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Cross-reacting antibacterial auto-antibodies are produced within coronary atherosclerotic plaques of acute coronary syndrome patients.

Canducci F, Saita D, Foglieni C, Piscopiello MR, Chiesa R, Colombo A, Cianflone D, Maseri A, Clementi M, Burioni R - PLoS ONE (2012)

Bottom Line: In vitro fibrocytes obtained by differentiating CD14+ cells isolated from peripheral blood mononuclear cells also interacted with Fab7816, thus supporting the hypothesis of a specific recognition of fibrocytes into the atherosclerotic lesions.Interestingly, the same antibody, cross-reacted with the outer membrane proteins of Proteus mirabilis and Klebsiella pneumoniae (and possibly with homologous proteins of other enterobacteriaceae present in the microbiota).From all the other three libraries, we were able to clone, by immunoaffinity selection, human monoclonal antibodies cross-reacting with bacterial outer membrane proteins and with transgelin.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Microbiology and Virology, Ospedale San Raffaele, Milan, Italy. Canducci.filippo@hsr.it

ABSTRACT
Coronary atherosclerosis, the main condition predisposing to acute myocardial infarction, has an inflammatory component caused by stimuli that are yet unknown. We molecularly investigated the nature of the immune response within human coronary lesion in four coronary plaques obtained by endoluminal atherectomy from four patients. We constructed phage-display libraries containing the IgG1/kappa antibody fragments produced by B-lymphocytes present in each plaque. By immunoaffinity, we selected from these libraries a monoclonal antibody, arbitrarily named Fab7816, able to react both with coronary and carotid atherosclerotic tissue samples. We also demonstrated by confocal microscopy that this monoclonal antibody recognized human transgelin type 1, a cytoskeleton protein involved in atherogenesis, and that it co-localized with fibrocyte-like cells transgelin+, CD68+, CD45+ in human sections of coronary and carotid plaques. In vitro fibrocytes obtained by differentiating CD14+ cells isolated from peripheral blood mononuclear cells also interacted with Fab7816, thus supporting the hypothesis of a specific recognition of fibrocytes into the atherosclerotic lesions. Interestingly, the same antibody, cross-reacted with the outer membrane proteins of Proteus mirabilis and Klebsiella pneumoniae (and possibly with homologous proteins of other enterobacteriaceae present in the microbiota). From all the other three libraries, we were able to clone, by immunoaffinity selection, human monoclonal antibodies cross-reacting with bacterial outer membrane proteins and with transgelin. These findings demonstrated that in human atherosclerotic plaques a local cross-reactive immune response takes place.

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Related in: MedlinePlus

Cross reactivity of commercial available monoclonal antibodies with TAGLN and OMPs.A) WB of OmpK36 with five commercial monoclonal mouse anti-human TAGLN antibodies (used at 1 or 5 µg/ml). Three commercial mouse monoclonal antibodies were used as negative controls. Anti-6xHIS antibody (Roche) was used as positive control B) ELISA with all representative and five commercial monoclonal mouse anti-human TAGLN antibodies on purified human TAGLN or C) bacterial OMPs. Reactivity against bovine serum albumin (BSA) used as blocking antigen, is also shown.
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pone-0042283-g011: Cross reactivity of commercial available monoclonal antibodies with TAGLN and OMPs.A) WB of OmpK36 with five commercial monoclonal mouse anti-human TAGLN antibodies (used at 1 or 5 µg/ml). Three commercial mouse monoclonal antibodies were used as negative controls. Anti-6xHIS antibody (Roche) was used as positive control B) ELISA with all representative and five commercial monoclonal mouse anti-human TAGLN antibodies on purified human TAGLN or C) bacterial OMPs. Reactivity against bovine serum albumin (BSA) used as blocking antigen, is also shown.

Mentions: Five different commercial monoclonal mouse anti-human TAGLN IgG antibodies (Abnova, H00006876-M01, M02, M03, M04, M05) and a monoclonal mouse anti-human TAGLN IgM antibody (Abnova, H00006876-M06A) were tested for their potential cross-reactivity against human TAGLN in WB (Figure 11a) and in ELISA (Figure 11b) and against bacterial OMPs (both OmpK36 and OmpF) in ELISA (Figure 11c). While all commercial monoclonal antibodies recognized TAGLN and purified OmpK36 and OmpF in ELISA (although apparently with distinct efficiencies), three of them, M03, M04 and M06, recognized TAGLN also in WB further con confirming the existence of a cross-mimicry between human TAGLN and the bacterial OMPs.


Cross-reacting antibacterial auto-antibodies are produced within coronary atherosclerotic plaques of acute coronary syndrome patients.

Canducci F, Saita D, Foglieni C, Piscopiello MR, Chiesa R, Colombo A, Cianflone D, Maseri A, Clementi M, Burioni R - PLoS ONE (2012)

Cross reactivity of commercial available monoclonal antibodies with TAGLN and OMPs.A) WB of OmpK36 with five commercial monoclonal mouse anti-human TAGLN antibodies (used at 1 or 5 µg/ml). Three commercial mouse monoclonal antibodies were used as negative controls. Anti-6xHIS antibody (Roche) was used as positive control B) ELISA with all representative and five commercial monoclonal mouse anti-human TAGLN antibodies on purified human TAGLN or C) bacterial OMPs. Reactivity against bovine serum albumin (BSA) used as blocking antigen, is also shown.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3412836&req=5

pone-0042283-g011: Cross reactivity of commercial available monoclonal antibodies with TAGLN and OMPs.A) WB of OmpK36 with five commercial monoclonal mouse anti-human TAGLN antibodies (used at 1 or 5 µg/ml). Three commercial mouse monoclonal antibodies were used as negative controls. Anti-6xHIS antibody (Roche) was used as positive control B) ELISA with all representative and five commercial monoclonal mouse anti-human TAGLN antibodies on purified human TAGLN or C) bacterial OMPs. Reactivity against bovine serum albumin (BSA) used as blocking antigen, is also shown.
Mentions: Five different commercial monoclonal mouse anti-human TAGLN IgG antibodies (Abnova, H00006876-M01, M02, M03, M04, M05) and a monoclonal mouse anti-human TAGLN IgM antibody (Abnova, H00006876-M06A) were tested for their potential cross-reactivity against human TAGLN in WB (Figure 11a) and in ELISA (Figure 11b) and against bacterial OMPs (both OmpK36 and OmpF) in ELISA (Figure 11c). While all commercial monoclonal antibodies recognized TAGLN and purified OmpK36 and OmpF in ELISA (although apparently with distinct efficiencies), three of them, M03, M04 and M06, recognized TAGLN also in WB further con confirming the existence of a cross-mimicry between human TAGLN and the bacterial OMPs.

Bottom Line: In vitro fibrocytes obtained by differentiating CD14+ cells isolated from peripheral blood mononuclear cells also interacted with Fab7816, thus supporting the hypothesis of a specific recognition of fibrocytes into the atherosclerotic lesions.Interestingly, the same antibody, cross-reacted with the outer membrane proteins of Proteus mirabilis and Klebsiella pneumoniae (and possibly with homologous proteins of other enterobacteriaceae present in the microbiota).From all the other three libraries, we were able to clone, by immunoaffinity selection, human monoclonal antibodies cross-reacting with bacterial outer membrane proteins and with transgelin.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Microbiology and Virology, Ospedale San Raffaele, Milan, Italy. Canducci.filippo@hsr.it

ABSTRACT
Coronary atherosclerosis, the main condition predisposing to acute myocardial infarction, has an inflammatory component caused by stimuli that are yet unknown. We molecularly investigated the nature of the immune response within human coronary lesion in four coronary plaques obtained by endoluminal atherectomy from four patients. We constructed phage-display libraries containing the IgG1/kappa antibody fragments produced by B-lymphocytes present in each plaque. By immunoaffinity, we selected from these libraries a monoclonal antibody, arbitrarily named Fab7816, able to react both with coronary and carotid atherosclerotic tissue samples. We also demonstrated by confocal microscopy that this monoclonal antibody recognized human transgelin type 1, a cytoskeleton protein involved in atherogenesis, and that it co-localized with fibrocyte-like cells transgelin+, CD68+, CD45+ in human sections of coronary and carotid plaques. In vitro fibrocytes obtained by differentiating CD14+ cells isolated from peripheral blood mononuclear cells also interacted with Fab7816, thus supporting the hypothesis of a specific recognition of fibrocytes into the atherosclerotic lesions. Interestingly, the same antibody, cross-reacted with the outer membrane proteins of Proteus mirabilis and Klebsiella pneumoniae (and possibly with homologous proteins of other enterobacteriaceae present in the microbiota). From all the other three libraries, we were able to clone, by immunoaffinity selection, human monoclonal antibodies cross-reacting with bacterial outer membrane proteins and with transgelin. These findings demonstrated that in human atherosclerotic plaques a local cross-reactive immune response takes place.

Show MeSH
Related in: MedlinePlus