title: Activation of inflammatory cells and cytokines by peptide epitopes in vitro: a simple in-vitro screening assay for prioritizing them for in-vivo studies.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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 ApoB(tm2Sgy)/Ldlr(tm1Her/J) knockout mice.
  
  
  CONCLUSIONS
  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.
authors: Thrombosis Research Institute, Narayana Hrudayalaya, 258/A, Bommasandra Industrial Area, Anekal Taluk, Bangalore, India. lakshmi.mundkur@triindia.org.in
authors: Lakshmi A
authors: LA
authors: Mundkur
authors: Meenakshi
authors: M
authors: Varma
authors: Hemapriya
authors: H
authors: Shivanandan
authors: Dhanush
authors: D
authors: Krishna
authors: Kiran
authors: K
authors: Kumar
authors: Xinjie
authors: X
authors: Lu
authors: Vijay V
authors: VV
authors: Kakkar
mesh_terms: D000328:Adult; D000818:Animals; D001055:Apolipoproteins B; D050197:Atherosclerosis / Q000188:drug therapy / Q000276:immunology; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D018834:Chaperonin 60; D016207:Cytokines / Q000378:metabolism; D003713:Dendritic Cells / Q000187:drug effects / Q000276:immunology; D059305:Diet, High-Fat; D000939:Epitopes / Q000494:pharmacology* / Q000627:therapeutic use; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D010455:Peptides / Q000494:pharmacology* / Q000627:therapeutic use; D011951:Receptors, Complement; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology
chemical_list: D001055:Apolipoproteins B; D018834:Chaperonin 60; D016207:Cytokines; D000939:Epitopes; D010455:Peptides; D011951:Receptors, Complement
references: Clin Immunol. 2008 Apr;127(1):89-97
references: 18294918
references: Circulation. 2006 Oct 31;114(18):1968-76
references: 17060383
references: Atherosclerosis. 2010 Oct;212(2):472-80
references: 20609438
references: Nat Rev Immunol. 2006 Jul;6(7):508-19
references: 16778830
references: Eur J Immunol. 2007 Mar;37(3):686-95
references: 17301951
references: Arterioscler Thromb Vasc Biol. 2012 Oct;32(10):2358-71
references: 22837469
references: Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6393-8
references: 8692825
references: Immunol Rev. 2011 May;241(1):241-59
references: 21488901
references: Arterioscler Thromb Vasc Biol. 2007 Dec;27(12):2677-83
references: 17901374
references: Curr Opin Immunol. 2001 Jun;13(3):291-8
references: 11406360
references: J Cardiovasc Pharmacol. 2005 Jun;45(6):591-8
references: 15897787
references: Circulation. 2002 Sep 24;106(13):1708-15
references: 12270867
references: Nat Immunol. 2000 Sep;1(3):199-205
references: 10973276
references: Cytometry B Clin Cytom. 2007 Jul;72(4):265-75
references: 17328032
references: Immunol Today. 1999 Dec;20(12):561-7
references: 10562707
references: Nature. 1998 Mar 19;392(6673):245-52
references: 9521319
references: Nat Rev Immunol. 2003 Dec;3(12):984-93
references: 14647480
references: Trends Immunol. 2001 Jul;22(7):394-400
references: 11429324
references: Arterioscler Thromb Vasc Biol. 2010 May;30(5):946-52
references: 20167655
references: Nat Immunol. 2005 Apr;6(4):345-52
references: 15785760
references: Circulation. 2009 Jun 23;119(24):3133-41
references: 19546396
references: Autoimmunity. 2005 Mar;38(2):171-9
references: 16040338
references: Semin Immunopathol. 2009 Jun;31(1):95-101
references: 19468734
references: Clin Immunol. 2010 Jan;134(1):33-46
references: 19635683
references: Curr Opin Immunol. 2000 Dec;12(6):676-83
references: 11102772
references: Arterioscler Thromb Vasc Biol. 2007 May;27(5):1095-100
references: 17303776
references: J Intern Med. 2009 Sep;266(3):221-31
references: 19702790
references: Swiss Med Wkly. 2007 Nov 17;137(45-46):625-34
references: 18027108
references: Circulation. 2004 Oct 5;110(14):2047-52
references: 15451805
references: Arterioscler Thromb Vasc Biol. 2001 Jan;21(1):108-14
references: 11145941
references: J Exp Med. 1994 Apr 1;179(4):1109-18
references: 8145033
references: Nat Protoc. 2007;2(9):2057-67
references: 17853861
references: Nat Protoc. 2007;2(9):2049-56
references: 17853860
references: Nat Immunol. 2005 Apr;6(4):338-44
references: 15785759
references: Nature. 2007 Sep 27;449(7161):419-26
references: 17898760
references: Eur Heart J. 2005 Sep;26(17):1700-4
references: 15855191
references: Blood. 2006 Oct 15;108(8):2655-61
references: 16763205