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Role of repetitive antigen patterns for induction of antibodies against antibodies.

Fehr T, Bachmann MF, Bucher E, Kalinke U, Di Padova FE, Lang AB, Hengartner H, Zinkernagel RM - J. Exp. Med. (1997)

Bottom Line: Experience shows that they are usually difficult to induce experimentally.Why and how such anti-antibodies are induced during autoimmune diseases, has remained largely unclear.The results indicate a novel link between anti-antibody responses and infectious agents, and suggest a similar role for repetitive self-antigens such as DNA or collagen involved in chronic immunopathological diseases.

View Article: PubMed Central - PubMed

Affiliation: Institute of Experimental Immunology, University of Zürich, CH-8091 Zürich, Switzerland.

ABSTRACT
Antibody responses against antibodies, such as rheumatoid factors, are found in several immunopathological diseases and may play a role in disease pathogenesis. Experience shows that they are usually difficult to induce experimentally. Antibodies specific for immunoglobulin constant regions (anti-allotypic) or for variable regions (anti-idiotypic) have been investigated in animal models; the latter have even been postulated to regulate antibody and T cell responses via network-like interactions. Why and how such anti-antibodies are induced during autoimmune diseases, has remained largely unclear. Because repetitively arranged epitopes in a paracrystalline structure of a viral envelope cross-link B cell receptors efficiently to induce a prompt T-independent IgM response, this study used immune complexes containing viruses or bacteria to evaluate the role of antigen pattern for induction of anti-antibody responses. We present evidence that antibodies bound to strictly ordered, but not to irregularly arranged, antigens dramatically enhance induction of anti-antibodies, already after a single immunization and without using adjuvants. The results indicate a novel link between anti-antibody responses and infectious agents, and suggest a similar role for repetitive self-antigens such as DNA or collagen involved in chronic immunopathological diseases.

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IgG anti-IgM response after immunization with immune complexes versus uncomplexed antigen. (A) BALB/c or (B) A/J  mice were immunized twice (interval, 14 d)  with 1 μg of the monoclonal anti-VSV-G IgM  antibody M1 complexed with 108 PFU UV- inactivated VSV (closed squares), 10 μg of recombinant VSV-G (closed triangles), or 4 μg of rat  anti–mouse Cκ antibody (closed circles), with IgM  alone (open diamonds) or with VSV alone (open  squares). 6 d after secondary immunization, IgG  anti-IgM titers were determined by ELISA as  described in Materials and Methods. (C) IgG  anti-IgM titers of 20-fold prediluted sera were  determined over a time period of 80 d after primary immunization as described in A. The same symbols are used. (D) Dose dependence of IgG anti-IgM induction shown by immunization of BALB/c  mice with IC made of 108 PFU UV-inactivated VSV and titrated amounts of two different monoclonal anti-VSV-G IgM antibodies M1 (closed bars) and  M3 (open bars). (E) C57BL/6 mice were immunized once with 5 μg of the monoclonal anti-VSV-G IgM antibody M5 complexed with 108 PFU UV- inactivated VSV (closed squares), with 5 μg of M5 alone (open diamonds) or with 108 PFU VSV alone (open squares). 4 d later, IgMa-specific IgMb antibodies  were determined by ELISA as described in Materials and Methods.
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Figure 1: IgG anti-IgM response after immunization with immune complexes versus uncomplexed antigen. (A) BALB/c or (B) A/J mice were immunized twice (interval, 14 d) with 1 μg of the monoclonal anti-VSV-G IgM antibody M1 complexed with 108 PFU UV- inactivated VSV (closed squares), 10 μg of recombinant VSV-G (closed triangles), or 4 μg of rat anti–mouse Cκ antibody (closed circles), with IgM alone (open diamonds) or with VSV alone (open squares). 6 d after secondary immunization, IgG anti-IgM titers were determined by ELISA as described in Materials and Methods. (C) IgG anti-IgM titers of 20-fold prediluted sera were determined over a time period of 80 d after primary immunization as described in A. The same symbols are used. (D) Dose dependence of IgG anti-IgM induction shown by immunization of BALB/c mice with IC made of 108 PFU UV-inactivated VSV and titrated amounts of two different monoclonal anti-VSV-G IgM antibodies M1 (closed bars) and M3 (open bars). (E) C57BL/6 mice were immunized once with 5 μg of the monoclonal anti-VSV-G IgM antibody M5 complexed with 108 PFU UV- inactivated VSV (closed squares), with 5 μg of M5 alone (open diamonds) or with 108 PFU VSV alone (open squares). 4 d later, IgMa-specific IgMb antibodies were determined by ELISA as described in Materials and Methods.

Mentions: We used a sandwich ELISA with the following steps: (a) coating with isotype-specific goat anti– mouse antibody (1 μg/ml; Southern Biotechnologies, Birmingham, AL), (b) blocking with 2% BSA (Fluka, Buchs, Switzerland) in PBS, (c) mAb supernatant (0.2 μg/ml), (d) 20-fold prediluted mouse serum, titrated 1:2 over 11 dilution steps, (e) isotype-specific horseradish peroxidase (HRPO)–labeled goat anti–mouse antibodies(0.5 μg/ml, Southern Biotechnologies), (f) substrate ABTS (2.2′-azino-di-[3-ethylbenzthiazolin-sulfonate (6)], Boehringer Mannheim) and H2O2 (Fluka). Plates were coated over night at 4°C, all other incubations were for 60 to 90 min at room temperature. Between incubations, plates were washed three times with PBS containing 0.5 ml Tween 20 per liter. OD was measured at 405 nm in an ELISA reader. All anti-antibody titers are indicated as -log2 of 20-fold prediluted sera. For Fig. 1, C and D, the dilution step at half maximal OD was determined as shown in Fig. 1 A and then taken as anti-antibody titer. For Fig. 1 E, allotype-specific antiIgM antibodies (Southern Biotechnologies) were used for coating and detection at the same concentrations as described above.


Role of repetitive antigen patterns for induction of antibodies against antibodies.

Fehr T, Bachmann MF, Bucher E, Kalinke U, Di Padova FE, Lang AB, Hengartner H, Zinkernagel RM - J. Exp. Med. (1997)

IgG anti-IgM response after immunization with immune complexes versus uncomplexed antigen. (A) BALB/c or (B) A/J  mice were immunized twice (interval, 14 d)  with 1 μg of the monoclonal anti-VSV-G IgM  antibody M1 complexed with 108 PFU UV- inactivated VSV (closed squares), 10 μg of recombinant VSV-G (closed triangles), or 4 μg of rat  anti–mouse Cκ antibody (closed circles), with IgM  alone (open diamonds) or with VSV alone (open  squares). 6 d after secondary immunization, IgG  anti-IgM titers were determined by ELISA as  described in Materials and Methods. (C) IgG  anti-IgM titers of 20-fold prediluted sera were  determined over a time period of 80 d after primary immunization as described in A. The same symbols are used. (D) Dose dependence of IgG anti-IgM induction shown by immunization of BALB/c  mice with IC made of 108 PFU UV-inactivated VSV and titrated amounts of two different monoclonal anti-VSV-G IgM antibodies M1 (closed bars) and  M3 (open bars). (E) C57BL/6 mice were immunized once with 5 μg of the monoclonal anti-VSV-G IgM antibody M5 complexed with 108 PFU UV- inactivated VSV (closed squares), with 5 μg of M5 alone (open diamonds) or with 108 PFU VSV alone (open squares). 4 d later, IgMa-specific IgMb antibodies  were determined by ELISA as described in Materials and Methods.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196322&req=5

Figure 1: IgG anti-IgM response after immunization with immune complexes versus uncomplexed antigen. (A) BALB/c or (B) A/J mice were immunized twice (interval, 14 d) with 1 μg of the monoclonal anti-VSV-G IgM antibody M1 complexed with 108 PFU UV- inactivated VSV (closed squares), 10 μg of recombinant VSV-G (closed triangles), or 4 μg of rat anti–mouse Cκ antibody (closed circles), with IgM alone (open diamonds) or with VSV alone (open squares). 6 d after secondary immunization, IgG anti-IgM titers were determined by ELISA as described in Materials and Methods. (C) IgG anti-IgM titers of 20-fold prediluted sera were determined over a time period of 80 d after primary immunization as described in A. The same symbols are used. (D) Dose dependence of IgG anti-IgM induction shown by immunization of BALB/c mice with IC made of 108 PFU UV-inactivated VSV and titrated amounts of two different monoclonal anti-VSV-G IgM antibodies M1 (closed bars) and M3 (open bars). (E) C57BL/6 mice were immunized once with 5 μg of the monoclonal anti-VSV-G IgM antibody M5 complexed with 108 PFU UV- inactivated VSV (closed squares), with 5 μg of M5 alone (open diamonds) or with 108 PFU VSV alone (open squares). 4 d later, IgMa-specific IgMb antibodies were determined by ELISA as described in Materials and Methods.
Mentions: We used a sandwich ELISA with the following steps: (a) coating with isotype-specific goat anti– mouse antibody (1 μg/ml; Southern Biotechnologies, Birmingham, AL), (b) blocking with 2% BSA (Fluka, Buchs, Switzerland) in PBS, (c) mAb supernatant (0.2 μg/ml), (d) 20-fold prediluted mouse serum, titrated 1:2 over 11 dilution steps, (e) isotype-specific horseradish peroxidase (HRPO)–labeled goat anti–mouse antibodies(0.5 μg/ml, Southern Biotechnologies), (f) substrate ABTS (2.2′-azino-di-[3-ethylbenzthiazolin-sulfonate (6)], Boehringer Mannheim) and H2O2 (Fluka). Plates were coated over night at 4°C, all other incubations were for 60 to 90 min at room temperature. Between incubations, plates were washed three times with PBS containing 0.5 ml Tween 20 per liter. OD was measured at 405 nm in an ELISA reader. All anti-antibody titers are indicated as -log2 of 20-fold prediluted sera. For Fig. 1, C and D, the dilution step at half maximal OD was determined as shown in Fig. 1 A and then taken as anti-antibody titer. For Fig. 1 E, allotype-specific antiIgM antibodies (Southern Biotechnologies) were used for coating and detection at the same concentrations as described above.

Bottom Line: Experience shows that they are usually difficult to induce experimentally.Why and how such anti-antibodies are induced during autoimmune diseases, has remained largely unclear.The results indicate a novel link between anti-antibody responses and infectious agents, and suggest a similar role for repetitive self-antigens such as DNA or collagen involved in chronic immunopathological diseases.

View Article: PubMed Central - PubMed

Affiliation: Institute of Experimental Immunology, University of Zürich, CH-8091 Zürich, Switzerland.

ABSTRACT
Antibody responses against antibodies, such as rheumatoid factors, are found in several immunopathological diseases and may play a role in disease pathogenesis. Experience shows that they are usually difficult to induce experimentally. Antibodies specific for immunoglobulin constant regions (anti-allotypic) or for variable regions (anti-idiotypic) have been investigated in animal models; the latter have even been postulated to regulate antibody and T cell responses via network-like interactions. Why and how such anti-antibodies are induced during autoimmune diseases, has remained largely unclear. Because repetitively arranged epitopes in a paracrystalline structure of a viral envelope cross-link B cell receptors efficiently to induce a prompt T-independent IgM response, this study used immune complexes containing viruses or bacteria to evaluate the role of antigen pattern for induction of anti-antibody responses. We present evidence that antibodies bound to strictly ordered, but not to irregularly arranged, antigens dramatically enhance induction of anti-antibodies, already after a single immunization and without using adjuvants. The results indicate a novel link between anti-antibody responses and infectious agents, and suggest a similar role for repetitive self-antigens such as DNA or collagen involved in chronic immunopathological diseases.

Show MeSH
Related in: MedlinePlus