Effects of Thymoglobulin on Human B Cells

NCT ID: NCT02137876

Last Updated: 2018-03-29

Basic Information

• Recruitment Status: WITHDRAWN
• Study Classification: OBSERVATIONAL
• Study Start Date: 2014-04-30
• Study Completion Date: 2018-03-31

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.

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.

Conditions

Acute Humoral Rejections; Chronic Humoral Rejections; Transplant

Study Design

• Observational Model Type: CASE_ONLY
• Study Time Perspective: PROSPECTIVE

Interventions

Blood Draws

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.

Intervention Type: PROCEDURE

Eligibility Criteria

Inclusion Criteria

The patient:

1. should be more than 18 years old,

2. should weigh more than 110 lbs and

3. should not be pregnant at the time of blood donation.

• Minimum Eligible Age: 18 Years
• Eligible Sex: ALL
• Accepts Healthy Volunteers: Yes

Sponsors

Sanofi

INDUSTRY

Sponsor Role: collaborator

Northwestern University

OTHER

Sponsor Role: lead

Responsible Party

Lorenzo Gallon

Professor

Responsibility Role: PRINCIPAL_INVESTIGATOR

Principal Investigators

Lorenzo Gallon, MD

Role: PRINCIPAL_INVESTIGATOR

Northwestern University

Locations

Northwestern University Comprehensive Transplant Center

Chicago, Illinois, United States

Countries

United States

Other Identifiers

STU00092183

Identifier Type: -

Identifier Source: org_study_id