白血病细胞膜相关因子（MAF—J6—1）单克隆抗体的制备及性质

用部分纯化的白血病细胞膜相关因子（ＭＡＦ－Ｊ６－１）对ＢＡＬＢ／Ｃ小鼠脾细胞进行体外免疫，然后与ＳＰ２／０小鼠骨髓瘤细胞融合获杂交瘤，经３次再克隆得到能产生ＭＡＦ－Ｊ６－１单克隆抗体（单抗）的杂交瘤细胞系。用ＡＢＣ免疫酶标法检测，该单抗在Ｊ６－１细胞上产生强阳性反应，并与ｒｈ－Ｍ－ＣＳＦ单抗对ＭＡＦ－Ｊ６－１及ｒｈ－Ｍ－ＣＳＦ产生交叉中和反应。ＭＡＦ－Ｊ６－１单抗还对Ｊ６－２、ＪＣＬ、Ｋ５６２等细     

人白血病细胞系(J6-1)膜相关调节因子(MAF-J6-1)的分离与性质

我们由人白血病细胞系J6-1发现了对自身有增殖刺激作用的膜相关生长调节因子(MAF-J6-1)。经Sephadex G-100凝胶过滤及连续两次快速蛋白液相层析(FPLC,Mono Q)将其分离。理化性质研究的结果表明,它是一种对酸、碱和热相对稳定的蛋白,分子量约50kD。抗M-CSF单抗与MAF-J6-1的中和试验结果表明,MAF-J6-1属M-CSF类。     

八种细胞因子对人白血病细胞系（J6－1，J6－2）增殖的调节

测定了Ｍ－ＣＳＦ，Ｃ－ＣＳＦ，ＧＭ－ＣＳＦ，ＩＬ－３，ＩＬ－６，ＭＧＦ，ＬＩＦ和ＴＧＦ－β＿１８种重组人生长因子单个或不同组合对白血病细胞系Ｊ６－１，Ｊ６－２集落形成率的影响。单个因子除ＴＧＦ－β＿１和ＬＩＦ表现出明显抑制效应外，其它因子均有不同程度的刺激活性。而在双因子协同下，ＴＧＦ－β＿１除与Ｍ－ＣＳＦ协同外，仍表现出明显的抑制作用，表明ＴＧＦ－β是Ｊ６－１，Ｊ６－２细胞系重要的负调节因子。从三种因子组合结果看出Ｍ－ＣＳＦ是Ｊ６－１细胞必需的细胞因子。值得注意的是，ＴＧＦ－β＿１与Ｍ－ＣＳＦ的组合对Ｊ６－１细胞有很强的增殖刺激效应。另外由Ｊ６－１细胞膜分离出的膜相关细胞因子（ＭＡＦ－Ｊ６－１）单独和与ＴＧＦ－β＿１联合，对Ｊ６－１，Ｊ６－２细胞的作用与Ｍ－ＣＳＦ的作用相似     

巨噬细胞集落刺激因子及其受体在造血细胞中的表达

目的研究异常造血时巨噬细胞集落刺激因子（ＭＣＳＦ）及其受体（ＭＣＳＦＲ）在细胞中的分布并探讨其临床意义。方法用ＭＣＳＦ、ＭＣＳＦＲ的单抗以卵白素生物素辣根过氧化物酶复合物免疫标记法（ＡＢＣ免疫酶标法）,测定其在６株血源细胞系,１４４例患者骨髓和１６０例患者外周血细胞中的表达。结果６株血源细胞系均有细胞ＭＣＳＦ和ＭＣＳＦＲ的表达。正常人骨髓和外周血细胞中未测出细胞ＭＣＳＦ和ＭＣＳＦＲ的表达。多种恶性和少数良性血液病患者骨髓和外周血细胞中可测出ＭＣＳＦ和ＭＣＳＦＲ的表达,其中急性非淋巴细胞白血病组的表达频率和积分值明显高于其他组（Ｐ＜０．０５）。ＭＣＳＦ、ＭＣＳＦＲ阳性细胞主要为粒系细胞（Ｍ４,Ｍ５亦以粒细胞为主）,单核系也较多见,红系和巨核系少见,未见阳性的淋巴细胞。阳性反应主要见于细胞膜和细胞浆,细胞核少见。结论异常造血时细胞ＭＣＳＦ、ＭＣＳＦＲ分布甚广,髓系白血病表达尤高,值得深入研究。     

粒细胞集落刺激因子受体在急性白血病中的表达及临床意义

目的探讨粒细胞集落刺激因子受体(GCSFR)在急性白血病(AL)中的表达及临床意义。方法选初诊或难治复发的急性髓性白血病(AML)患者30例,急性淋巴细胞白血病(ALL)患者20例,正常对照20例。化疗前留取骨髓5ml,用GCSFR、CD34单抗,采用流式细胞技术(FCM)检测GCSFR、CD34在AL细胞的表达情况。同时制备骨髓单个核细胞(MNC)悬液,并分别加入不同浓度的GCSF(5、10、15、20和25ng/ml),培养24h后用FCM检测其DNA倍体的量。结果GCSFR、CD34的表达率:AML为(76.5±12.8)%和(45.15±4.22)%;ALL为(6.12±1.98)%和(46.75±3.15)%;对照组为(80.5±10.8)%和(3.15±0.22)%。骨髓MNC培养24h后DNA倍体量在AML随着GCSF浓度的增加有上升的趋势,在ALL和对照组无明显变化。结论GCSFR主要表达于AML细胞,并促进其增殖;不表达于ALL细胞,不促进其增殖;也表达于成熟粒细胞,但不促进其增殖。     

Survivin在白血病细胞中的表达及GM-CSF对其影响的研究

本研究旨在探讨凋亡存活蛋白survivin在白血病的原代细胞和白血病细胞系中的表达情况，并观察粒-巨噬细胞细胞集落刺激因子（GM-CSF）对其的影响。用RT-PCR方法检测37例白血病患者、10例正常成人骨髓和3种白血病细胞系（K562、HL-60、U937）中survivin的表达。以HL-60细胞为对象，加入合适浓度的GM-CSF，用RT-PCR和Western blot分别检测加药前后surivin mRNA和蛋白水平的变化。结果显示：37例白血病患者中有25例表达survivin，阳性率为67．6％，其中急性淋巴细胞白血病（ALL）细胞中survivin mRNA表达阳性率为73．3％，高于急性髓系白血病（AML），但均高于正常对照组（20．0％）（P〈0．05）。3种细胞系K562、HL-60和U937全部表达survivin。合适浓度的GM-CSF作用2天后，HL-60细胞中survivin的表达明显升高，其中mRNA水平升高了26％，蛋白水平升高了49％。结论：suvivin在白血病的原代细胞和细胞系中均有较高表达，而GM-CSF能显著提高HL-60细胞中survivin的水平。     

应用联合化疗加人粒细胞集落刺激因子治疗急性白血病

目的:探讨联合化疗加人粒细胞集落刺激因子治疗急性白血病的临床效果.方法:治疗组42例急性白血病患者在常规联合化疗基础上加用重组人粒细胞集落刺激因子(升白灵)1～2μg@kg-1@d-1,皮下注射或静滴,从上一化疗疗程结束用至下一疗程开始前停药.对照组28例单用联合化疗.结果:治疗组完全缓解率80.9%,对照组71.4%(P＜0.05);治疗组缓解期4～52个月,中数缓解期20.5个月,生存期7～53个月,中数生存期24个月;对照组缓解期4～31个月,中数缓解期14个月,生存期5.5～35个月,中数生存期14.2个月.结论:应用联合化疗加人粒细胞集落刺激因子治疗急性白血病比单用联合化疗临床效果更好,能更进一步改善其预后.     

白细胞介素6基因转染的白血病细胞克隆株的建立及其生物学特性的观察

作者将人白细胞介素６（ＩＬ－６）基因转染至ＦＢＬ－３红白血病细胞，建立了高分泌ＩＬ６的ＦＢＬ－３细胞克隆株，并观察了其生物学特性。采用磷酸钙ＤＮＡ共沉淀法将ＩＬ－６表达载体ＢＣＭＧＮｅｏ－ＩＬ－６转移至ＦＢＬ－３细胞中，通过Ｇ４ｌ８抗性筛选、有限稀释法和上清中ＩＬ－６活性的测定，从多株阳性克隆中筛选到一株高分泌ＩＬ－６（２２５．６Ｕ／ｍｌ）的克隆株。体外观察表明，ＩＬ－６基因转染的ＦＢＬ－３细胞体外生长能力、集落形成能力均减弱，且其生长抑制程度与分泌ＩＬ－６水平呈正相关。给小鼠皮下接种后肿瘤结节形成率降低，生长速度减慢，荷瘤小鼠存活期延长。以上结果表明，ＩＬ－６基因转染的ＦＢＬ－３细胞致瘤性下降，这为将该细胞制备成新型瘤苗治疗白血病打下了基础。     

PREPARATION AND ANTIGENICITY OF M PROTEIN RELEASED FROM GROUP A, TYPE 1 STREPTOCOCCAL CELL WALLS BY PHAGE-ASSOCIATED LYSIN

The lysis of Group A type 1 streptococcal cell walls by phage-associated lysin has been described. In the preparation of lysin, a new semisynthetic non-protein media to support growth of the propagating strain of Group C streptococci was employed. Following lysis of the cell walls, the resulting digest was partially purified by ion-exchange chromatography and ammonium sulfate precipitation. In addition to M protein, the resulting preparation (called lysin M protein) contained the group-specific carbohydrate and the T protein—but did not contain antigens, detectable by precipitin tests, which cross-reacted with absorbed heterologous group or type antisera. Capillary precipitin reactions between the lysin M protein and type-specific antiserum did not occur in the presence of high ionic strength buffers; these buffers did not similarly affect precipitin reactions of acid M protein. Type 1 lysin M protein is shown to be a good antigen. A total of 1.5 mg. injected intradermally in saline produced bactericidal antibody in eight of nine rabbits; when injected in adjuvant in one rabbit, protective serum antibodies developed. Streptococci grown in sera from seven of nine rabbits immunized with lysin M protein demonstrated significantly longer chains than when grown in normal rabbit serum. Antibody as demonstrated by each of these three tests was shown to be type-specific. No local or systemic toxicity was noted following intradermal injection in rabbits of lysin M protein.     

A NEUTROPHIL-DEPENDENT PATHWAY FOR THE GENERATION OF A NEUTRAL PEPTIDE MEDIATOR : PARTIAL CHARACTERIZATION OF COMPONENTS AND CONTROL BYα-1-ANTITRYPSIN

A biologically active neutral peptide mediator is cleaved from a plasma protein substrate by an α-1-antitrypsin-inhibitable serine protease apparently residing on the membrane of the human neutrophil. The peptide mediator has an approximate mol wt of 1,000, and is distinguished from the kinin peptides by a neutral isoelectric point, susceptibility to inactivation by trypsin as well as chymotrypsin and activity on the isolated, atropinized, and antihistamine-treated guinea pig ileum with relatively little action on the estrous rat uterus. The neutrophil protease is fully inhibitable by DFP, trypsin inhibitors from lima or soy bean, and α-1-antitrypsin and is associated with the high mol wt fragments of the neutrophil and not the nuclear, lysosomal, or cytoplasmic subcellular fraction. The substrate has an approximate mol wt of 90,000 and is chromatographically separable from kininogen. The exquisite sensitivity of the neutrophil protease to α-1-antitrypsin was established both by inhibition with highly purified α-1-antitrypsin and by the inability of the protease to generate detectable neutral peptide in a homozygous (ZZ) α-1-antitrypsin-deficient patient without heat inactivation of the residual inhibitor. On the other hand, plasma from a (null) α-1-antitrypsin-deficient patient supported neutral peptide generation and revealed an additional factor which inactivated neutral peptide.