sj-pdf-1-opp-10.1177_10781552231167824 - Supplemental material for Immunosuppressive therapy management in cancer patients with autoimmune diseases treated with immune checkpoint inhibitors: A case series and systematic literature review

Supplemental material, sj-pdf-1-opp-10.1177_10781552231167824 for Immunosuppressive therapy management in cancer patients with autoimmune diseases treated with immune checkpoint inhibitors: A case series and systematic literature review by Stephanie C.M. Wuyts, Charlotte A.H. Cappelle, Marthe Verhaert, Bert Bravenboer and Sandrine Aspeslagh in Journal of Oncology Pharmacy Practice</p

α-GalCer treatment decreases viral titers in the spleen and liver.

<p>Spleen (A) and liver (B) were harvested at 48 hr post-infection and MCMV titers were determined by standard plaque assay. *p-value≤0.007.</p

α-GalCer does not alter the overall number of MCMV specific CD8⁺ T cells in the spleen or liver.

<p>At 9 weeks post-infection, cells were stained with M45 and M57 tetramers. Frequency of tetramer positive cells in the spleen (A) or liver (B). Absolute number of tetramer positive cells in the spleen (C) or liver (D). p-value = 0.003.</p

α-GalCer treatment modulates MCMV-dependent cytokine profile.

<p>Blood sera were collected at 38 hours post-infection and cytokines measured using the BD CBA Flex Kit. Cytokines not detected under any condition tested: IL-10, IL-4, IL-21, IL-17A, GM-CSF. *p-value≤0.002.</p

α-GalCer promotes Th1 and Th2 cytokine production by 4 hours.

<p>IFN-γ (A) and IL-4 (B) levels were determined at 4 hours in blood serum after α-GalCer treatment and/or MCMV infection by ELISA. *p-value<0.005.</p

PCA score plot with the two first principal component vectors t(1) and t(2).

<p>Each PCA vector represents a specific combination of the 1656 chemical descriptors. The three most outlying groups can be observed. An in depth study was performed with HCA analysis.</p

Comparison of the different test-models.

<p>(A) Intra-variability. Box-plots of relative standard deviations are shown. Each value represents a compound with its relative standard deviation. The human test-model shows a lower median than the mice test-models, which means that in the human test-model more compounds have uniform cytokine-responses between different research groups (IL-4 p = 0.015, IFN-γ p = 0.13, Kruskal-Wallis test). (B) Inter-variability. The height of the bars represents the discriminating power of a specific test-model for a specific cytokine. This is calculated by a relative standard deviation of the biological responses in a specific test-model (corrected for the intrinsic intra-variability). (C + D) Relation between <i>mice/in vitro</i> and <i>human/in vitro</i> assay for (C) IFN-γ (ρ_s = 0.56, p = 0.002) and (D) IL-4 (ρ_s = 0.38, p = 0.18).</p

Th1/Th2 polarization in the different test-models.

<p>The graph has a color-gradient with the darker parts representing stronger polarizers. α-GalCer is shown in red on the y = x line.</p

Distribution of methodologies used in α-GalCer immunological studies.

<p>Distribution of methodologies used in α-GalCer immunological studies.</p

iNKT numbers in MM patients.

<p>Human iNKT (number/mL) were analyzed in blood samples of 51 Patients (mean age is 64 years old) and 62 healthy donors (mean age is 66 years old) by flow cytometry. Total number of iNKT cells were calculated by determining the % iNKT cells (CD3, TCRVα24 and TCRVβ11+ cells) on total T lymphocyte number (* and ** indicate p<0.05 and p<0.01, Student t-test, ns =  not significant).</p