Non Randomized Comparative Study With Control (Allo-NK-CMV)

Study of Natural Killer Immunity During Infections With CMV or AdV After Allogeneic Hematopoietic Stem Cells

NK cells are lymphocytes who play a role in the control of viral infections , tumor and fœtal tolerance. They belong to innate immune cells but they have a link with adaptative immunity. Indeed, after some viral infections such as CMV, Chikungunya, B hepatitis etc, a subset of NKG2C+ NK cells expands and can transfer, in murine models, a " memory " that can better control CMV infections. CMV reactivation is a major cause of morbidity and mortality after allogeneic hematopoietic ste cell transplantation in humans. The aim of this prospective study is to evaluate the role of NK cells, in particular NKG2C+ NK cells in the control of CMV but also Adenovirus after allo HSCT. Peripheral NK cells from 30 and 10 patients who reactivated respectively CMV and AdV are prospectively studied (extensive phenotyping and functional studies before and after administration of anti viral drugs) and compared with 30 allotransplanted patients who didn't reactivate CMV in a pair analysis, and 30 healthy donors serologically + for CMV.

Study Overview

• Status: Unknown
• Conditions: Allogeneic Hematopoietic Stem Cell Transplantation
• Intervention / Treatment: Other: blood sample

Detailed Description

For both groups: 28ml of peripheral blood samples are collected at different points. Group 1 : before and after anti viral treatment . Control group 2: one point after allo-HSCT; Control group 3: 1 point before transplant. Phenotypical study of NK cells: activating and inhibitory receptors, activation and differentiation's markers. Phenotypical studies of the ligands on infected cells. Genotypic study of KIR receptors (Kirotype); functional studies: polyfunctionality essay (flow cytometry): degranulation CD107a, IFNg, TNFa production).

• Study Type: Observational
• Enrollment (Anticipated): 90

Contacts and Locations

• Study Contact: Name: Stephanie Nguyen Quoc, Doctor; Phone Number: 33142162823; Email: stephanie.nguyen-quoc@aphp.fr
• Study Contact Backup: Name: Laetitia Souchet, Doctor; Phone Number: 33142162823

Study Locations

• France
  • Paris, France, 75651
    • Recruiting
    • Hopital Pitie Salpetriere

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

Allogeneic HSCT for malignant or non malignant hematological disease in adult patients

Description

Inclusion Criteria:

1. patients who reactivate CMV or AdV after allogeneic HSCT ;

   • Age>18 years; indication for a antiviral treatment:
   • at least 1 PCR CMV>1000 copies/ml or 1 PCR ADV>1000 copies/ml or at least 2 ADV PCR positive - sites (stools, throat, urines);
   • signed informed consent;

2. Control group: allogeneic HSC transplanted patients;

   • Age>18 years; no CMV or AdV reactivation ;
   • signed informed consent;

3. Healthy donors group: HSC donor;

   • Age>18 years;
   • signed informed consent

Exclusion Criteria:

none

Study Plan

How is the study designed?

Number of groups / cohorts

3

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
patients who reactivate CMV or AdV after allogeneic HSCT; allotransplanted patients who reactivated respectively CMV (n=30) and AdV (n=10)	Other: blood sample; peripheral blood samples are collected
Control group: allogeneic HSC transplanted patients; allotransplanted patients who didn't reactivate CMV	Other: blood sample; peripheral blood samples are collected
Healthy donors group; healthy donors serologically + for CMV	Other: blood sample; peripheral blood samples are collected

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Nk cells phenotype (activation, differentiation, memory NK cell) and function (cytoxicity and cytokines production) measured by flow cytometry	CD3-CD56+ NK cells will be analyzed by Flow cytometry with an appropriate monoclonal antibodies (mAb) cocktail: anti-CD3 ,CD56, CD16, CD159a/NKG2A , CD85J; HLA-DR ; CD62L , CD161; KIR2DL1 and KIR3DL1, and KIR2DL2/KIR2DL3. The state of NK cells differentiation and maturation will be assessed by the analyze of thoses phenotypic markers and will be compared with healthy donors.	Change from "before antiviral treatment", " half treatment" (1 week or 2 week after the beginning of the treatment), 1 month "post treatment" and 3 month "post treatment"
NK cells cytoxicity and cytokines production when incubated with standard HLA class I negative K562 target cells	Polyfunctional assay will test the capacity of NK cells degranulation and production of cytokines when incubated with standard HLA class I negative K562 target cells in the presence of anti-CD107a ,IFN-g, or TNF-a mAb. The state of NK cells differentiation and maturation will be assessed by the analyze of thoses phenotypic and will be compared with healthy donors.	Change from "before antiviral treatment", " half treatment" (1 week or 2 week after the beginning of the treatment), 1 month "post treatment" and 3 month "post treatment"

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Effect of CMV and AdV infection compared with healthy donors	The state of NK cells differentiation and maturation will be assessed by the analyze of thoses phenotypic and functional markers and will be compared with healthy donors.	Change from "before antiviral treatment", " half treatment" (1 week or 2 week after the beginning of the treatment), 1 month "post treatment" and 3 month "post treatment"
In vitro Nk cells ligands analyzes on infected cells by CMV	Monocyte are differentiated in macrophage cells and infected by the CMV strain TB40/E. Nk cell ligands will be analyzed on infected cells. NK cells from infected patients post allogenic transplantation will be tested against in vitro infected cells. Being a model in vitro, it is not possible to determine in advance which ligands will be identified.	approximately 18 months after Study Completion Date (last participant's last visit)

Collaborators and Investigators

• Sponsor: Assistance Publique - Hôpitaux de Paris
• Investigators: Principal Investigator: Stephanie Nguyen Quoc, Doctor, Assistance Publique - Hôpitaux de Paris

Study record dates

Study Major Dates

• Study Start (Actual): September 27, 2013
• Primary Completion (Anticipated): June 1, 2018
• Study Completion (Anticipated): June 1, 2018

Study Registration Dates

• First Submitted: February 15, 2016
• First Submitted That Met QC Criteria: September 21, 2017
• First Posted (Actual): September 27, 2017

Study Record Updates

• Last Update Posted (Actual): May 29, 2018
• Last Update Submitted That Met QC Criteria: May 25, 2018
• Last Verified: May 1, 2018

More Information

Terms related to this study

• Keywords: adenovirus; Cytomegalovirus; NK cells; Allogeneic transplantation
• Other Study ID Numbers: P111107; IDRCB (Registry Identifier: 2022-A02601-42)

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO