The proposed experiments will test the hypothesis: The separation of leukocytes by their intrinsic properties using Dean Flow Fractionation will enable analyses of innate and adaptive immune responses in microliter quantities of whole blood. Acute and chronic inflammatory responses are integral to the clinical expression of many diseases, yet routine clinical assessment of immune function is based solely on leukocyte count. This is typically measured with a complete blood count and leukocyte differential count. In current clinical practice, critical clinical decisions regarding immune modulation are made based on the count of leukocytes and leukocyte subclasses without assessment of leukocyte function. In addition, the measurement of a peripheral venous complete blood count uses 3-5 mL. Based on the significant blood volume required for the complete blood count and other routine laboratory testing, patients can develop iatrogenic anemia. There is a critical need to develop assays that enable assessment of immune function and require significantly smaller volumes of blood. In preliminary results, we have developed an inertial microfluidic approach to selectively separate diverse cellular targets that is based on non-labeled, cell type specific physical features. From peripheral venous blood, this new approach can separate leukocytes from erythrocytes and platelets, leukocyte subclasses and distinguish activated neutrophils from unactivated cells by tandem iso-dielectric separation. This multi-functional, extracorporeal separation system is high throughput and capable of preparatory separation of leukocytes prior to coupled analyses of systemic and cellular immune function. To address our hypothesis, two specific aims are proposed: 1. Design microliter scale preparatory inertial microfluidic separation of leukocytes in peripheral blood; 2. Determine microliter scale assays of systemic and cell-based immune function. Recent advances in high-throughput microfluidic engineering enable this project, paving the way for a new approach to complex immune monitoring.

Public Health Relevance

Some of the most common and important conditions are secondary to allergy, infection, disordered immune responses and transplantation. There is a critical need for assays to monitor immunological changes associated with disease and responses to immunomodulatory treatment that are feasible with only small volumes of precious blood. In this application, we will prepare a novel microfluidic device to provide leukocytes and plasma for systemic and cell-based diagnostic measures of immune function.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Resource-Related Research Projects--Cooperative Agreements (U24)
Project #
5U24AI118656-03
Application #
9278111
Study Section
Special Emphasis Panel (ZAI1-MM-I (M1))
Program Officer
Bourcier, Katarzyna
Project Start
2015-06-23
Project End
2020-05-31
Budget Start
2017-06-01
Budget End
2018-05-31
Support Year
3
Fiscal Year
2017
Total Cost
$383,060
Indirect Cost
$75,187
Name
Brigham and Women's Hospital
Department
Type
Independent Hospitals
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
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