Design methods for some dose-response models.

A recently described design method for one-parameter biomedical models such as limiting or serial dilution assays is generalized to two-parameter models for which the dose-response relationship can be expressed as a linear regression model with parameters alpha (intercept) and beta (slope). Design formulae are proposed for three different cases in which prior information about the unknown regression parameters alpha and beta is available (alpha known, beta known and neither known, respectively). A suitable transformation of the two-parameter model enables the direct application of the one-parameter design method to the first two cases, while the third needs more advanced considerations.

Using jackknife methods for estimating the parameter in dilution series.

Dilution assays are quantal dose-response assays that detect a positive or negative response in each individual culture within groups of replicate cultures that vary in the dose of cells/organisms tested. We propose three jackknife versions of the maximum likelihood estimator of the unknown parameter, i.e.

Computer aided design and evaluation of limiting and serial dilution experiments.

Two computer programs are presented which can be used for the determination of the design and the statistical evaluation of data in the context of limiting and serial dilution analysis. The first program (DESIGN) gives the experimenter the opportunity to set up different designs, to improve a design according to several suggestions, to make a picture of a design and to evaluate the results of artificial data, obtained from a random experiment corresponding with the chosen design, until (s)he is convinced of having a suitable design for a particular experiment. The second program (EVALUATE) evaluates the experimental data.

Limiting dilution assays. Experimental design and statistical analysis.

Two issues in limiting dilution analysis are considered. The first concerns the experimental design: a mathematical algorithm has been developed which calculates the number of replicate culture groups, and the (mean) number of cells per well to be used on the basis of the experimenter's a priori information about the unknown frequency. The procedure guarantees useful data if the a priori interval estimate of the frequency to be determined is correct and the cells are willing to grow.

Analysis of cytotoxic T lymphocyte response in rejecting allografted canine kidneys.

The involvement of cytotoxic T lymphocytes (CTL) in the rejection process of allografted canine kidneys was studied. The frequency of donor-specific (precursor) CTL was determined with a sensitive limiting dilution assay. Longitudinal sampling of peripheral blood and kidney aspiration biopsies were used to obtain information on the CTL response toward the graft.