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Determinant spreading of T helper cell 2 (Th2) responses to pancreatic islet autoantigens.

Tian J, Lehmann PV, Kaufman DL - J. Exp. Med. (1997)

Bottom Line: Surprisingly, induction of antiinflammatory Th2 responses to a single beta cell antigen (betaCA) resulted in the spreading of Th2 cellular and humoral immunity to unrelated betaCAs in an infectious manner and protection from IDDM.The data suggest that both Th1 and Th2 autoimmunity evolve in amplificatory cascades by generating site-specific, but not antigen-specific, positive feedback circuits.Determinant spreading of Th2 responses may be a fundamental mechanism underlying antigen-based immunotherapeutics, explaining observations of infectious tolerance and providing a new theoretical framework for therapeutic intervention.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Medical Pharmacology, University of California, Los Angeles, California 90095-1735, USA.

ABSTRACT
The nature (Th1 versus Th2) and dynamics of the autoimmune response during the development of insulin-dependent diabetes mellitus (IDDM) and after immunotherapy are unclear. Here, we show in nonobese diabetic (NOD) mice that the autoreactive T cell response starts and spreads as a pure Th1 type autoimmunity, suggesting that a spontaneous Th1 cascade underlies disease progression. Surprisingly, induction of antiinflammatory Th2 responses to a single beta cell antigen (betaCA) resulted in the spreading of Th2 cellular and humoral immunity to unrelated betaCAs in an infectious manner and protection from IDDM. The data suggest that both Th1 and Th2 autoimmunity evolve in amplificatory cascades by generating site-specific, but not antigen-specific, positive feedback circuits. Determinant spreading of Th2 responses may be a fundamental mechanism underlying antigen-based immunotherapeutics, explaining observations of infectious tolerance and providing a new theoretical framework for therapeutic intervention.

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Propagation of IgG1 responses to βCAs. NOD mice were  treated with HEL, GAD, or insulin B chain, as described in Materials and  Methods. GAD (a) and insulin (b) antibodies were characterized at 12 wk  of age using antigen-specific ELISA assays (18). The background OD was  ∼0.05 ± 0.01 for all samples. Serial dilutions of sera showed a linear relationship with resulting OD. The data are represented as the mean absorbance values over background of triplicate samples from individual mice.  Experimental and control sera were tested simultaneously in two separate  assays (n = 5 for each group). The variance in absorbance values between  triplicate samples from the two sets of experiments was <8%. Humoral  responses to GAD and insulin in control NOD mice treated with HEL  were similar to those of unmanipulated NOD mice. BALB/c mice treated  with βCAs developed antibodies only against the injected antigen (data  not shown), consistent with the observed lack of Th2 spreading in these  mice (Table 1B). Antibodies to GAD and insulin in sera from untreated  BALB/c and AKR mice were at background levels (data not shown).
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Figure 1: Propagation of IgG1 responses to βCAs. NOD mice were treated with HEL, GAD, or insulin B chain, as described in Materials and Methods. GAD (a) and insulin (b) antibodies were characterized at 12 wk of age using antigen-specific ELISA assays (18). The background OD was ∼0.05 ± 0.01 for all samples. Serial dilutions of sera showed a linear relationship with resulting OD. The data are represented as the mean absorbance values over background of triplicate samples from individual mice. Experimental and control sera were tested simultaneously in two separate assays (n = 5 for each group). The variance in absorbance values between triplicate samples from the two sets of experiments was <8%. Humoral responses to GAD and insulin in control NOD mice treated with HEL were similar to those of unmanipulated NOD mice. BALB/c mice treated with βCAs developed antibodies only against the injected antigen (data not shown), consistent with the observed lack of Th2 spreading in these mice (Table 1B). Antibodies to GAD and insulin in sera from untreated BALB/c and AKR mice were at background levels (data not shown).

Mentions: At the time of sacrifice, sera was collected and the isotype of GAD and insulin autoantibodies were characterized using an ELISA assay as described in the legend to Fig. 1 and in reference 18. In brief, GAD (Synectics Biomedical, Stockholm) or insulin B chain at 10 μg/ml were bound to 96-well plates (Nunc), in 0.1 M NaHCO3, pH 8.5 (GAD) or pH 9.6 (insulin B chain), at 4°C overnight. The wells were rinsed with PBS and then blocked with 3% BSA in PBS for 1 h. Mouse sera was added (0.1 ml of a 1:500 dilution) and incubated for 1 h at 37°C. After washing, bound Ig was characterized using affinity purified HRP-coupled goat anti–mouse IgG+A+M (H+L) (Pierce Chemical Co., Rockford, IL), or HRP-coupled goat anti– mouse isotype-specific antibodies for IgG1 and IgG2a (Southern Biotechnology Associates, Birmingham, AL) and ABTS. Sera from untreated BALB/c and AKR mice were used as negative controls.


Determinant spreading of T helper cell 2 (Th2) responses to pancreatic islet autoantigens.

Tian J, Lehmann PV, Kaufman DL - J. Exp. Med. (1997)

Propagation of IgG1 responses to βCAs. NOD mice were  treated with HEL, GAD, or insulin B chain, as described in Materials and  Methods. GAD (a) and insulin (b) antibodies were characterized at 12 wk  of age using antigen-specific ELISA assays (18). The background OD was  ∼0.05 ± 0.01 for all samples. Serial dilutions of sera showed a linear relationship with resulting OD. The data are represented as the mean absorbance values over background of triplicate samples from individual mice.  Experimental and control sera were tested simultaneously in two separate  assays (n = 5 for each group). The variance in absorbance values between  triplicate samples from the two sets of experiments was <8%. Humoral  responses to GAD and insulin in control NOD mice treated with HEL  were similar to those of unmanipulated NOD mice. BALB/c mice treated  with βCAs developed antibodies only against the injected antigen (data  not shown), consistent with the observed lack of Th2 spreading in these  mice (Table 1B). Antibodies to GAD and insulin in sera from untreated  BALB/c and AKR mice were at background levels (data not shown).
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199172&req=5

Figure 1: Propagation of IgG1 responses to βCAs. NOD mice were treated with HEL, GAD, or insulin B chain, as described in Materials and Methods. GAD (a) and insulin (b) antibodies were characterized at 12 wk of age using antigen-specific ELISA assays (18). The background OD was ∼0.05 ± 0.01 for all samples. Serial dilutions of sera showed a linear relationship with resulting OD. The data are represented as the mean absorbance values over background of triplicate samples from individual mice. Experimental and control sera were tested simultaneously in two separate assays (n = 5 for each group). The variance in absorbance values between triplicate samples from the two sets of experiments was <8%. Humoral responses to GAD and insulin in control NOD mice treated with HEL were similar to those of unmanipulated NOD mice. BALB/c mice treated with βCAs developed antibodies only against the injected antigen (data not shown), consistent with the observed lack of Th2 spreading in these mice (Table 1B). Antibodies to GAD and insulin in sera from untreated BALB/c and AKR mice were at background levels (data not shown).
Mentions: At the time of sacrifice, sera was collected and the isotype of GAD and insulin autoantibodies were characterized using an ELISA assay as described in the legend to Fig. 1 and in reference 18. In brief, GAD (Synectics Biomedical, Stockholm) or insulin B chain at 10 μg/ml were bound to 96-well plates (Nunc), in 0.1 M NaHCO3, pH 8.5 (GAD) or pH 9.6 (insulin B chain), at 4°C overnight. The wells were rinsed with PBS and then blocked with 3% BSA in PBS for 1 h. Mouse sera was added (0.1 ml of a 1:500 dilution) and incubated for 1 h at 37°C. After washing, bound Ig was characterized using affinity purified HRP-coupled goat anti–mouse IgG+A+M (H+L) (Pierce Chemical Co., Rockford, IL), or HRP-coupled goat anti– mouse isotype-specific antibodies for IgG1 and IgG2a (Southern Biotechnology Associates, Birmingham, AL) and ABTS. Sera from untreated BALB/c and AKR mice were used as negative controls.

Bottom Line: Surprisingly, induction of antiinflammatory Th2 responses to a single beta cell antigen (betaCA) resulted in the spreading of Th2 cellular and humoral immunity to unrelated betaCAs in an infectious manner and protection from IDDM.The data suggest that both Th1 and Th2 autoimmunity evolve in amplificatory cascades by generating site-specific, but not antigen-specific, positive feedback circuits.Determinant spreading of Th2 responses may be a fundamental mechanism underlying antigen-based immunotherapeutics, explaining observations of infectious tolerance and providing a new theoretical framework for therapeutic intervention.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Medical Pharmacology, University of California, Los Angeles, California 90095-1735, USA.

ABSTRACT
The nature (Th1 versus Th2) and dynamics of the autoimmune response during the development of insulin-dependent diabetes mellitus (IDDM) and after immunotherapy are unclear. Here, we show in nonobese diabetic (NOD) mice that the autoreactive T cell response starts and spreads as a pure Th1 type autoimmunity, suggesting that a spontaneous Th1 cascade underlies disease progression. Surprisingly, induction of antiinflammatory Th2 responses to a single beta cell antigen (betaCA) resulted in the spreading of Th2 cellular and humoral immunity to unrelated betaCAs in an infectious manner and protection from IDDM. The data suggest that both Th1 and Th2 autoimmunity evolve in amplificatory cascades by generating site-specific, but not antigen-specific, positive feedback circuits. Determinant spreading of Th2 responses may be a fundamental mechanism underlying antigen-based immunotherapeutics, explaining observations of infectious tolerance and providing a new theoretical framework for therapeutic intervention.

Show MeSH
Related in: MedlinePlus