title: Statistical estimation & inference of cell counts from ELISPOT limiting dilution assays.
abstract: The ELISPOT assay is often used for cell count determination in immunological studies. Automated methods are needed to estimate cell concentrations from spot counts obtained from the assay. Three major distributions are assumed for observational cell counts. For each assumed distribution, individual least squares (LS)/ maximum likelihood and/or individual robust least squares (RLS) are applied for parameter estimation. Distributions of study endpoints (derived variables), defined as percentages of antigen-specific memory cell per total immunoglobulin G (IgG), are investigated to provide a basis for hypothesis testing. We show, under some weak conditions, that the distribution of endpoint estimates across subjects is approximately the same within a group. Thus, the t -test or the Wilcoxon Rank Sum test can be applied to detect group differences. These methods are compared through simulations and application to real data.
authors.affiliation: Department of Biostatistics & Computational Biology, University of Rochester Medical Center, Rochester, NY 14642, USA. Hongmei_Yang@urmc.rochester.edu
authors.forename: Hongmei
authors.initials: H
authors.lastname: Yang
authors.forename: David J
authors.initials: DJ
authors.lastname: Topham
authors.forename: Jeanne
authors.initials: J
authors.lastname: Holden-Wiltse
authors.forename: Hulin
authors.initials: H
authors.lastname: Wu
mesh_terms: D000938:Antigen-Presenting Cells / Q000166:cytology / Q000276:immunology; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology; D003198:Computer Simulation; D003627:Data Interpretation, Statistical; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods / Q000706:statistics & numerical data*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007156:Immunologic Memory; D009980:Influenza A virus; D007252:Influenza Vaccines / Q000276:immunology; D016018:Least-Squares Analysis; D015233:Models, Statistical; D009010:Monte Carlo Method; D016008:Statistical Distributions
chemical_list: D007074:Immunoglobulin G; D007252:Influenza Vaccines