在无血清组织培养系统中由流感病毒诱导的人类二次免疫反应。

Human secondary immune responses induced by influenza in a serum-free tissue culture system.

作者信息

Weiler J M, Rinderknecht S B, Sharath M D, Yarchoan R

出版信息

J Immunol Methods. 1985 Oct 10;82(2):333-9. doi: 10.1016/0022-1759(85)90365-5.

Abstract

Peripheral blood lymphocytes are capable of producing antibody in response to antigenic stimulation by a recall antigen, influenza, during culture for 10 days in medium consisting of 1 part Ham's F-12 and 1 part Iscove's modified Dulbecco's medium supplemented with sodium bicarbonate, bovine crystalline insulin, human transferrin, 2-mercaptoethanol, progesterone, and bovine serum albumin. Anti-influenza antibody levels in the supernatants were determined by enzyme-linked immunosorbent assay. Optimal conditions for production of anti-influenza antibody in this serum-free medium were: influenza concentration, 0.032-0.125 HAU/ml; day of harvest, 10; and cell concentration, 3.0-4.0 X 10(5) cells in 200 microliter of medium per well. Use of serum-free medium will allow examination of the effects of various additives to tissue culture without concern for unknown factors or potential interaction with serum.

摘要

外周血淋巴细胞在由1份Ham's F - 12和1份添加了碳酸氢钠、牛结晶胰岛素、人转铁蛋白、2 - 巯基乙醇、孕酮和牛血清白蛋白的Iscove改良杜尔贝科培养基组成的培养基中培养10天时，能够对回忆抗原流感病毒的抗原刺激产生抗体。通过酶联免疫吸附测定法测定上清液中的抗流感抗体水平。在这种无血清培养基中产生抗流感抗体的最佳条件是：流感病毒浓度为0.032 - 0.125血凝单位/毫升；收获天数为10天；细胞浓度为每孔200微升培养基中含3.0 - 4.0×10⁵个细胞。使用无血清培养基将允许在不考虑未知因素或与血清潜在相互作用的情况下，研究各种添加剂对组织培养的影响。