Abstract
Results obtained from hypothesis-driven research may not represent the complex interactions between cells and cytokines required for the development of an acquired immune response. We applied a novel algorithm to measure change in immune response over time in a mouse model of anti-Factor VIII (FVIII) antibody formation in hemophilia A. Splenocytes isolated from FVIII injected mice treated with PBS or an experimental tolerogen (CFA/1) were cultured for 72 hours and cytokine concentrations determined via multiplex analysis at days 7 and 14. The concentrations of 22 individual cytokines were compared between the control and CFA/1-treated groups, and 3 cytokines statistically differentiated the two groups at day 7 and 5 cytokines at day 14. Median cytokine concentrations/group of each significance set were converted to pmol/L and Ingenuity Pathway Analysis was used to generate likely biological network pathways. Network graphs were converted to matrices and linear algebra was used to compare treatment groups. Sixteen different molecules were predicted to occur in PBS vs. CFA/1 groups at day 7. We validated the presence of two predicted outcomes - expression of TLR 4 and secretion of IL-25. At day 14, the predicted outcomes were similar between the treatment groups, despite being evoked from a larger significance set, suggesting that statistical differences in cytokine secretion do not consistently represent biologically relevant changes in immune response over time.
- Copyright © 2012 by The American Association of Immunologists, Inc.
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