ABSTRACT
TGF-beta is a well-described immunoregulatory molecule that is produced by most cell types. Many studies have been aimed at investigating the role played by TGF-beta in different cell types and situations. Most methods of measuring TGF-beta have previously relied on enzyme-linked immunosorbent assay (ELISA) or assays of its anti-proliferative effects on various cell lines. Both assays, though useful, cannot be used to effectively identify the cells that are producing TGF-beta in a mixture of cells. It is especially important to know the source and dynamics of TGF-beta secretion in cell culture studies since most cell types are known to be capable of producing TGF-beta. We describe here a technique of qualitative and quantitative measurement of TGF-beta production using flow cytometry. Previous work by others has led to the production of polyclonal and monoclonal antibodies to human and other mammalian TGF-beta. We have developed an intracellular cytokine staining for human TGF-beta using a monoclonal antibody, TB21.
Subject(s)
Staining and Labeling/methods , Transforming Growth Factor beta/analysis , Animals , Antibodies, Monoclonal , Cell Line , Flow Cytometry/methods , Humans , Leukocytes, Mononuclear/immunology , Transforming Growth Factor beta/biosynthesis , Transforming Growth Factor beta/immunologyABSTRACT
A competitive, chaos-free, implicit, finite-difference method is developed and used for a novel deterministic model for the perturbation of HIV by combination antiretroviral therapy. The compartmental model monitors the interaction between HIV and CD4(+) T cells, its principal target and site of replication in vivo, in the presence of reverse transcription inhibitors and protease inhibitors. The model exhibits two steady states, an uninfected (trivial) steady state (with no virus present) and an endemically infected state (with virus and infected T cells present). Stability and bifurcation analyses together with numerical simulations of the resulting dynamical system are reported.
