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Effect of IFN-γ on Innate Immune Cells

Primary Purpose

Chronic Granulomatous Disease

Status
Completed
Phase
Phase 1
Locations
United States
Study Type
Interventional
Intervention
Administration of drug (Interferon-gamma 1-b) subcutaneously
Sponsored by
University of Colorado, Denver
About
Eligibility
Locations
Arms
Outcomes
Full info

About this trial

This is an interventional basic science trial for Chronic Granulomatous Disease focused on measuring Interferon-gamma, innate immunity

Eligibility Criteria

18 Years - 60 Years (Adult)All SexesAccepts Healthy Volunteers

Inclusion Criteria:

  1. Healthy adults over the age of 18 years up to 60 years.
  2. At time of screening subject is well and healthy;
  3. Acute infections resolved;
  4. Subject off treatment medications;
  5. No diagnosis of chronic conditions or active health care issues for which the subject is actively followed by a health care provider or is on chronic medications.
  6. Non-prescription medications for mild inter-current illnesses will be allowed at the discretion of the principal investigator.

Exclusion Criteria:

  1. Pregnancy.
  2. History of current infection;
  3. Two weeks from most recent intercurrent infection;
  4. History of recurrent infections or immunodeficiency.

Sites / Locations

  • University of Colorado Denver, Anschutz Medical Campus

Arms of the Study

Arm 1

Arm Type

Other

Arm Label

SD or SS

Arm Description

In this study, IFN- γ-1b will be subcutaneously administered a total of 30 subjects in one of two cohorts; Single Dose (SD) or Steady State (SS) dosing. Dosing of IFN- γ-1b will be based upon the time subject became eligible and started study. In this non-randomized, open-label study, subjects will be enrolled on the SD cohort first, and once that cohort has been filled, enrollment to the SS cohort will begin. Although not required, subjects in the SD cohort may also volunteer to participate in the SS cohort if they still meet eligibility criteria. Separate consents will be used for the SD and SS cohorts. In the event not all the SD subjects choose to continue onto the SS cohort, we will plan to recruit new participants from our local campus community.

Outcomes

Primary Outcome Measures

Change in Neutrophil Nox2 activity.
Nox2 activity will be measured by DHR oxidation or Sod inhibitable cytochrome c reduction.
Change in Plasma IL-10 and Neuropterin concentration.
IL-10nd Neuropterin will be measured by ELISA.
Change in Neutrophil Gene Expression Analysis.
RNA will be extracted and gene expression will be determined by Affimetrix Gene Chip assay.
Change in IFN concentration and detection of anti-IFN antibody
IFN levels and anti-drug antibody will be completed by standard assays.

Secondary Outcome Measures

Change in Neutrophil Function studies.
Neutrophil chemotaxis, bactericidal activity,ingestion, degranulation, f-Actin expression, CD11b/18 expression, Nox2 activity to a variety of agonists.
Change in Anti-IFN antibody.
Anti-drug antibody to be determined by standard assay.
Change in Monocyte function studies.
Monocyte chemotaxis, bactericidal activity,ingestion, Expression of monocyte specific surface determinants, CD11b/18 expression, Nox2 activity to a variety of agonists, cell content of specific proteins, and antibody dependent cellular cytotoxicity.
Change in Neutrophil and Monocyte Gene Expression Analysis.
RNA will be extracted and gene expression will be determined by Affimetrix Gene Chip assay.

Full Information

First Posted
November 11, 2015
Last Updated
February 11, 2019
Sponsor
University of Colorado, Denver
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1. Study Identification

Unique Protocol Identification Number
NCT02609932
Brief Title
Effect of IFN-γ on Innate Immune Cells
Official Title
Effect of Interferon-gamma 1-b on Innate Immune Cells
Study Type
Interventional

2. Study Status

Record Verification Date
February 2019
Overall Recruitment Status
Completed
Study Start Date
July 2016 (undefined)
Primary Completion Date
August 2018 (Actual)
Study Completion Date
February 2019 (Actual)

3. Sponsor/Collaborators

Responsible Party, by Official Title
Sponsor
Name of the Sponsor
University of Colorado, Denver

4. Oversight

Data Monitoring Committee
No

5. Study Description

Brief Summary
The investigators hypothesize that neutrophils and monocytes developed under the influence of Interferon- gamma-1b (IFN-γ-1b, Actimmune*) in vivo will display enhanced function across a broad range of activities related in large part to the transcriptional activation effects of this cytokine. The investigators will evaluate the effects of IFN-γ in healthy human subjects in vivo on gene expression, biologic activity markers, and functional activity of myeloid cells in single dose studies and in steady state studies.
Detailed Description
Named for their potent ability to interfere and protect against viral infections, interferons (IFNs) have many regulatory effects on the immune system.1 Of the members of the two classes of these compounds, IFN-γ has the most diverse and powerful immune effects. Studies have mostly evaluated IFN-γ interactions with cells of adaptive immunity, including macrophages and lymphocytes. Effects on innate immunity, particularly polymorphonuclear leukocytes or neutrophils and monocytes are less well studied. However, investigations have suggested that IFN-γ may be involved in signal transduction, gene expression, the respiratory burst and neutrophil NADPH oxidase (Nox2) activity, phagocytosis, motility, microbicidal activity, and apoptosis. Not all of these functions are enhanced by IFN-γ; but the clinical use of this cytokine has been driven, in part by these results. For example, the primary motivation for initiating investigation of its beneficial clinical effects in Chronic Granulomatous Disease (CGD) was its effects on Nox2 activity.2 Most data in this area was based on studies using differentiated neutrophils from peripheral blood.1 However, the phenotype of neutrophils developed under the influence of this cytokine, not just changes expressed by exposure of differentiated cells to IFN-γ, is critical to understanding the physiologic effects of IFN-γ and the broad applications for its use in treatment of a range of human diseases. To expand their understanding of the role of IFN-γ in the development and functional integrity of the neutrophil, the investigators have completed a series of studies with PLB-985 cells in an in vitro culture system of myeloid cells. In this proposal, the investigators will evaluate innate immune activation and phagocyte function in healthy adult volunteers who are receiving IFN-γ.

6. Conditions and Keywords

Primary Disease or Condition Being Studied in the Trial, or the Focus of the Study
Chronic Granulomatous Disease
Keywords
Interferon-gamma, innate immunity

7. Study Design

Primary Purpose
Basic Science
Study Phase
Phase 1
Interventional Study Model
Single Group Assignment
Masking
None (Open Label)
Allocation
N/A
Enrollment
20 (Actual)

8. Arms, Groups, and Interventions

Arm Title
SD or SS
Arm Type
Other
Arm Description
In this study, IFN- γ-1b will be subcutaneously administered a total of 30 subjects in one of two cohorts; Single Dose (SD) or Steady State (SS) dosing. Dosing of IFN- γ-1b will be based upon the time subject became eligible and started study. In this non-randomized, open-label study, subjects will be enrolled on the SD cohort first, and once that cohort has been filled, enrollment to the SS cohort will begin. Although not required, subjects in the SD cohort may also volunteer to participate in the SS cohort if they still meet eligibility criteria. Separate consents will be used for the SD and SS cohorts. In the event not all the SD subjects choose to continue onto the SS cohort, we will plan to recruit new participants from our local campus community.
Intervention Type
Drug
Intervention Name(s)
Administration of drug (Interferon-gamma 1-b) subcutaneously
Other Intervention Name(s)
Actimmune
Intervention Description
SD = group who has received a single dose of IFN-gamma (10, 25, 50, and 100 mcg/m2) given once with subsequent analysis of effects (serum IL-10, Neuropterin, and IFN levels as well as neutrophil Nox2 activity and gene expression by Affimetrics Chip analysis). One month is allowed between doses. SS = administration of four doses (50 mcg/m2) of IFN-gamma given on Monday, Wednesday Friday schedule with neutrophil or monocyte function studies performed before the first and after the fourth dose to determine steady state effects.
Primary Outcome Measure Information:
Title
Change in Neutrophil Nox2 activity.
Description
Nox2 activity will be measured by DHR oxidation or Sod inhibitable cytochrome c reduction.
Time Frame
Determine the change in Nox2 activity at baseline compared to results for 8,24,48,72,96 hours after each IFN dose for the SD cohort.
Title
Change in Plasma IL-10 and Neuropterin concentration.
Description
IL-10nd Neuropterin will be measured by ELISA.
Time Frame
Determine the change in IL-10 and neuropterin concentration at baseline compared to 4, 8, 12, 24, 36, 48, 72, and 96 hours after each IFN dose, SD cohort.
Title
Change in Neutrophil Gene Expression Analysis.
Description
RNA will be extracted and gene expression will be determined by Affimetrix Gene Chip assay.
Time Frame
Determine the change in gene expression at baseline compared to 4, 8, 12, 24, 36, 48, 72, and 96 hours after each IFN dose, SD cohort.
Title
Change in IFN concentration and detection of anti-IFN antibody
Description
IFN levels and anti-drug antibody will be completed by standard assays.
Time Frame
Determine the change in IFN level at baseline compared to 4, 8, 12, and 24 hours after administration of each dose of IFN-gamma in the SD cohort. Determine the change in IFN antibody at baseline compared to day 7-10 and day 30 after IFN
Secondary Outcome Measure Information:
Title
Change in Neutrophil Function studies.
Description
Neutrophil chemotaxis, bactericidal activity,ingestion, degranulation, f-Actin expression, CD11b/18 expression, Nox2 activity to a variety of agonists.
Time Frame
Determine the change in neutrophil function at baseline compared to results on Day 8 after the 4th dose of IFN.
Title
Change in Anti-IFN antibody.
Description
Anti-drug antibody to be determined by standard assay.
Time Frame
Determine the change in IFN antibody at baseline compared to results for 7-10 da. and 30 da. after IFN.
Title
Change in Monocyte function studies.
Description
Monocyte chemotaxis, bactericidal activity,ingestion, Expression of monocyte specific surface determinants, CD11b/18 expression, Nox2 activity to a variety of agonists, cell content of specific proteins, and antibody dependent cellular cytotoxicity.
Time Frame
Determine the change in monocyte function at baseline compared to results on Day 8 after the 4th dose of IFN.
Title
Change in Neutrophil and Monocyte Gene Expression Analysis.
Description
RNA will be extracted and gene expression will be determined by Affimetrix Gene Chip assay.
Time Frame
Determine the change in monocyte function at baseline compared to results on Day 8 after the 4th dose of IFN.

10. Eligibility

Sex
All
Minimum Age & Unit of Time
18 Years
Maximum Age & Unit of Time
60 Years
Accepts Healthy Volunteers
Accepts Healthy Volunteers
Eligibility Criteria
Inclusion Criteria: Healthy adults over the age of 18 years up to 60 years. At time of screening subject is well and healthy; Acute infections resolved; Subject off treatment medications; No diagnosis of chronic conditions or active health care issues for which the subject is actively followed by a health care provider or is on chronic medications. Non-prescription medications for mild inter-current illnesses will be allowed at the discretion of the principal investigator. Exclusion Criteria: Pregnancy. History of current infection; Two weeks from most recent intercurrent infection; History of recurrent infections or immunodeficiency.
Overall Study Officials:
First Name & Middle Initial & Last Name & Degree
Daniel R. Ambruso, MD
Organizational Affiliation
University of Colorado, Denver
Official's Role
Principal Investigator
Facility Information:
Facility Name
University of Colorado Denver, Anschutz Medical Campus
City
Aurora
State/Province
Colorado
ZIP/Postal Code
80045
Country
United States

12. IPD Sharing Statement

Plan to Share IPD
No
Citations:
PubMed Identifier
26317224
Citation
Ellison MA, Thurman G, Gearheart CM, Seewald RH, Porter CC, Ambruso DR. INF-gamma Enhances Nox2 Activity by Upregulating phox Proteins When Applied to Differentiating PLB-985 Cells but Does Not Induce Nox2 Activity by Itself. PLoS One. 2015 Aug 28;10(8):e0136766. doi: 10.1371/journal.pone.0136766. eCollection 2015.
Results Reference
background
PubMed Identifier
35113934
Citation
Ambruso DR, Briones NJ, Baroffio AF, Murphy JR, Tran AD, Gowan K, Sanford B, Ellison M, Jones KL. In vivo interferon-gamma induced changes in gene expression dramatically alter neutrophil phenotype. PLoS One. 2022 Feb 3;17(2):e0263370. doi: 10.1371/journal.pone.0263370. eCollection 2022.
Results Reference
derived

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Effect of IFN-γ on Innate Immune Cells

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