巢及肝功能恢复的影响

目的 探讨移植途径对骨髓间充质干细胞（MSCs）归巢及促进肝切除大鼠肝再生的影响.方法 建立肝切除大鼠模型,随机分为3 组,即肝切除对照组、尾静脉移植组和门静脉移植组.移植组分别经尾静脉和门静脉注射DAPI 标记的MSCs 约1.5×106/ 只,分别于第3 天和第9 天后采血清检测肝功能,第9 天处死大鼠取肝脏标本,并通过荧光显微镜观察两种移植途径对MSCs 向肝脏迁移的影响.结果 门静脉移植组（18.1 ± 3.4）个细胞/100 倍视野到肝脏归巢及定植的 MSCs 多于尾静脉移植组（7.6 ± 2.0）个细胞/100 倍视野,差异有统计学意义（P 〈 0.01）.术后第9 天各组大鼠肝功能均有好转,丙氨酸氨基转移酶（ALT）及天冬氨酸氨基转移酶（AST）3 组之间对比差异无统计学意义（F = 2.822,1.046,P = 0.057,0.365,P 〉 0.05）;但两移植组与单纯肝切除组比较血浆白蛋白（ALB）均有明显升高,差异具有统计学意义（F = 6.259,P = 0.006）;尾静脉移植组与门静脉移植组两移植组之间相比,差异无统计学意义（P 〉 0.05）.结论 移植途径对 MSCs 归巢、定植到肝脏有一定影响,门静脉途径优于外周静脉,MSCs 移植对肝大部切除大鼠肝功能恢复具有促进作用.     

Monosomy 7 in donor cell-derived leukemia after bone marrow transplantation for severe aplastic anemia: Report of a new case and review of the literature

Monosomy 7 arises as a recurrent chromosome aberration in donor cell leukemia after hematopoietic stem cell transplantation. We report a new case of donor cell leukemia with monosomy 7 following HLA-identical allogenic bone marrow transplantation for severe aplastic anemia (SAA). The male patient received a bone marrow graft from his sister, and monosomy 7 was detected only in the XX donor cells, 34 months after transplantation. The patient’s bone marrow microenvironment may have played a role in the leukemic transformation of the donor hematopoietic cells.     

骨髓间充质干细胞移植对急性肝功能衰竭大鼠肝组织miRNA-155和TNF-α表达的影响

目的探讨骨髓间充质干细胞（BMSCs）移植对急性肝功能衰竭（ALF）大鼠肝组织中miRNA-155和TNF-α表达的影响,以及与BMSCs疗效间的关系。方法将SD大鼠随机分为健康对照组、ALF组、BMSCs治疗组和BMSCs预防组,其中ALF组予以900 mg/kg D-GalN＋10μg/kg脂多糖腹腔注射建立模型;BMSCs治疗组在900 mg/kg D-GalN＋10μg/kg脂多糖腹腔注射后2 h,予以尾静脉注射BMSCs 5.0×10^6;BMSCs预防组在900 mg/kg D-GalN＋10μg/kg脂多糖腹腔注射前予以尾静脉注射BMSCs 5.0×10^6;健康对照组予以0.9﹪氯化钠溶液1 ml腹腔注射。给药7 h后每组处死大鼠,检测大鼠血清ALT和AST,ELISA法检测TNF-α水平,实时定量PCR检测肝组织miRNA-155、TNF-αmRNA。各组间肝功指标差异采用方差分析,同时观察每组大鼠的24 h生存率,并用卡方检验比较各组生存率的差异。结果 D-GalN/脂多糖诱导7 h后,与ALF组相比,BMSCs预防和BMSCs治疗组大鼠ALT、AST、TNF-α水平均有所降低（P〈0.01）;同时两组肝组织TNF-αmRNA和miRNA-155表达水平均有下调（P〈0.01）;但两组间相比较差异无统计学意义。ALF组大鼠肝组织miRNA-155上调和TNF-αmRNA诱导呈正相关（r=0.734,P=0.001）。BMSCs预防组和BMSCs治疗组miRNA-155和TNF-αmRNA的部分逆转亦呈正相关（r值分别为0.687和0.590,P值分别为0.004和0.006）。给药后24 h,健康对照组、ALF组、BMSCs治疗组和BMSCs预防组大鼠死亡率组间比较差异有统计学意义（c2=19.078,P〈0.01）。结论在BMSCs干预大鼠ALF发病过程中,可以部分逆转上调的肝组织miRNA-155和TNF-α,且存在协同性,提示BMSCs治疗ALF可能通过对肝组织miRNA-155和TNF-α的调控发生作用。