Additional file 1 of Incidence and severity of immune-related hepatitis after dual checkpoint therapy is linked to younger age independent of herpes virus immunity
Creators: name: Zhang, Zhen
name: Rafei-Shamsabadi, David
name: Lehr, Saskia
name: Buettner, Nico
name: Diehl, Rebecca
name: Huzly, Daniela
name: Pinato, David J
name: Thimme, Robert
name: Meiss, Frank
name: Bengsch, Bertram
Publisher: name: figshare
Year: 2024
Subjects: subject: Immunology
subject: FOS: Clinical medicine

description: Additional file 1: Fig. S1. Characteristics of discovery cohort. (A) Binned histogram depicting age distribution of patients (n = 39, min–max [median]: 19–73 [54]). (B) and (C) Pie charts depicting cohort gender distribution and tumor response in the first 3 months. Fig. S2. Workflow of patient selection. Melanoma patients treated with anti-PD1 &amp; anti-CTLA4 combination therapy in Dermatology, University Hopsital of Freiburg between 01/2016–09/2021 were identified in clinical records, with n = 40 in the prospective discovery cohort and n = 71 in retrospective validation cohort. Patients with untraceable clinical data after therapy initiation and patients with hepatitis with other etiology in parallel were further excluded. A total of 106 patients were evaluated for irAE, tumor response and other clinical information. N = 96 plasma was used for IgG serostatus detection against CMV/EBV/HSV. N = 29 PBMCs were analyzed for immune populations at baseline. Fig. S3. Baseline AST and ALT values are associated with Immune-checkpoint hepatitis. Baseline AST and ALT values are identified from medical records. Slight elevation of baseline AST (A, left) and ALT (A, right) values were observed in patients who developed hepatitis within 100 days after therapy initiation. (B) ROC analysis showed potential role of baseline AST and ALT values in predicting hepatitis incidence. (C) Incidence of hepatitis in patients stratified by baseline liver transaminases levels. P value was determined by Fisher’s exact test. (D) and (E) Sub-cohort analysis of baseline AST, ALT values stratified by CMV IgG serostatus or age. Fig. S4. EBV/HSV/CMV IgG serostatus does not correlate with Immune-checkpoint hepatitis. Kaplan–Meier analysis of hepatitis events in patients grouped by IgG positivity against EBV and HSV in pooled cohort (A), observation cohort (B) and validation cohort (C). (D) Incidence of hepatitis in patients stratified by anti-viral IgG serostatus. (E) Sub-cohort analysis on patients with normal baseline liver transaminase levels stratified by CMV serostatus with or without age subgroups. Fig. S5. Gating strategies for CD4 and CD8 subpopulations. CD4 and CD8 T cells are gated on the basis of live CD3 + T cells. Naïve CD8 T cells are gated as CCR7 + CD45RA + CD27 + CD8, central memory (Tcm) gated as CCR7 + CD45RA-CD27 + CD8, early effector memory (early Tem) gated as CCR7-CD45RA-CD27 + CD8, late effector memory (late Tem) gated as CCR7-CD45RA-CD27-CD8, Temra gated as CCR7-CD45RA + CD27-CD8. Effector memory CD4 T cells (Tem CD4) are gated as CCR7-CDRA-CD4, as in Hutchinson et al. Fig. S6. Age-associated T cell populations are not related to incidence of Hepatitis. Peripheral T cell populations were quantified in 29 melanoma patients before cCBI. (A) Spearman correlation analysis of T cell sub-population distribution with increasing age. Correlation strength and directionality (left: negative correlation, right: positive correlation) is shown for each T cell sub-population as indicated. Correlation significance for each population is denoted by shading (p &lt; 0.01, black, p &lt; 0.05, gray). (B) linear regression between age and %Tn CD8 (left) or %TIGIT + CD8 (right). Patients without hepatitis are represented in black dots, patients with mild (CTC-AE I-II) hepatitis in purple dots and patients with severe (CTC-AE III-IV) hepatitis in blue dots. (C) Comparisons of %Tn CD8 (left) and %TIGIT + CD8 T cells (right) between patients without hepatitis and patients with mild or severe hepatitis. Statistics was performed with Mann–Whitney test. Fig. S7. Tumor progression-free survival analysis according to ICB-hepatitis, age and irAE. Kaplan–Meier analysis of tumor progression free survival (PFS) was performed based on (A) incidence of hepatitis, (B) age (cutoff 55 years) and (C) incidence of at least one of the top 5 most frequent (hepatitis, enteritis, thyroiditis, hypophysitis and skin abnormality) irAE in our cohort.