EXPRESSION OF PARATHYROID HORMONE-RELATED PROTEIN mRNA IN TUMORS OBTAINED FROM PATIENTS WITH HUMORAL HYPERCALCEMIA OF MALIGNANCY

Human parathyroid hormone-related protein (PTHrP) mRNA was detected in all fresh cancer tissues consecutively obtained from 6 patients with humoral hypercalcemia of malignancy (HHM). The primary sites of the cancers in these cases were distributed among different organs including the lung, esophagus, kidney and ovary. PTHrP mRNA was undetectable in all 10 fresh cancer tissues obtained from normocalcemic patients. These results suggest PTHrP as a major cause of HHM.     

Urinary excretion of parathyroid hormone-related protein as a predictor of hypercalcemia in patients with adult T-cell leukemia.

Hypercalcemia with adult T-cell leukemia (ATL) is chiefly caused by an excessive production by tumor cells of parathyroid hormone-related protein (PTHrP). We have previously reported hypercalcemic patients with solid tumors to excrete a large amount of the C-terminal fragments of PTHrP (C-PTHrP) into their urine. To elucidate whether PTHrP production correlates with or predicts the development of hypercalcemia, we studied the urinary excretion of C-PTHrP in 36 ATL patients. The urinary excretion of C-PTHrP was in the normal range (< 0.40 nmol equivalent to PTHrP (109-141)/g creatinine) in HTLV-1-positive carriers (n 3), ATL patients in complete remission (n 2) and chronic type ATL patients (n 2). It was marginally increased in seven patients in partial remission, and gradually increased as the disease progressed. In 20 patients who died without or with hypercalcemia, it was increased to 1.98 +/- 0.69 (n 9) and 7.6 +/- 2.1 nmol/g creatinine (mean +/- SD, n 11, P < 0.01), respectively. Urinary C-PTHrP excretion was significantly correlated with serum calcium and LDH levels as well as with CD25-positive cells in the peripheral blood. In four patients whose urinary excretion had been serially determined, it increased prior to the development of hypercalcemia. The findings suggest the urinary excretion of C-PTHrP to be of use as a predictor of the development of hypercalcemia in ATL patients. In ATL patients whose urinary excretion of C-PTHrP is progressively increasing, the serum calcium concentration should be carefully monitored to prevent hypercalcemic crisis.     

成人T细胞白血病/淋巴瘤外周血异形淋巴细胞超微结构特征

目的进一步了解中国人嗜人T细胞病毒Ⅰ型（HTLV－Ⅰ）相关性成人T细胞白血病／淋巴瘤（ATLL）外周血异形淋巴细胞超微结构特征。方法用透射电镜对最近发现的12例ATLL中4例急性型患者外周血异形淋巴细胞的超微结构进行了观察。结果主要特点为：细胞大小基本一致,与正常淋巴细胞大小相似,偶见较大的异形淋巴细胞。核为多形性,表现为切迹、凹陷、扭曲、折叠、脑回状、分叶或花瓣状。细胞器减少和发育不良,线粒体肿胀和变性。1例见类病毒颗粒。结论对ATLL细胞超微结构的认识有助于了解和探索ATLL的致病过程以及结构与功能的关系。     

A SOLUBLE-FACTOR(S) SECRETED BY A HUMAN SKIN CANCER CELL LINE SUPPORTS CLONAL GROWTH OF ADULT T-CELL LEUKEMIA CELLS

Leukemic cells from four out of eight patients with adult T-cell leukemia (ATL) were successfully grown by cocultivation with HSC-I cells, a human skin cancer cell line, in the presence of interleukin-2. Three of these four cultures of growing cells showed rearrangement of the T-cell receptor β-chain gene like the original leukemic cells in vivo , and also showed conservation of the patterns of HTLV-I integration of the original leukemic cells in vivo . Cell-to-cell contact between HSC-I cells and leukemic cells was not necessary for growth of the leukemic cells. The results indicate that some soluble growth factor secreted by HSC-I cells and interleukin-2 are required for the in vitro growth of leukemic cells from some patients with adult T-cell leukemia.     

Enhancing Effect of Interleukin-2 on Production of Parathyroid Hormone-related Protein by Adult T-Cell Leukemia Cells

Leukemic cells from patients with adult T-cell leukemia (ATL) can produce a calcium-regulating protein, parathyroid hormone-related protein (PTHrP). Moreover, it has been reported that ATL cells produce some cytokines besides PTHrP and that these cells respond to the T-cell growth factors, interleukin-2 (IL-2) and interIenkin-4 (IL-4). To elucidate whether PTHrP produced by ATL cells is regulated by IL-2 or IL-4, we investigated the in vitro effects of IL-2 and IL-4 on the release of PTHrP. IL-2 increased the release of PTHrP into the conditioned medium from leukemic cells in some, but not all, ATL patients; however, IL-4 did not affect the PTHrP release. PTHrP messenger RNA (mRNA) levels were increased in ATL cells cultured in the presence of IL-2. These data suggest that IL-2 plays a role in the regulation of hypercalcemia by enhancing the production of PTHrP in ATL patients.     

Circulating Parathyroid Hormone-related Protein (109-141) in Malignancy-associated Hypercalcemia

For differential diagnosis between hypercalcemia-induced bonemetastasis and humoral hypercalcemia of malignancy (HHM), serumparathyroid hormone-related protein (PTHrP) concentrations weremeasured in normal subjects and patients with malignancy-associatedhypercalcemia according to the presence or absence of bone metastasis,using a new sensitive PTHrP(109–141) radioimmunoassaysystem. The serum PTHrP(109–141) levels in all of 14 patientswithout bone metastasis were significantly higher than thosein normal subjects. However, in four patients with hypercalcemiaassociated with bone metastasis the levels were nearly the sameas those in normal subjects. The time course in two hypercalcemicpatients with esophageal carcinoma revealed that serum PTHrP(109–141)levels were elevated before hypercalcemia developed and thatchanges in PTHrP(109–141) and corrected serum calciumlevels were significantly correlated. These findings suggestthat determination of serum PTHrP(109–141) may be clinicallyimportant not only for differential diagnosis of HHM but alsoas a useful predictive marker of hypercalcemia.     

长期联合化疗对成人白血病患者外周血细胞姐妹染色单体交换率的影响

目的 :探讨长期联合化疗对缓解期成人白血病患者是否有潜在的诱变性。方法 :应用姐妹染色单体交换 (SCE)技术检测 44例白血病患者外周血细胞 SCE率的改变。结果 :初治、复发的急性白血病 (AL)和 CML组患者 SCE频率明显高于 CR- AL 组和正常对照组 ;CR- AL 仍有少部分高 SCE病例 (5 / 16 ) ,而此类病例主要分布在接受联合化疗在 9个疗程以上的 CR- AL 的患者中 (3/ 6 )。结论 :初治、复发 AL 和 CML 患者存在着 DNA的损伤或 DNA修复机制缺陷 ,长期反复联合化疗可能引起部分病例 DNA的不稳定。     

Serum Soluble Interleukin-2 Receptor Levels in Patients with Adult T-cell Leukemia and Human T-cell Leukemia/Lymphoma Virus Type-I Seropositive Healthy Carriers

Using an enzyme-linked immunosorbent assay (ELISA) technique, we measured the soluble interleukin 2 receptor (s-IL-2R) levels in the sera of patients with adult T-cell leukemia (ATL) in Japan. The s-IL-2R levels in the sera of the ATL patients were markedly higher (range 540-310, 400 U/ml, mean ±SD=62,800 ±81,000 U/ml, n = 42) than those in normal individuals (range 42-950 U/ml, mean ±SD=322 ±198 U/ml, n = 35, P<0.01). The patients with acute-type or lymnhoma-type ATL had high s-IL-2R levels (range 11,900-310,400 U/ml, mean ±SD= 110,340 ± 370 U/ml, n = 15; range 26,400-214,400 U/ml, mean ±SD=90,170 ±59,040 U/ml, n = 7, respectively). All of the patients with hypercalcemia (Ca>10 mg/dl) or elevated serum LDH levels (LDH > 500 IU/liter) also had s-IL-2R levels above 10,000 U/ml. The high s-IL-2R levels in the sera of ATL patients indicate abnormal IL-2 receptor production and its release from the leukemic cells in vivo . Thus, the serum s-IL-2R level may be a sensitive and useful marker to monitor the total amount of tumor cells in ATL, especially in the lymphoma type. We next examined the serum s-IL-2R levels in human T-cell leukemia/lymphoma virus type-I (HTLV-I) seropositive healthy carriers to investigate whether there might he abnormal IL-2 receptor expression in such individuals. However, there was no statistically significant difference between the S-IL-2R level of 71 HTLV-I seropositive healthy carriers (range 65-880 U/ml, mean±SD =394±212 U/ml) and that of 71 age- and sex-matched normal individuals (range 33-950 U/ml, mean ±SD=357 ±224 U/ml) who lived in Okinawa Prefecture.     

急性白血病及其细胞株bcl-2蛋白表达

目的　检测 60例急性白血病患者和 6种白血病细胞株的 bcl- 2蛋白表达水平。方法　应用抗 bcl- 2单抗 ,免疫细胞化学染色方法。结果　急性白血病患者有较高 bcl- 2蛋白阳性表达 ,与 FAB分型无一定关系。AL L初治组 bcl- 2蛋白阳性细胞平均百分率、平均染色强度及平均染色系数均高于复发 /难治组 ,AML 初治组与复发 /难治组 bcl- 2蛋白阳性细胞平均百分率、平均染色强度及平均染色系数之间无明显相关性。 6株白血病细胞对 bcl- 2蛋白的阳性表达亦较高。结论　 bcl- 2蛋白检测作为判断 AL 临床化疗敏感性或药物耐受的指标有一定意义。     

EXPRESSION OF CR2 (C3d RECEPTOR) ON THE CELL MEMBRANES OF ADULT T CELL LEUKEMIA

MT - 2 cells, which produce human T-cell lymphotropic virus type I (HTLV-I), are known to have a complement receptor. We have established that the complement receptor is CR2 which binds C3d on immune complexes but not CR1. CR2 was also detected on ATL-3I cells but no complement receptor was detected on ATL-1K cells which lack ATL antigen (ATLA). Since CR2 is not detectable on normal T lymphocytes, the presence of CR2 on some ATL cells might suggest that ATL cells were derived from a particular minor lineage of T cells, or HTLV-I has a capacity to induce CR2, which has been demonstrated to be an α-type growth factor for B lymphocytes and to be a receptor for Epstein-Barr virus.     

急性单核细胞白血病细胞核基质蛋白单克隆抗体的制备及初步鉴定

目的 :制备抗急性单核细胞白血病细胞核基质蛋白的特异性单抗。方法 :SDS- PAGE分析正常骨髓细胞与急性单核细胞白血病细胞核基质蛋白的差异。用急性单核细胞白血病细胞核基质免疫 Balb/ c小鼠 ,利用杂交瘤技术制备了一株能稳定分泌单克隆抗体的杂交瘤细胞株 8E7,并对其作了初步鉴定。结果 :免疫组化结果显示 ,8E7单抗与急性单核细胞白血病细胞核仁和胞质反应 ,与正常骨髓单核细胞胞核及红白血病细胞株 K5 6 2细胞胞质反应。Western Blotting结果表明 ,8E7单抗与电泳图谱上急单白血病细胞核基质 94k D的蛋白反应 ,与正常骨髓细胞核基质 2 7k D的蛋白反应。结论 :对 8E7单抗反应蛋白的进一步研究有助于阐明白血病的发病机制。     

XENOGENIZATION OF TUMOR CELLS BY TRANSFECTION WITH PLASMID CONTAINING env GENE OF FRIEND LEUKEMIA VIRUS

A rat hepatocellular carcinoma cell line (cKDH-8 cl-11) showed decreased tumorigenicity after transfection with an envelope gene derived from a Friend leukemia virus (FV- env gene). FV- env gene product was found by indirect immunofluorescence staining to be expressed on the cell surface of the FV- env gene-transfected cells. The FV- env -transfected cells (FV- env cKDH-8), however, grew well in X-irradiated immunosuppressed rats, indicating that the reduction in tumorigenicity of the transfected cells is based on immunological reaction in the host. The rats which rejected FV-env cKDH-8 cells showed resistance to rechallenge with the parent cKDH-8 cl-11 tumor cells. These results suggest that the FV- env gene product may elicit antitumor immunity against FV-env cKDH-8 cells in a host with a resultant reduction in the tumorigenicity of these cells.     

非亲缘性双份脐血移植治疗成人慢性髓系白血病

目的 :探索非亲缘性双份脐血移植 (CBT)治疗成人恶性血液病的可行性及并发症的防治。方法 :1例 2 7岁体重为 6 0 kg的慢性髓系白血病急变期 (CML- BP)患者 ,由北京大学人民医院脐血库提供 HL A配型部分相合的双份脐血。予改良的环磷酰胺 +6 0 Co全身照射 (TBI)方案进行预处理 ,经股动脉同时回输双份脐血 ,有核细胞数 (MNC)分别为 1.5 4× 10 7/ kg和 1.2 5× 10 7/ kg。移植物抗宿主病 (GVHD)的预防用环胞菌素 A(Cs A)、甲氨蝶呤 (MTX)及霉酚酸酯 (MMF)、白细胞介素 2(IL - 2 )及甲泼尼龙 ,肝静脉闭塞病 (VOD)的预防用前列腺素 E1 (PGE1 ) ,以更昔洛韦预防巨细胞病毒(CMV)感染。结果 :移植后第 2 2天 (+2 2天 )中性粒细胞计数 >0 .5× 10 9/ L ,+2 8天血小板计数 >2 .0× 10 9/ L。移植 +40天骨髓血 DNA位点检测与其中一份编码为 980 2 180 2的脐血完全相符。移植过程仅出现 度急性移植物抗宿主病 (a GVHD) ,+30天出现出血性膀胱炎。+44天出现低热 ,血尿 CMV检查 (+) ,加用更昔洛韦 2周 ,CMV病毒转阴。移植过程共输血小板 7次。随访 2 5个月 ,患者一般情况好 ,骨髓细胞 Ph染色体和 bcr/ abl融合基因阴性。结论 :同一受者接受 HL A配型部分相合的两份脐血移植在临床上是可行的 ,其造血功能获得及时恢复     

APPEARANCE OF ANTIBODIES AGAINST PHOSPHOTYROSINE AND TYROSINE PHOSPHORYLATION OF LYMPHOCYTE PROTEINS IN HUMAN T CELL LEUKEMIA VIRUS TYPE-I INFECTION

Antibodies reactive with O-phosphotyrosine (Ptyr) were detected in all of 56 patients with adult T cell leukemia (ATL) with high titers ranging from 1:1,600 to 1:25,000 in 91% of the patients, whereas 97% of 70 healthy subjects showed titers of 1:400 or less. Ptyr-containing proteins (molecular weights of 70k, 45k and 30k) were detected in peripheral lymphocytes from patients with ATL. These proteins might be involved as antigens in the induction of such antibodies.     

BC369对白血病造血祖细胞的体外药敏实验研究

目的研究ＢＣ３６９直接抑制或杀伤白血病造血祖细胞的作用。方法采用白血病造血祖细胞体外培养，分别设立ＢＣ３６９实验组和生理盐水对照组，然后同时放入ＣＯ２孵育箱中培养，６天后对比计算ＢＣ３６９的杀伤率。结果急性淋巴细胞白血病平均杀伤率为４９．３５％（Ｐ＜０．０１）；急性粒细胞白血病平均杀伤率为５７．４３％（Ｐ＜０．０１）；慢性粒细胞白血病平均杀伤率为４９．４８％（Ｐ＜０．０１）；多发性骨髓瘤杀伤率为１９．０３％（Ｐ＞０．０５）。结论ＢＣ３６９对１４例白血病患者骨髓造血祖细胞集落形成单位（Ｌ－ＣＦＵ）进行体外药敏实验；ＢＣ３６９对白血病细胞具有直接杀伤作用，其平均杀伤率为５１．２６％（Ｐ＜０．０１）。     

急性髓系白血病的bcl-2基因表达及其临床意义

目的：探讨bcl—2基因在急性髓系白血病(AML)中的表达与预后的关系。方法：应用链亲和素-胶体金原位杂交(ISH—SAG)法检测57例AML患者单个核细胞的bcl—2基因表达水平。结果：57例AML患者不同程度表达bcl—2基因，范围从0—98％，其中初治组的阳性率为46．2％(24／52)，缓解组的阳性率为40．7％(11／27)，难治复发组的阳性率为100％(15／15)；难治复发组与初治组及缓解组之间差异均具有显著性(P＜0．01)，AML各亚型之间表达无显著性差异(P＞0．05)；疗效分析发现bcl-2基因表达与临床缓解密切相关，阴性组缓解率(76．2％)显著高于阳性组(42．1％)(P＜0．05)。结论：bcl—2基因过度表达对AML的预后有重要关系，可作为判断预后和合理制定治疗方案的重要依据。     

急性白血病端粒酶活性与p53蛋白表达规律的研究

目的：探讨争性白血病（AL）患者中端粒酶活性与突变型P53蛋白的表达规律,方法：采用聚合酶链反应一酶联免疫吸附法（PCR－ELISA）对端粒酶活性进行定半量测定,用流式细胞仪检测突变型P53蛋白表达,结果：AL患者端粒酶性显著高于对照组＊（P＜0．001）,端粒酶活性在AL的不同病期有统计学差异,其中初治组,复发组均显著高于完全缓解CR组（P＜0．01）,在治疗敏感组与不敏感组间无统计学差异（P＞0．05）,AL患者中突变型P53蛋白阳性率为19．4％（7／36）,在AL的不同病期P53阳性率无统计学差别。在初治敏感组与不敏感组之间亦无统计学差别（P均＞0．05）,端粒酶活性与突变型P53蛋白的表达无关。结论：端粒酶活性与AL的病期和发展密切相关,突变型P53蛋白在AL中表达较低,对临床预后的判断价值较小,端粒酶的激活与P53蛋白表达无相关性。     

PATTERN OF T-CELL RECEPTOR DELTA GENE REARRANGEMENT BY SOUTHERN BLOTTING AND POLYMERASE CHAIN REACTION TECHNIQUE IN HONG KONG CHINESE PATIENTS WITH NON-T-CELL HEMATOLOGICAL MALIGNANCIES

It has been recognized that clonal T-cell receptor delta (TCRδ) gene rearrangement is present in both T- and B-cell malignancies. The highly sensitive polymerase chain reaction (PCR) technique may be applicable to cases of leukemia and lymphoma of non-T-cell origin for detection of minimal residual disease (MRD). A PCR technique was used in this study to investigate the pattern of clonal TCRδ gene rearrangement in Hong Kong Chinese patients with non-T-cell hematological malignancies. Seventy-three patients with the diagnosis of acute leukemia and non-Hodgkin's lymphoma of non-T-cell origin were included in this study. There were 20 patients with common ALL (cALL), seven precursal B-cell ALL (PreB-ALL), 23 acute myeloid leukemia (AML), and 23 non-Hodgkin's lymphoma of B-lineage (B-NHL). Clonal rearrangement was detectable by Southern analysis using a Jδ1 probe in 41 per cent of ALL of B-lineage but in none of the B-NHL or AML. The samples were also studied further by monoclonal PCR amplification for TCRδ gene rearrangement. Three different sets of primers were employed to detect clonal rearrangement of the TCRδ gene. The Vδ1(D)Jδ1 recombination typically seen in T-cell malignancies were not seen in any of the of the non-T-cell malignancies. The Vδ2(D)Dδ3 recombination was found exclusively in ALL of B-lineage and was seen in 73 per cent of the Southern positive cases. Although clonal TCRδ gene rearrangement was undetectable by Southern analysis in our AML cases, 26 per cent had a Vδ2(D)Jδ1 recombination found by the PCR technique. Sensitivity of the PCR technique was determined by serial mixing and was up to 5–10 leukemic cells per 10⁴ nucleated cells. It was apparent from this study that it was feasible to detect clonal TCRδ gene rearrangement by the PCR technique in a proportion of the cases of non-T-cell hematological malignancies. The PCR technique can be applied to detect residual leukemic cells in marrow of patients in an apparent morphological complete remission. The value of this application requires further clinical evaluation and correlation.     

Basic Investigations EXPRESSION OF GAP JUNCTION PROTEIN Cx43 IN CULTURED HUMAN NORMAL AND MALIGNANT LUNG CELLS

ＢａｓｉｃＩｎｖｅｓｔｉｇａｔｉｏｎｓＥＸＰＲＥＳＳＩＯＮＯＦＧＡＰＪＵＮＣＴＩＯＮＰＲＯＴＥＩＮＣｘ４３ＩＮＣＵＬＴＵＲＥＤＨＵＭＡＮＮＯＲＭＡＬＡＮＤＭＡＬＩＧＮＡＮＴＬＵＮＧＣＥＬＬＳＺｈａｎｇＺｈｉｑｉａｎ；张志谦；ＬｉｎＺｈｏｎｇｘｉａｎｇ...