Gating strategy for CD4+ T cell subsets - Investigation of systemic inflammation in aneurysmal

10. Appendices

10.6. Gating strategy for CD4+ T cell subsets

Gating strategy was similar to monocytes until the identification of CD45+ leucocytes.

Lymphocytes were selected from these CD45+ events by their low side scatter and high CD45 expression (1). These lymphocytes were then displayed as CD3 vs CD4 plot and a quadrant gate was applied (2). Then, from this quadrant gate CD3+ CD4+ events were selected as CD4+ T cells. CD4+ T cells were then displayed on a CD25 vs CD127 plot and CD25hiCD127lo events were gated as Tregs and this gating strategy was based on CD25 FMO control (3). Then, by applying a NOT gating function, events other than Tregs were selected and were named as Th cells. These Th cells were further displayed on a bivariate plot of CXCR3 vs CCR6 (4). Then, a quadrant gate was used based on FMO controls for CXCR3 and CCR6 to identify CXCR3+ CCR6- cells as Th1, CXCR3- CCR6- cells as Th2 and CXCR3- CCR6+ Th17 cells. Then, HLA-DR+ and CD38+ cells were identified among these subsets such as Tregs (5), Th1, Th2, and Th17 cells (6) by using FMO controls and applying a quadrant gate.

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List of Publications underlying thesis

Chaudhry, SR., Güresir, E., Vatter, H., Kinfe, TM., Dietrich, D., Lamprecht, A., Muhammad, S. Aneurysmal Subarachnoid Hemorrhage lead to systemic upregulation of IL-23/IL-17 inflammatory axis. Cytokine, 2017, 97: 96 – 103.

Chaudhry, SR., Stoffel-Wagner, B., Kinfe, TM., Güresir, E., Vatter, H., Dietrich, D., Lamprecht, A., Muhammad, S. Elevated systemic IL-6 levels in patients with aneurysmal subarachnoid hemorrhage is an unspecific marker for post-SAH complications. International Journal of Molecular Sciences, 2017, 18: 2580, doi: 10.3390/ijms18122580.

Muhammad, S*., Chaudhry, SR*., Güresir, A., Stoffel-Wagner, B., Fimmers, R., Kinfe, TM., Dietrich, D., Lamprecht, A., Vatter, H., Güresir, E. Systemic HMGB1, a novel predictive biomarker for cerebral vasospasm in Subarachnoid Hemorrhage. Critical Care Medicine (Submitted). *Equally contributed.

Kinfe, TM., Muhammad, S., Link, C., Roeske, S., Chaudhry, SR., Yearwood, TL. Burst Spinal Cord stimulation increases peripheral antineuroinflammatory Interleukin 10 levels in Failed Back Surgery Syndrome patients with predominant back pain. Neuromodulation, 2017, DOI: 10.1111/ner.12586.

Muhammad, S., Chaudhry, SR., Yearwood, T., Krauss, J., Kinfe, TM. Changes of metabolic disorders associated peripheral cytokine/adipokine traffic in non-obese chronic back patients responsive to burst spinal cord stimulation. Neuromodulation, 2017, DOI:

10.1111/ner.12708.

Chaudhry, SR., Dietrich, D., Vatter, H., Lamprecht, A., Muhammad, S. A review on the role of danger associated molecular pattern molecules (DAMPs) in aSAH. (In preparation).

Chaudhry, S., Frede, S., Seifert, G., Vatter, H., Dietrich, D., Lamprecht, A., Muhammad, S.

Temporal profile of serum mitochondrial DNA (mtDNA) in patients with aneurysmal Subarachnoid Hemorrhage (aSAH). (In preparation).

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Curriculum Vitae (CV) Shafqat Rasul Chaudhry

08.10.1985, Mianwali, Pakistan Education

HEC-DAAD Scholarship for Ph.D students: April 2013 – March 2017, Department of Neurosurgery, University Hospital Bonn, University of Bonn, Bonn, Germany

M.Phil (Pharmacology): Session 2010 – 12, The Islamia University of Bahawalpur, Pakistan

Research work title: “Investigation of Indigenous Medicinal Plants for their Antidiabetic and Antihyperlipidemic Activities”. 1st Division, CGPA 3.71/4.00.

Pharm.D: Session 2004 – 2009, 1st Division 2009, The Islamia University of Bahawalpur, Pakistan

HSSC/Intermediate (F.Sc Premedical): 1st Division 2004 (FBISE Islamabad, Pakistan) SSC/Matriculation: 1st Division 2002 (BISE Sargodha, Pakistan)

Work Experience

-Worked as visiting faculty part time lecturer in Pharmacology at Faculty of Pharmacy &

Alternative Medicine, The Islamia University of Bahawalpur, Pakistan. (From 10th January, 2012 to 13th June, 2012)

-Worked as a scientific assistant at The University of Applied Sciences Bonn-Rhein-Sieg, Rheinbach. (From 04th December, 2013 to 28th February, 2014)

-Worked as scientific assistant at the Dept. of Neurosurgery, University Hospital Bonn.

(From 20th June, 2017 to 31st August, 2017 and 1st November to 31st December) List of publications

Review: Chaudhry, SR., Muhammad, S., Eidens, M., Klemm, M., Khan, D., Efferth, T., Weisshaar, MP. Pharmacogenetic prediction of individual variability in drug response based on CYP2D6, CYP2C9 and CYP2C19 genetic polymorphisms. Current Drug Metabolism, 2014, 15 (7): 711-718.

Chaudhry, SR., Akram, A., Aslam, N., Asif, M., Wajid, M., Kinfe, TM., Jabeen, Q., Muhammad, S. Antidiabetic and antidyslipidemic effects of Heliotropium strigosum in rat models of type I and type II diabetes. Acta Poloniae Pharmaceutica – Drug Research, 2016, 73 (6): 1575 – 1586.

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Muhammad, S., Roeske, S., Chaudhry, SR., Kinfe, TM. Burst or high frequency (10 kHz) spinal cord stimulation in failed back surgery syndrome patients with predominant back pain:

one year comparative data. Neuromodulation, 2017, DOI:10.1111/ner.12611.

Chaudhry, SR., Akram, A., Aslam, N., Wajid, M., Iqbal, Z., Nazir, I., Jabeen, Q., Muhammad, S. Antidiabetic and antidyslipidemic potential of Echinops echinatus Roxb. in rat models of type I and type II diabetes. Pakistan Journal of Pharmaceutical Sciences, 2018 (accepted).

10.1111/ner.12708.

Boström, A., Scheele, D., Roeske, S., Stoffel-Wagner, B., Hoenig, F., Muhammad, S., Chaudhry, SR., Hurlemann, R., Krauss, J., Kinfe, TM. Inter-ictal saliva oxytocin and interleukin IL-1β signaling in therapy-resistant migraine patients responsive to adjunctive cervical non-invasive vagus nerve stimulation: The MOXY feasibility study.

Neuromodulation (submitted).

Chaudhry, SR., Dietrich, D., Vatter, H., Lamprecht, A., Muhammad, S. A review on the role of danger associated molecular pattern molecules (DAMPs) in aSAH. (In preparation).

Chaudhry, S., Frede, S., Seifert, G., Vatter, H., Dietrich, D., Lamprecht, A., Muhammad, S.

Temporal profile of serum mitochondrial DNA (mtDNA) in patients with aneurysmal Subarachnoid Hemorrhage (aSAH). (In preparation)

169 Posters

Shafqat Rasul Chaudhry, Stilla Frede, Gerald Seifert, Rolf Fimmers, Dirk Dietrich, Alf Lamprecht, Erdem Güresir, Hartmut Vatter and Sajjad Muhammad. Temporal profile of mitochondrial DNA (mtDNA) in the serum of patients with aneurysmal Subarachnoid Hemorrhage (aSAH). 5^th Venusberg meeting on Neuroinflammation, Bonn, Germany, 2017.

Shafqat Rasul Chaudhry, Erdem Güresir, Dirk Dietrich, Alf Lamprecht, Hartmut Vatter and Sajjad Muhammad. Aneurysmal Subarachnoid Hemorrhage leads to elevated systemic levels of proinflammatory cytokine IL-23. 68^th Annual Meeting of the German Society of Neurosurgery; 7^th Joint Meeting with the Society of British Neurological Surgeons, Magdeburg, Germany, 2017.

Sajjad Muhammad, Shafqat Rasul Chaudhry, Jasmin Scorzin, Birgit Stoffel Wagner, Rolf Fimmers, Hartmut Vatter, Erdem Güresir. Danger Signal Molecule HMGB1 is differentially regulated in patients with Aneurysmal Subarachnoid Hemorrhage developing Cerebral Vasospasm. 7th Int. Symposium on DAMPS and HMGB1, Bonn, Germany, 2015.

Maria Weisshaar, Eslam Shehata, Shadi Amin, Stephan Irsen, Julio Reinecke, Peter Wehling and Shafqat Rasul Chaudhry. Comparison of the traditional ultracentrifugation techniques and PEG precipitation for isolating exosomes from ACS (autologous conditioned serum): a kinetic study. 4^th Int. Meeting of ISEV, Washington, USA, 2015.

Conferences:

5th Venusberg Meeting on “Neuroinflammation”. Privileged or not: basic, translational and clinical aspects of immune activation in the brain. Bonn, 11 – 13 May, 2017.

4th Venusberg Meeting on “Neuroinflammation”: Privileged or not: basic, translational and clinical aspects of immune activation in the brain. Bonn, 7– 9 May, 2015.

Im Dokument Investigation of systemic inflammation in aneurysmal subarachnoid hemorrhage (aSAH) and its impact on post-aSAH complications (Seite 176-181)