IgA类鼠抗人A血型单抗的研制和临床应用

应用Ｂ淋巴细胞杂交瘤技术，以Ａ血型物质ＭＳＳ１０％２×（Ａ１）为抗原，经ＰＥＧ细胞融合，获得两株分泌ＩｇＡ类鼠抗人Ａ血型单抗杂交瘤细胞，细胞培养上清液效价为１０２４～２０４８，亲和力２～３ｓ。其中２Ｈ１０Ｅ１１杂交瘤细胞经血凝特异性试验、血凝抑制试验、吸收放散试验、红细胞标准谱细胞试验，表明２Ｈ１０Ｅ１１单抗仅对Ａ血型红细胞特异。２Ｈ１０Ｅ１１细胞置液氮中贮存３年，复苏传代后分泌抗体能力不变。单抗置５６℃水浴３０ｍｉｎ，－２０℃和３７℃反复冻融３次，效价不变。临床试用近百万人次，无一例错型，准确率１００％。证明可以替代标准血清，用于临床血型定型。     

A reliable method for recovering cryopreserved hybridoma cells for cloning

Summary A method is described for the reliable recovery of hybridoma cell lines from cryogenic storage. This procedure provides for (a) the efficient retrieval of cells secreting antibody for rapid confirmation of specific antibody production; (b) cells for cloning; (c) cells for restocking the liquid nitrogen storage banks; and (d) cultures to detect and possibly eliminate contamination.     

分泌本周氏蛋白单克隆抗体杂交瘤细胞系的建立

本文报告了以人的本周氏蛋白(BJK)免疫BALB/C小鼠脾细胞与SP2/o—Ag14小鼠骨髓瘤细胞在聚乙二醇(PEG)MW400作用下融合,并经7次克隆后获得二株抗人的BJK杂交瘤细胞系(E_6,H_4)。经染色体核型分析证实为杂交瘤细胞。本单克隆抗体经标准兔抗小鼠IgG亚类血清鉴定,属小鼠IgG_2a,IgG_2b。两株细胞经液氮冻存、复苏仍能正常分泌抗体。用酶联免疫吸附试验(ELISA)测定植瘤小鼠的腹水及其血清的抗体滴度分别为10~4—10~5,10~3—10~4。     

抗间日疟原虫红内期单克隆抗体的研制和鉴定

用Ｐｅｒｃｏｌｌ密度梯度离心法从志愿者血液中分离出感染间日疟原虫的红细胞。以此作为免疫原，免疫Ｂａｌｂ／ｄ小鼠，获得５株杂交瘤细胞系。融合率平均为４５％，抗体阳性率为０．６６％。用ＩＦＡｆｏＩＰＡ染色进行免疫组化分析，４Ｂ２和８Ｅ３分泌的单克隆抗体呈较强的阳性反应，与约氏疟原虫红内期无交叉反应。     

Isolation, characterization, and application of monoclonal antibodies to rat tyrosine hydroxylase

Tyrosine hydroxylase (TH, tyrosine 3-monooxygenase; EC 1.14.16.2) activity in crude extracts of rat pheochromocytoma, rat brain, and bovine adrenal medulla can be immunoprecipitated in an indirect assay by monoclonal antibodies prepared against partially purified rat pheochromocytoma TH. One of these monoclonal antibodies, TH-2D8-2, can be used for immunocytochemical localization of TH in cell bodies, dendrites, and axons in catecholaminergic neurons (e.g., cells in the substantia nigra) of rat brain and in the cell body, neurites, and growth cones of rat pheochromocytoma cells after treatment with nerve growth factor. When linked to Affi-gel 10, this monoclonal antibody can also be used for immunoaffinity purification of rat and bovine TH. These results suggest that TH-2D8-2 is a valuable reagent with which to investigate the localization, physiological regulation, and function of this important enzyme.     

The use of somatic antigen of Haemophilus influenzae for the monitoring of T cell-mediated skin test reactivity in man

To investigate its usefulness as a skin test antigen, Haemophilus influenzae somatic antigen was tested in 28 healthy individuals, both in soluble and aggregated form. All subjects were found to possess specific antibodies against H. influenzae of both IgG and IgM subclass, thus showing their previous exposure to this commensal micro-organism. The somatic antigen in solution was found to be a poor antigen for eliciting a delayed hypersensitivity skin response: only 2 out of 16 subjects reacted with a positive DTH pattern. In contrast, 25 out of 28 persons showed a positive DTH pattern when somatic antigen was used in aggregated form. Two types of DTH reaction patterns could be detected (in a ratio of approximately 3:2), viz. those with an early (24 h) and those with a late (48 h) maximal swelling. Histology of 3 early and 1 late DTH reaction showed perivascular infiltrates of mainly Thelper/Tinducer lymphocytes. Hardly any basophils were seen. One negative skin test, biopsied at 6 h, showed no signs of Arthus reactivity. It can be concluded that skin tests using the aggregated form of the somatic antigen of H. influenzae are useful for assaying specific T-cell-mediated reactivity in man.     

抗粘放菌5519Ⅰ型菌毛单克隆抗体的制备和鉴定

目的制备抗粘放菌5519Ⅰ型菌毛单克隆抗体。方法采用BALB/C小鼠免疫后取脾细胞与SP2/0小鼠骨髓瘤细胞杂交融合,制备单克隆抗体,并以ELISA法和双向琼脂扩散法进行鉴定。结果共得到6株杂交瘤细胞,抗体效价达110～100万,类型为IgG,亚类为IgG1。结论经BALB/C小鼠免疫可得到高纯度的抗粘放菌Ⅰ型菌毛单克隆抗体,而且效价高,细胞株稳定分泌抗体。     

Immunization of human lymphocytes with Epstein-Barr virus capsid antigens in vitro

Research Center for Development and Introduction of Modern Methods of Molecular Diagnosis, Ministry of Health of the USSR, Moscow. (Presented by Academician of the academy of Medical Sciences of the USSR A. I. Borob'ev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 111, No. 1, pp. 62–64, January, 1991.     

Plasmodium falciparum-inhibitory monoclonal antibodies produced by human hybridomas

Stable human hybridomas were generated that produced inhibitory anti-Plasmodium falciparum monoclonal antibodies. Peripheral blood lymphocytes, obtained from adults in Liberia, a malaria endemic area, were immortalized with Epstein-Barr virus and then fused with KR4, a human, lymphoblastoid cell line. Stable hybridomas that produced anti-P. falciparum monoclonal antibody were identified by an ELISA assay that used the trophozoite and schizont antigens of both the Honduras I and FCR3 parasite strains. Monoclonal antibodies produced by selected hybridomas derived from lymphocytes of two individuals were subsequently studied. The anti-parasite antibodies were produced at 1-3 micrograms/ml in culture supernatants. All of the monoclonal antibodies bound specifically to trophozoites and schizonts of both strains of parasite in an indirect immunofluorescence assay and inhibited production of ring stage parasites by more than 90% when added to trophozoite or schizont containing erythrocytes in culture. Western immunoblot analysis of antigens obtained from trophozoites and schizonts (parasite age span of 36 to 48 h) was performed using either affinity purified or ammonium sulfate-concentrated monoclonal antibody. Antibody from three hybridomas which bound primarily to antigens of the Honduras 1 strain had Mr of approximately 140,000, 130,000 and 123,000.