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Monocytic Expression of Heme Oxidase-1 (HO-1) in Sickle Cell Patients and Correlation With the Humoral Immune Response to Vaccine and With Allo-immunization.

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ClinicalTrials.gov Identifier: NCT03111589
Recruitment Status : Completed
First Posted : April 13, 2017
Last Update Posted : October 18, 2018
Sponsor:
Information provided by (Responsible Party):
Francis Corazza, Brugmann University Hospital

Brief Summary:

Sickle cell disease (SCD) is an autosomal recessive disorder resulting from a substitution in the β chain of hemoglobin (Hb) which causes hemoglobin S to polymerize when deoxygenated. SCD patients present immune abnormalities that have always been attributed to functional asplenia. It it is now being recognized that patients with SCD have a pro-inflammatory condition with altered immune system activation contributing to the pathology of SCD. Increased levels of neutrophils, monocytes or cytokines have been reported in SCD patients.

SCD is associated with many acute and chronic complications requiring immediate support. Actual strongly recommended therapies include chronic blood transfusions (CT) and hydroxyurea (HU). In addition, episodic transfusions are recommended and commonly used to manage many acute SCD complications.There is strong evidence to support the use of HU in adults with 3 or more severe vaso-occlusive crises during any 12-month period, with SCD pain or chronic anemia, or with severe or recurrent episodes of acute chest syndrome. HU use is now also common in children with SCD. Some patients receive chronic monthly RBC transfusion with the objective to reduce the proportion of HbS to < 30 %. Long-term RBC transfusions prevent and treat complications of SCD decreasing the risk of stroke and the incidence of acute chest syndrome (ACS).

Therapeutic complications, such as alloimmunization against RBC in 20-50% of patients or hematopoietic stem cell transplantation (HSCT) graft rejection, constitute an immune-based clinical issue in SCD. Poorly understood RBC alloimmunization is responsible for serious hemolytic transfusion reaction associated with severe mortality and morbidity underlying the need for a better understanding of the immunology of SCD to improve SCD transfusion support/outcome. Little evidence exists about HU effects on immune functions in SCD. HU treatment doesn't appear to have deleterious effects on immune function and appears to decrease the abnormally elevated number of total WBC and lymphocytes, while CT does not.

Patients with SCD are at higher risk of infections and prophylactic vaccination is strongly recommended. Recent data suggest that vaccinal response to pneumococcal antigens in SCD patients is identical to healthy control while controversy concern the stability of the immune protection after vaccination of SCD patient. Antibody levels declined over the year and the need for more frequent vaccination in SCD patient should be investigated. Currently, there is no evidence whether HU may interfere with pneumococcal immune response. Purohit showed that immune response to inactivated influenza A (H1N1) virus vaccine was altered in patient with SCD receiving CT but little is known on immune response to vaccination in patients with SCD receiving HU.

Recent data suggest that not only inflammatory status but also humoral immune response to antigens in SCD patients may differ according to treatment. Yazdanbakhsh reported an imbalance between regulatory T cell (Treg) and effector T cell (Teff) in alloimmunized SCD patients with as consequence an increase in antibody production. In a model proposed by the authors, the balance between Treg and Teff is dictated by the monocyte control of cytokines expression. Altered activity of monocyte heme oxidase-1 (HO-1) would be responsible of a decrease in IL-12 and an increase in IL-10 cytokines secretion impacting the Treg/Teff cells ratio and promoting antibody production by B cells.

The objectives of the project are to assess whether different humoral immune responses to vaccines or to erythrocyte alloantigens are related to the type of treatment administered to patients with SCD. We also aim to study if these differences might be related to different expressions of HO-1 by monocytes.


Condition or disease Intervention/treatment Phase
Sickle Cell Disease Biological: Inactivated influenza A (H1N1) virus vaccine Diagnostic Test: Blood sampling Not Applicable

Study Type : Interventional  (Clinical Trial)
Actual Enrollment : 102 participants
Allocation: Non-Randomized
Intervention Model: Parallel Assignment
Masking: None (Open Label)
Primary Purpose: Basic Science
Official Title: Monocytic Expression of Heme Oxidase-1 (HO-1) in Sickle Cell Patients and Correlation With the Humoral Immune Response to Vaccine and With Allo-immunization
Actual Study Start Date : October 2016
Actual Primary Completion Date : October 2018
Actual Study Completion Date : October 2018

Resource links provided by the National Library of Medicine


Arm Intervention/treatment
Experimental: SCD patients under regular chronic exchange transfusion
Sickle cell disease patients (SCD) under regular chronic exchange transfusions. Pediatric and adult patients from the HUDERF and CHU-Brugmann Hospitals.
Biological: Inactivated influenza A (H1N1) virus vaccine
All groups of patients and the control group will receive the new annually recommended inactivated influenza A (H1N1) virus vaccine.

Diagnostic Test: Blood sampling
Testing of the different humoral immune responses to vaccines or to erythrocyte alloantigens.

Experimental: SCD patients under HU treatment alone
Sickle cell disease patients (SCD) under hydroxyurea (HU) alone. Pediatric and adult patients from the HUDERF and CHU-Brugmann Hospitals.
Biological: Inactivated influenza A (H1N1) virus vaccine
All groups of patients and the control group will receive the new annually recommended inactivated influenza A (H1N1) virus vaccine.

Diagnostic Test: Blood sampling
Testing of the different humoral immune responses to vaccines or to erythrocyte alloantigens.

Experimental: SCD patients under HU treatment+sporadic transfusion
Sickle cell disease patients (SCD) under hydroxyurea (HU) and receiving sporadic transfusions.Pediatric and adult patients from the HUDERF and CHU-Brugmann Hospitals.
Biological: Inactivated influenza A (H1N1) virus vaccine
All groups of patients and the control group will receive the new annually recommended inactivated influenza A (H1N1) virus vaccine.

Diagnostic Test: Blood sampling
Testing of the different humoral immune responses to vaccines or to erythrocyte alloantigens.

Active Comparator: Control group
Pediatric and adult patients from the HUDERF and CHU-Brugmann Hospitals.
Biological: Inactivated influenza A (H1N1) virus vaccine
All groups of patients and the control group will receive the new annually recommended inactivated influenza A (H1N1) virus vaccine.

Diagnostic Test: Blood sampling
Testing of the different humoral immune responses to vaccines or to erythrocyte alloantigens.




Primary Outcome Measures :
  1. Intracellular HO-1 expression in monocytes [ Time Frame: 1 month post vaccination ]
    Intracellular monocyte heme oxidase-1 (HO-1) expression will be measured by flow cytometry.The protein expression of HO-1 will be confirmed by Western blot. A commercial ELISA kit will be used in parallel to assess HO-1 levels in PBMC cell lysate.

  2. HO-1 level in serum [ Time Frame: 1 month post vaccination ]
    Monocyte heme oxidase-1 (HO-1) level in serum will be measured by a commercial ELISA kit

  3. Cytokines levels measurement [ Time Frame: 1 month post vaccination ]
    Pro-inflammatory cytokine (IL-12) and anti-inflammatory cytokine (IL-10) levels will be evaluated in serum and in IL-1 stimulated whole blood supernatants using an ELISA assay.

  4. Identification of T regulatory cells [ Time Frame: 1 month post vaccination ]
    Evaluation of Treg cells in peripheral blood mononuclear cells (PBMC) will be performed by flow cytometry using appropriate fluorochrome conjugated monoclonal antibodies for CD25 and FoxP3 markers

  5. Immune response to vaccination [ Time Frame: 1 month post vaccination ]
    Post-vaccination serum H1N1 antibodies titers (IgG and IgM) will be measured by an ELISA kit


Secondary Outcome Measures :
  1. Intracellular HO-1 expression in monocytes [ Time Frame: Baseline: at vaccination ]
    Intracellular HO-1 expression will be measured by flow cytometry.The protein expression of HO-1 will be confirmed by Western blot. A commercial ELISA kit will be used in parallel to assess HO-1 levels in PBMC cell lysate.

  2. Intracellular HO-1 expression in monocytes [ Time Frame: 3 months post vaccination ]
    Intracellular HO-1 expression will be measured by flow cytometry.The protein expression of HO-1 will be confirmed by Western blot. A commercial ELISA kit will be used in parallel to assess HO-1 levels in PBMC cell lysate.

  3. Intracellular HO-1 expression in monocytes [ Time Frame: 6 months post vaccination ]
    Intracellular HO-1 expression will be measured by flow cytometry.The protein expression of HO-1 will be confirmed by Western blot. A commercial ELISA kit will be used in parallel to assess HO-1 levels in PBMC cell lysate.

  4. HO-1 level in serum [ Time Frame: Baseline: at vaccination ]
    HO-1 level in serum will be measured by a commercial ELISA kit

  5. HO-1 level in serum [ Time Frame: 3 months post vaccination ]
    HO-1 level in serum will be measured by a commercial ELISA kit

  6. HO-1 level in serum [ Time Frame: 6 months post vaccination ]
    HO-1 level in serum will be measured by a commercial ELISA kit

  7. Cytokines levels measurement [ Time Frame: Baseline: at vaccination ]
    Pro-inflammatory cytokine (IL-12) and anti-inflammatory cytokine (IL-10) levels will be evaluated in serum and in IL-1 stimulated whole blood supernatants using an ELISA assay.

  8. Cytokines levels measurement [ Time Frame: 3 months post vaccination ]
    Pro-inflammatory cytokine (IL-12) and anti-inflammatory cytokine (IL-10) levels will be evaluated in serum and in IL-1 stimulated whole blood supernatants using an ELISA assay.

  9. Cytokines levels measurement [ Time Frame: 6 months post vaccination ]
    Pro-inflammatory cytokine (IL-12) and anti-inflammatory cytokine (IL-10) levels will be evaluated in serum and in IL-1 stimulated whole blood supernatants using an ELISA assay.

  10. Identification of T regulatory cells [ Time Frame: Baseline: at vaccination ]
    Evaluation of Treg cells in PBMC will be performed by flow cytometry using appropriate fluorochrome conjugated monoclonal antibodies for CD25 and FoxP3 markers

  11. Identification of T regulatory cells [ Time Frame: 3 months post vaccination ]
    Evaluation of Treg cells in PBMC will be performed by flow cytometry using appropriate fluorochrome conjugated monoclonal antibodies for CD25 and FoxP3 markers

  12. Identification of T regulatory cells [ Time Frame: 6 months post vaccination ]
    Evaluation of Treg cells in PBMC will be performed by flow cytometry using appropriate fluorochrome conjugated monoclonal antibodies for CD25 and FoxP3 markers

  13. Immune response to vaccination [ Time Frame: Baseline: at vaccination ]
    Post-vaccination serum H1N1 antibodies titers (IgG and IgM) will be measured by an ELISA kit

  14. Immune response to vaccination [ Time Frame: 3 months post vaccination ]
    Post-vaccination serum H1N1 antibodies titers (IgG and IgM) will be measured by an ELISA kit

  15. Immune response to vaccination [ Time Frame: 6 months post vaccination ]
    Post-vaccination serum H1N1 antibodies titers (IgG and IgM) will be measured by an ELISA kit



Information from the National Library of Medicine

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Ages Eligible for Study:   Child, Adult, Older Adult
Sexes Eligible for Study:   All
Accepts Healthy Volunteers:   No
Criteria

Inclusion Criteria:

- Pediatric and adult patients with sickle cell disease from the HUDERF and CHU-Brugmann Hospital

Exclusion Criteria:


Information from the National Library of Medicine

To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor.

Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT03111589


Locations
Belgium
CHU Brugmann
Brussels, Belgium, 1020
HUDERF
Brussels, Belgium, 1020
Sponsors and Collaborators
Francis Corazza
Investigators
Principal Investigator: Francis Corazza, MD CHU Brugmann

Publications:

Responsible Party: Francis Corazza, Head of clinic, Brugmann University Hospital
ClinicalTrials.gov Identifier: NCT03111589     History of Changes
Other Study ID Numbers: CHUB-HO1 sickle cell
First Posted: April 13, 2017    Key Record Dates
Last Update Posted: October 18, 2018
Last Verified: October 2018
Individual Participant Data (IPD) Sharing Statement:
Plan to Share IPD: No

Studies a U.S. FDA-regulated Drug Product: No
Studies a U.S. FDA-regulated Device Product: No

Keywords provided by Francis Corazza, Brugmann University Hospital:
Sickle Cell Disease
Heme oxidase-1 (HO-1)

Additional relevant MeSH terms:
Anemia, Sickle Cell
Anemia, Hemolytic, Congenital
Anemia, Hemolytic
Anemia
Hematologic Diseases
Hemoglobinopathies
Genetic Diseases, Inborn
Vaccines
Immunologic Factors
Physiological Effects of Drugs