分泌抗HSV—2型特异McAb的B_5杂交瘤细胞株的建立和初步应用

本文应用小鼠骨髓瘤细胞SP2/0—Ag14与用HSV—2 SaV株病毒免疫的BALB/C小鼠脾细胞融合,建立了一株分泌特异性抗HSV—2病毒的单克隆抗体(McAb)的杂交瘤细胞株。冻存20个月后复苏传代,仍能稳定分泌McAb。经ELISA间接法检测,该细胞培养上清滴度为1∶120,制备腹水的滴度为10~(-7),定名为B_5细胞株。B_5细胞的染色体数目为92～101,B_5McAb经鉴定属于IgG_1亚类,与HSV—2有高效价的结合反应,而与HSV-1、CMV、EHFV、Ad_3、RSV_(1387)等病毒不发生反应。应用微量细胞培养中和试验.鸡胚绒毛尿囊膜中和试验和家兔皮内感染HSV—2病毒保护试验证明,B_5McAb在试管内或机体内(兔)都能中和HSV—2病毒。B_5McAb与临床分离的19株HSV-2病毒中的14株起反应,鉴定符合率为73.6%、与5株HSV—1病毒株都不起反应。

ESTABLISHMENT AND INITIAL USE OF B_5 HYBRIDOMA CELL STRAIN SECRETING SPECIFIC MONOCLONAL ANTIBODY AGAINST THE HERPES SIMPLEX VIRUS-2

A hybridoma cell strain secreting specific monoclonal antibody(McAb) against the herpes simplex virus-2(HSV-a) was established by fusing the mouse lmyeloma cell line SP 2/0-Ag 14 with the spleen cell of Balb/c mouse immunized by the HSV-2 Sav strain. The hybridoma cell strain can steadily secret McAb after being refrigarated for twenty months. The McAb titer in culture supernatants was 1: 120 and in mouse ascites was 10~(-7) by the indirect method ELISA. It is termed B_5 cell strain. The chromosome amount of B_5 cell was92-101; B_5 McAb was of IgGl subtype. It can react highly with HSV-2, and can not react with HSV-1, CMV, EHFV, Ad_3, RSV_(1387). Using the "microplate neutralization test", "Chick embryo chorioallantoic membrane neutralazation test" and "McAb for rabbit inoculated intracutaneously protectivity test", we demontrated that B_5 McAb has neutralization activity in vitro or in vivo (rabbit). 50% neutralization endpoint log is -1.7; B_5 McAb may react with fourteen of nineteen HSV-2 strains isolated from clinical patients. The indentification coincidence rate is 73.6% and can not react with five HSV-1 strains.