感染小鼠白血病病毒的L783V／3T3细胞系的建立及鉴定

L783是上海医科大学病理生理教研室七十年代建立的一株小鼠可移植性淋巴细胞型白血病。用L783发病小鼠的血液、脾脏等组织的无细胞提取液,分别感染NIH-3T3培养细胞,建立了L783小鼠白血病病毒感染的NIH-3T3细胞系(L783V/3T3)。实验结果表明:L783V/3T3细胞的XC合胞形成试验阳性,电镜下可观察到典型的A型和C型病毒颗粒。制备L783V/3T3细胞无细胞提取液,接种新生昆明种乳鼠,能诱发淋巴细胞型白血病。     

单克隆抗体对肾综合征出血热病毒感染乳属保护作用的研究

本文进一步观察了自制的24株抗HFRS病毒血凝素McAbs对感染HFRS病毒新生乳鼠的保护作用。将HFRS病毒陈株100LD_(50)腹腔感染昆明种小鼠的新生乳鼠后24h,腹腔分别接种24株抗HFRS病毒McAb,观察McAb对感染新生乳鼠的保护作用。结果发     

病毒性L6565白血病细胞克隆株的建立及其特性研究进展

Ｔ638和Ｌ6565模型是目前国内仅有的两株ＲＮＡ病毒诱发的白血病 ,均属于我国天津 (Ｔ) -上海 (Ｓ) -遵义 (Ｚ)小鼠白血病系统之内[1 ,2 ] 。近年来未见Ｔ638白血病研究的进一步报道 ,情况不详。Ｌ6565病毒性淋巴细胞白血病是由程立等于 1 965年建立的瘤株[3～ 6] 。 1 986年张雷等[7] 应用Ｌ6565白血病小鼠外周血液无细胞提取液感染ＮＩＨ/3Ｔ3细胞 ,在体外建成了Ｌ6565-Ｂ1 细胞系 ,惜已失传。最近 ,殷莲华等[8] 将Ｌ6565白血病小鼠脾脏在灭菌条件下制成细胞悬液后 ,用ＲＰＭＩ 1 640液和 1 0 %小牛血清中培养 2 0代后 ,再用有限稀释法置…     

淋巴瘤细胞（系）克隆和白血病病毒感染细胞系的建立及其分化诱导研究

为了进行白血病病毒病因学研究,本室已先后建成了小鼠L6565病毒性白血病、SRS淋巴瘤和L783移植性白血病等瘤株,对它们的生物学特性已进行了详细的研究。近年来,在动物模型基础上,在体外建立了小鼠SRS-82细胞系及其SAC系列克隆,并应用分离的白     

用移植方法对L6565、SRS及L783三瘤株抗原性的研究

用移植方法对L6565、SRS及L783三瘤株的抗原性所作的实验研究表明,经丝裂霉素处理的SRS瘤细胞(SRSm)免疫的昆明种小鼠及经丝裂霉素处理的L733白血病细胞(L783m)免疫的SW1系小鼠,分别对SRS瘤细胞及L783白血病细胞的攻击能产生保护作用,提示这两株瘤细胞具有肿瘤相关性移植抗原(TATA);经L6565及L783白血病细胞免疫的昆明种小鼠,也能产生抗SRS瘤细胞攻击的作用,经L6565白血病细胞免疫的SW1系小鼠可保护其对L783白血病细胞的攻击,说明L6565、SRS及L783三瘤株间具有交叉性TATA。实验还发现L6565白血病组织的无细胞提取液及SRS瘤细胞的3M KC1提取物也具有一定的TATA活性。     

SARS病毒对乳鼠和鸡胚致病性的初步观察

目的　建立SARS病毒的动物模型。方法　选用 1～ 5日龄的BALB c乳鼠和昆明种乳鼠 ,通过脑内、腹腔和鼻腔 3种途径 ,分别接种自广州和北京地区非典型肺炎病人中分离的SARS病毒BJ0 1和GZ0 1株 ,观察该病毒对不同种系小鼠的致病性。同时 ,选用 8～ 10日龄鸡胚经尿囊腔接种这 2株病毒 ,观察对鸡胚的致病性。结果与结论　采用SARS病毒BJ0 1和GZ0 1株经脑内接种 ,均可使BALB c乳鼠和昆明种乳鼠发病或死亡 ,但这种致病性不规律 ;而从腹腔和鼻腔途径接种病毒均不能使乳鼠发病。鸡胚对该病毒不敏感。     

SRS白血病病毒的核糖核酸和蛋白质及其致白血病作用研究

1971年我校病理生理学教研室建立了小鼠SRS腹水瘤模型,研究表明这是一株不带有T、B标记的淋巴干细胞瘤株,电镜观察见到A型和C型病毒颗粒(命名为SRSV),注射新生乳鼠有致白血病活性。程立等     

小鼠SRS腹水瘤SAC克隆系列的建立及其鉴定

应用有限稀释法,由小鼠SRS-82腹水瘤细胞系分离得到SAC克隆株系列。实验结果表明SRS-82细胞系是一个异质的群体,亲系及其SAC-ⅡB2,-ⅡC3克隆在细胞生长特性、染色体核型和致瘤性等方面都存在明显的差异。XC检测阳性。电镜观察证实上述细胞均有A型和C型病毒颗粒存在。ABC免疫组织化学方法和免疫荧光染色检测结果提示它们属于小鼠T淋巴细胞类型。     

不同种鼠对实验脑型疟发生的影响研究

目的比较昆明小鼠和C57BL/6小鼠作为种鼠对实验脑型疟模型的影响。方法分别以伯氏疟原虫ANKA株感染C57BL/6小鼠和昆明小鼠作为传代用种鼠,当种鼠原虫率为5%～15%时接种子代C57BL/6小鼠,观察两组小鼠原虫率、脑型疟发生率以及死亡率。同时,通过脑组织切片和脑部淋巴细胞的流式检测,观察两组发生脑型疟小鼠的脑部微血管中感染疟原虫红细胞和CD8＋T细胞的粘附情况,另外,通过感染CD8＋TKO小鼠,证实CD8＋T细胞在两组小鼠发生脑型疟中的作用。结果用昆明小鼠作为种鼠的实验组的原虫率和脑型疟发生率均明显高于用C57BL/6小鼠作为种鼠的实验组,发生脑型疟小鼠的脑部组织切片发现,脑部微血管可见明显的感染疟原虫红细胞的粘附和CD8＋T淋巴细胞浸润;而用昆明小鼠作为种鼠感染CD8＋T细胞缺失的C57BL/6小鼠并不能诱导实验脑型疟的发生。结论与C57BL/6小鼠相比,昆明小鼠作为种鼠的实验组的脑型疟发病率更高,而且感染疟原虫红细胞和CD8＋T细胞在脑部微血管内的粘附也是该脑型疟发生的主要因素,因此,更适合用于实验脑型疟模型的建立及其机制的探讨。     

重组结核抗原痘苗病毒Ankara株的构建及其免疫原性研究

目的 构建5种不同类型的表达结核杆菌特异抗原的重组痘苗病毒,并研究其特异免疫原性.方法 运用同源重组技术将含结核分泌抗原Ag85A和ESAT-6的基因片段插入痘苗病毒表达质粒p18中.重组质粒导入痘苗病毒Ankara(MVA)后构建重组痘苗病毒,经筛选和Western blot鉴定,得到5个种类的带有结核抗原基因的重组病毒.用构建的5种重组病毒免疫小鼠,MTT法检测免疫后小鼠脾淋巴细胞对特异结核抗原的增殖反应;ELISA检测小鼠脾淋巴细胞培养上清液中IFN-γ的含量;结核菌素纯蛋白衍化物(PPD)皮内试验以检测重组病毒引发的针对结核抗原的特异细胞免疫应答.结果 构建的5种蘑组病毒介导的细胞表达产物经Western blot鉴定确认相对分子质量与结核抗原一致.免疫小鼠两次后,5种重组病毒免疫组脾淋巴细胞体外与Ag85A-ESAT-6融合蛋白共培养后表现出明显的增殖活性(P<0.01),培养上清液中IFN-γ的浓度均较同组细胞经生理盐水刺激明显增高(P<0.05);与空痘苗病毒或生理盐水免疫后小鼠相比,5种重组MVA免疫组脾淋巴细胞与AgB5A.ESAT-6融合蛋白共培养后同样表现出明显的增殖活性(P<0.01),与Ag85A-ESAT-6融合蛋白共培养的细胞上清液中IFN-γ的浓度均升高(P<0.01).与空痘苗病毒或生理盐水免疫后小鼠相比,5种重组MVA免疫组小鼠对PPD都表现出显著的迟发型超敏反应应答(P<0.05).结论 成功构建了5种不同类型的表达结核杆菌抗原的重组痘苗病毒疫苗,其免疫小鼠后可激发针对结核杆菌抗原的特异性细胞免疫.     

Etiologic role of lactic dehydrogenase virus infection in an age-dependent neuroparalytic disease in C58 mice

Lactic dehydrogenase virus (LDV) associated with transplantable line Ib lymphocytic leukemia in C58/Wm mice, K36 lymphocytic leukemia in AKR/J mice, and the Gardner lymphosarcoma in C3H/HeJ mice elicited a fatal neuroparalytic disease when injected ip into 7- to 9-month-old X-irradiated indicator C58 mice. LDV associated with the WEHI-3B line of transplantable myelomonocytic leukemia or the Harding-Passey transplantable myeloma in BALB/c mice failed to elicit the disease. Recipients of such tumor extracts were immune to rechallenge by line Ib-associated LDV. Tumor lines free of LDV failed to elicit the disease or immunize recipient mice to line Ib LDV challenge. The Plagemann (P-LDV), Riley (R-LDV), and Notkins (N-LDV) strains of LDV were less neuropathogenic than the line Ib-derived strain (Ib-LDV). Indicator C58 mice that survived infection by the P-LDV, R-LDV, and N-LDV strains were immune to rechallenge by Ib-LDV. Antiserum prepared in young C58 mice to Ib-LDV or R-LDV protected indicator C58 mice from Ib-LDV challenge. These results show that a common viral contaminant of transplantable tumors and virus stocks that ordinarily is not pathogenic elicits a fatal neurologic disease in genetically susceptible, immunosuppressed, C58 mice.     

DNA病毒与RNA病毒相互作用小鼠模型的建立

用Ｃ型病毒（ＲＮＡ病毒）和／或紫外线灭活的（疱疹病毒）ＨＳＶ－２（Ｗｕ）或ＨＳＶ－２（３３３）接种新生小鼠，结果表明接种了ＨＳＶ－２（Ｗｕ）、ＨＳＶ－２（３３３）和作为对照的小鼠均未发生白血病，在ＨＳＶ－２（Ｗｕ）组中接种了Ｃ型病毒加ＨＳＶ－２（Ｗｕ）的小鼠白血病发生率是７５００％，只接种了Ｃ型病毒的小鼠中有４４．７４％发生白血病，两者之间有高度显著性差异（Ｐ＜０．０１）。另外，在ＨＳＶ－２（３３３）组中用Ｃ型病毒加ＨＳＶ－２（３３３）接种小鼠以后有７９．１６％发生了白血病，只接种了Ｃ型病毒的小鼠中有４１．０２％发生白血病，其差异也有高度显著性（Ｐ＜０．０１）。由此可见Ｃ型病毒分别与ＨＳＶ－２（Ｗｕ）或ＨＳＶ－２（３３３）在诱发小鼠白血病过程中存在相互作用。此动物模型可用于研究两类不同病毒相互作用的致癌原理。     

A glycoprotein of molecular weight 85,000 on human cells of B-lineage: detection with a family of monoclonal antibodies

Immunization of BALB/c mice with glycoproteins purified from a detergent extract of human chronic lymphocytic leukemia (CLL) cells by affinity to Lens culinaris lectin led to the production of several monoclonal antibodies with similar reactivity. One of the antibodies, 50B4, was purified and the corresponding antigen was isolated from a B-lymphoblastoid cell line extract by affinity chromatography to the 50B4-IgG immunoadsorbent. Co-purification of the antigenic activities associated with five other monoclonal antibodies was achieved. Purified and radiolabelled 50B4 antigen could be specifically immunoprecipitated not only by 50B4 but also by the other five antibodies. SDS-PAGE analysis revealed that all antibodies precipitated the same component, a polypeptide chain of apparent mol. wt 85,000 under reducing conditions. Competitive-binding studies between the purified antibodies indicated the presence of two distinct epitopes on the antigen. The epitopes, each recognized by three different antibodies, were equally accessible on the cell surface of either a B-CLL (3 X 10(5) molecules/cell), a B-lymphoblastoid cell line (11 X 10(5) molecules/cell) or two acute lymphocytic leukemia (ALL) cell lines of pre-B phenotype (5 X 10(5) and 0.8 X 10(5) molecules/cell respectively). Although the antigens purified from the strongly positive ALL cell line gave a gel pattern identical to that of the B-lymphoblastoid cell line, the antigens purified from the B-CLL extract were resolved into two distinct glycosylated polypeptides of mol. wts 85,000 and 77,000 under reducing conditions. The distribution of the antigen(s) is not restricted to cells of the B-lineage as mature T-cells and a variety of non-hematopoietic cell types express both epitopes of the antigen(s).     

Histopathologic studies on myeloproliferative sarcoma virus (MPSV) induced leukemias and hemangiosarcoma in Jar-2 rats

It is well known that MPSV induces myeloproliferative syndrome (MPS) in mice. Intravenous one shot inoculation of myeloproliferative sarcoma virus (MPSV) with Friend murine leukemia virus (F-MuLV) as a helper in newborn Jar-2 rats (on the second neonatal day) yielded hematopoietic malignancies in all the treated rats (25/25 rats) after 2 weeks' latency. MPS appeared from the 14th day in 14 rats. In the midst of the myeloproliferative field of the spleen and bone marrow, myeloblastic or myeloblastic-erythroblastic foci were observed. From 19th day, acute myeloblastic leukemia occurred in 3 rats and erythroleukemia in 8 rats. MPSV induced first MPS which remained as such or later developed into acute leukemia. Myelofibrosis as seen in mice was not observed. In addition, hemangiosarcoma of the brain, spinal cord and spleen appeared in 15 rats from the 24th day, and were often multiple. MPSV can yield the tumor only in newborn rats, and target cells of MPSV are not only hematopoietic cells but also endothelial cells of the brain, spinal cord and occasionally spleen.     

Human B cells express membrane-bound and soluble forms of the CD14 myeloid antigen.

The expression of the myeloid differentiation antigen CD14 on the B lineage was analyzed. A CD14-specific monoclonal antibody was used to isolate the antigen from normal B, B-type chronic lymphocytic leukemia cells, and a representative Epstein-Barr virus-transformed B lymphoblastoid cell line (EBVLCL). A soluble form of this protein was detected in the culture supernatant of all the B cell types tested. The molecule expressed in the normal B and B-type chronic lymphocytic leukemia cells was identical in size to the 52,000 mol. wt monocyte-isolated CD14 glycoprotein. A 64,000 mol. wt antigen was isolated from the lymphoblastoid cell line. Similar 2-D gel electrophoretic patterns to that of the monocyte-derived CD14 were obtained from the normal B and B-type chronic lymphocytic leukemia cell-isolated molecules. These similarities were reflected in minor isoelectric point (pI) differences between the polypeptide spots (pI 4.8), in the first dimension, and identical molecular weight (52,000) in the second dimension. The EBVLCL-isolated polypeptide, when analyzed by 2-D gel electrophoresis, showed a pI identical to that of the myeloid antigen (pI 4.6). The isolated soluble form was of smaller (47,000 mol. wt, normal B and B-type chronic lymphocytic leukemia cells) or similar size (64,000 mol. wt, lymphoblastoid cell line) compared with their corresponding membrane-bound forms. Interestingly, two-colour immunofluorescence analysis showed that only two out of four CD14-specific mAb tested bound to the B cells. We conclude that the CD14 antigen is, in fact, expressed in the B lineage. Its cell surface expression and serum level in the prognosis of B-type chronic lymphocytic leukemia patients needs to be evaluated.     

On cysts in mice of the low cancer line C57BL

Summary Autopsy findings revealed cystic changes in the lymph glands observed in old mice of the C57BL line formerly subjected to the test of leukemogenic activity of the brain filtrate of a man who had died of acute leukemia.The cystic fluid inoculated to newborn mice of the same line gave negative results. In a series of consecutive passages of cellular suspensions there was a development of tumors and cysts in young mice of homologous C57BL line as well as in mice of the CC57BR and CC57W lines. In transplantation of a cell-free material to newborn mice of four different lines (C57BL, CC57BR, CC57W, and C3HA) the cysts usually developed only at a more mature age, whereas in subsequent serial passages of suspensions of cystic walls and tumor from these mice — at the age from 4.5 mo to 18 days.Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 55, No. 4, pp. 82–91, April, 1963     

Effect of murine host genotype on MCF virus expression, latency, and leukemia cell type of leukemias induced by Friend murine leukemia helper virus

Leukemias induced by neonatal inoculations of several mouse strains with different strains of Friend murine leukemia helper virus (F-MuLV) were followed for time of disease onset, cytochemical analysis of predominant cell types in leukemic organs, and expression of infectious mink cell focus-inducing (MCF) viruses detected by mink cell foci or MCF-specific monoclonal antibodies. Most BALB.B and IRW mice had a rapidly appearing, severe anemia and hepatosplenomegaly consisting of erythroid cells. MCF viruses were usually isolated from enlarged spleens of IRW mice. In contrast, C57BL/10 mice had a lower incidence of disease and much slower course. Splenomegaly and lymphadenopathy with mild anemia were seen, and the predominant cell types were either myeloid (chloroleukemia) or lymphoid. MCF viruses were never isolated from this mouse strain. (C57BL/10 X IRW)F1 mice were intermediate in latency, but all mice had disease by 8 months. Myeloid, lymphoid, and some mixed leukemias with an erythroid component were observed, but in no case did we see the severe anemia or pure erythroid involvement typical of IRW and BALB.B mice. MCF viruses were, however, isolated from 22% of these mice regardless of leukemia cell type. DBA/2 mice had a disease pattern similar to the (C57BL/10 X IRW)F1 mice, and MCF viruses were isolated from three of six mice tested. Inoculation of IRW mice with the low virulence B3 strain of F-MuLV produced disease with a longer latency than F-MuLV 57, but similar cell types were transformed by both viruses. In vitro cell lines were derived from 14 mice, and most were tumorigenic in vivo. Three lines released infectious MCF virus, and three others expressed MCF-specific cell surface antigens but did not release virus. Eight lines expressed no MCF infectious virus or viral antigens. Several lines released infectious xenotropic viruses and/or expressed xenotropic MuLV cell surface antigens recognized by monoclonal antibodies reactive with xenotropic viruses. The lack of MCF expression in many primary leukemic tissues as well as in in vitro derived leukemia cell lines of C57BL/10 and (B10 X IRW)F1 mice suggested that MCF virus generation and expression may not be required for leukemogenesis in some mouse strains or in some hemopoietic lineages.     

L7212白血病小鼠IL-2活性、IL-2 mRNA表达及复方中药清毒饮对其影响的研究

目的：探讨Ｌ７２１２白血病小鼠及其受体鼠──近交系６１５小鼠ＩＬ－２活性和ＩＬ－２ｍＲＮＡ的表达及复方中药清毒饮对其影响。方法：采用ＩＬ－２依赖株ＣＴＬＬ－２及细胞原位杂交的方法检测了由ＣｏｎＡ诱导的脾细胞培养上清中ＩＬ－２活性及脾细胞ＩＬ－２ｍＲＮＡ表达。结果：发现由ＣｏｎＡ诱导的Ｌ７２１２的白血病小鼠脾细胞培养上清中ＩＬ－２活性明显低于６１５小鼠,Ｐ＜０．０５;其脾细胞中ＩＬ－２ｍＲＮＡ表达也明显低于６１５小鼠,Ｐ＜０．０１。清毒饮显著提高ＣｏｎＡ诱导的Ｌ７２１２白血病小鼠脾细胞培养上清中的ＩＬ－２活性,Ｐ＜０．００１,并能显著提高其ｍＲＮＡ表达水平,Ｐ＜０．００１。结论：清毒饮可以作为生物反应调节剂用于白血病的临床治疗。