湖泊浮游细菌与浮游植物的溶原病毒诱导率检测方法优化

溶原性感染作为浮游病毒生态学效应表达方式之一,常以24 h化学诱导得到的溶原性宿主可诱导率来量化。为了获得适合淡水环境的溶原性宿主可诱导率研究方法,本研究采集6个淡水湖泊水样,对每个水样采用直接法、滤膜法、滤膜振荡法和直接振荡法4种诱导培养方法,通过宿主诱导致死率计算6个淡水湖泊浮游细菌和浮游植物溶原诱导率。结果表明:无论是浮游细菌还是浮游植物,直接法获得的诱导结果普遍高于其他方法,表明游离病毒的裂解性感染能够显著影响溶原诱导率的测定结果;直接振荡法获得的6个湖泊溶原诱导率与滤膜法及滤膜振荡法获得结果间均无显著差异,说明实验中振荡培养能有效抑制裂解性病毒对宿主的裂解性感染。本研究通过直接诱导结合振荡培养能够获得与滤膜法一致的诱导效果,但相比于滤膜法,直接振荡法实验过程更加简单,是一种更为高效的测定方法。

In vitro Induction of primary antibody responses to particulate and soluble protein antigens in T cell-replaced murine spleen cell cultures

Specific antibody responses could be induced in serumfree condition.Specific anti-SRBC or anti-SRBC ghost antibody were induced from anti-Thy treated (T-depleted) murine spleen cells in serum-free culture in the presence of Con A conditioned medium.This induction system may facilitate the study of lymphokine functions on antigen triggered B cells. In T cell-replaced cultures,the antibody responses of B cells could be successfully induced when soluble SRBC membrane proteins were used as antigens.It thus indicates that antigen together with lymphokines are sufficient to drive B cells to become antibody secreting cells in the absence of T cells.The T cell-replaced system provides a more stable way for in vitro immunization and may be applied to monoclonal antibody production when in vivo immunization is difficult to be carried out.