Agrup G, Källén B, Nilsson O
Int Arch Allergy Appl Immunol. 1980;63(1):105-12. doi: 10.1159/000232615.
A two-step modification of Clausen's technique of leukocyte migration under agarose is described. Blood leukocytes from a subject are incubated for approximately 4 days with or without antigen. The cultures, after reconstitution with antigen followed by repeated washing of the cultured cells, are added to fresh, dextran-separated blood leukocytes, and migration tests are performed in the absence of the antigen. Cells preincubated with antigen give a very marked reduction of the migration of the target leukocytes which must not necessarily be autologous. An inhibition of target cell migration can be seen with cells cultured without antigen for up to 3 days; with longer culture time, this effect is reduced or disappears. The effect of cells cultured with antigen, however, is equally strong irrespective of the culture time up to 1 week, and the most clearcut results will, therefore, be obtained after approximately 4 days culture. Also migration enhancement has been seen. Treatment of cells during the incubation with antigen and cycloheximide gives a dose-dependent blocking of the inhibiting effect. 5 patients were studied both before and after sensitization with DNCB: low concentrations of DNCB gave an inhibition and high concentrations an enhancement of the migration when the patients were sensitized. The technique is more sensitive and reproducible than the original Clausen technique. It is also suitable in studies where shortage of cells is a problem.
本文描述了对克劳森琼脂糖下白细胞迁移技术的两步改进方法。将受试者的血液白细胞在有或无抗原的情况下孵育约4天。在用抗原重构培养物并对培养细胞进行反复洗涤后，将培养物添加到新鲜的经葡聚糖分离的血液白细胞中，并在无抗原的情况下进行迁移试验。预先用抗原孵育的细胞会使靶白细胞的迁移显著减少，靶白细胞不一定必须是自体的。在无抗原培养长达3天的细胞中可观察到靶细胞迁移受到抑制；培养时间延长，这种效应会减弱或消失。然而，用抗原培养的细胞的效应，无论培养时间长达1周，都同样强烈，因此，在大约4天培养后将获得最明确的结果。还观察到迁移增强现象。在用抗原和环己酰亚胺孵育期间对细胞进行处理，会产生剂量依赖性的抑制作用阻断。对5名患者在用二硝基氯苯致敏前后进行了研究：当患者致敏时，低浓度的二硝基氯苯会抑制迁移，高浓度则会增强迁移。该技术比原始的克劳森技术更灵敏且可重复。它也适用于细胞短缺是一个问题的研究。
