B Lymphocyte “Original Sin” in the Bone Marrow Enhances Islet Autoreactivity in Type 1 Diabetes–Prone Nonobese Diabetic Mice

Bonami, Rachel H.; Williams, Jonathan M.; Rachakonda, Amita; Karamali, Mariam; Kendall, Peggy L.; Thomas, James W.

doi:10.4049/jimmunol.1201359

Cited by 28 publications

(70 citation statements)

References 40 publications

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“…7C–D). Vκ4–74 clones were paired with all Jκ, forming different CDR3 amino acid junctions associated with previously recognized low/moderate (P-L) and high (P-P) affinity for rodent insulin, as deduced from sequences identified in studies of anti-insulin hybridomas (35). The 5’ degenerate primer sequence was removed, and Vκ sequences were deposited into GenBank (http://www.ncbi.nlm.nih.gov/genbank/) with the following accession numbers: unimmunized (KP790058-KP790071), non-GC (KP790072-KP790083), IgM + GC (KP790084-KP790092), and IgM − GC (KP790093-KP790098).…”

Section: Resultsmentioning

confidence: 98%

“…MAb123 recognizes a different insulin epitope and binds insulin-occupied BCRs (25, 35). Flow cytometry staining with mAb123 confirms that BCRs are occupied by endogenous insulin in V H 125 SD B6 mice (0.4 ± 0.1%, Fig.…”

Section: Resultsmentioning

confidence: 99%

“…7). Prior studies show that Vκ4–74 pairs with several Jκ to form autoreactive insulin-binding BCRs when combined with V H 125 (35). Thus, insulin-binding B cells from the pre-immune repertoire, and not obscure clonally ignorant B cells, are selected for GC entry during the insulin-BRT response.…”

Section: Discussionmentioning

confidence: 99%

“…Abs reactive to murine B220 (RA3–6B2), IgM a (DS-1), IgM b (AF6–78), CD21 (7G6), CD23 (B3B4), IgD (11–26), GL7, or Fas/CD95 (Jo2) (BD Pharmingen), or IgM (µ-chain specific, Invitrogen) were used, along with biotinylated human insulin (35) or biotinylated anti-insulin mAb 123 (36) with streptavidin-conjugated fluorochrome to detect insulin-binding B cells, and 7-aminoactinomycin D for cell viability. Flow cytometry acquisition was performed after cell suspension in FACS buffer (1X PBS, 10% FBS, 1% sodium azide, 1% EDTA) using an LSR II (BD Biosciences), and sorting experiments were performed using a FACSAria I or II cell sorter in the Vanderbilt University Shared Resource Facility.…”

Section: Methodsmentioning

confidence: 99%

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Reversing Tolerance in Isotype Switch–Competent Anti-Insulin B Lymphocytes

Williams

Bonami

Hulbert

et al. 2015

The Journal of Immunology

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B lymphocytes that escape central tolerance and mature in the periphery are a liability for developing autoimmunity. IgG insulin autoAbs that predict type 1 diabetes and complicate insulin therapies indicate that mechanisms for tolerance to insulin are flawed. To examine peripheral tolerance in anti-insulin B cells, we generated C57BL/6 mice that harbor anti-insulin VDJH-125 site-directed to the native Ig H chain locus (VH125SD). Class switch-competent anti-insulin B cells fail to produce IgG Abs following T cell-dependent (TD) immunization of VH125SD mice with heterologous insulin, and they exhibit markedly impaired proliferation to anti-CD40 plus insulin in vitro. In contrast, co-stimulation with LPS plus insulin drives robust anti-insulin B cell proliferation. Further, VH125SD mice produce both IgM and IgG2a anti-insulin Abs following immunization with a conjugate of insulin to type 1 T cell-independent Brucella abortus ring test Ag (BRT). Anti-insulin B cells undergo clonal expansion in vivo and emerge as IgM+ and IgM− GL7+ Fas+ germinal center (GC) B cells following immunization with insulin-BRT, but not BRT alone. Analysis of Igκ genes in VH125SD mice immunized with insulin-BRT reveals that anti-insulin Vκ from the pre-immune repertoire are selected into GCs. These data demonstrate that class switch-competent anti-insulin B cells remain functionally silent in TD immune responses, yet these B cells are vulnerable to reversal of anergy following combined BCR/TLR engagement that promotes Ag-specific GC responses and Ab production. Environmental factors that lead to infection and inflammation could play a critical yet under-appreciated role in driving loss of tolerance and promoting autoimmune disease.

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Section: Resultsmentioning

confidence: 98%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Reversing Tolerance in Isotype Switch–Competent Anti-Insulin B Lymphocytes

Williams

Bonami

Hulbert

et al. 2015

The Journal of Immunology

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“…Components of the extracellular matrix newly discovered to contribute to MZ development are also overexpressed in the MZ of NOD mice (20). Our group has shown that polymorphisms in Igκ genes contribute to increased propensity for autoreactivity in NOD B cells and that these polymorphisms are shared with SLE-prone mice (21, 22). B cell intrinsic tolerance defects in the NOD strain also allow increased numbers of autoreactive B cells to reach maturity (22-24).…”

Section: Introductionmentioning

confidence: 99%

Bruton’s Tyrosine Kinase Synergizes with Notch2 To Govern Marginal Zone B Cells in Nonobese Diabetic Mice

Case

Bonami

Nyhoff

et al. 2015

The Journal of Immunology

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Expansion of autoimmune-prone marginal zone (MZ) B cells has been implicated in type 1 diabetes (T1D). To test disease contributions of MZ B cells in NOD mice, Notch2 haploinsufficiency (Notch2+/−) was introduced, but failed to eliminate the MZ, as it does in C57BL/6 mice. Notch2+/−/NOD have MZ B cell numbers similar to WT C57BL/6, yet still develop diabetes. To test whether BCR-signaling supports Notch2+/−/NOD MZ B cells, Bruton's tyrosine kinase (Btk)-deficiency was introduced. Surprisingly, MZ B cells failed to develop in Btk-deficient Notch2+/−/NOD mice. Expression of Notch2 and its transcriptional target, Hes5, were increased in NOD MZ B cells compared with C57BL/6 MZ B cells. Btk-deficiency reduced Notch2+/− signaling exclusively in NOD B cells, suggesting that BCR-signaling enhances Notch2 signaling in this autoimmune model. The role of BCR-signaling was further investigated using an anti-insulin transgenic BCR (125Tg). Anti-insulin B cells in 125Tg/Notch2+/−/NOD mice populate an enlarged MZ, suggesting that low level BCR signaling overcomes reliance on Notch2. Tracking clonotypes of anti-insulin B cells in H chain only VH125Tg/NOD mice showed that BTK-dependent selection into the MZ depends on strength of antigenic binding, while Notch2-mediated selection does not. Importantly, anti-insulin B cell numbers were reduced by Btk-deficiency, but not Notch2-haploinsufficiency. These studies show that: 1) Notch2-haploinsufficiency limits NOD MZ B cell expansion without preventing T1D, 2) BTK supports the Notch2 pathway in NOD MZ B cells, and 3) autoreactive NOD B cell survival relies on BTK more than Notch2, regardless of MZ location, which may have important implications for disease-intervention strategies.

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Silencing of high-affinity insulin-reactive B lymphocytes by anergy and impact of the NOD genetic background in mice

et al. 2018

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Aims/hypothesis Previous studies have demonstrated that high-affinity insulin-binding B cells (IBCs) silenced by anergy in healthy humans lose their anergy in islet autoantibody-positive individuals with recent-onset type 1 diabetes, and in autoantibody-negative first-degree relatives carrying certain risk alleles. Here we explore that hypothesis that IBCs are found in the immune periphery of disease-resistant C57BL/6-H2g7 mice, where, as in healthy humans, they are anergic, but that in disease-prone genetic backgrounds (NOD) become activated and migrate to the pancreas and pancreatic lymph nodes, where they participate in the development of type 1 diabetes. Methods We compared the status of high-affinity IBCs in disease-resistant VH125.C57BL/6-H2g7 and disease-prone VH125.NOD mice. Results Consistent with findings in healthy humans, high-affinity IBCs reach the periphery in disease-resistant mice and are anergic, as indicated by a reduced expression of membrane IgM, unresponsiveness to antigen and failure to become activated or accumulate in the pancreatic lymph nodes or pancreas. In NOD mice, high-affinity IBCs reach the periphery early in life and increase in number prior to the onset of hyperglycaemia. These cells are not anergic; they become activated, produce autoantibodies and accumulate in the pancreas and pancreatic lymph nodes prior to disease development. Conclusions/interpretation These findings are consistent with genetic determination of the escape of high-affinity IBCs from anergy and their early contribution to the development of type 1 diabetes.

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B Lymphocyte “Original Sin” in the Bone Marrow Enhances Islet Autoreactivity in Type 1 Diabetes–Prone Nonobese Diabetic Mice

Cited by 28 publications

References 40 publications

Reversing Tolerance in Isotype Switch–Competent Anti-Insulin B Lymphocytes

Reversing Tolerance in Isotype Switch–Competent Anti-Insulin B Lymphocytes

Bruton’s Tyrosine Kinase Synergizes with Notch2 To Govern Marginal Zone B Cells in Nonobese Diabetic Mice

Silencing of high-affinity insulin-reactive B lymphocytes by anergy and impact of the NOD genetic background in mice

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