CTLA\4, cytotoxic T\lymphocyteCassociated protein 4; Foxp3, forkhead protein 3; GARP, glycoprotein A repetitions predominant; HLA\DR, human leukocyte antigen\DR isotype; nR, non\responder; PD\1, programmed cell death protein 1; R, responder; Treg, regulatory T cells To quantify the T cell\to\Treg composition in the peripheral blood, we calculated the corresponding T cell\to\Treg ratios. non\responders that benefit from ICI immunotherapies. value and asterisks. Plots without asterisks indicate that there were no significant differences. Statistical significance was defined as p?0.05 (*), p?0.01 (**), and p?0.001 (***). Since the graphs show the results of pooled data, Kobe2602 different ICI treatments were color\coded: anti\CTLA\4 (Ipilimumab) in black, anti\PD\1 (Nivolumab, Pembrolizumab) in blue, and anti\CTLA\4/PD\1 combination therapy in red. The number of patients per column may differ because parameters (i.e., Ki\67) were analyzed toward the end of the study, patient samples were used up, or the complete blood count of patients was not available. 3.?RESULTS 3.1. Patient characteristics 45 patients with unresectable, late\stage, malignant melanoma were recruited, of which 60% were about to begin anti\CTLA\4, 24% anti\PD\1, and 16% anti\CTLA\4/PD\1 combination therapy (Table?1). The average study participant was male, older than 60?years, diagnosed with stage IV melanoma BRAF wild type (73% of patients), and had received no previous systemic treatments prior to the study. TABLE 1 Patient characteristics and treatments
Patients45100GenderMale2760Female1840Median age, years (range)70 (27C86)<65?years194265?years2658Melanoma stageUnresectable melanoma stage IIIC37Unresectable melanoma stage IV4293Treatments during the studyIpilimumab2760Ipilimumab/Nivolumab716Pembrolizumab920Nivolumab24Response to treatments during the studyTherapy success1329Complete response12Partial response920Stable disease37Therapy failure2964Unknown outcome37Treatments prior to the studySystemic treatmentChemotherapy12BRAF and MEK inhibitors920Checkpoint inhibitors37Number of systemic treatments032711920249RadiotherapyCerebral radiation37Peripheral radiation511Adjuvant immunotherapyAdjuvant interferon immunotherapy24Mutanome Engineered RNA Immunotherapy (MERIT)37Transarterial chemoembolization (TACE)12Electro cancer therapy (ECT)12 Open in a separate window NoteThe percentages refer to the total number of patients (n?=?45) and are rounded. Prior to the study, patients received up to two previous systemic treatments. Three patients had already been treated with different immune checkpoint Kobe2602 inhibitors: One patient received Nivolumab, the other Ipilimumab, and the third one received Ipilimumab followed later by Pembrolizumab. Abbreviations: BRAF, B\Raf proto\oncogene kinase; MEK, mitogen\activated protein kinase kinase. This study confirmed that about one in three patients responds to ICI therapy. Rabbit polyclonal to ITLN1 12 Most often, the response observed was a partial response. Only one patient responded completely. Taken together, patients with clinical benefit of ICI therapy (responders) lived significantly longer than non\responders (Physique?1A, p?=?0.0253?*). The median OS of non\responders was 336?days. The median OS of responders was undefined. The Kobe2602 1\year and 2\year OS of responders were about twice and three times higher than that of non\responders, respectively. After 684?days, 50% of all study participants had died or were lost to follow\up regardless of response to ICI therapy. In this study, immune\related adverse events (irAE) were not associated with ICI therapy response (Table?S1). 13 One third of patients developed irAE, most commonly colitis. 50% of these patients discontinued therapy. Ipilimumab displayed the highest irAE incidence and discontinuation percentage. Open in a separate window FIGURE 1 Immunomonitoring of lymphocytes, monocytes, platelets, and the melanoma biomarkers, S100 and LDH. (A) Overall survival of responders and non\responders to immune checkpoint inhibitor (ICI) therapy (Log\rank (Mantel\Cox) test, p?=?0.0253?*). (B) Lymphocytes. The bar diagrams depict the percentage of lymphocytes in viable cells. Lymphocytes were identified in FSC\A/SSC\A plots via flow cytometry. Doublets and dead cells were excluded from analysis. The dashed line marks the median non\responder value (two\tailed Mann\Whitney test, p?=?0.0486?*). (C) Platelets, platelet\to\lymphocyte ratio (PLR), and survival analysis. Complete blood count (CBC) provided lymphocyte and platelet count to calculate PLR. The dashed lines mark the normal range (two\tailed Mann\Whitney test, p(platelets)?=?0.0395?*, p(PLR)?=?0.0552; Log\rank (Mantel\Cox) test, survival analysis, p(platelets, nR >271 vs. nR <271)?=?0.0251?*, p(platelets, nR >271 vs. R)?=?0.0006?***, p(PLR, nR >289 vs. R)?=?0.0028?**, p(PLR, nR >289 vs. nR <289)?=?0.0353?*). (D) LDH, S100, and survival analysis. CBC provided data on LDH and S100 levels. The dashed lines mark the normal range (two\tailed Mann\Whitney test, p(LDH)?=?0.0569, p(S100)?=?0.2532); Log\rank (Mantel\Cox) test, survival analysis, p(LDH, nR >333 units/L vs. nR <333 units/L)?=?0.0063?**, p(LDH, nR >333 units/L vs. R)?=?0.0002?***; p(S100, nR >0.1?g/L vs. nR <0.1?g/L)?=?0.0188?*, p(S100, nR >0.1?g/L vs. R)?=?0.0004?***). Medians with interquartile range. CTLA\4, cytotoxic T\lymphocyteCassociated protein 4; ICI, immune checkpoint inhibitors; LDH, lactate dehydrogenase; nR, non\responder; PD\1, programmed cell death protein 1; R, responder; S100, S100 protein Altogether, these data highlight that ICI therapy response is the key factor for long\term OS. Since ICI target T cells, we hypothesized that immune cell composition in the peripheral blood of.