Spectratyping analysis of the islet-reactive T cell repertoire in diabetic NOD Igμnull mice after polyclonal B cell reconstitution

Journal of Translational Medicine

Claudia Raja Gabaglia¹  Eli E Sercarz¹  Todd A Braciak¹  Huiming Sheng¹  Patricia Y Norori¹  Nazila Daneshjou¹  Allen M Vong¹ 

[1] Laboratory of Vaccine Research, Torrey Pines Institute for Molecular Studies. 3550 General Atomics Court. San Diego, 92121, CA, USA

关键词: IL-6;    B cells;    T cell receptor;    immunoscope;    diabetes;    NOD.Igμ^null;    NOD;   

Others  :  1207917
DOI  :  10.1186/1479-5876-9-101

 received in 2011-03-23, accepted in 2011-07-02,  发布年份 2011

【 摘 要 】

Background

Non Obese Diabetic mice lacking B cells (NOD.Igμ^null mice) do not develop diabetes despite their susceptible background. Upon reconstitution of B cells using a chimera approach, animals start developing diabetes at 20 weeks of age.

Methods

We have used the spectratyping technique to follow the T cell receptor (TCR) V beta repertoire of NOD.Igμ^null mice following B cell reconstitution. This technique provides an unbiased approach to understand the kinetics of TCR expansion. We have also analyzed the TCR repertoire of reconstituted animals receiving cyclophosphamide treatment and following tissue transplants to identify common aggressive clonotypes.

Results

We found that B cell reconstitution of NOD.Igμ^null mice induces a polyclonal TCR repertoire in the pancreas 10 weeks later, gradually diversifying to encompass most BV families. Interestingly, these clonotypic BV expansions are mainly confined to the pancreas and are absent from pancreatic lymph nodes or spleens. Cyclophosphamide-induced diabetes at 10 weeks post-B cell reconstitution reorganized the predominant TCR repertoires by removing potential regulatory clonotypes (BV1, BV8 and BV11) and increasing the frequency of others (BV4, BV5S2, BV9, BV16-20). These same clonotypes are more frequently present in neonatal pancreatic transplants under the kidney capsule of B-cell reconstituted diabetic NOD.Igμ^null mice, suggesting their higher invasiveness. Phenotypic analysis of the pancreas-infiltrating lymphocytes during diabetes onset in B cell reconstituted animals show a predominance of CD19⁺ B cells with a B:T lymphocyte ratio of 4:1. In contrast, in other lymphoid organs (pancreatic lymph nodes and spleens) analyzed by FACS, the B:T ratio was 1:1. Lymphocytes infiltrating the pancreas secrete large amounts of IL-6 and are of Th1 phenotype after CD3-CD28 stimulation in vitro.

Conclusions

Diabetes in NOD.Igμ^null mice appears to be caused by a polyclonal repertoire of T cell accumulation in pancreas without much lymphoid organ involvement and is dependent on the help by B cells.

【 授权许可】

   
2011 Vong et al; licensee BioMed Central Ltd.

【 图 表 】

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