In vivo and in vitro post-translational polymorphism of chicken natural auto-antibodies

T.J. Bergstra, K. Smeets, M.G.B. Nieuwland, H.K. Parmentier

Research output: Contribution to journalArticleAcademicpeer-review

3 Citations (Scopus)

Abstract

Natural antibodies (NAb) perform many important functions in various immune responses and are often polyreactive of nature with low binding affinity. Natural auto-antibodies (N(A)Ab) are NAb binding at least one auto-antigen. Polyreactivity of N(A)Ab has been proposed to rest on post-translational polymorphism of the immunoglobulin F(ab)2 fragment caused by various locally present oxidizing agents, salts and lower or higher pH. Challenge with pathogen-associated molecular patterns (PAMP), such as lipopolysaccharide (LPS) or lipoteichoic acid (LTA), respectively, may underlie N(A)Ab polymorphism by the activation of inflammatory cells whose products affect the three-dimensional structure of N(A)Ab F(ab)2 fragments. We evaluated by Western blotting the effects of subcutaneous administered LPS and LTA, respectively, on binding characteristics of chicken N(A)Ab towards the ‘auto-antigen’ chicken-liver-cell-lysate (CCL) in situ prior to (day 0) and 3 days after subcutaneous challenge, as well as the effect of different in vitro maltreatments in the form of oxidizing agents: 5 mM hydrogen peroxide, 10 mM hydrogen peroxide, pH 2.6, and pH 2.0, aqua dest, and phosphate buffered saline (PBS) as control, respectively, on chicken N(A)Ab polymorphism. On both days 0 and 3 after challenge, N(A)Ab in plasma from all chickens bound to CCL. No significant differences of in vivo or in vitro maltreatments were found on the number of CCL fragments bound by the N(A)Ab. However, significant differences in the staining patterns of individual CCL molecular weight-identified fragments (MWIF) were found. The sum (S) of newly stained fragments and disappeared fragments (SMWIF) that were bound by plasma samples was significantly different between in vivo LPS or in vivo LTA challenged birds. Also, significant differences in the percentages of extinction intensity of these SMWIF were found. In addition, the plasma samples obtained at day 0 and day 3 from both LTA and LPS challenged birds were all similarly prone to in vitro maltreatment. In vitro maltreatment with pH 2.0 had the highest effect on chicken N(A)Ab polymorphism, whereas aqua dest had the lowest effect. The change of CCL fragments recognized by chicken N(A)Ab was not caused by unmasking immune complexes. The present findings suggest that (1) N(A)Ab are present in chicken plasma, (2) chicken N(A)Ab are prone to irreversible post-translational polymorphism in vitro, and (3) post-translational polymorphism of chicken N(A)Ab can be initiated by PAMP-induced inflammatory agents in situ. The consequences of these finding are discussed
Original languageEnglish
Pages (from-to)821-827
JournalDevelopmental and Comparative Immunology
Volume34
Issue number8
DOIs
Publication statusPublished - 2010

Fingerprint

Chickens
Antibodies
Lipopolysaccharides
Liver
In Vitro Techniques
Oxidants
Hydrogen Peroxide
Birds
Antigens
Antigen-Antibody Complex
Immunoglobulins
Salts
Molecular Weight
Western Blotting
Phosphates
Staining and Labeling

Keywords

  • redox-reactive autoantibodies
  • igg antibody repertoire
  • broiler-chickens
  • immunoglobulin-g
  • antigen-binding
  • specificity
  • responses
  • immune
  • lipopolysaccharide
  • poultry

Cite this

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title = "In vivo and in vitro post-translational polymorphism of chicken natural auto-antibodies",
abstract = "Natural antibodies (NAb) perform many important functions in various immune responses and are often polyreactive of nature with low binding affinity. Natural auto-antibodies (N(A)Ab) are NAb binding at least one auto-antigen. Polyreactivity of N(A)Ab has been proposed to rest on post-translational polymorphism of the immunoglobulin F(ab)2 fragment caused by various locally present oxidizing agents, salts and lower or higher pH. Challenge with pathogen-associated molecular patterns (PAMP), such as lipopolysaccharide (LPS) or lipoteichoic acid (LTA), respectively, may underlie N(A)Ab polymorphism by the activation of inflammatory cells whose products affect the three-dimensional structure of N(A)Ab F(ab)2 fragments. We evaluated by Western blotting the effects of subcutaneous administered LPS and LTA, respectively, on binding characteristics of chicken N(A)Ab towards the ‘auto-antigen’ chicken-liver-cell-lysate (CCL) in situ prior to (day 0) and 3 days after subcutaneous challenge, as well as the effect of different in vitro maltreatments in the form of oxidizing agents: 5 mM hydrogen peroxide, 10 mM hydrogen peroxide, pH 2.6, and pH 2.0, aqua dest, and phosphate buffered saline (PBS) as control, respectively, on chicken N(A)Ab polymorphism. On both days 0 and 3 after challenge, N(A)Ab in plasma from all chickens bound to CCL. No significant differences of in vivo or in vitro maltreatments were found on the number of CCL fragments bound by the N(A)Ab. However, significant differences in the staining patterns of individual CCL molecular weight-identified fragments (MWIF) were found. The sum (S) of newly stained fragments and disappeared fragments (SMWIF) that were bound by plasma samples was significantly different between in vivo LPS or in vivo LTA challenged birds. Also, significant differences in the percentages of extinction intensity of these SMWIF were found. In addition, the plasma samples obtained at day 0 and day 3 from both LTA and LPS challenged birds were all similarly prone to in vitro maltreatment. In vitro maltreatment with pH 2.0 had the highest effect on chicken N(A)Ab polymorphism, whereas aqua dest had the lowest effect. The change of CCL fragments recognized by chicken N(A)Ab was not caused by unmasking immune complexes. The present findings suggest that (1) N(A)Ab are present in chicken plasma, (2) chicken N(A)Ab are prone to irreversible post-translational polymorphism in vitro, and (3) post-translational polymorphism of chicken N(A)Ab can be initiated by PAMP-induced inflammatory agents in situ. The consequences of these finding are discussed",
keywords = "redox-reactive autoantibodies, igg antibody repertoire, broiler-chickens, immunoglobulin-g, antigen-binding, specificity, responses, immune, lipopolysaccharide, poultry",
author = "T.J. Bergstra and K. Smeets and M.G.B. Nieuwland and H.K. Parmentier",
year = "2010",
doi = "10.1016/j.dci.2010.03.002",
language = "English",
volume = "34",
pages = "821--827",
journal = "Developmental and Comparative Immunology",
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}

In vivo and in vitro post-translational polymorphism of chicken natural auto-antibodies. / Bergstra, T.J.; Smeets, K.; Nieuwland, M.G.B.; Parmentier, H.K.

In: Developmental and Comparative Immunology, Vol. 34, No. 8, 2010, p. 821-827.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - In vivo and in vitro post-translational polymorphism of chicken natural auto-antibodies

AU - Bergstra, T.J.

AU - Smeets, K.

AU - Nieuwland, M.G.B.

AU - Parmentier, H.K.

PY - 2010

Y1 - 2010

N2 - Natural antibodies (NAb) perform many important functions in various immune responses and are often polyreactive of nature with low binding affinity. Natural auto-antibodies (N(A)Ab) are NAb binding at least one auto-antigen. Polyreactivity of N(A)Ab has been proposed to rest on post-translational polymorphism of the immunoglobulin F(ab)2 fragment caused by various locally present oxidizing agents, salts and lower or higher pH. Challenge with pathogen-associated molecular patterns (PAMP), such as lipopolysaccharide (LPS) or lipoteichoic acid (LTA), respectively, may underlie N(A)Ab polymorphism by the activation of inflammatory cells whose products affect the three-dimensional structure of N(A)Ab F(ab)2 fragments. We evaluated by Western blotting the effects of subcutaneous administered LPS and LTA, respectively, on binding characteristics of chicken N(A)Ab towards the ‘auto-antigen’ chicken-liver-cell-lysate (CCL) in situ prior to (day 0) and 3 days after subcutaneous challenge, as well as the effect of different in vitro maltreatments in the form of oxidizing agents: 5 mM hydrogen peroxide, 10 mM hydrogen peroxide, pH 2.6, and pH 2.0, aqua dest, and phosphate buffered saline (PBS) as control, respectively, on chicken N(A)Ab polymorphism. On both days 0 and 3 after challenge, N(A)Ab in plasma from all chickens bound to CCL. No significant differences of in vivo or in vitro maltreatments were found on the number of CCL fragments bound by the N(A)Ab. However, significant differences in the staining patterns of individual CCL molecular weight-identified fragments (MWIF) were found. The sum (S) of newly stained fragments and disappeared fragments (SMWIF) that were bound by plasma samples was significantly different between in vivo LPS or in vivo LTA challenged birds. Also, significant differences in the percentages of extinction intensity of these SMWIF were found. In addition, the plasma samples obtained at day 0 and day 3 from both LTA and LPS challenged birds were all similarly prone to in vitro maltreatment. In vitro maltreatment with pH 2.0 had the highest effect on chicken N(A)Ab polymorphism, whereas aqua dest had the lowest effect. The change of CCL fragments recognized by chicken N(A)Ab was not caused by unmasking immune complexes. The present findings suggest that (1) N(A)Ab are present in chicken plasma, (2) chicken N(A)Ab are prone to irreversible post-translational polymorphism in vitro, and (3) post-translational polymorphism of chicken N(A)Ab can be initiated by PAMP-induced inflammatory agents in situ. The consequences of these finding are discussed

AB - Natural antibodies (NAb) perform many important functions in various immune responses and are often polyreactive of nature with low binding affinity. Natural auto-antibodies (N(A)Ab) are NAb binding at least one auto-antigen. Polyreactivity of N(A)Ab has been proposed to rest on post-translational polymorphism of the immunoglobulin F(ab)2 fragment caused by various locally present oxidizing agents, salts and lower or higher pH. Challenge with pathogen-associated molecular patterns (PAMP), such as lipopolysaccharide (LPS) or lipoteichoic acid (LTA), respectively, may underlie N(A)Ab polymorphism by the activation of inflammatory cells whose products affect the three-dimensional structure of N(A)Ab F(ab)2 fragments. We evaluated by Western blotting the effects of subcutaneous administered LPS and LTA, respectively, on binding characteristics of chicken N(A)Ab towards the ‘auto-antigen’ chicken-liver-cell-lysate (CCL) in situ prior to (day 0) and 3 days after subcutaneous challenge, as well as the effect of different in vitro maltreatments in the form of oxidizing agents: 5 mM hydrogen peroxide, 10 mM hydrogen peroxide, pH 2.6, and pH 2.0, aqua dest, and phosphate buffered saline (PBS) as control, respectively, on chicken N(A)Ab polymorphism. On both days 0 and 3 after challenge, N(A)Ab in plasma from all chickens bound to CCL. No significant differences of in vivo or in vitro maltreatments were found on the number of CCL fragments bound by the N(A)Ab. However, significant differences in the staining patterns of individual CCL molecular weight-identified fragments (MWIF) were found. The sum (S) of newly stained fragments and disappeared fragments (SMWIF) that were bound by plasma samples was significantly different between in vivo LPS or in vivo LTA challenged birds. Also, significant differences in the percentages of extinction intensity of these SMWIF were found. In addition, the plasma samples obtained at day 0 and day 3 from both LTA and LPS challenged birds were all similarly prone to in vitro maltreatment. In vitro maltreatment with pH 2.0 had the highest effect on chicken N(A)Ab polymorphism, whereas aqua dest had the lowest effect. The change of CCL fragments recognized by chicken N(A)Ab was not caused by unmasking immune complexes. The present findings suggest that (1) N(A)Ab are present in chicken plasma, (2) chicken N(A)Ab are prone to irreversible post-translational polymorphism in vitro, and (3) post-translational polymorphism of chicken N(A)Ab can be initiated by PAMP-induced inflammatory agents in situ. The consequences of these finding are discussed

KW - redox-reactive autoantibodies

KW - igg antibody repertoire

KW - broiler-chickens

KW - immunoglobulin-g

KW - antigen-binding

KW - specificity

KW - responses

KW - immune

KW - lipopolysaccharide

KW - poultry

U2 - 10.1016/j.dci.2010.03.002

DO - 10.1016/j.dci.2010.03.002

M3 - Article

VL - 34

SP - 821

EP - 827

JO - Developmental and Comparative Immunology

JF - Developmental and Comparative Immunology

SN - 0145-305X

IS - 8

ER -