Effect of myelopeptides on DNA and protein synthesis and on antibody secretion by mouse B-cell hybridomas

Institute of Immunology, Ministry of Health of the USSR, Moscow. (Presented by Academician of the Academy of the Medical Sciences of the USSR, I. P. Ashmarin.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 107, No. 6, pp. 724–726, June, 1989.     

分泌血吸虫抗原杂交瘤的制备

目的 制备分泌血吸虫抗原的杂交瘤。方法 采用杂交瘤技术，将血吸虫成虫细胞与Ｓｐ２／０骨髓瘤细胞融合，ＨＡＴ选择性培养，结果 获得了１株培养上 有与血吸虫病人血清起反应的杂交瘤。其染色体数为７２－８４，该杂交瘤细胞可持续培养１１周以上，但其生长缓慢，在培养８周后，出现细菌逐渐死亡。结论 杂交瘤技术不适于远缘杂交，细胞死亡可能与ＤＮＡ隔离一染色体物质丢失有关。     

一株IgG3抗异麦芽四糖—硬脂酰胺单克隆抗体的制备及免疫化学研究

以异麦芽四糖—硬脂酰胺(S-IM_4)免疫C_(57)BL/6J小鼠,取其脾细胞与骨髓瘤细胞融合,从4次细胞融合中,筛选出1株分泌IgG_3K抗S-IM_4杂交瘤。用定量沉淀试验和ELISA定量抑制试验测定此单克隆抗体结合部位的特异性与大小,发现其互补于异麦芽七糖(IM_7)。用亲和电泳法测得此单克隆抗体对葡聚糖B_(512)和IM_7的亲和常数分别为1.4×10~5ml/g和3.1×10~4M~(-1)。     

Establishment of stable human T-T hybridomas

Human T-T hybridomas potentially provide an invaluable resource for a variety of immunoregulatory molecules that modulate the immune response. To date, success in this technology, using human cell populations, has been hampered by several problems associated with proliferative and functional instability of the hybrid cells. These forms of instability are the result of a multifactorial process, with 1 parameter of importance being the chromosome number of the malignant parent cell line used for fusion. The present studies describe the production of a stable human T-T hybridoma generated by fusing a near diploid (modal chromosome number of 48) aminopterin-sensitive T cell line, CEM TG E11, and lectin-stimulated human peripheral blood lymphocytes. The rapidly growing hybrid cells have been clonally selected for the production of a B cell growth factor. Hybridization was documented by the presence of HLA phenotypes reflecting the combined antigens of the fusion partners. Fusions with 4 other partners besides CEM TG E11, where the majority of the cells had modal chromosome numbers ranging from 78 to 94, were proliferatively unstable. To date, hybrid cells derived from the CEM TG E11 fusion have been doubling approximately every 48 h for greater than 12 months, and selected clones constitutively produce B cell growth factor.     

A simple,rapid method for freezing hybridomas

Summary A simple, rapid method for freezing hybridomas in bio-freeze vials and in situ in 96-well microculture cluster plates is described. The method offers a convenient means of preserving hybridomas and a more manageable system for distributing the workload necessitated by the exponential increase in the number of wells after fusion or cloning or both.     

分泌抗B型流感病毒抗体杂交瘤细胞的制备及鉴定

目的 制备抗B型流感病毒的单克隆抗体，用以建立特异、灵敏、简便的B型流感金标试纸检测方法。方法 将B型流感病毒为完全抗原免疫BALB/c小鼠，得到的脾细胞与SP2／0细胞融合，获得稳定分泌抗流感B单抗的杂交瘤细胞株，对抗体进行鉴定。结果 用杂交瘤技术，得到了3株可稳定分泌抗流感B单抗的杂交瘤细胞株，能分泌高滴度的针对B型流感病毒的特异单抗。结论 所获抗B型流感病毒杂交瘤细胞株单克隆抗体，为金标法快速诊断B型流感感染奠定了基础。     

丙型肝炎病毒单克隆抗体的制备及其鉴定

目的：以基因工程表达的丙型肝炎病毒抗原蛋白（HCAg）为抗原，建立分泌单克隆抗体（McAb）的杂交瘤细胞系．并对其分泌的McAh进行鉴定。方法：用纯化HCV基因工程表达抗原（HCAg）免疫Balb／c小鼠，取脾细胞及骨髓瘤细胞按常规方法融合、筛选、克隆化及腹水注射制备McAb。采用ELISA及分片断鉴定技术进行鉴定。结果：筛选出5株能稳定分泌特异性抗-HC的McAb杂交瘤细胞株，经多次复苏传代仍能稳定分泌抗体，特异性强，效价高。与不同抗原蛋白的ELISA反应呈特异性阳性反应。分片断鉴定结果显示，所获5株McAb主要为抗HCV核心蛋白和非结构蛋白NS3。结论：此5株丙肝杂交瘤细胞株分泌的抗体对丙肝抗原具有特异亲和性，为丙型肝炎病毒抗原诊断试剂的研制提供了关键技术基础。