Herpes virus reactivation in brain-injured patients : The IBIS virus study
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1 Herpes virus reactivation in brain-injured patients : role and mechanisms of CD4 T cell defects. The IBIS virus study Antoine Roquilly University Hospital of Nantes, France UPRES 386 Medical University of Nantes, France
2 Déclaration d intérêts de 14 à 17 Intérêts financiers : néant Liens durables ou permanents : néant Interventions ponctuelles : néant Intérêts indirects : MSD
3 Critical-illness related immunosuppression / Virome Hotchkiss et al. Nature Immunol 13 De Vlaminck et al. Cell 13
4 The IBIS Virus project Herpes simplex Virus 49 patients 19 Traumatic brain injuries 1 SubArachnoid Hemorrhage 39 Stroke 77 (3%) serology negative 17 (71%) serology positive (3%) in-icu acquisition blood lungs 14 (6%) without reactivation NA blood NA lungs 68 (4%) with reactivation 13 blood 66 lungs 11 (14%) in-icu death 1 (16%) death (6 months) 18 (17%) in-icu death (19%) death (6 months) 17 (%) in-icu death (9%) death (6 months)
5 Risk of HSV and CMV reactivations in 49 brain-injured patients
6 Association of HSV reactivation with poor in-icu outcomes
7 Association of CMV reactivation with outcomes
8 HSV reactivations are associated with poor neurological outcome
9 Working hypothesis : BI-induced immunosuppression is responsible for Herpes Simplex Virus reactivation WP 1 : Increased viral replication or loss of immune control?
10 TT viral load (log/ml) Decreased circulating viral load of Torque Teno Virus in BI patients without HSV reactivation 3 1 volunteer HSV reactivation no HSV reactivation controls day 1 day 4 day 7 6 months
11 Working hypothesis : BI-induced T cell defects are responsible for Herpes Simplex Virus reactivation WP : Does the number of T cells matter?
12 Spleen volume (cm 3 ) CD4 T cells CD4 T cells Lymphopenia after brain injury IBIS - biocollection Essai clinique randomisé ** Modeles murins Sham day 1 day 7 TBI 4 *** * Sham Day 1 Day 8 Sham day 1 TBI day 7
13 No proof of apoptosis / necrosis of lymphocytes in Spleen Tissue after brain-injury cleaved-caspase-3 + cells per field (counts) Mode IBIS - biocollection Sham Brain injury human Activated Caspase control sham trauma Brain-dead patient white pulp red pulp Classical histology Activated Caspase 3
14 Lymphopenia is not associated with HSV reactivation, just a marker of severity? Lymphocytes (cells/mm 3 ) Lymphocytes (cells/mm 3 ) 1 1 p= No Yes HSV reactivation Survivors Non survivors
15 Working hypothesis : BI-induced T cell defects are responsible for the severity of HSV reactivation WP 3 : Do functions of T cells matter?
16 % of IFNg + EM Tcell % of IFNg + CM Tcell IFN-γ + CD8 T cells (%) IFN-γ + CD4 T cells (%) % of IFNg + Effe Tcell Defective production of IFN-g by memory CD4 T cells HD Day 1 Day 7 Month HD ** * J1 J7 M6 1 1 HD Day1 Day 7 Month 6 1 HD Day1 Day 7 Month * HD Day1 Day 7 Month 6
17 PD1 MFI Altered T cells are not exhausted CD4 CFSE en % Patient CD4 CFSE en % Patient Number of T cell division CD8 CFSE en % CD8 CFSE en % * * 1 * * Number of T cell division Patient Patient -4-6 IBIS d1 IBIS d4-7 CTL IBIS d1 IBIS d4-7 CTL IBIS d1 IBIS d4-7 CTL CD4 CD8 CD3
18 % of FoxP3+ CD4 % of RORgT+ CD4 % of T-bet + CD4 T cells % of GATA-3+ CD4 Brain injury alters the transcriptional factor program of CD4 T ** 1 ** * Day 1 Day 7 Month 6 Day 1 Day 7 Month Day 1 Day 7 Month 6 Day 1 Day 7 Month 6
19 TT viral load (log/ml) % IL-4 T CD4+ % IL- T CD4+ CD4 T cells re-programming is associated with immunosuppression and HSV reactivation R =.1, p=.8 Immune competence ** ** 4 6 %IL4 T cells IL4 re-programming No reactivation HSV reactivation Re-programming No reactivation HSV reactivation
20 Working hypothesis : BI-induced T cell defects are responsible for the severity of HSV reactivation WP 4 : Several mechanisms the CD4 T cells reprogramming?
21 CD4 T cells reprogramming and soluble circulating mediators? IBIS - biocollection Essai clinique randomisé IBIS - biocollection Essai clinique randomisé serum PBMCs Modeles murins serum Modeles murins
22 CD4 T cells reprogramming is continuously maintained by CD14+ cells, and other mediators are involved % of IFNg + CD4 Tcell % of IFNg + CD4 Tcell % of IL4+ CD4 % of IL4+ CD4 Tcell IFN-γ+ (% of CD4 T cells) IFN-γ+ (% of CD4 T cells) IBIS - biocollection Essai clinique randomisé PBMCs ** PBMCs Modeles murins CD4 T cells + CD14 cells 1 1 NT CD4 + PBMC ** CD4 CD NT CD4 + PBMC *** CD4 CD4 1 PBMC CD4 CD4+CD14 PBMC - CD PBMC CD4 CD4+cDC PBMC - cdc
23 %IFN-γ+ CD4 T cells IL4+ CD4 T cells (%) CD4 T cells reprogramming is partially IL-4 dependent, but not TGF-b dependent IBIS - biocollection Essai clinique randomisé Blocking antibody Y Y Modeles murins 3 1 ** Brain injury PBMCs Vehicle Anti IL-4 Vehicle anti-il4 isotypique Ctrl Anti TGFb Brain injury Vehicle anti-il4 isotypique Ctrl Anti TGFb
24 Conclusions HSV reactivations are common in brain injured patients, associated with poor outcome Long-lasting lymphopenia No apoptosis (caspase ko mice?) Not associated with HSV reactivation Long lasting TF reprogramming of CD4 T cells after BI No exhaustion Altered TF reprogramming of CD4 T cells is associated with HSV reactivation Microenvironment : CD14, IL-4. Other mechanisms under the scope : epigenetic modifications? CD14 cells? Role of direct activation by brain-released DAMPs?
25 Acknowledgments C. Bressollette (UMR 164) M. Burelle A. Irimia C. Bossard (URM 187) T. Woodberry (Darwin Uni) Pr Asehnoune lab Tanguy Roman (PhD) Barbara Misme-Aucouturier (PhD) Cédric Jacqueline (PhD) Alexis Broquet (PhD) Intensive Care Unit - CHU Nantes Raphael Cinotti (MD) Mickael Vourc h (MD) Delphine Flattres (TEC)
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