ABSTRACT
The leucocyte migration inhibition test in agarose as described by Clausen (1971) was modified into a statistically designed assay of LIF activity using human polymorphonuclear leucocytes from single blood donors. Individual assays included a laboratory standard of lymphokine with LIF activity prepared from the culture supernatants of the RPMI 1788 human lymphoblastoid cell line (LCL-LK). Analysis of 157 LIF assays revealed simple criteria by which the acceptability of an individual assay could be judged before subjecting it to statistical analysis. The failure of LIF assays to meet these criteria of acceptability was particularly associated with low areas of control polymorph migration in the absence of lymphokine ('spontaneous migration'). We demonstrate that the statistically designed assay permits the measurement, with precision, of LIF activity in units/ml by reference to a working standard of LCL-LK. We illustrate the use of this assay in the measurement of LIF activity generated by tuberculin-stimulated human peripheral blood lymphocytes.
Subject(s)
Leukocyte Migration-Inhibitory Factors/analysis , Lymphokines/analysis , Lymphokines/standards , Animals , Cell Line , Cell Migration Inhibition , Dose-Response Relationship, Immunologic , Guinea Pigs , Humans , Leukocyte Migration-Inhibitory Factors/pharmacology , Lymphocyte Activation , Lymphokines/biosynthesis , Neutrophils/immunology , Reference Standards , Tuberculin/immunologyABSTRACT
The migration inhibiton test in agarose as described by Clausen (Acta Allergol 26:56, 1971) was modified into a statistically designed assay comprising a laboratory standard in each assay, and using buffy coat cells or peripheral polymorphonuclear leukocytes from pigs as target cells. The precision of the assay was improved by combining cells from several pigs. Fifty-5000 microliters of lymphokine-containing test fluid could be used in the assay provided that this fluid contains the essential nutrients. In our hands, the capillary tests was unsuitable for a quantitative assessment of migration inhibition activity.
Subject(s)
Cell Migration Inhibition , Leukocytes/immunology , Lymphokines/pharmacology , Animals , Biological Assay , Capillary Action , Guinea Pigs , Sepharose , SwineABSTRACT
Treatment of NZB/NZW F1 (B/W) female and castrated male mice with testosterone or 19-nortestosterone (nandrolone), either by implantation in silastic tubing or by subcutaneous injections of their decanoate esters, reduced in a dose-dependent manner symptoms associated with murine lupus (proteinuria, IgG antibodies to DNA) and prolonged survival. These phenomena were observed under both prophylactic (start at 3-4 weeks) and therapeutic treatments (start 27-29 weeks). Nandrolone and its decanoate ester were at least as potent as testosterone and testosterone decanoate. As the unwanted androgenic properties of nandrolone and its ester are significantly less pronounced than those of testosterone and its ester, also in these NZB/NZW mice, the beneficial effect on murine lupus does not seem to be associated with these properties.
