Limits...
Severe anaphylactic reactions to glutamic acid decarboxylase (GAD) self peptides in NOD mice that spontaneously develop autoimmune type 1 diabetes mellitus.

Pedotti R, Sanna M, Tsai M, DeVoss J, Steinman L, McDevitt H, Galli SJ - BMC Immunol. (2003)

Bottom Line: Accordingly, various protocols of "peptide immunotherapy" of T1DM are under investigation.These findings document severe anaphylaxis to self peptide preparations used in an attempt to devise immunotherapy for a spontaneous autoimmune disease.Taken together with the findings in EAE, these results suggest that peptide therapies designed to induce a TH1 to TH2 shift carry a risk for the development of anaphylactic reactivity to the therapeutic peptides.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, Stanford University School of Medicine, Stanford, California, USA. rpedotti@stanford.edu

ABSTRACT

Background: Insulin dependent (i.e., "type 1") diabetes mellitus (T1DM) is considered to be a T cell mediated disease in which TH1 and Tc autoreactive cells attack the pancreatic islets. Among the beta-cell antigens implicated in T1DM, glutamic acid decarboxylase (GAD) 65 appears to play a key role in the development of T1DM in humans as well as in non-obese diabetic (NOD) mice, the experimental model for this disease. It has been shown that shifting the immune response to this antigen from TH1 towards TH2, via the administration of GAD65 peptides to young NOD mice, can suppress the progression to overt T1DM. Accordingly, various protocols of "peptide immunotherapy" of T1DM are under investigation. However, in mice with experimental autoimmune encephalomyelitis (EAE), another autoimmune TH1 mediated disease that mimics human multiple sclerosis, anaphylactic shock can occur when the mice are challenged with certain myelin self peptides that initially were administered with adjuvant to induce the disease.

Results: Here we show that NOD mice, that spontaneously develop T1DM, can develop fatal anaphylactic reactions upon challenge with preparations of immunodominant GAD65 self peptides after immunization with these peptides to modify the development of T1DM.

Conclusions: These findings document severe anaphylaxis to self peptide preparations used in an attempt to devise immunotherapy for a spontaneous autoimmune disease. Taken together with the findings in EAE, these results suggest that peptide therapies designed to induce a TH1 to TH2 shift carry a risk for the development of anaphylactic reactivity to the therapeutic peptides.

Show MeSH

Related in: MedlinePlus

Peptide specific IgG1 and IgG2a antibodies in NOD mice immunized with G7 (A), PD (B), or HEL/OVA (C) peptides. Serum was collected 2–3 days before challenge with G7, PD, HEL/OVA or saline. IgG1 and IgG2a antibody responses specific for the G7, PD or HEL/OVA peptide epitopes were analyzed by ELISA. Each mouse was tested individually at a serum dilution of 1:20,000 for IgG1, and 1:25 for IgG2a for mice immunized against G7 peptides and 1:500 for IgG1 and 1:200 for IgG2a for mice immunized against PD or HEL/OVA peptides. Data are shown as mean +/- s. e. m. "Saline" = mice injected with saline/IFA; none = non-injected naïve mice.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC153530&req=5

Figure 2: Peptide specific IgG1 and IgG2a antibodies in NOD mice immunized with G7 (A), PD (B), or HEL/OVA (C) peptides. Serum was collected 2–3 days before challenge with G7, PD, HEL/OVA or saline. IgG1 and IgG2a antibody responses specific for the G7, PD or HEL/OVA peptide epitopes were analyzed by ELISA. Each mouse was tested individually at a serum dilution of 1:20,000 for IgG1, and 1:25 for IgG2a for mice immunized against G7 peptides and 1:500 for IgG1 and 1:200 for IgG2a for mice immunized against PD or HEL/OVA peptides. Data are shown as mean +/- s. e. m. "Saline" = mice injected with saline/IFA; none = non-injected naïve mice.

Mentions: As demonstrated in our study, immunization of NOD mice with PD peptides can induce both a specific IgG1 response and also anaphylactic reactivity. On the other hand, as might be predicted, PD peptides induced a less robust IgG1 response (Figure 2) and also a lower incidence and severity of anaphylaxis (see Table 1 and Figure 1) when injected into NOD mice than did G7 peptides. In an attempt to induce anaphylactic reactivity to peptides known to induce TH2 responses associated with allergic reactions, NOD mice were immunized using the same protocol with hen egg lysozyme and ovalbumin peptides (HEL 81–96, OVA 323–339) [21-23]. As a negative control, NOD mice received 3 weekly injections of saline emulsified in IFA. Four weeks after the last of the 3 i.p. injections of peptides/IFA or saline/IFA, mice injected with peptides/IFA were challenged i.p. with the same peptides used for the immunizations dissolved in saline, whereas mice that had been injected with saline/IFA were challenged with saline alone. By the day of challenge, 10–15% of all mice had developed diabetes, with the exception of the mice in the saline group (0%).


Severe anaphylactic reactions to glutamic acid decarboxylase (GAD) self peptides in NOD mice that spontaneously develop autoimmune type 1 diabetes mellitus.

Pedotti R, Sanna M, Tsai M, DeVoss J, Steinman L, McDevitt H, Galli SJ - BMC Immunol. (2003)

Peptide specific IgG1 and IgG2a antibodies in NOD mice immunized with G7 (A), PD (B), or HEL/OVA (C) peptides. Serum was collected 2–3 days before challenge with G7, PD, HEL/OVA or saline. IgG1 and IgG2a antibody responses specific for the G7, PD or HEL/OVA peptide epitopes were analyzed by ELISA. Each mouse was tested individually at a serum dilution of 1:20,000 for IgG1, and 1:25 for IgG2a for mice immunized against G7 peptides and 1:500 for IgG1 and 1:200 for IgG2a for mice immunized against PD or HEL/OVA peptides. Data are shown as mean +/- s. e. m. "Saline" = mice injected with saline/IFA; none = non-injected naïve mice.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC153530&req=5

Figure 2: Peptide specific IgG1 and IgG2a antibodies in NOD mice immunized with G7 (A), PD (B), or HEL/OVA (C) peptides. Serum was collected 2–3 days before challenge with G7, PD, HEL/OVA or saline. IgG1 and IgG2a antibody responses specific for the G7, PD or HEL/OVA peptide epitopes were analyzed by ELISA. Each mouse was tested individually at a serum dilution of 1:20,000 for IgG1, and 1:25 for IgG2a for mice immunized against G7 peptides and 1:500 for IgG1 and 1:200 for IgG2a for mice immunized against PD or HEL/OVA peptides. Data are shown as mean +/- s. e. m. "Saline" = mice injected with saline/IFA; none = non-injected naïve mice.
Mentions: As demonstrated in our study, immunization of NOD mice with PD peptides can induce both a specific IgG1 response and also anaphylactic reactivity. On the other hand, as might be predicted, PD peptides induced a less robust IgG1 response (Figure 2) and also a lower incidence and severity of anaphylaxis (see Table 1 and Figure 1) when injected into NOD mice than did G7 peptides. In an attempt to induce anaphylactic reactivity to peptides known to induce TH2 responses associated with allergic reactions, NOD mice were immunized using the same protocol with hen egg lysozyme and ovalbumin peptides (HEL 81–96, OVA 323–339) [21-23]. As a negative control, NOD mice received 3 weekly injections of saline emulsified in IFA. Four weeks after the last of the 3 i.p. injections of peptides/IFA or saline/IFA, mice injected with peptides/IFA were challenged i.p. with the same peptides used for the immunizations dissolved in saline, whereas mice that had been injected with saline/IFA were challenged with saline alone. By the day of challenge, 10–15% of all mice had developed diabetes, with the exception of the mice in the saline group (0%).

Bottom Line: Accordingly, various protocols of "peptide immunotherapy" of T1DM are under investigation.These findings document severe anaphylaxis to self peptide preparations used in an attempt to devise immunotherapy for a spontaneous autoimmune disease.Taken together with the findings in EAE, these results suggest that peptide therapies designed to induce a TH1 to TH2 shift carry a risk for the development of anaphylactic reactivity to the therapeutic peptides.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, Stanford University School of Medicine, Stanford, California, USA. rpedotti@stanford.edu

ABSTRACT

Background: Insulin dependent (i.e., "type 1") diabetes mellitus (T1DM) is considered to be a T cell mediated disease in which TH1 and Tc autoreactive cells attack the pancreatic islets. Among the beta-cell antigens implicated in T1DM, glutamic acid decarboxylase (GAD) 65 appears to play a key role in the development of T1DM in humans as well as in non-obese diabetic (NOD) mice, the experimental model for this disease. It has been shown that shifting the immune response to this antigen from TH1 towards TH2, via the administration of GAD65 peptides to young NOD mice, can suppress the progression to overt T1DM. Accordingly, various protocols of "peptide immunotherapy" of T1DM are under investigation. However, in mice with experimental autoimmune encephalomyelitis (EAE), another autoimmune TH1 mediated disease that mimics human multiple sclerosis, anaphylactic shock can occur when the mice are challenged with certain myelin self peptides that initially were administered with adjuvant to induce the disease.

Results: Here we show that NOD mice, that spontaneously develop T1DM, can develop fatal anaphylactic reactions upon challenge with preparations of immunodominant GAD65 self peptides after immunization with these peptides to modify the development of T1DM.

Conclusions: These findings document severe anaphylaxis to self peptide preparations used in an attempt to devise immunotherapy for a spontaneous autoimmune disease. Taken together with the findings in EAE, these results suggest that peptide therapies designed to induce a TH1 to TH2 shift carry a risk for the development of anaphylactic reactivity to the therapeutic peptides.

Show MeSH
Related in: MedlinePlus