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. 2010 Feb;134(2):121-9.
doi: 10.1016/j.clim.2009.09.005.

Monitoring of antigen-specific CD8 T cells in patients with type 1 diabetes treated with antiCD3 monoclonal antibodies

Simona Cernea  1 , Kevan C Herold
Affiliations

Monitoring of antigen-specific CD8 T cells in patients with type 1 diabetes treated with antiCD3 monoclonal antibodies

Simona Cernea et al. Clin Immunol. 2010 Feb.

Abstract

The way in which anti-CD3 monoclonal antibodies (mAbs) modify human immune responses in type 1 diabetes (T1DM) is not known. We prepared a panel of Class I HLA-A2.1 tetramers with peptides from diabetes-associated antigens and studied the frequency and phenotype of the cells in patients with T1DM and blood donors and in patients treated with anti-CD3 mAb (Teplizumab). More patients with T1DM showed positive staining for at least 1 tetramer using frozen and fresh samples (p<0.05). Three months following treatment with anti-CD3 mAb, the proportion of GAD65- and InsB-peptide reactive CD8+ T cells increased (p<0.05). The phenotype of these cells was modulated from naïve to effector memoryRA+. We concludethat Class I MHC tetramers can identify antigen specific CD8+ T cells in patients with T1DM. The frequency of certain specificities increases after treatment with anti-CD3 mAb. Their modulated phenotype may have functional consequences for their pathogenicity.

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Figures

Figure 1
Figure 1. Identification of antigen specific CD8+ T cells with Class I MHC tetramers
The representative staining of fresh cells from a representative patient with T1DM and a blood donor (both HLA-A2+) are shown. The X axis shows staining with anti-CD8 mAb and the Y axis represents staining with tetramer. Electronic gates were placed around lymphocytes on the basis of forward and side scatter and around CD8+ cells. The numbers in each contour plot represents the percentage of CD8+ T cells that are tetramer+ minus the background staining with the negative tetramer (% negative tetramer: 0.06% [T1DM] and 0.03% [Blood donor], respectively). Bolded numbers represent positive staining (above threshold for positivity from staining of HLA-A2− control subjects (see Supplemental Table 1)).
Figure 2
Figure 2. Phenotype of tetramer+ cells
A: PBMC were stained with mAbs to CD8, CCR7, CD45RA, and the indicated tetramers. Electronic gates were placed around the CD8+tetramer+ cells and the expression of CCR7 (X axis) and CD45RA (Y axis) was analyzed. B: The functional phenotypes, on the basis of expression of CCR7 and CD45RA are shown for diabetes antigen specific CD8+ T cells (DT) and EBV specific T cells (EBV) from patients with T1DM. There was a significant difference in the distribution of phenotypes among the DT+ vs EBV+ T cells (p<0.001). EMRA: terminally differentiated effector memory RA+; N: naïve-like; EM: effector memory; CM: central memory; C: The expression of CCR7 and CD45RA among diabetes antigen specific T cells after treatment with anti-CD3 mAb is shown. D: The frequency of phenotypes of CD8+diabetes tetramer+ T cells before and after treatment with anti-CD3 mAb are shown. There was a significant change in the distribution of phenotypes after treatment (p<0.001).
Figure 3
Figure 3. Changes in the numbers of lymphocytes during and following treatment with Teplizumab
A: The absolute lymphocyte (ALC) (n=6), CD4, CD8 (L axis), cell counts, and CD4:CD8 ratios (R axis)(n=6) were calculated for subjects receiving Teplizumab for 14 days. B. The absolute numbers of tetramer+ cells are shown.
Figure 4
Figure 4. Changes in diabetes antigen specific CD8+ T cells with treatment with Teplizumab
A Representative staining for CD8 and tetramers from a single participant is shown before (day 0) and 3 months after (day 90) treatment with Teplizumab. The numbers refer to the percentage tetramer+ of the CD8+ T cells. Bolded numbers are considered positive staining based on threshold values shown in Supplemental Figure 1. B: Changes in the frequency of diabetes antigen specific T cells in individual patients treated with Teplizumab and untreated patients with T1DM. (tetramers: InsA1 ▼;, InsA2 ▲, Ins B ■ PPI ◆ GAD ● IGRP □, Flu ○).
Figure 5
Figure 5. Changes in EBV reactive T cells and anti-EBV immunoglobulins after treatment with Teplizumab
The changes in EBV reactive CD8+ T cells were monitored in parallel with the diabetes-antigen specific T cells. The EBV IgG and IgM titers were measured at the indicated study days (the mAb was given from days 0–14).
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References

    1. Atkinson MA. ADA Outstanding Scientific Achievement Lecture 2004. Thirty years of investigating the autoimmune basis for type 1 diabetes: why can’t we prevent or reverse this disease? Diabetes. 2005;54:1253–63. - PubMed
    1. Miller BJ, Appel MC, O’Neil JJ, Wicker LS. Both the Lyt-2+ and L3T4+ T cell subsets are required for the transfer of diabetes in nonobese diabetic mice. J Immunol. 1988;140:52–8. - PubMed
    1. Christianson SW, Shultz LD, Leiter EH. Adoptive transfer of diabetes into immunodeficient NOD-scid/scid mice. Relative contributions of CD4+ and CD8+ T-cells from diabetic versus prediabetic NOD. NON-Thy-1a donors. Diabetes. 1993;42:44–55. - PubMed
    1. Katz JD, Wang B, Haskins K, Benoist C, Mathis D. Following a diabetogenic T cell from genesis through pathogenesis. Cell. 1993;74:1089–100. - PubMed
    1. Amrani A, Verdaguer J, Serra P, Tafuro S, Tan R, Santamaria P. Progression of autoimmune diabetes driven by avidity maturation of a T-cell population. Nature. 2000;406:739–42. - PubMed

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