Development of an in vivo model of human multiple myeloma bone disease
Development of an in vivo model of human multiple myeloma bone disease
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DOI：
10.1182/blood.v87.4.1495.bloodjournal8741495
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发表时间：
1996-02-15
期刊：
影响因子：
20.3
通讯作者：
Roodman, GD
中科院分区：
文献类型：
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作者：
Alsina, M;Boyce, B;Roodman, GD
溶骨性骨破坏及其并发症、骨痛、病理性骨折和高钙血症是多发性骨髓瘤患者发病率和死亡率的主要来源。多发性骨髓瘤中的骨破坏是由于破骨细胞（OCL）活性增加和骨髓瘤细胞附近骨区域的骨形成减少。体内多发性骨髓瘤骨质溶解的机制尚不清楚。我们使用人浆细胞白血病细胞系ARH-77作为人多发性骨髓瘤的模型，该细胞系在患有严重联合免疫缺陷（SCID）的小鼠中具有播散性生长并表达IgG κ。用400拉德照射SCID小鼠，并在照射后24小时静脉内给小鼠注射10（6）个ARH-77细胞（ARH-77小鼠）或载体。通过血液离子钙水平、X射线和组织学评估骨疾病的发展。所有ARH-77小鼠，但没有一只对照小鼠在照射后存活，在注射后28至35天发生后肢麻痹，并在截瘫后平均5天发生高钙血症（1.35至1.46 mmol/ L）。在所有检查的高钙血症小鼠中，使用X射线检测到溶解性骨病变。对照组无溶解性病变或高钙血症。对照组或ARH-77小鼠在出现高钙血症后，处死，从长骨中获得骨髓血浆，浓缩，并测定骨吸收活性。与对照组相比，ARH-77小鼠的骨髓血浆在胎鼠长骨吸收试验中诱导了显著的骨吸收（释放的总Ca-45百分比= 35% +/- 4% v 11% +/- 1%）。ARH-77小鼠组织的组织学检查显示，肝脏和脾脏中骨髓瘤细胞浸润，椎骨和长骨中有明显浸润，骨小梁丢失，OCL数量增加。有趣的是，ARH-77小鼠骨髓的早期OCL前体（集落形成单位-粒细胞-巨噬细胞[CFU-GM]）培养物显示，ARH-77骨髓的CFU-GM比对照组增加了3倍（185 +/- 32 v40 +/- 32 × 10（5）接种细胞）。与对照小鼠相比，ARH-77小鼠血清或骨髓血浆中骨吸收性人和鼠细胞因子，如白细胞介素-6（IL-6）、IL-1 α或β、TGF α、骨光素和TNF α均未显著增加，尽管ARH-77细胞在体外产生IL-6和骨光素。与未处理的培养基相比，来自ARH-77细胞的条件培养基在胎鼠长骨吸收测定中诱导显著的骨吸收（释放的总Ca-45的百分比= 22% +/-2%v 11% +/- 1%）。这种作用并未被抗IL-6或抗淋巴毒素阻断（释放的总Ca-45百分比分别= 19% +/- 1%和22% +/-1%）。因此，我们开发了一种人类多发性骨髓瘤骨病模型，该模型对于剖析多发性骨髓瘤骨破坏的发病机制非常有用。(C)1996年，美国血液学会。
Osteolytic bone destruction and its complications, bone pain, pathologic fractures, and hypercalcemia, are a major source of morbidity and mortality in patients with multiple myeloma. The bone destruction in multiple myeloma is due to increased osteoclast (OCL) activity and decreased bone formation in areas of bone adjacent to myeloma cells. The mechanisms underlying osteolysis in multiple myeloma in vivo are unclear. We used a human plasma cell leukemia cell line, ARH-77, that has disseminated growth in mice with severe combined immunodeficiency (SCID) and expresses IgG kappa, as a model for human multiple myeloma. SCID mice were irradiated with 400 rads and mice were injected either with 10(6) ARH-77 cells intravenously (ARH-77 mice) or vehicle 24 hours after irradiation. Development of bone disease was assessed by blood ionized calcium levels, x-rays, and histology. All ARH-77, but none of control mice that survived irradiation, developed hind limb paralysis 28 to 35 days after injection and developed hypercalcemia (1.35 to 1.46 mmol/ L) a mean of 5 days after becoming paraplegic. Lytic bone lesions were detected using x-rays in all the hypercalcemic mice examined. No lytic lesions or hypercalcemia developed in the controls. Controls or ARH-77 mice, after developing hypercalcemia, were then killed and bone marrow plasma from the long bones was obtained, concentrated, and assayed for bone-resorbing activity. Bone marrow plasma from ARH-77 mice induced significant bone resorption in the fetal rat long bone resorption assay when compared with controls (percentage of total Ca-45 released = 35% +/- 4% v 11% +/- 1%). Histologic examination of tissues from the ARH-77 mice showed infiltration of myeloma cells in the liver and spleen and marked infiltration in vertebrae and long bones, with loss of bony trabeculae and increased OCL numbers. Interestingly, cultures of ARH-77 mouse bone marrow for early OCL precursors (colony-forming unit-granulocyte-macrophage [CFU-GM]) showed a threefold increase in CFU-GM from ARH-77 marrow versus controls (185 +/- 32 v 40 +/- 3 per 2 x 10(5) cells plated). Bone-resorbing human and murine cytokines such as interleukin-6 (IL-6), IL-1 alpha or beta, TGF alpha, lymphotoxin, and TNF alpha were not significantly increased in ARH-77 mouse sera or marrow plasma, compared with control mice, although ARH-77 cells produce IL-6 and lymphotoxin in vitro. Conditioned media from ARH-77 cells induced significant bone resorption in the fetal rat long bone resorption assay when compared with untreated media (percentage of total Ca-45 released = 22% +/- 2% v 11% +/- 1%). This effect was not blocked by anti-IL-6 or antilymphotoxin (percentage of total Ca-45 released = 19% +/- 1% and 22% +/- 1%, respectively). Thus, we have developed a model of human multiple myeloma bone disease that should be very useful to dissect the pathogenesis of the bone destruction in multiple myeloma. (C) 1996 by The American Society of Hematology.