PMC:3537549 / 26401-29780
Annnotations
{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/3537549","sourcedb":"PMC","sourceid":"3537549","source_url":"https://www.ncbi.nlm.nih.gov/pmc/3537549","text":"Prototype CpG-A, ODN 2216, induce proliferation of primary feline immune cells and enhance their expression of costimulatory surface molecules\nProliferation of PBMCs in response to treatment with CpG ODN gives strong indications about the biological activity of the stimulatory molecule and has been used to screen ODN in many species[25]. In relation to this, the potential of CpG-A to induce proliferation of feline PBMCs were assessed by measurement of 3 H-thymidine incorporation after stimulation. Although considerable variability was observed between individuals, a 2-fold increase in proliferation was observed after stimulation with ODN 2216 when compared to stimulation with inactive control ODN 2243 (p = 0.0281, Figure1A). The cells of three cats (c08, c09, c12) could be stimulated to particularly high proliferative rates by ODN 2216 (values above the mean of eight cats illustrated in Figure1A) in the following order: c08 \u003e c12 \u003e c09.\nAnother characteristic feature of stimulatory ODN is their ability to enhance interactions between various immune cell populations by upregulation of cell surface costimulatory molecules. With the objective to test whether ODN 2216 could exert such properties in feline immune cells, the expression of B7.1 and MHCII was measured by flow cytometry in stimulated PBMCs of the same eight cats as above. The expression of both co-stimulatory molecules was evaluated in gates defined to contain a PBMC population, a lymphocyte population and a non-lymphocyte population of cells. The observed effects varied considerably between the cells of individual cats, ranging from no alterations to an increase of 400% stained cells in some cellular subpopulations after ODN 2216 stimulation. Also, in some animals, stimulation with the control ODN 2243 indicated similar staining patterns as PBS, whereas in other cats the induction of effects comparable to those of ODN 2216 could be observed. The response of cells originating from a particular cat to stimulation with ODN 2243 did not however correlate with the influence of ODN 2216 on the expression of either surface molecule on these same cells. Overall, an increased expression of both B7.1 and MHCII was measured in gated PBMCs upon a 24 h stimulation with ODN 2216 when overlaid with the expression of these molecules after ODN 2243 or PBS stimulation (Figure1B and C). Considering all eight cats, lymphocytic cells expressed significantly higher levels of B7.1 and MHCII when exposed to ODN 2216 than after treatment with PBS (p = 0.0195 and p = 0.039 respectively, Figure1D and F). Feline MHCII expression on lymphocytes also seemed to be affected by ODN 2216 and ODN 2243 in a similar manner (Figure1F). For non-lymphocytic cells, ODN 2216 more specifically affected the expression of B7.1 molecules, inducing significant upregulation of surface levels of this molecule when compared to stimulation with ODN 2243 (p = 0.0391) and PBS (p = 0.0039) for 24 h (Figure1E). In contrast, the expression of MHCII was not significantly altered in cells gated as non-lymphocytes at this time point after stimulation (Figure1G). Finally, the cells of the three cats (c08, c09 and c12) that exhibited the highest proliferation rates in response to ODN 2216 (Figure1A) also indicated the strongest increase in expression of both cell surface molecules in all cellular subpopulations tested.","divisions":[{"label":"Title","span":{"begin":0,"end":142}}],"tracks":[{"project":"2_test","denotations":[{"id":"22906110-11763350-143530979","span":{"begin":335,"end":337},"obj":"11763350"}],"attributes":[{"subj":"22906110-11763350-143530979","pred":"source","obj":"2_test"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"2_test","color":"#ad93ec","default":true}]}]}}