Christopher Stephen Inchley, 2015

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2 Christopher Stephen Inchley, 2015 Series of dissertations submitted to the Faculty of Medicine, University of Oslo No ISBN All rights reserved. No part of this publication may be reproduced or transmitted, in any form or by any means, without permission. Cover: Hanne Baadsgaard Utigard. Printed in Norway: AIT Oslo AS. Produced in co-operation with Akademika Publishing. The thesis is produced by Akademika Publishing merely in connection with the thesis defence. Kindly direct all inquiries regarding the thesis to the copyright holder or the unit which grants the doctorate.

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52 Cord blood from 2108 newborns 70 Confirmed RSV disease 6 excluded Asthma Cleft palate 64 Included Down syndrome Recurrent Pneumonia 47 Sufficient RNA 25 severe disease 22 mild disease 37 age < 12months 10 age > 12 months

55 139 with suspected viral respiratory infection 120 RSV positive 110 Included 105 RNA samples 40 mild 25 moderate 40 severe 24 mild 11 moderate 25 severe 16 mild 7 moderate 19 severe 19 RSV negative 10 excluded: 3 Co-infections 4 Premature 1 Previous bronchial obstruction 1 Previous lower respiratory tract infection 1 Chronic liver disease 80 control infants 5 Only nasal cytology smear taken 38 individual samples 21 paired samples Failed post-rna isolation quality control 59 control samples 45 RSV positive samples 31 control samples Failed post-pcr quality control 28 control samples 18 RSV positive samples 9 control samples 19 control samples Inclusion, clinical data and sample collection RNA isolation qpcr Statistical Analysis

66 mir-23b mir-27b 2 2 Fold Change (Log 2) Fold Change (Log 2) /230 ratio /230 ratio Control p = 0.3 RSV positive p < Control RSV positive p = 0.4 p < Fold Change (Log 2) Fold Change (Log 2) /230 ratio Control 0.6 p = 0.6 RSV positive 0.6 p = /230 ratio Control 0.6 p = 0.9 RSV positive 0.6 p = 0.7

67 mir-130a let-7d 4 4 Fold Change (Log 2) Fold Change (Log 2) /230 ratio /230 ratio Control RSV positive p = 0.1 p < Control RSV positive p = 0.2 p < Fold Change (Log 2) Fold Change (Log 2) /230 ratio Control 0.6 p = 0.9 RSV positive 0.6 p = /230 ratio Control 0.6 p = 0.8 RSV positive 0.6 p = 0.6

72 let-7d mir-30b mir-34c Control RSV positive p=0.6 p= Control RSV positive p=0.3 p= RIN RIN mir-125b Control RSV positive p=0.5 p= RIN Fold Change (Log 2) Fold Change (Log 2) Fold Change (Log 2) RIN mir-200b Control RSV positive p=0.5 p=0.4 Fold Change (Log 2) Fold Change (Log 2) mir RIN RIN Control RSV positive Control RSV positive p=0.5 p=0.7 p=0.9 p=0.6 Fold Change (Log 2)

75 AgiMiRNA AgiMiRNA

83 A B Change in light intensity 10 4 Reference gene Cq ΔCq Target gene Cq Target threshold 10 1 Reference threshold PCR Cycle Target gene Non-specific Logarithmic Plateau Phase: Reference gene Target gene

90 12 Age (months) Control RSV positive Mild Moderate Severe

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112 mirna Expression in this study Functions in the immune system Both innate and adaptive immune functions mir-29c Downregulation Positive regulation of NFκB signaling [198] InfluenzaUpregulation. Promotes apoptosis mir-31 Upregulation Proliferation of myeloid cells [78] mir-155 Upregulation Proliferation of myeloid cells [78] Dendritic cell maturation [77, 201] Positive regulation of TLR signaling [202] Required for dendritic, B and T-cell function [204] let-7d Upregulation Promotes Th1 polarization [203] EndotoxinUpregulation (TLR4 dep.) [191] Specific targets TNFAIP3 [198] BCL2L2 [199] E-selectin [200] SHIP1 [78] PU.1 [201], SOCS1, CD115, KPC1 [77] SOCS1 [202] IL13 [203] Disease associations (Human studies) Tuberculosis upregulated [195] Asthma downregulated [196] Influenza downregulated [104] Influenza downregulated [104] Asthma downregulated [197]

113 mirna Expression in this study Primarily innate immune functions mir-16 Upregulation mir-27b Downregulation mir-34b Downregulation mir-34c Downregulation Functions in the immune system Negative regulation of inflammation [206] Positive regulation of NFκB signaling [207] Negative regulation of NFκB signaling [192] EndotoxinUpregulation (TLR4 dep) [191] TLR4/NFκB induced nitric oxide production [208] EndotoxinUpregulation (NFκB dep) [209] EndotoxinUpregulation (TLR4 dep) [191] Negative regulation of NFκB signaling; DAMPs*Upregulation [189] In cord blood monocytes: IFNγUpregulation ; Endotoxinno regulation [190] Specific targets TNFα [206] SMRT [207] IKKα [192] KSRP [208] IKKγ [189] Disease associations (Human studies) Sepsis improved survival [205] Asthma downregulated [196] Asthma downregulated [196] Asthma downregulated [196]

114 mirna Expression in this study Primarily innate immune functions mir-125a Downregulation mir-125b Downregulation mir-203a Upregulation Functions in the immune system Positive regulation of NFκB signaling [212] Inhibits CCL5 production [213] Macrophage polarization [220, 221] Positive regulation of NFκB signaling [212] Negative regulation of NFκB signaling [202] Activates Macrophages [214] Modulates dendritic cell differentiation [215] Maintains naïve state in CD4+ T-cells [216] Negative regulation of inflammation [217] Negative regulation of NFκB signaling [218] Negative regulation of inflammation [219] Specific targets TNFAIP3 [212] KLF13 [213] TNFAIP3 [212] MyD88 [202] IRF4 [214] PRDM1 [215] IFNγ, IL2RB, IL10RA, PRDM1 [216] TNFα [217] Myd88 [218] TNFα, IL24 [219] Disease associations (Human studies) CERS** upregulated [210] Asthma upregulated [211]

115 mirna Expression in this study Functions in the immune system Specific targets Disease associations (Human studies) No known immune functions mir-429 Downregulation :

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Copyright is owned by the Author of the thesis. Permission is given for a copy to be downloaded by an individual for the purpose of research and

Copyright is owned by the Author of the thesis. Permission is given for a copy to be downloaded by an individual for the purpose of research and private study only. The thesis may not be reproduced elsewhere