Acute Humoral RejectionsNCT02137876
Effects of Thymoglobulin on Human B Cells
An observational study of Acute Humoral Rejections and Chronic Humoral Rejections, run by Northwestern University at 1 site in United States.
Trial summary
This study is being done to better understand how immunosuppressive medications (anti-rejection medications that may or may not be approved by the Food and Drug Administration (FDA)) affect the alloimmune response (how a person's immune system reacts against another person's immune system). Read more about this study
- Who is it for?
- Ages 18 years and olderHealthy volunteers can take part
- Conditions the study is for
- Acute Humoral Rejections, Chronic Humoral Rejections, Transplant
- What is being tested?
- Blood Draws
- Type of treatment
- Surgery or procedure
- How many people
- 0 participants
- Where
- 1 siteUnited States
- Study dates
- Apr 2014to Mar 2018
- Phase
- Observational
- Are there results?
- None posted in the registry
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 years and older
- All sexes
- Healthy volunteers accepted
- Adult, Older adult
Study population: Study population will consist of healthy volunteers who meet the inclusion/exclusion criteria.
Who may be able to join
- The patient:
- should be more than 18 years old,
- should weigh more than 110 lbs and
- should not be pregnant at the time of blood donation.
Who cannot join
- Not fitting within inclusion criteria
Trial locations and contacts
1 site in 1 country.
United States1 site
Northwestern University Comprehensive Transplant Center
Chicago, Illinois, 60611
About this study
Official title: Effects of Thymoglobulin on Proliferation, Activation, and Differentiation of Human B Cells
This study is being done to better understand how immunosuppressive medications (anti-rejection medications that may or may not be approved by the Food and Drug Administration (FDA)) affect the alloimmune response (how a person's immune system reacts against another person's immune system). This will be determined by the blood tests run on the patients donated blood cells.
Specifically, the investigators will test different immunosuppression medications using in-vitro assays (in the laboratory, in test tubes) alone and/or in combination to test how they can affect B-cell proliferation and differentiation, leading to an altered distribution among defined B-cell subsets and to study if exposure of B cells to thymoglobulin can have effects on subsequent interaction between B and T cell in vitro.
To enable the investigators to understand how immunosuppressant medications affect immune systems in transplant patients, the investigators need to understand how they affect immune systems in healthy people. To do this, the investigators will need to study blood collected from healthy volunteers.
Read the detailed description
This is a research study of healthy subjects who will furnish blood samples for various assays over a period of one year.
Peripheral blood mononuclear cells (PBMC) will be isolated from healthy volunteers and CD19+ B cells will be selected using CD19 magnetic micro-beads.
B-cell cultures will be performed in culture medium consisting of Iscove's MDM (IMDM) medium (Gibco, Paisley, UK) supplemented with 10% human serum (HNS- GemCell), 0.5 ml Human Insulin (Sigma), 200mM L-Glutamine (Sigma) and 0.5% Gentamycin (Gibco).
Total CD19+B cells will be cultured in the presence of anti-IgM (3μg/ml), anti-CD40 (100ng/ml), IL-21 (100ng/ml) (BCR/CD40-stimulation) for 6 days in culture plates 96 wells round bottom (1.0x105 cells/well), and Thymoglobulin at different concentrations (10 to 100 μg/ml) with or without calcineurin inhibitor (Tacrolimus, TAC) and mTOR inhibitor (Sirolimus, SRL) will be added at day 0.
A variety of parameters of B cell activity including proliferation, activation, differentiation and cytokine production will be monitored by flow cytometry.These experiments are further explained in the outcome measures section below.
The investigators will conduct univariate and bivariate analyses for all variables. The investigators will evaluate continuous variables using t-tests or F-tests and categorical variables using Chi-square or Fisher Exact tests. The investigators will examine data distributions and test all variables for linear relationships or non-linear relationships. Descriptive statistics (means, standard deviation, frequency) and correlations (both Pearson and Spearman) among variables will be conducted to assess the data. Necessary transformation and imputations will be conducted based on the raw data distribution. All data analyses will be performed using SAS 9.3 statistical software (SAS Inc., Cary, NC). All tests will be two-sided and an error rate of α <0.05 will be considered statistically significant.
- Being tested
- Blood Draws
- Keywords
- B cells, Thymoglubulin, humoral rejections, transplant, rATG
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
Blood DrawsProcedure or surgery
Patients will be asked to provide blood for immune tests.Patients will not take any study medications, all the research will be performed on blood donated for the study.Patients will donate about 20mls of blood (2 tbls)to be used to determine the subject's blood group and tissue typing (genetic).Results and the patient's contact information will be maintained in a database that is accessible only by the investigators and study personnel conducting the study.For the duration of this study, the investigator will contact patients by phone to request additional samples when needed. Study staff may ask patients donate more blood on short notice (1 day). This will consist of 60-120 ml (6-12 tbls) of blood.The frequency of blood collection will not exceed more than 2 times per week.The total amount of blood collected from subjects will not exceed 360 ml (1.5 cups) within any eight week period throughout the two year timespan patients may be asked to participate in this study.
What is being measured
The primary outcome is the main question the study is designed to answer.
CD19+B cell division
3-6x106 cells of interest will be labeled with 1.5μM CFSE before stimulation. After 6 days, B cells will be recovered, washed, counted and used to define marker settings. The cell phenotype will be analyzed by five-color flow cytometry (FC500, Beckman-Coulter) and by 13-color flow cytometry (Fortessa) and data will be analyzed using CXP software. The supernatant of these B-cell cultures will be using for the cytokines detection, and to determine the number of IgM and IgG producing cells by Luminex.At day 6, cells will be re-stimulated with phorbol 12-myristate 13-acetate (PMA) and Ionomycin, and analyzed by intracellular cytokine staining. Total CD19+ B cells will be sorted into a CD27- (naïve) and CD27+ (memory) B-cell fraction and the phenotypic characterization (assessed by flow cytometry) of these cells will be showed after 6 days of culture with anti-IgM (5μg/ml), anti-CD40 (100ng/ml), and IL-21 (100ng/ml) in the presence or absence of the drugs.
Measured: 12 days
Secondary outcomes (1)
proliferated T-cells
The investigators will co-culture B cells (pre-stimulated with α-IgM and α-CD40 mAb plus IL-21 for 6 days in the presence and absence of Thymoglobulin) with allo-CD4+CD25- T cells (ratio 2:1). After 6 days the investigators will analyze the % of proliferated T-cells and the cytokine production.
Measured: 6 days
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted14 May 2014 (estimated)
- Study startApr 2014
- Primary completionMar 2018
- Study completionMar 2018
Why it stopped: Subject recruitment was unsuccessful.
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Other trials for Acute Humoral RejectionsStudies of the same condition, open and under way.
- More trials run by Northwestern UniversityThe sponsor's other registered studies.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Case only
- Time perspective
- Prospective
- Enrolment
- 0 participants (actual)
Sponsor and investigators
- Lead sponsor
- Northwestern University
- Sponsor type
- University, hospital or other
- Collaborators
- Sanofi
- Responsible party
- Principal investigator, Lorenzo Gallon, Northwestern University
- Investigators
- Lorenzo Gallon, MD, Principal investigator, Northwestern University
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT02137876
- Sponsor's protocol ID
- STU00092183
- Status last verified
- Mar 2018
- First submitted
- 30 Apr 2014
- Last updated
- 29 Mar 2018
- Results posted
- No
- Data monitoring committee
- No
Study at a glance
From the registry record, last updated by the sponsor on 29 Mar 2018. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Effects of Thymoglobulin on Proliferation, Activation, and Differentiation of Human B Cells
- ClinicalTrials.gov ID
- NCT02137876
- Acronym / sponsor ID
- STU00092183
- Keywords
- B cells, Thymoglubulin, humoral rejections, transplant, rATG
- Study type
- Observational
- Recruitment status
- WithdrawnReason stopped: Subject recruitment was unsuccessful.
- Enrolment
- 0 participants (actual)
- Lead sponsor
- Northwestern UniversityUniversity, hospital or other
- Collaborators
- Sanofi
- Study start
- Apr 2014 (actual)
- Primary completion
- Mar 2018 (actual)
- Study completion
- Mar 2018 (actual)
- First posted
- 14 May 2014 (estimated)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
This study is being done to better understand how immunosuppressive medications (anti-rejection medications that may or may not be approved by the Food and Drug Administration (FDA)) affect the alloimmune response (how a person's immune system reacts against another person's immune system). This will be determined by the blood tests run on the patients donated blood cells.
Specifically, the investigators will test different immunosuppression medications using in-vitro assays (in the laboratory, in test tubes) alone and/or in combination to test how they can affect B-cell proliferation and differentiation, leading to an altered distribution among defined B-cell subsets and to study if exposure of B cells to thymoglobulin can have effects on subsequent interaction between B and T cell in vitro.
To enable the investigators to understand how immunosuppressant medications affect immune systems in transplant patients, the investigators need to understand how they affect immune systems in healthy people. To do this, the investigators will need to study blood collected from healthy volunteers.
Detailed description
This is a research study of healthy subjects who will furnish blood samples for various assays over a period of one year.
Peripheral blood mononuclear cells (PBMC) will be isolated from healthy volunteers and CD19+ B cells will be selected using CD19 magnetic micro-beads.
B-cell cultures will be performed in culture medium consisting of Iscove's MDM (IMDM) medium (Gibco, Paisley, UK) supplemented with 10% human serum (HNS- GemCell), 0.5 ml Human Insulin (Sigma), 200mM L-Glutamine (Sigma) and 0.5% Gentamycin (Gibco).
Total CD19+B cells will be cultured in the presence of anti-IgM (3μg/ml), anti-CD40 (100ng/ml), IL-21 (100ng/ml) (BCR/CD40-stimulation) for 6 days in culture plates 96 wells round bottom (1.0x105 cells/well), and Thymoglobulin at different concentrations (10 to 100 μg/ml) with or without calcineurin inhibitor (Tacrolimus, TAC) and mTOR inhibitor (Sirolimus, SRL) will be added at day 0.
A variety of parameters of B cell activity including proliferation, activation, differentiation and cytokine production will be monitored by flow cytometry.These experiments are further explained in the outcome measures section below.
The investigators will conduct univariate and bivariate analyses for all variables. The investigators will evaluate continuous variables using t-tests or F-tests and categorical variables using Chi-square or Fisher Exact tests. The investigators will examine data distributions and test all variables for linear relationships or non-linear relationships. Descriptive statistics (means, standard deviation, frequency) and correlations (both Pearson and Spearman) among variables will be conducted to assess the data. Necessary transformation and imputations will be conducted based on the raw data distribution. All data analyses will be performed using SAS 9.3 statistical software (SAS Inc., Cary, NC). All tests will be two-sided and an error rate of α <0.05 will be considered statistically significant.
Study design
- Observational model
- Case only
- Time perspective
- Prospective
Who may qualify
- Ages
- 18 years and older
- Sex
- All
- Healthy volunteers
- Accepted
- Age groups
- Adult, Older adult
- Study population
- Study population will consist of healthy volunteers who meet the inclusion/exclusion criteria.
Key inclusion criteria (4 listed)
- should be more than 18 years old,
- should weigh more than 110 lbs and
- should not be pregnant at the time of blood donation.
Key exclusion criteria (1 listed)
- Not fitting within inclusion criteria
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Interventions
| Intervention | Type | Arms |
|---|---|---|
| Blood DrawsPatients will be asked to provide blood for immune tests.Patients will not take any study medications, all the research will be performed on blood donated for the study.Patients will donate about 20mls of blood (2 tbls)to be used to determine the subject's blood group and tissue typing (genetic).Results and the patient's contact information will be maintained in a database that is accessible only by the investigators and study personnel conducting the study.For the duration of this study, the investigator will contact patients by phone to request additional samples when needed. Study staff may ask patients donate more blood on short notice (1 day). This will consist of 60-120 ml (6-12 tbls) of blood.The frequency of blood collection will not exceed more than 2 times per week.The total amount of blood collected from subjects will not exceed 360 ml (1.5 cups) within any eight week period throughout the two year timespan patients may be asked to participate in this study. | Procedure or surgery | – |
Primary outcome
CD19+B cell division
Time frame: 12 days
3-6x106 cells of interest will be labeled with 1.5μM CFSE before stimulation. After 6 days, B cells will be recovered, washed, counted and used to define marker settings. The cell phenotype will be analyzed by five-color flow cytometry (FC500, Beckman-Coulter) and by 13-color flow cytometry (Fortessa) and data will be analyzed using CXP software. The supernatant of these B-cell cultures will be using for the cytokines detection, and to determine the number of IgM and IgG producing cells by Luminex.At day 6, cells will be re-stimulated with phorbol 12-myristate 13-acetate (PMA) and Ionomycin, and analyzed by intracellular cytokine staining. Total CD19+ B cells will be sorted into a CD27- (naïve) and CD27+ (memory) B-cell fraction and the phenotypic characterization (assessed by flow cytometry) of these cells will be showed after 6 days of culture with anti-IgM (5μg/ml), anti-CD40 (100ng/ml), and IL-21 (100ng/ml) in the presence or absence of the drugs.
1 secondary outcome
proliferated T-cells
Time frame: 6 days
Results and safety
The study was withdrawn before enrolling participants, so there are no results.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 29 Mar 2018.
Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part. This study is not recruiting at the moment.
Lorenzo Gallon, MD principal investigator
Northwestern University
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
An observational study of Acute Humoral Rejections and Chronic Humoral Rejections, run by Northwestern University at 1 site in United States.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 4 inclusion and 1 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: withdrawn, as of the sponsor's last update on 29 Mar 2018. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in United States. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
The study was withdrawn before enrolling participants, so there are no results.
How can a doctor refer a patient or contact the research team?
The record lists no contact. The official record names the sponsor, who can be asked directly. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
CD19+B cell division (time frame: 12 days). It is what the study was designed to measure, not a finding.
Where can I read the original study information?
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 29 Mar 2018.