Research ArticleBasic Science Investigation

Noninvasive PET Detection of CD69-Positive Immune Cells Before Signs of Clinical Disease in Inflammatory Arthritis

Emmi Puuvuori, Yunbing Shen, Gry Hulsart-Billström, Bogdan Mitran, Bo Zhang, Pierre Cheung, Olivia Wegrzyniak, Sofie Ingvast, Jonas Persson, Stefan Ståhl, Olle Korsgren, John Löfblom, Fredrik Wermeling and Olof Eriksson
Journal of Nuclear Medicine February 2024, 65 (2) 294-299; DOI: https://doi.org/10.2967/jnumed.123.266336

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  • Figure
  • FIGURE 1.
    FIGURE 1.

    (A) Quantification of joint inflammation (clinical score, 0–12; 0–3 points per paw) as part of characterizing induced joint inflammation in KRN T-cell adoptive transfer model. (B) Weight change presented as percentage of weight at day 0. (C) Serum IgG anti-GPI levels comparing day 0 and day 12. **P < 0.01, by unpaired t test (n = 5). GPI = glucose phosphate isomerase; OD = optical density.

  • FIGURE 2.
    FIGURE 2.

    (A) PET images of [68Ga]Ga-DOTA-ZCAM241 uptake at baseline and 3, 7, and 12 d after injection as inflammatory arthritis developed in single representative individual mouse. Images are normalized to SUV of 0.5 for direct comparison between time points. (B) CD69 immunofluorescence Sytox (Thermo Fisher Scientific) staining of joints of representative animals during matching time points.

  • FIGURE 3.
    FIGURE 3.

    [68Ga]Ga-DOTA-ZCAM241 uptake expressed as SUVmean (each point represents average and SD of 5 scans) over time in joints either at group level (A) or in individuals (B). Error bars for day 0 time point in A are too small to be visualized. Asterisks indicate significance compared with baseline at day 0. (C) Uptake of [68Ga]Ga-DOTA-ZCAM241 expressed as ratio at each time point compared with baseline in individuals. (D) Correlation of uptake of [68Ga]Ga-DOTA-ZCAM241 in joints with clinical score. ***P < 0.001. ****P < 0.0001. M1–M5 = mouse 1 to mouse 5.

  • FIGURE 4.
    FIGURE 4.

    CD69 expression analysis in different cell populations on day 12. (A) Representative histogram plot of CD69 expression in different lymph organs (axillary, brachial, inguinal, mesenteric, and popliteal lymph nodes, as well as spleen and joints). Gate shows viable singlets. (B) Quantification of plots in A, with CD69-positive percentage of viable singlets (n = 3). (C) Representative plot showing frequency of B cells and T cells in CD69-positive viable singlet gate. (D) Quantification of plots in C, with percentage of B cells and T cells in CD69-positive viable singlet gate (n = 3). (E) SUVmean uptake of left and right axillary lymph nodes and muscle at day 12. ***P < 0.001, by ANOVA (n = 5). Axi = axillary; Bra = brachial; Freq. = frequency; Ingu = inguinal, LN = lymph node; Mes = mesenteric; Pop = popliteal; TCRβ = T-cell receptor β.

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