Mast Cell Activation Test in Allergic Disease

Assessment of Mast Cell Activation Diagnostic Test in Patients With Allergic Disease

Activation of mast cells in the immune system is known to cause allergic reactions sometimes with severe systemic symptoms. The investigators have recently developed a blood-based mast cell activation diagnostic test in which levels of functional activation in-vitro in primary cultured mast cells generated from the peripheral blood of single individuals can be assessed. It is the hypothesis that the test can be used to predict the potential state of in-vivo mast cell activation in any individual based on the functional activation profiles exhibited by their cultured mast cells. The investigators now wish to translate their in-vitro findings in a pilot study to disease groups where mast cell activation is expected to be high. These include highly allergic individuals; those with chronic idiopathic urticaria; those with mastocytosis; and those with the mast cell activation syndrome. Furthermore, they will use the functional genomics approach to identify gene expression biomarkers that are correlated with such diseases. The results will be compared with data that have been collected from a cohort of healthy control blood donors.

Study Overview

• Status: Completed
• Conditions: Asthma; Eczema; Food Allergy; Mastocytosis; Chronic Idiopathic Urticaria; Anaphylaxis; Mast Cell Activation Syndrome

Detailed Description

The objective of this study is to assess whether patients with severe allergic diseases exhibit high levels of mast cell activation as determined by the mast cell activation diagnostic test the investigators have developed. They anticipate that patients with history of severe allergic reactions will show up as high responders in the blood-based mast cell activation diagnostic test. Moreover, using microarray analysis as the approach for gene expression studies, they anticipate further that genomics biomarkers that are correlated with the high functional activation of the in-vitro mast cells derived from these patients can be readily identified.

About 100 ml of peripheral venous blood from individual patients will be drawn into heparinized syringes and collected in a blood-collecting bag/tube containing silica. The bag/tube will be promptly processed for the generation of primary human mast cell cultures using the protocol that has previously been developed [Inflammation Research 66: 25 (2017). After culturing for 9 weeks, the resulting human mast cells will be analyzed for both their functional activity in terms of histamine release in response to the activation of high-affinity Immunoglobulin E (IgE) receptors in these cells and their gene expression profiles using microarray analysis. The in-vitro functional and genomics data will be correlated with specific diagnoses and measurements of blood prostaglandin D2 (PGD2), PGD2 metabolites, tryptase and histamine levels in blood. For mediator assays, plasma from each subject will be collected after the cluster of differentiation 34 positive (CD34+) isolation step and the samples will be aliquoted and frozen at -80 degrees centigrade for subsequent analysis.

• Study Type: Observational
• Enrollment (Actual): 19

Contacts and Locations

Study Locations

• Hong Kong
  • Hong Kong, Hong Kong
    • Hong Kong Sanatorium & Hospital

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 16 years and older (Adult, Older Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

• Sampling Method: Non-Probability Sample

Study Population

Highly allergic individuals

Description

Inclusion Criteria:

Any patients with the above diagnoses -

Exclusion Criteria:

Children < 18 years old; those who are unwilling or unable to donate blood; pregnant mothers.

-

Study Plan

How is the study designed?

Design Details

• Observational Models: Cohort
• Time Perspectives: Cross-Sectional

Number of groups / cohorts

7

Cohorts and Interventions

Group / Cohort
urticaria; Patients with this condition
asthma; Patients with this condition
eczema; Patients with this condition
food allergy; Patients with this condition
anaphylaxis; Patients with this condition
mastocytosis; Patients with this condition
mast cell activating syndrome; Patients with this condition

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Mast Cell Activation Test Results for Validation Group	Mast Cell Activation Test results of high/low level of house dust mite sensitivity by using normal donor cultured mast cells for validation.	Mast cells studied after 9 weeks in culture
Evaluation of Patient's Serum for Autoreactivity on Normal Average Responder Mast Cell Activation	Patient's serum was incubated with mast cells derived from normal donors for detection of mast cell activation as evidenced by histamine release.	Mast cells studied after 9 weeks in culture
Percentage Histamine Release by Normal Average Responder Mast Cells + Patient's Serum to Assess Autoreactivity	percentage of total histamine release by normal average responder mast cells that had been activated with patient's serum	Mast cells studied after 9 weeks in culture
Percentage Histamine Release by Patient's Mast Cells + Immunoglobulin E (IgE)/Anti-IgE	percentage of total histamine release by patient's mast cells that had been activated with anti-IgE	Mast cells studied after 9 weeks in culture
Percentage of Histamine Release by Patient's Mast Cells + Patient's Serum + Allergen	Percentage of total histamine release when patient's mast cells were activated with patient's own serum and Der p2. Each patient's results are reported individually.	Mast cells studied after 9 weeks in culture
Cell Yield Per 100ml Blood	Cell yield of patient's mast cell culture after 9 weeks in culture as an index of cell growth and differentiation.	Mast cells studied after 9 weeks in culture

Collaborators and Investigators

• Sponsor: Hong Kong Sanatorium & Hospital
• Collaborators: Chinese University of Hong Kong; Stanford University
• Investigators: Principal Investigator: Tak H Lee, Hong Kong Sanatorium and Hospital

Publications and helpful links

General Publications

• Tam IYS, Ng CW, Tam SY, Lau HYA. Novel six-week protocol for generating functional human connective tissue-type (MCTC) mast cells from buffy coats. Inflamm Res. 2017 Jan;66(1):25-37. doi: 10.1007/s00011-016-0989-z. Epub 2016 Sep 15. Erratum In: Inflamm Res. 2017 Nov 13;:

Study record dates

Study Major Dates

• Study Start (Actual): February 1, 2018
• Primary Completion (Actual): December 1, 2019
• Study Completion (Actual): December 1, 2019

Study Registration Dates

• First Submitted: January 14, 2018
• First Submitted That Met QC Criteria: January 14, 2018
• First Posted (Actual): January 23, 2018

Study Record Updates

• Last Update Posted (Actual): April 9, 2020
• Last Update Submitted That Met QC Criteria: March 27, 2020
• Last Verified: March 1, 2020

More Information

Terms related to this study

• Keywords: biomarkers; allergy; genomic; mast cell
• Additional Relevant MeSH Terms: Skin Diseases; Immune System Diseases; Neoplasms, Connective and Soft Tissue; Neoplasms by Histologic Type; Neoplasms; Hypersensitivity, Immediate; Skin Diseases, Vascular; Hypersensitivity; Neoplasms, Connective Tissue; Immune Complex Diseases; Food Hypersensitivity; Urticaria; Mastocytosis; Chronic Urticaria; Anaphylaxis
• Other Study ID Numbers: REC-2017-17

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: No

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No