Effects of Thymoglobulin on Human B Cells

NCT02137876 | Withdrawn

• 1 other identifier: STU00092183
• Study Type: observational
• Target: N/A
• Locations: 1 country
• Sites: 1

Brief 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). 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.

Conditions

Acute Humoral Rejections; Chronic Humoral Rejections; Transplant

Keywords

B cells; Thymoglubulin; humoral rejections; transplant; rATG

Outcome Measures

Primary Outcomes (1)

• 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.

  12 days

Secondary Outcomes (1)

• proliferated T-cells

  6 days

Interventions

Blood Draws PROCEDURE

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.

Eligibility Criteria

• Age: 18 Years+
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)
• Sampling Method: Non-Probability Sample

Study Population

Study population will consist of healthy volunteers who meet the inclusion/exclusion criteria.

You may qualify if:

• 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.

Sponsors & Collaborators

• Northwestern University: lead
• Sanofi: collaborator

Study Sites (1)

Northwestern University Comprehensive Transplant Center

Chicago, Illinois, 60611, United States

Location

Biospecimen

Retention: SAMPLES WITH DNA

20ml of whole blood will be drawn from each subject each time they are asked to have blood drawn. 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.

MeSH Terms

Interventions

Blood Specimen Collection

Intervention Hierarchy (Ancestors)

Specimen Handling; Clinical Laboratory Techniques; Diagnostic Techniques and Procedures; Diagnosis; Punctures; Surgical Procedures, Operative; Investigative Techniques

Study Officials

• Lorenzo Gallon, MD

  Northwestern University

  PRINCIPAL INVESTIGATOR

Study Design

• Study Type: observational
• Observational Model: CASE ONLY
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Professor

Study Record Dates

• First Submitted: April 30, 2014
• First Posted: May 14, 2014
• Study Start: April 1, 2014
• Primary Completion: March 1, 2018
• Study Completion: March 1, 2018
• Last Updated: March 29, 2018

Record last verified: 2018-03

Locations

US (1)