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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-IgG titers after immunization with antibodies complexed to Salmonella typhi (S.t.) and Pseudomonas aeruginosa (P.a.). (A) BALB/c  mice were immunized twice (interval 14 d) with 108 CFU of S.t. complexed with 20 μg (closed squares) or 2 μg (closed triangles) of the anti-LPS antibody  WN1 222-5 (IgG2a) or with 108 (open squares) or 106 (open triangles) CFU of S.t. alone or with 20 μg of antibody WN1 222-5 alone (open diamonds). 6 d  after the second injection, IgG2b plus IgG1 anti-IgG2a titers in 20-fold prediluted sera were determined in a sandwich ELISA. (B) BALB/c mice were  immunized twice with 108 PFU of P.a. complexed with 7 μg of the anti-LPS antibody 99-T2 (IgG2b) (closed squares), with 108 CFU of P.a. alone (open  triangles) or with 7 μg of 99-T2 alone (open diamonds). IgG2a plus IgG1 anti-IgG2b titers of 20-fold prediluted sera were determined in a sandwich  ELISA. (C) Sera of BALB/c mice immunized twice with IC or with bacteria alone as described in A or B were tested on ELISA plates coated with different antibodies of the corresponding isotype, and anti-antibody titers were determined as described in Fig 1. (D) BALB/c, LPS-responsive C3H/HeN,  and LPS-nonresponsive C3H/HeJ mice were immunized twice with IC, and anti-antibody titers were determined 6 d after booster injection as described  in the legend to Fig 1. Bars in C and D represent geometrical means of 2–3 animals per group. Standard deviation was within ± one dilution step. Results of one out of three comparable experiments are shown.
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Figure 3: IgG anti-IgG titers after immunization with antibodies complexed to Salmonella typhi (S.t.) and Pseudomonas aeruginosa (P.a.). (A) BALB/c mice were immunized twice (interval 14 d) with 108 CFU of S.t. complexed with 20 μg (closed squares) or 2 μg (closed triangles) of the anti-LPS antibody WN1 222-5 (IgG2a) or with 108 (open squares) or 106 (open triangles) CFU of S.t. alone or with 20 μg of antibody WN1 222-5 alone (open diamonds). 6 d after the second injection, IgG2b plus IgG1 anti-IgG2a titers in 20-fold prediluted sera were determined in a sandwich ELISA. (B) BALB/c mice were immunized twice with 108 PFU of P.a. complexed with 7 μg of the anti-LPS antibody 99-T2 (IgG2b) (closed squares), with 108 CFU of P.a. alone (open triangles) or with 7 μg of 99-T2 alone (open diamonds). IgG2a plus IgG1 anti-IgG2b titers of 20-fold prediluted sera were determined in a sandwich ELISA. (C) Sera of BALB/c mice immunized twice with IC or with bacteria alone as described in A or B were tested on ELISA plates coated with different antibodies of the corresponding isotype, and anti-antibody titers were determined as described in Fig 1. (D) BALB/c, LPS-responsive C3H/HeN, and LPS-nonresponsive C3H/HeJ mice were immunized twice with IC, and anti-antibody titers were determined 6 d after booster injection as described in the legend to Fig 1. Bars in C and D represent geometrical means of 2–3 animals per group. Standard deviation was within ± one dilution step. Results of one out of three comparable experiments are shown.

Mentions: To evaluate whether these findings hold true not only for VSV, but also for bacteria exhibiting highly repetitive antigens on their surface, IC formed with gram negative bacteria (exhibiting regularly spaced LPS molecules) and anti-LPS antibodies were tested. 108 CFU of Salmonella typhi (S.t.) (Fig. 3 A) or Pseudomonas aeruginosa (P.a.) (Fig. 3 B) were UV-inactivated and complexed with a monoclonal IgG2a (origin: NZB) or IgG2b (origin: BALB/c) anti-LPS antibody, respectively, to immunize BALB/c mice. After 14 d, these mice were boosted once, and 6 d later IgG anti-IgG antibodies were found in both situations. The induction of these anti-antibodies was again dependent on the antibody dose used to form IC, as shown for S.t. (Fig. 3 A). With the highest dose (108 CFU) immunization with bacteria alone also induced some anti-antibodies, but at much lower titers than with IC. To test whether these results reflected polyclonal B cell activation by the bacterial LPS, we immunized LPS-responder (C3H/HeN, BALB/c) and LPS-nonresponder (C3H/HeJ) mice with IC or noncomplexed antibodies (Fig. 3 D) and tested the BALB/c sera on different antibodies of the same isotype (Fig 3 C). The results indicate: (a) Efficient LPS-mediated stimulation of B cells in the case of P.a., which, however, depended on the presence of IC; the induced anti-antibodies were of rheumatoid factor type because they recognized any autologous IgG2b of BALB/c origin (Fig. 3 C). (b) Successful induction of specific anti-allotypic antibodies by IC with S.t., which recognized only heterologous NZB-derived IgGs (Fig. 3 C) and were independent of B cell activation by LPS (Fig. 3 D). In the case of P.a., 38% (18 of 48) of the mice immunized twice with identical complexes died upon the second immunization from a shock-like syndrome compared with only 4% (2 of 48) of the control groups that were immunized with bacteria, purified LPS, or antibody only; this in vivo finding confirmed the presence of antiantibodies measured in vitro and, in addition, suggested a pathogenic function of RF-like anti-antibodies.


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-IgG titers after immunization with antibodies complexed to Salmonella typhi (S.t.) and Pseudomonas aeruginosa (P.a.). (A) BALB/c  mice were immunized twice (interval 14 d) with 108 CFU of S.t. complexed with 20 μg (closed squares) or 2 μg (closed triangles) of the anti-LPS antibody  WN1 222-5 (IgG2a) or with 108 (open squares) or 106 (open triangles) CFU of S.t. alone or with 20 μg of antibody WN1 222-5 alone (open diamonds). 6 d  after the second injection, IgG2b plus IgG1 anti-IgG2a titers in 20-fold prediluted sera were determined in a sandwich ELISA. (B) BALB/c mice were  immunized twice with 108 PFU of P.a. complexed with 7 μg of the anti-LPS antibody 99-T2 (IgG2b) (closed squares), with 108 CFU of P.a. alone (open  triangles) or with 7 μg of 99-T2 alone (open diamonds). IgG2a plus IgG1 anti-IgG2b titers of 20-fold prediluted sera were determined in a sandwich  ELISA. (C) Sera of BALB/c mice immunized twice with IC or with bacteria alone as described in A or B were tested on ELISA plates coated with different antibodies of the corresponding isotype, and anti-antibody titers were determined as described in Fig 1. (D) BALB/c, LPS-responsive C3H/HeN,  and LPS-nonresponsive C3H/HeJ mice were immunized twice with IC, and anti-antibody titers were determined 6 d after booster injection as described  in the legend to Fig 1. Bars in C and D represent geometrical means of 2–3 animals per group. Standard deviation was within ± one dilution step. Results of one out of three comparable experiments are shown.
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Figure 3: IgG anti-IgG titers after immunization with antibodies complexed to Salmonella typhi (S.t.) and Pseudomonas aeruginosa (P.a.). (A) BALB/c mice were immunized twice (interval 14 d) with 108 CFU of S.t. complexed with 20 μg (closed squares) or 2 μg (closed triangles) of the anti-LPS antibody WN1 222-5 (IgG2a) or with 108 (open squares) or 106 (open triangles) CFU of S.t. alone or with 20 μg of antibody WN1 222-5 alone (open diamonds). 6 d after the second injection, IgG2b plus IgG1 anti-IgG2a titers in 20-fold prediluted sera were determined in a sandwich ELISA. (B) BALB/c mice were immunized twice with 108 PFU of P.a. complexed with 7 μg of the anti-LPS antibody 99-T2 (IgG2b) (closed squares), with 108 CFU of P.a. alone (open triangles) or with 7 μg of 99-T2 alone (open diamonds). IgG2a plus IgG1 anti-IgG2b titers of 20-fold prediluted sera were determined in a sandwich ELISA. (C) Sera of BALB/c mice immunized twice with IC or with bacteria alone as described in A or B were tested on ELISA plates coated with different antibodies of the corresponding isotype, and anti-antibody titers were determined as described in Fig 1. (D) BALB/c, LPS-responsive C3H/HeN, and LPS-nonresponsive C3H/HeJ mice were immunized twice with IC, and anti-antibody titers were determined 6 d after booster injection as described in the legend to Fig 1. Bars in C and D represent geometrical means of 2–3 animals per group. Standard deviation was within ± one dilution step. Results of one out of three comparable experiments are shown.
Mentions: To evaluate whether these findings hold true not only for VSV, but also for bacteria exhibiting highly repetitive antigens on their surface, IC formed with gram negative bacteria (exhibiting regularly spaced LPS molecules) and anti-LPS antibodies were tested. 108 CFU of Salmonella typhi (S.t.) (Fig. 3 A) or Pseudomonas aeruginosa (P.a.) (Fig. 3 B) were UV-inactivated and complexed with a monoclonal IgG2a (origin: NZB) or IgG2b (origin: BALB/c) anti-LPS antibody, respectively, to immunize BALB/c mice. After 14 d, these mice were boosted once, and 6 d later IgG anti-IgG antibodies were found in both situations. The induction of these anti-antibodies was again dependent on the antibody dose used to form IC, as shown for S.t. (Fig. 3 A). With the highest dose (108 CFU) immunization with bacteria alone also induced some anti-antibodies, but at much lower titers than with IC. To test whether these results reflected polyclonal B cell activation by the bacterial LPS, we immunized LPS-responder (C3H/HeN, BALB/c) and LPS-nonresponder (C3H/HeJ) mice with IC or noncomplexed antibodies (Fig. 3 D) and tested the BALB/c sera on different antibodies of the same isotype (Fig 3 C). The results indicate: (a) Efficient LPS-mediated stimulation of B cells in the case of P.a., which, however, depended on the presence of IC; the induced anti-antibodies were of rheumatoid factor type because they recognized any autologous IgG2b of BALB/c origin (Fig. 3 C). (b) Successful induction of specific anti-allotypic antibodies by IC with S.t., which recognized only heterologous NZB-derived IgGs (Fig. 3 C) and were independent of B cell activation by LPS (Fig. 3 D). In the case of P.a., 38% (18 of 48) of the mice immunized twice with identical complexes died upon the second immunization from a shock-like syndrome compared with only 4% (2 of 48) of the control groups that were immunized with bacteria, purified LPS, or antibody only; this in vivo finding confirmed the presence of antiantibodies measured in vitro and, in addition, suggested a pathogenic function of RF-like anti-antibodies.

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