Obtaining Samples from Human Subjects to Facilitate Basic, Translational and Clinical Research

Clinical Center

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Abstract

This protocol defines in general terms the purposes for which blood, urine and ultrafiltrate will be collected from adult research subjects and healthy volunteers by members of the NIH Clinical Center's Critical Care Medicine Department and collaborating institutions and establishes general conditions under which sampling will be performed. Blood drawing will be consistent with NIH Clinical Center guidelines. The Critical Care Medicine Department and our associated investigators are involved in the development of new translational research protocols and maintenance of existing IRB approved research protocols. This protocol will support the establishment of new translational research protocols through assay development and provide information to guide the effective implementation of new protocols as well as maintain efficiency in operations of existing protocols. The protocol was approved by the National Heart, Lung, and Blood Institutional Review Board (subsequently changed to General Medicine 1 IRB) in August 2017. Over the course of the protocol, research subjects have provided blood samples for inflammatory marker studies, to characterize low density granulocytes as a novel therapeutic target for patients with sepsis, DNA gene sequencing for Pulmonary Arterial Hypertension (PAH), immunophenotyping for relapsing polychondritis, isolation of B cells for flow cytometry, cell free DNA isolation for PAH, thromboelastography (TEG) and other blood markers of coagulation and fibrinolysis, RNA isolation for sequencing, vitamin C levels, PBMCs isolated from whole blood for reprogramming and generation of induced pluripotent stem cell (iPSC) lines, NK cells isolation to study their cell physiology and function in the context of COVID-19, study the interaction between immune cells and Ebola virus proteins, neutrophil isolation to detect SYK using flow cytometry and western blot, analyzing Neutrophil Extracellular Traps (NETosis) and the role of an enzyme, Spleen Tyrosine Kinase, in this inflammatory cascade, and characterizing neutrophil bioenergetics, and to isolate cells and plasma to either 1) serve as a comparator for COVID-19 patients or 2) perform in-vitro stimulations to study several COVID-19 molecular mechanisms. In addition, recently samples from the protocol have been used to evaluate SARS-CoV-2 Immune Complex Mediated Neutrophil Activation, normal density neutrophil isolation, comparison studies of venous blood vs dried blood spots as relates to Liquid Chromatography/Mass Spectrometry proteomic and metabolomic studies, stimulation of neutrophils and endothelial cells, the novel role of Semaphorin 5A, a neuroimmune protein, on neutrophil function, generation of extracellular vesicles, protocol validation for phagocytoses, and standardizing IncuCyte NETosis assay. During the current reporting period 4 subjects were newly enrolled (4 healthy volunteers and 0 patients). Over the life of the protocol (as of September 2025) a total of 125 subjects have enrolled (76 healthy volunteers and 49 patients). Over the current reporting period 70 blood samples were provided to investigators and over the life of the protocol 901 samples have been provided to investigators. The following abstract have been presented using samples obtained from this protocol. Brusca SB, Elinoff JM, Jang MK, Demirkale CY, Valantine HA, Solomon MA, and Agbor-Enoh ST. Plasma Cell-free DNA as a Novel Marker of Disease Severity in Pulmonary Arterial Hypertension. American College of Cardiology 68thAnnual Scientific Session, J Am Coll Cardiol, 73(9, S1): S1897, 2019. Lu M, Blaine K, Cullinane A, Hall C, Dulau-Florea ,A, Sun J, Chenwi H, Graninger G, Harper B, Elinoff JM, and Solomon MA. Pulmonary Arterial Hypertension Patients Display Normal Kinetics of Clot Formation. American Heart Association Scientific Sessions, 140: A10714, 2019 Becicka W, Garrad E, Berreby G, Wang J, Nghiem K, Ramos-Benitez M, Cowling B, Moitra J, Huffstutler R, Carney K, Ferrante E, Cudrici C, Brofferio A, Tourdot B, Jacobson K, Knight J, Kanthi Y, and Boehm M. Genetic Deficiency of the Ectoenzyme CD73 Increases Neutrophil Extracellular Trap Formation in Patients with ACDC. Arteriosclerosis, Thrombosis, and Vascular Biology: Vascular Discovery Conference 2022 Becicka W, Berreby G, Wang J, Moitra J, Panicker S, Tourdot BE, Norris P, Kelly K, Zuo Y, Dumont L, Solomon MA, Knight JS and Kanthi Y. Antiphospholipid antibodies in COVID-19 Convalescent Plasma. Arteriosclerosis, Thrombosis, and Vascular Biology: AHA Vascular Discovery Conference 2022 Strich J, Chertow D, Ramos-Benitez, Marcos, and Warner S. SYK Inhibition Abrogates Neutrophil Hyperactivation in Response to Lipopolysaccharides. Critical Care Medicine, 51(1): 562, 2023. The following papers have been published using samples obtained from this protocol. 1. Lu M, Blaine KP, Cuillinane A, Hall C, Dulau-Florea A, Sun J, Graninger GM, Harper BJ, Brusca SB, Elinoff JM, and Solomon MA. Pulmonary Arterial Hypertension Patients Display Normal Kinetics of Clot Formation. Pulm Circ, 11(3): 1-9, 2021 2. Strich RJ, Ramos-Benitez MJ, Randazzo D, Stein SR, Babyak A, Davey RT, Suffredini AF, Childs RW, Chertow DS. Fostamatinib Inhibits Neutrophils Extracellular Traps Induced by COVID-19 Patient Plasma: A Potential Therapeutic. The Journal of Infectious Diseases, 223 (6): 981984, 2021. 3. Perez-Valencia LJ, Vannella KM, Ramos-Benitez MJ, Sun J, Abu-Asab M, Dorward DW, Awad KS, Platt A, Jacobson E, Kindrachuk J, Chertow DS. Ebola virus shed glycoprotein is toxic to human T, B, and natural killer lymphocytes. iScience 26 (8): 107323. https://doi.org/10.1016/j.isci.2023.107323, 2023. 4. Warner S, Teague HL, Ramos-Benitez MJ, Panicker S, Allen K, Gaihre S, Moyer T, Parachalil Gopalan B, Douagi I, Shet A, Kanthi Y, Suffredini AF, Chertow DS, Strich JR. R406 reduces lipopolysaccharide-induced neutrophil activation. Cell Immunol, DOI: 10.1016/j.cellimm.2024.104860, 2024. 5. Allen KC, Warner S, Teague HL, Ramos-Benitez MJ, Miao R, Tian X, Reger R, Burbelo PD, Pang CWJ, Kanthi Y, Cohen JI, Gopalan BP, Suffredini AF, King C, Nathan SD, Childs RW, Chertow DS, Strich JR. SARS-CoV-2 Immune Complex-Mediated Neutrophil Activation. Open Forum Infect Dis, 12(4) DOI: 10.1093/ofid/ofaf199, 2025.