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. 2018 Jun 18;9:1355. doi: 10.3389/fimmu.2018.01355

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Copyright © 2018 Ozkaynak, Gilman, London, Naranjo, Diccianni, Tenney, Smith, Messer, Seeger, Reynolds, Smith, Shulkin, Parisi, Maris, Park, Sondel and Yu.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Cytokine and nitrate levels during course 1 and 4 of immunotherapy: (A) changes in serum cytokine and nitrate levels during Immunotherapy. (B) Relationship between allergic reaction of any grade and serum cytokine levels on immunotherapy. Serum samples were collected before starting immunotherapy and just before the fourth doses of ch14.18 infusions on courses 1 (days 1 and 6) and 4 (days 80 and 90), and analyzed as described for cytokine and nitrate levels. Box represents 25–75% distribution, error bars represent SD, solid line is the median, dashed line is the mean. Boxes and values shown include any not-shown outliers unless otherwise mentioned. (A) Comparisons are by Wilcoxon signed-rank test of paired samples: day −1 versus day 6 and day 80 versus day 90. A p-value of < 0.0036 is considered statistically significant at level 0.05 and are Bonferroni corrected for the 14 cytokine comparisons at each course, or p < 0.05 for nitrate. Values of outliers excluded from the graph are: d-1: IL-6 (447.4 and 416.7 pg/ml), tumor necrosis factor alpha (268.5 pg/ml), IL-4 (0.78 pg/ml), IL-12p70 (5.48 pg/ml), IL-17A (22.4 pg/ml), IL-8 (31,818 pg/ml and also excluded from the mean, median and statistical determinations); d6: IL-6 (618 pg/ml), IL1-Ra (11,489.1 pg/ml), CCL22 (39,956.7 pg/ml), IL-15 (19.2 pg/ml); d80: IL-6 (862.0 pg/ml), IL-8 (2,185.5 pg/ml), IL-15 (33.7 pg/ml and 17.1 pg/ml); d90: IL-6 (187.1 pg/ml), IL1-Ra (13,329.9 pg/ml), IFNγ (9,591.9 pg/ml), IL-10 (193.7 pg/ml), IL-4 (0.47 pg/ml), IL-5 (7,500.9 pg/ml), IL-17A (20.7 pg/ml), CXCL9 (24,172.7 and 19,116.1 pg/ml), IL-15 (33.4 pg/ml). (B) Two-sided Mann–Whitney U-test comparison of no AR versus any AR; values are not adjusted for multiple comparisons. Values of outliers excluded from the graph are: d6, no AR: IL1-Ra (11,489.1 pg/ml), CCL22 (39,956.7 pg/ml), IL-15 (19.2 pg/ml); d6 AR: IL-6 (618 pg/ml); d90, no AR: IL1-Ra (13,329.9 pg/ml), IFNγ (9,591.9 pg/ml), IL-4 (0.47 pg/ml), IL-5 (7,500.9 pg/ml), IL-15 (33.4 pg/ml); d90 AR: IL-10 (193.7 pg/ml), IL-17A (20.7 pg/ml), CXCL9 (24,172.7 and 19,116.1 pg/ml). Notes: IL-1Ra: interleukin-1 receptor antagonist; many CCL22 values were out of the range of the multiplex assay and were extrapolated. Thus, the CCL22 data and analyses should be considered qualitative.