Activation of inflammatory cells and cytokines by peptide epitopes in vitro: a simple in-vitro screening assay for prioritizing them for in-vivo studies
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Antigen-specific immune modulation is an attractive approach to atherosclerosis treatment. The aim of this study was to develop an in-vitro assay to screen peptide molecules for their inflammatory propensity.
Human dendritic cells derived from CD14+ monocytes were activated using peptides derived from apolipoprotein B100 (ApoB), heat shock protein 60 (HSP60) and complement cascade (peptide A) in vitro, and used for priming autologous T cells. Proliferation of T cells, their differentiation to regulatory cells (Treg) and their cytokine profile were studied. The efficacy of the peptides in preventing atherosclerosis was studied in ApoBtm2Sgy/Ldlrtm1Her/J knockout mice.
Results and conclusion
ApoB and HSP60 peptides induced T-cell proliferation and expansion of regulatory T cells with interleukin-10 and transforming growth factor-β secretion. In comparison, peptide A was a poor stimulator of T cells and was found to induce tumor necrosis factor-α secretion by activated T cells. ApoB and HSP60 peptides were found to reduce early atherosclerotic lesion formation in mice by 32.1 and 33.5 %, respectively, while the reduction with peptide A was 5.7 %. Thus the in-vitro assay shows an apparent correlation with in-vivo activity and can be developed as a screening assay to prioritize the candidate molecules for animal efficacy testing.
KeywordsAtherosclerosis Inflammation Dendritic cells T-regulatory cells Cytokines Vaccine
We gratefully acknowledge the support of the trustees of Thrombosis Research Institute, London and Bangalore, Tata Social Welfare Trust, India (TSWT/IG/SNB/JP/Sdm) and Department of Biotechnology, Ministry of Science and Technology, Government of India (BT/01/CDE/08/07). The sponsors did not participate in the design, conduct, sample collection analysis and interpretation of the data or in the preparation, review or approval of the manuscript. The authors have no conflicts of interest to disclose.
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