Immune Cells Phenotypes During COVID-19 (IMMUNO-COVID)

Alterations of Innate and Adaptive Immune Cells During the Course of SARS CoV-2 Pneumonia

The ongoing pandemic caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS CoV-2) has infected more than one hundred twenty million peoples worldwide one year after its onset with a case-fatality rate of almost 2%. The disease due to the coronavirus 2019 (i.e., COVID-19) is associated with a wide range of clinical symptoms. As the primary site of viral invasion is the upper respiratory airways, lung infection is the most common complication. Most infected patients are asymptomatic or experience mild or moderate form of the disease (80 %). A lower proportion (15%) develop severe pneumonia with variable level of hypoxia that may required hospitalization for oxygen therapy. In the most severe cases (5%), patients evolve towards critical illness with organ failure such as the acute respiratory distress syndrome (ARDS). At this stage, invasive mechanical ventilation is required in almost 70 % and the hospital mortality rises to 37 %.

Immune cells are key players during SARS CoV-2 infection and several alterations have been reported including lymphocytes (T, B and NK) and monocytes depletion, and cells exhaustion. Such alterations were much more pronounced in patients with the most severe form of the disease. Beside, a dysregulated proinflammatory response has also been pointed out as a potential mechanism of lung damage. Finally, COVID-19 is associated with an unexpectedly high incidence of thrombosis which probably results from the viral invasion of endothelial cells.

The investigators aim to explore prospectively the alterations of innate and adaptive immune cells during both the acute and the recovery phase of SARS CoV-2 pneumonia. Flow and Spectral cytometry will be used to perform deep subset profiling focusing on T, B, NK, NKT, gamma-gelta T, monocytes and dendritic cells. Each specific cell type will be further characterized using markers of activation/inhibition, maturation/differenciation and senescence as well as chemokines receptors.

T-cell memory specificity will be explore using specific SARS CoV-2 pentamer. Platelet activation and circulating microparticles will be explore using flow cytometry. Serum SARS CoV-2 antibodies (IgA, IgM, IgG), serum cytokines, and serum biomarkers of alveolar epithelial and endothelial cells will be analyze using ELISA and correlate with the severity of the disease.

Study Overview

• Status: Unknown
• Conditions: Endothelial Dysfunction; Sars-CoV2; Immunization; Infection; Innate Immunity; Alveolar Lung Disease
• Intervention / Treatment: Biological: Peripheral blood samples

• Study Type: Observational
• Enrollment (Anticipated): 100

Contacts and Locations

• Study Contact: Name: Line MEDDEB; Phone Number: 33 0413732347; Email: line.meddeb@ap-hm.fr
• Study Contact Backup: Name: Joana VITTE, MD, PhD; Email: joana.vitte@ap-hm.fr

Study Locations

• France
  • Marseille, France, 13003
    • Recruiting
    • Hôpital Européen Marseille
    • Contact: Jérôme ALLARDET-SERVENT, MD; Phone Number: 33 0413427450; Email: j.allardetservent@hopital-europeen.fr
    • Contact: Philippe HALFON, MD, PhD; Phone Number: 33 0413428120; Email: philippe.halfon@alphabio.fr
  • Marseille, France, 13015
    • Recruiting
    • Hopital Nord
    • Contact: Marc LEONE, MD, PhD; Email: m.leone@aph-hm.fr
    • Sub-Investigator: Amélie MENARD, MD

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years and older (Adult, Older Adult)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: All

• Sampling Method: Non-Probability Sample

Study Population

Patients presenting with a first episode of SARS CoV-2 pneumonia and requiring hospitalization either in a ward or an Intensive Care Unit will constitute the COVID-19 group.

Healthy blood donors from the Etablissement Français du Sang (EFS) will constituted the control group

Description

Inclusion Criteria:

• Age > 18 y
• Laboratory confirmed SARS CoV-2 infection (positive RT-PCR).
• Ground-glass opacity on chest computed-tomography
• Time from hospital admission to inclusion < or equal to 72 h

Exclusion Criteria:

• Pregnant
• Under legal restriction

Study Plan

How is the study designed?

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Profiling of innate and adaptive immune cells during SARS CoV-2 infection.	Determination of cells population using spectral cytometry of PBMCs.	Day 0
Profiling of innate and adaptive immune cells during SARS CoV-2 infection.	Determination of cells population using spectral cytometry of PBMCs.	Day 7
Profiling of innate and adaptive immune cells during SARS CoV-2 infection.	Determination of cells population using spectral cytometry of PBMCs.	Day 14
Profiling of innate and adaptive immune cells during SARS CoV-2 infection.	Determination of cells population using spectral cytometry of PBMCs.	Day 28
Profiling of innate and adaptive immune cells during SARS CoV-2 infection.	Determination of cells population using spectral cytometry of PBMCs.	Day 90
Profiling of innate and adaptive immune cells during SARS CoV-2 infection.	Determination of cells population using spectral cytometry of PBMCs.	Day 180
Functional state of innate and adaptive immune cells during SARS CoV-2 infection.	Determination of the functional state of immune cells using spectral cytometry	Day 0
Functional state of innate and adaptive immune cells during SARS CoV-2 infection.	Determination of the functional state of immune cells using spectral cytometry	Day 7
Functional state of innate and adaptive immune cells during SARS CoV-2 infection.	Determination of the functional state of immune cells using spectral cytometry	Day 14
Functional state of innate and adaptive immune cells during SARS CoV-2 infection.	Determination of the functional state of immune cells using spectral cytometry	Day 28
Functional state of innate and adaptive immune cells during SARS CoV-2 infection.	Determination of the functional state of immune cells using spectral cytometry	Day 90
Functional state of innate and adaptive immune cells during SARS CoV-2 infection.	Determination of the functional state of immune cells using spectral cytometry	Day 180
Serum IgA, IgM and IgG antibodies during SARS CoV-2 infection.	Measurement of serum SARS CoV-2 IgA, IgM and IgG antibodies using Elisa.	Day 0
Serum IgA, IgM and IgG antibodies during SARS CoV-2 infection.	Measurement of serum SARS CoV-2 IgA, IgM and IgG antibodies using Elisa.	Day 7
Serum IgA, IgM and IgG antibodies during SARS CoV-2 infection.	Measurement of serum SARS CoV-2 IgA, IgM and IgG antibodies using Elisa.	Day 14
Serum IgA, IgM and IgG antibodies during SARS CoV-2 infection.	Measurement of serum SARS CoV-2 IgA, IgM and IgG antibodies using Elisa.	Day 28
Serum IgA, IgM and IgG antibodies during SARS CoV-2 infection.	Measurement of serum SARS CoV-2 IgA, IgM and IgG antibodies using Elisa.	Day 90
Serum IgA, IgM and IgG antibodies during SARS CoV-2 infection.	Measurement of serum SARS CoV-2 IgA, IgM and IgG antibodies using Elisa.	Day 180
Platelet activation and circulating microparticles assessment during SARS CoV-2 infection.	Determination of platelet activation and circulating microparticles levels using flow cytometry.	Day 0
Platelet activation and circulating microparticles assessment during SARS CoV-2 infection.	Determination of platelet activation and circulating microparticles levels using flow cytometry.	Day 7
Platelet activation and circulating microparticles assessment during SARS CoV-2 infection.	Determination of platelet activation and circulating microparticles levels using flow cytometry.	Day 14
Platelet activation and circulating microparticles assessment during SARS CoV-2 infection.	Determination of platelet activation and circulating microparticles levels using flow cytometry.	Day 28

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Serum concentration of Pro-inflammatory and Anti-inflammatory cytokines in response to SARS CoV-2 infection.	Measurement of IL1β, IL-6, IL-10, IL-17A, IL-18, TNFα, IFNγ, CRTP-6 using Elisa.	Day 0
Serum concentration of Pro-inflammatory and Anti-inflammatory cytokines in response to SARS CoV-2 infection.	Measurement of IL1β, IL-6, IL-10, IL-17A, IL-18, TNFα, IFNγ, CRTP-6 using Elisa.	Day 7
Serum concentration of Pro-inflammatory and Anti-inflammatory cytokines in response to SARS CoV-2 infection.	Measurement of IL1β, IL-6, IL-10, IL-17A, IL-18, TNFα, IFNγ, CRTP-6 using Elisa.	Day 14
Serum concentration of Pro-inflammatory and Anti-inflammatory cytokines in response to SARS CoV-2 infection.	Measurement of IL1β, IL-6, IL-10, IL-17A, IL-18, TNFα, IFNγ, CRTP-6 using Elisa.	Day 28
Serum concentration of Pro-inflammatory and Anti-inflammatory cytokines in response to SARS CoV-2 infection.	Measurement of IL1β, IL-6, IL-10, IL-17A, IL-18, TNFα, IFNγ, CRTP-6 using Elisa.	Day 90
Serum concentration of Pro-inflammatory and Anti-inflammatory cytokines in response to SARS CoV-2 infection.	Measurement of IL1β, IL-6, IL-10, IL-17A, IL-18, TNFα, IFNγ, CRTP-6 using Elisa.	Day 180
Serum alveolar epithelial and endothelial cells biomarkers during SARS CoV-2 infection.	Measurement of KL-6, CC-16, S-RAGE, ANG-2 using ELISA.	Day 0
Serum alveolar epithelial and endothelial cells biomarkers during SARS CoV-2 infection.	Measurement of KL-6, CC-16, S-RAGE, ANG-2 using ELISA.	Day 7
Serum alveolar epithelial and endothelial cells biomarkers during SARS CoV-2 infection.	Measurement of KL-6, CC-16, S-RAGE, ANG-2 using ELISA.	Day 14
Serum alveolar epithelial and endothelial cells biomarkers during SARS CoV-2 infection.	Measurement of KL-6, CC-16, S-RAGE, ANG-2 using ELISA.	Day 28
Kinetic of surface biomarkers expression on neutrophils (C64) and monocytes (CD169, HLA-DR) during SARS CoV-2 infection.	Measurement of nCD64, mCD169 and mHLA-DR using the VersaPOC one-step rapid flow cytometry method.	Day 0
Kinetic of surface biomarkers expression on neutrophils (C64) and monocytes (CD169, HLA-DR) during SARS CoV-2 infection.	Measurement of nCD64, mCD169 and mHLA-DR using the VersaPOC one-step rapid flow cytometry method.	Day 1
Kinetic of surface biomarkers expression on neutrophils (C64) and monocytes (CD169, HLA-DR) during SARS CoV-2 infection.	Measurement of nCD64, mCD169 and mHLA-DR using the VersaPOC one-step rapid flow cytometry method.	Day 2
Kinetic of surface biomarkers expression on neutrophils (C64) and monocytes (CD169, HLA-DR) during SARS CoV-2 infection.	Measurement of nCD64, mCD169 and mHLA-DR using the VersaPOC one-step rapid flow cytometry method.	Day 3
Kinetic of surface biomarkers expression on neutrophils (C64) and monocytes (CD169, HLA-DR) during SARS CoV-2 infection.	Measurement of nCD64, mCD169 and mHLA-DR using the VersaPOC one-step rapid flow cytometry method.	Day 5
Kinetic of surface biomarkers expression on neutrophils (C64) and monocytes (CD169, HLA-DR) during SARS CoV-2 infection.	Measurement of nCD64, mCD169 and mHLA-DR using the VersaPOC one-step rapid flow cytometry method.	Day 7
Kinetic of surface biomarkers expression on neutrophils (C64) and monocytes (CD169, HLA-DR) during SARS CoV-2 infection.	Measurement of nCD64, mCD169 and mHLA-DR using the VersaPOC one-step rapid flow cytometry method.	Day 9
Kinetic of surface biomarkers expression on neutrophils (C64) and monocytes (CD169, HLA-DR) during SARS CoV-2 infection.	Measurement of nCD64, mCD169 and mHLA-DR using the VersaPOC one-step rapid flow cytometry method.	Day 11
Kinetic of surface biomarkers expression on neutrophils (C64) and monocytes (CD169, HLA-DR) during SARS CoV-2 infection.	Measurement of nCD64, mCD169 and mHLA-DR using the VersaPOC one-step rapid flow cytometry method.	Day 14
Kinetic of surface biomarkers expression on neutrophils (C64) and monocytes (CD169, HLA-DR) during SARS CoV-2 infection.	Measurement of nCD64, mCD169 and mHLA-DR using the VersaPOC one-step rapid flow cytometry method.	Day 17
Kinetic of surface biomarkers expression on neutrophils (C64) and monocytes (CD169, HLA-DR) during SARS CoV-2 infection.	Measurement of nCD64, mCD169 and mHLA-DR using the VersaPOC one-step rapid flow cytometry method.	Day 21
Kinetic of surface biomarkers expression on neutrophils (C64) and monocytes (CD169, HLA-DR) during SARS CoV-2 infection.	Measurement of nCD64, mCD169 and mHLA-DR using the VersaPOC one-step rapid flow cytometry method.	Day 28

Collaborators and Investigators

• Sponsor: Institut Hospitalo-Universitaire Méditerranée Infection
• Collaborators: Assistance Publique Hopitaux De Marseille; Institut Paoli-Calmettes; Hôpital Européen Marseille; Beckman Coulter, Inc.
• Investigators: Principal Investigator: Jean-Louis MEGE, MD, PhD, Institut Hospitalo-Universitaire Méditérranée Infection

Publications and helpful links

General Publications

• Wu Z, McGoogan JM. Characteristics of and Important Lessons From the Coronavirus Disease 2019 (COVID-19) Outbreak in China: Summary of a Report of 72 314 Cases From the Chinese Center for Disease Control and Prevention. JAMA. 2020 Apr 7;323(13):1239-1242. doi: 10.1001/jama.2020.2648. No abstract available.
• Ackermann M, Verleden SE, Kuehnel M, Haverich A, Welte T, Laenger F, Vanstapel A, Werlein C, Stark H, Tzankov A, Li WW, Li VW, Mentzer SJ, Jonigk D. Pulmonary Vascular Endothelialitis, Thrombosis, and Angiogenesis in Covid-19. N Engl J Med. 2020 Jul 9;383(2):120-128. doi: 10.1056/NEJMoa2015432. Epub 2020 May 21.
• Qin C, Zhou L, Hu Z, Zhang S, Yang S, Tao Y, Xie C, Ma K, Shang K, Wang W, Tian DS. Dysregulation of Immune Response in Patients With Coronavirus 2019 (COVID-19) in Wuhan, China. Clin Infect Dis. 2020 Jul 28;71(15):762-768. doi: 10.1093/cid/ciaa248.
• Zheng M, Gao Y, Wang G, Song G, Liu S, Sun D, Xu Y, Tian Z. Functional exhaustion of antiviral lymphocytes in COVID-19 patients. Cell Mol Immunol. 2020 May;17(5):533-535. doi: 10.1038/s41423-020-0402-2. Epub 2020 Mar 19. No abstract available.
• Giamarellos-Bourboulis EJ, Netea MG, Rovina N, Akinosoglou K, Antoniadou A, Antonakos N, Damoraki G, Gkavogianni T, Adami ME, Katsaounou P, Ntaganou M, Kyriakopoulou M, Dimopoulos G, Koutsodimitropoulos I, Velissaris D, Koufargyris P, Karageorgos A, Katrini K, Lekakis V, Lupse M, Kotsaki A, Renieris G, Theodoulou D, Panou V, Koukaki E, Koulouris N, Gogos C, Koutsoukou A. Complex Immune Dysregulation in COVID-19 Patients with Severe Respiratory Failure. Cell Host Microbe. 2020 Jun 10;27(6):992-1000.e3. doi: 10.1016/j.chom.2020.04.009. Epub 2020 Apr 21.
• Kuri-Cervantes L, Pampena MB, Meng W, Rosenfeld AM, Ittner CAG, Weisman AR, Agyekum RS, Mathew D, Baxter AE, Vella LA, Kuthuru O, Apostolidis SA, Bershaw L, Dougherty J, Greenplate AR, Pattekar A, Kim J, Han N, Gouma S, Weirick ME, Arevalo CP, Bolton MJ, Goodwin EC, Anderson EM, Hensley SE, Jones TK, Mangalmurti NS, Luning Prak ET, Wherry EJ, Meyer NJ, Betts MR. Comprehensive mapping of immune perturbations associated with severe COVID-19. Sci Immunol. 2020 Jul 15;5(49):eabd7114. doi: 10.1126/sciimmunol.abd7114.
• Hue S, Beldi-Ferchiou A, Bendib I, Surenaud M, Fourati S, Frapard T, Rivoal S, Razazi K, Carteaux G, Delfau-Larue MH, Mekontso-Dessap A, Audureau E, de Prost N. Uncontrolled Innate and Impaired Adaptive Immune Responses in Patients with COVID-19 Acute Respiratory Distress Syndrome. Am J Respir Crit Care Med. 2020 Dec 1;202(11):1509-1519. doi: 10.1164/rccm.202005-1885OC.

Helpful Links

• WHO Coronavirus Disease (COVID-19) Dashboard

Study record dates

Study Major Dates

• Study Start (Actual): March 25, 2020
• Primary Completion (Anticipated): June 1, 2021
• Study Completion (Anticipated): December 1, 2021

Study Registration Dates

• First Submitted: February 12, 2021
• First Submitted That Met QC Criteria: March 24, 2021
• First Posted (Actual): March 25, 2021

Study Record Updates

• Last Update Posted (Actual): March 25, 2021
• Last Update Submitted That Met QC Criteria: March 24, 2021
• Last Verified: March 1, 2021

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Respiratory Tract Diseases; Lung Diseases
• Other Study ID Numbers: 2020-A00756-33

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: UNDECIDED

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No