Non Randomized Comparative Study With Control

Study Overview
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Brief Summary
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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.

Detailed Description
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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).

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

Conditions
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Allogeneic Hematopoietic Stem Cell Transplantation

Intervention / Treatment
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* Other: blood sample


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: 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; Control group: allogeneic HSC transplanted patients; Age>18 years; no CMV or AdV reactivation ; signed informed consent; Healthy donors group: HSC donor; Age>18 years; signed informed consent Exclusion Criteria: none

Ages Eligible for Study
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Minimum Age: 18 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

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

What is the study measuring?
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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) | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Allogeneic transplantation, Cytomegalovirus, adenovirus, NK cells