HIV infection due to a platelet transfusion after allogeneic bone marrow transplantation

A 30-yr-old man with chronic granulocytic leukaemia received a bone marrow transplant from his histocompatible sister in December 1982. His post-transplant course was complicated by Grade III graft-versus-host disease and multiple infectious episodes until his death from pneumonia on d + 190. He was later found to be seropositive for anti-HIV at the time of his death. Retrospective analysis of stored sera showed a transient period of seropositivity from d + 11 to d + 20 thought to reflect passive transfer of antibody from a blood product transfused prior to d + 11 when he was also exposed to infectious virus. He remained seronegative until d + 78 when anti-HIV was again found. Seropositivity persisted until his death and was attributed to endogenous antibody response. Although it is unclear whether his clinical course was due to AIDS, exposure of an immunosuppressed patient to HIV may be associated with more rapid development of clinical disease.     

Antiviral nucleoside toxicity in canine bone marrow progenitor cells and its relationship to drug permeation.

The most promising nucleoside analogs that are currently undergoing preclinical and clinical testing for anti-HIV activity belong to the dideoxynucleoside group. We have studied the toxicity of 3'-azido,3'-deoxythymidine (AZT), 2',3'-dideoxycytidine (DDC), and 2',3'-dideoxyinosine (DDI) in canine bone marrow progenitor cells in culture. AZT potently inhibited both canine CFU-GM and CFU-E with IC50 values of 2 and 8 mumol/l respectively, while DDC was relatively non-toxic to either progenitor with IC50 of > 200 mumol/l and 80 mumol/l respectively. DDI was mildly toxic to the bone marrow progenitors, with IC50 values of 62 mumol/l for CFU-GM and 70 mumol/l for CFU-E. Dipyridamole, a nucleoside transport inhibitor, did not influence the toxicity of these dideoxynucleosides in either progenitor at concentrations up to 10 mumol/l. Using uridine as the prototype endogenous nucleoside, we have demonstrated that there is a saturable "zero-trans" nucleoside transport system in canine bone marrow mononuclear cells, which is completely inhibited by 1 mumol/l dipyridamole (Ki = 0.02 mumol/l). None of the dideoxynucleosides appeared to be a substrate for this transport system, and dipyridamole did not alter their influx. Permeation of radiolabeled AZT into bone marrow mononuclear cells was slow and non-saturable, while the permeation of DDI was even slower. DDC did not permeate bone marrow cells well, with very little cell accumulation even after 2 hours of equilibration. Our toxicity data from canine bone marrow progenitor cells paralleled the clinical hematotoxicity profiles of these dideoxynucleosides in AIDS patients and suggest that the myelotoxicity of a nucleoside analog is related to its ability to permeate the progenitor cells in question. Canine bone marrow progenitor cultures may serve well as an in vitro model for drug hematotoxicity studies.     

Role of heme metabolism in AZT-induced bone marrow toxicity

We studied the effects of azidothymidine (AZT) on rat bone marrow heme metabolism and colony growth as determined by assays of granulocyte-macrophage (CFU-GM), erythroid (CFU-E), burst-forming erythroid (BFU-E), and alpha-aminolevulinic acid synthase (ALAS), the first enzyme in the heme pathway. In all cases, AZT (1-0.01 microM) was found to be toxic to bone marrow colony growth. When AZT was included in colony assays, 1 microM resulted in 98-100% inhibition, whereas lower concentrations (0.01 microM) inhibited growth by 58-76%. In addition, cultures from AZT-treated animals had a marked reduction in colony growth as compared with sham controls. In most cases, hemin (10(-5) M) was found to overcome some of the colony inhibitory effects of AZT. Analysis of heme metabolism indicated that ALAS activity was reduced by 71% in bone marrow cells from treated animals. ALAS activity for control was 204 +/- 33 pM ALA formed/4 X 10(6) cells/hr, whereas ALAS activity from AZT-treated animals was only 60 +/- 3 pM ALA formed/4 x 10(6) cells/hr. It is considered that AZT toxicity may be due to a depression in the pool of available heme, which is required for adequate hematopoiesis.     

The radiation response and recovery of bone marrow stroma with particular reference to long-term bone marrow cultures

There is evidence for long-term haematopoietic dysfunction in some patients treated with radiotherapy. Although the underlying mechanisms are unclear, both stem cell and environmental defects have been implicated. In the present article we review the evidence concerning the role of stromal cells. According to the endpoints used, a wide range of radiosensitivities for the stroma have been reported. Long-term bone marrow cultures provide a system in which both functional and regenerative aspects of the stroma can be studied. A dose of 5 Gy applied prior to the establishment of long-term bone marrow cultures decreases both the formation of a confluent adherent stromal layer and its capacity to support haematopoiesis. In contrast, in its fully established phase, the adherent layer displays a high radioresistance due to the low proliferative stress applied to its stromal populations. A dose of 10 Gy given to a fully established adherent layer does not prevent haematopoietic engraftment and sustained haematopoiesis. At doses above 100 Gy a macrophage-like and epithelioid cell-type become dominant, which preserve their ability of producing growth regulatory molecules at doses as high as 500 Gy. These data suggest that the main effect on the stroma is a delayed expression of irradiation damage due to the slow rate of turnover of stromal cells. So far, there is little evidence for persistent deficiencies in the functional roles of stromal cell populations.     

骨髓细胞参与甘油引起的小鼠急性肾小管坏死修复的实验观察

目的：观察骨髓来源的细胞能否分化成肾小管上皮细胞。方法：15只绿色荧光蛋白（GFP）标记的C57BL／6转基因小鼠提供骨髓细胞，64只同种无荧光标记的C57BL／6小鼠随机分为正常对照组（N组）、全身照射组（TBI组）、骨髓移植组（BMT组）、骨髓移植＋甘油注射组（B＋G组），每组16只。各组小鼠于不同时间点取血检测血常规、尿素氮及血肌酐，并取肾行H-E染色检查肾脏病理变化。流式细胞仪可以明确受体鼠骨髓细胞中CFP阳性细胞的比例，利用荧光显微镜及激光共聚焦显微镜采用荧光组织化学、免疫组织化学等方法观察GFP阳性细胞在受体鼠肾脏的分布及数量。结果：致死剂量^60Co照射虽然引起血常规三系减少，但并未对肾脏的组织结构及功能造成损伤。BMT组受体鼠在骨髓移植后第56天、84天时，肾小管中有少量GFP阳性细胞的存在，B＋G组受体鼠于上述同样时间点时肾小管中的GFP阳性细胞增多。激光共聚焦显微镜进一步证实了这些GFP阳性细胞位于肾小管上皮，并且荧光组织化学显示，这些GFP阳性细胞表达肾小管上皮细胞特异性功能蛋白Megalin。结论：骨髓细胞可以向肾小管上皮细胞分化．参与肾小管上皮细胞的更新。并且损伤可以使骨髓细胞的肾向分化率增加。     

A simple technique for processing of bone marrow for human marrow transplantation

Summary A simple stainless steel device is described for the processing of marrow for intravenous infusion to marrow graft recipients. The device can be cleaned easily and sterilized for re-use.     

Pancytopenia due to bone marrow necrosis in acute myelogenous leukemia: Role of reactive CD8 cells

Bone marrow necrosis is a rare clinical condition often associated with hematological malignancy. The mechanism by which malignant disease causes marrow necrosis is unknown. We present a case of a patient with newly diagnosed pancytopenia with bone marrow biopsy evidence of extensive marrow necrosis. Upon further work-up utilizing Tc bone scan directed bone marrow biopsy, a massive CD8+ T cell marrow infiltrate was discovered engulfing AML-M2 blasts. The role of Tc bone scans in the work-up of bone marrow necrosis as well as the potential mechanism of AML-M2 induced marrow necrosis in the setting of reactive CD8+ T cell infiltration is discussed. Am. J. Hematol. 59:74– 78, 1998. © 1998 Wiley-Liss, Inc.     

Bronchiolitis obliterans after bone marrow transplantation

Bronchiolitis obliterans occurred in the setting of chronic graft-versus-host disease 1 year after allogeneic bone marrow transplantation for chronic myelogenous leukemia. The severe obstructive pulmonary disease followed an episode of interstitial pneumonitis. The etiology and possible relationship to graft-versus-host disease of this rare pulmonary lesion following bone marrow transplantation are discussed.     

Continuous long-term culture of human bone marrow

Summary Human bone marrow was cultured with alpha medium, 10^?6m hydrocortisone and a mixture of horse and fetal calf serum in a long-term liquid culture system. An adherent layer was formed which contained fat cells, macrophages, fibroblast-like and epithelioid cells. The layer harboured myeloid cells and their presumptive precursors whilst the nonadherent cells were composed of immature myeloid elements plus mature macrophages and granulocytes whose numbers were maintained for periods of up to 12 weeks. Experiments showed that the use of serum mixtures was superior to horse or fetal calf serum alone and successful culture was accompanied by early growth of the hydrocortisone-dependent fat cells in the adherent layer.     

晚期非小细胞肺癌患者化疗骨髓抑制与生存相关性研究

目的探讨晚期非小细胞肺癌患者化疗过程中出现的骨髓抑制与远期疗效的相关性。方法回顾性分析102例晚期非小细胞肺癌患者采用GP方案全身联合化疗：吉西他滨：1250mg/m2静脉滴注,d1,8;顺铂：25mg/m2d1～3;21d为一个周期;化疗至少3周期,了解化疗期间出现的骨髓抑制对远期疗效的预测作用。结果 102例晚期非小细胞肺癌患者,平均中位生存时间151d。其中GO组中位生存时间124d,G1～2组中位生存时间195d、G3～4组中位生存时间203d（95%GI为148.9～267.0）。G0组、G1～2组、G3～4组三组生存时间水平间整体比较,差异有显著性（P=0.018）;G0组与G1～2组比较,差异有显著性（P=0.037）;G0组与G3～4组比较,差异有显著性（P=0.01）。而G1～2组与G3～4组比较,差异无显著性（P=0.26）。结论对于晚期非小细胞肺癌患者,化疗过程中出现中性粒细胞抑制与远期生存时间的延长明显相关,而未出现中性粒细胞抑制可能提示药物剂量的不足。     

Haemostatic and fibrinolytic responses to bone marrow transplantation

Thrombin and plasmin activation markers were serially measured in 80 patients undergoing bone marrow transplantation (BMT). There were prothrombotic and fibrinolytic responses observed during autograft and allograft BMTs. Thrombin-antithrombin and prothrombin fragment F1+2 levels increased from day -7 to -3 (P<0.0001) from 3.7 to 7 ng/ml and 1.2 to 1.63 nmol/l, respectively. A rise in plasmin-antiplasmin levels occurred between days 4 and 14 (P< 0.0004), from 393 ng/ml on day -7 to a peak of 795 ng/ml on day 11. No correlation between reduced protein C levels post-BMT and a prothrombotic state was observed.     

异种骨复合自体骨髓修复骨缺损的实验研究

目的　观察异种骨复合自体红骨髓对骨缺损的修复能力。方法　选健康纯系 ６ 月龄大耳白兔 ２０ 只,双侧桡骨中段制成 １５～１８ ｃｍ 的骨缺损模型,骨蜡封闭两断端骨髓腔,均植入等长度的异种骨,左侧注射红骨髓作为实验组,右侧不注射作为实验对照组。术后 ２、４、６、８ ｗ 拍 Ｘ 线片,行组织学检查。结果　实验组 ８ ｗ 时, Ｘ 线示骨缺损修复良好、髓腔基本相通,组织学检查见皮质骨形成、骨髓腔相通。而对照组 ８ ｗ 时 Ｘ 线下仍可见未完全吸收的移植骨,光镜示缺损区有残存的坏死骨,纤维组织充填缺损区。结论　异种骨复合自体红骨髓后可明显促进骨融合,提高修复骨缺损的能力     

Regeneration of CALLA (CD10+), TdT+ and double-positive cells in the bone marrow and blood after autologous bone marrow transplantation.

In this prospective study we investigated the frequency of CD10+, TdT+ and CD10+TdT+ mononuclear cells in the bone marrow (BM) and peripheral blood (PB) before and after autologous bone marrow transplantation (ABMT). 49 patients treated for acute lymphoblastic or myeloblastic leukaemia, malignant lymphoma or multiple myeloma were included. A significant increase in CD10+ cells occurred in BM in both children and adults after ABMT. In children, we also found a significant increase in CD10+ cells in PB. In individual patients remaining in remission, up to 34% CD10+ cells having a normal Ig kappa/lambda light chain ratio were recorded after ABMT. In children, the percentage of TdT+ and CD10+ TdT+ cells increased significantly in BM. In most cases the CD10/TdT-ratio was greater than 1.0, but during early regeneration after ABMT this ratio was less than 1.0 in several patients remaining in complete remission. In patients remaining in remission, CD10+TdT+ cells were detected in the blood in only 2 out of 140 samples tested, and the proportion of these cells never exceeded 0.03%. We conclude that quantitation of CD10+TdT+ cells in peripheral blood is helpful in the evaluation of complete remission in patients treated for pre-B-ALL.     

骨髓活检与涂片同步观察对诊断慢性淋巴细胞白血病的探讨

目的 :探讨骨髓活检和骨髓涂片同步观察在慢性淋巴细胞白血病 (CLL)诊断方面的重要性。方法 :针对 2 0例CLL患者 ,采用骨髓抽吸 活检双标本一步法取材 ,同时观察其骨髓涂片和活检塑料包埋切片。结果 :①骨髓活检切片增生度检出率为 10 0 % ,骨髓涂片检出率为 70 % ,两者差异有显著性意义 (P <0 .0 5 ) ;②切片中原始和幼稚淋巴细胞百分率略高于涂片 ;③ 85 %CLL患者伴有网状纤维增高 ,5 %CLL患者有胶原纤维增高。结论 :同步观察比常规涂片形态学观察的检出率、准确率均明显增高。