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Modulation of recombinant antigenic constructs containing multi-epitopes towards effective reduction of atherosclerotic lesion in B6;129S-Ldlr(tm1Her)Apob(tm2Sgy)/J mice.

Xia M, Chen D, Endresz V, Lantos I, Szabo A, Kakkar V, Lu X - PLoS ONE (2015)

Bottom Line: Our previous study has demonstrated that construct AHHC [ApoB100688-707 + hHSP60303-312 + hHSP60153-163 + Cpn derived peptide (C)] significantly reduced atherosclerotic lesion.Reduction of plaque size in the aortic sinus and descending aorta correlated with alterations in cellular immune responses when compared with controls.We conclude that a recombinant construct RPHC may provide new antigenic and structural features which are favorable for significant reduction in atherosclerotic lesion formation.

View Article: PubMed Central - PubMed

Affiliation: The Mary and Garry Weston Molecular Immunology Laboratory, Thrombosis Research Institute, London, United Kingdom.

ABSTRACT
Atherosclerosis is increasingly recognized as a complex chronic inflammatory disease. Many more studies have extended vaccination against atherosclerosis by using epitopes from self-antigens or beyond and demonstrated that vaccination with antigens or derivatives could reduce the extent of the lesions in atherosclerosis-prone mice. Our previous study has demonstrated that construct AHHC [ApoB100688-707 + hHSP60303-312 + hHSP60153-163 + Cpn derived peptide (C)] significantly reduced atherosclerotic lesion. The aim of this study was to investigate whether AHHC can be modulated towards increased lesion reduction in mice by creating two other derivatives with a sequential epitope-substitution named RHHC in which A was replaced by an "R" (C5aR1-31) and RPHC with a further "H" (hHSP60303-312) conversion into "P" (protease-activated receptor-142-55) in mice. Antigenic epitopes were incorporated into a dendroaspin scaffold. Immunization of B6;129S-Ldlrtm1HerApobtm2Sgy/J mice with three constructs elicited production of high levels of antibodies against each epitope (apart from hHSP60153-163 and P which induced a low antibody response). Histological analyses demonstrated that the mice immunized with either RPHC or RHHC showed significant reductions in the size of atherosclerostic lesions compared to those with AHHC (69.5±1.1% versus 55.7±3.4%, P<0.01 or 65.6±1.3% versus 55.7±3.4%, P<0.01). Reduction of plaque size in the aortic sinus and descending aorta correlated with alterations in cellular immune responses when compared with controls. We conclude that a recombinant construct RPHC may provide new antigenic and structural features which are favorable for significant reduction in atherosclerotic lesion formation. This approach offers a novel strategy for developing anti-atherosclerotic agents.

No MeSH data available.


Related in: MedlinePlus

Evaluation of the content of TLR4 and MyD88 contents at the lesion sites.(A) Photomicrographs showing IHC staining for TLR4 (red) (scale bar: 100 μm and 12.5μm for magnified ones). (B) Scatter plot showing means of anti-TLR4-stained area (N = 6). (C) Photomicrographs showing IHC staining for MyD88 (red) (scale bar: 100 μm and 12.5μm for magnified ones). (D) Scatter plot showing means of anti-MyD88-stained area (N = 6).
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pone.0123393.g008: Evaluation of the content of TLR4 and MyD88 contents at the lesion sites.(A) Photomicrographs showing IHC staining for TLR4 (red) (scale bar: 100 μm and 12.5μm for magnified ones). (B) Scatter plot showing means of anti-TLR4-stained area (N = 6). (C) Photomicrographs showing IHC staining for MyD88 (red) (scale bar: 100 μm and 12.5μm for magnified ones). (D) Scatter plot showing means of anti-MyD88-stained area (N = 6).

Mentions: We examined the impact of the treatment with these recombinant constructs on TLR4 and MyD88 contents in lesions. The reduction of atherosclerosis in mice treated with these constructs was associated with a decrease in both TLR4 and MyD88 contents. Anti-TLR4 stained area was significantly smaller in the plaques of mice immunized with AHHC, RHHC and RPHC, showing 4.6 ± 1.0%, 3.4 ± 0.5% and 4.2 ± 0.6%, respectively, compared with that in controls immunized with dendroaspin (8.1 ± 1.1%) (Fig 8A and 8B). Similarly, anti-MyD88 stained area was significantly smaller in the lesions of mice immunized with these three constructs, showing 9.7 ± 1.2%, 9.6 ± 1.6% and 10.2 ± 1.9%, respectively, compared with that in controls (18.6 ± 2.4%) (Fig 8C and 8D). Additionally, overlapping anti-CD11c and anti-TLR4 stained area showed significantly smaller in the lesions of mice immunized with all three constructs, showing 3.4 ± 0.6%, 2.9 ± 0.7% and 2.7 ± 0.8%, respectively, compared with that in controls (6.6 ± 0.8%) (S5A and S5B Fig).


Modulation of recombinant antigenic constructs containing multi-epitopes towards effective reduction of atherosclerotic lesion in B6;129S-Ldlr(tm1Her)Apob(tm2Sgy)/J mice.

Xia M, Chen D, Endresz V, Lantos I, Szabo A, Kakkar V, Lu X - PLoS ONE (2015)

Evaluation of the content of TLR4 and MyD88 contents at the lesion sites.(A) Photomicrographs showing IHC staining for TLR4 (red) (scale bar: 100 μm and 12.5μm for magnified ones). (B) Scatter plot showing means of anti-TLR4-stained area (N = 6). (C) Photomicrographs showing IHC staining for MyD88 (red) (scale bar: 100 μm and 12.5μm for magnified ones). (D) Scatter plot showing means of anti-MyD88-stained area (N = 6).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4382319&req=5

pone.0123393.g008: Evaluation of the content of TLR4 and MyD88 contents at the lesion sites.(A) Photomicrographs showing IHC staining for TLR4 (red) (scale bar: 100 μm and 12.5μm for magnified ones). (B) Scatter plot showing means of anti-TLR4-stained area (N = 6). (C) Photomicrographs showing IHC staining for MyD88 (red) (scale bar: 100 μm and 12.5μm for magnified ones). (D) Scatter plot showing means of anti-MyD88-stained area (N = 6).
Mentions: We examined the impact of the treatment with these recombinant constructs on TLR4 and MyD88 contents in lesions. The reduction of atherosclerosis in mice treated with these constructs was associated with a decrease in both TLR4 and MyD88 contents. Anti-TLR4 stained area was significantly smaller in the plaques of mice immunized with AHHC, RHHC and RPHC, showing 4.6 ± 1.0%, 3.4 ± 0.5% and 4.2 ± 0.6%, respectively, compared with that in controls immunized with dendroaspin (8.1 ± 1.1%) (Fig 8A and 8B). Similarly, anti-MyD88 stained area was significantly smaller in the lesions of mice immunized with these three constructs, showing 9.7 ± 1.2%, 9.6 ± 1.6% and 10.2 ± 1.9%, respectively, compared with that in controls (18.6 ± 2.4%) (Fig 8C and 8D). Additionally, overlapping anti-CD11c and anti-TLR4 stained area showed significantly smaller in the lesions of mice immunized with all three constructs, showing 3.4 ± 0.6%, 2.9 ± 0.7% and 2.7 ± 0.8%, respectively, compared with that in controls (6.6 ± 0.8%) (S5A and S5B Fig).

Bottom Line: Our previous study has demonstrated that construct AHHC [ApoB100688-707 + hHSP60303-312 + hHSP60153-163 + Cpn derived peptide (C)] significantly reduced atherosclerotic lesion.Reduction of plaque size in the aortic sinus and descending aorta correlated with alterations in cellular immune responses when compared with controls.We conclude that a recombinant construct RPHC may provide new antigenic and structural features which are favorable for significant reduction in atherosclerotic lesion formation.

View Article: PubMed Central - PubMed

Affiliation: The Mary and Garry Weston Molecular Immunology Laboratory, Thrombosis Research Institute, London, United Kingdom.

ABSTRACT
Atherosclerosis is increasingly recognized as a complex chronic inflammatory disease. Many more studies have extended vaccination against atherosclerosis by using epitopes from self-antigens or beyond and demonstrated that vaccination with antigens or derivatives could reduce the extent of the lesions in atherosclerosis-prone mice. Our previous study has demonstrated that construct AHHC [ApoB100688-707 + hHSP60303-312 + hHSP60153-163 + Cpn derived peptide (C)] significantly reduced atherosclerotic lesion. The aim of this study was to investigate whether AHHC can be modulated towards increased lesion reduction in mice by creating two other derivatives with a sequential epitope-substitution named RHHC in which A was replaced by an "R" (C5aR1-31) and RPHC with a further "H" (hHSP60303-312) conversion into "P" (protease-activated receptor-142-55) in mice. Antigenic epitopes were incorporated into a dendroaspin scaffold. Immunization of B6;129S-Ldlrtm1HerApobtm2Sgy/J mice with three constructs elicited production of high levels of antibodies against each epitope (apart from hHSP60153-163 and P which induced a low antibody response). Histological analyses demonstrated that the mice immunized with either RPHC or RHHC showed significant reductions in the size of atherosclerostic lesions compared to those with AHHC (69.5±1.1% versus 55.7±3.4%, P<0.01 or 65.6±1.3% versus 55.7±3.4%, P<0.01). Reduction of plaque size in the aortic sinus and descending aorta correlated with alterations in cellular immune responses when compared with controls. We conclude that a recombinant construct RPHC may provide new antigenic and structural features which are favorable for significant reduction in atherosclerotic lesion formation. This approach offers a novel strategy for developing anti-atherosclerotic agents.

No MeSH data available.


Related in: MedlinePlus