Device and method for analysis of blood components and identifying inhibitors and promoters of the inflammatory response

A solid phase having at least one surface containing a plurality of one or more substantially purified molecules, said molecules being capable of mediating rolling of a leukocyte on the solid phase surface, wherein the solid phase has a plurality of zones on said surface, each zone containing a different said molecule.

An apparatus comprising (a) a solid phase having at least one surface containing a plurality of one or more substantially purified molecules, said molecules being capable of mediating rolling of a leukocyte on the solid phase surface and (b) a housing enclosing the solid phase, said housing having an inlet end and an outlet end for respectively receiving and removing a sample containing or suspected of containing leukocytes wherein the solid phase is a microscope slide.

A solid phase having at least one surface containing a plurality of one or more substantially purified molecules, said molecules being capable of mediating rolling of a leukocyte on the solid phase surface, wherein the solid phase is a planar phase comprising glass, and a lipid bilayer is present on said surface, said lipid bilayer containing said molecules.

A solid phase having at least one surface containing a plurality of one or more substantially purified molecules, said molecules being capable of mediating rolling of a leukocyte on the solid phase surface, wherein said surface also contains a plurality of one or more substantially purified integrin binding partners immobilized on said surface.

A solid phase having at least one surface containing (a) a plurality of one or more substantially purified molecules, said molecules being capable of mediating rolling of a leukocyte on the solid phase surface;

• and (b) a plurality of one or more substantially purified integrin binding partners;
• said solid phase further comprising a plurality of leukocytes adhering to said surface of the solid phase.

A method of collecting or purifying leukocytes from a fluid sample comprising:

• (a) contacting a surface of a solid phase with a fluid sample containing leukocytes, said surface containing a plurality of one or more substantially purified molecules, said molecules being capable of mediating rolling of a leukocyte on said solid phase surface, in which the contacting is under conditions that provide relative movement between the solid phase and the leukocytes in the sample, thereby allowing a plurality of leukocytes in the sample which express a binding partner to said molecules to roll on the surface of the solid phase;
• and
• (b) recovering the leukocytes that roll on the surface.

A method of collecting or purifying leukocytes from a fluid sample comprising:

• (a) contacting a surface of a solid phase with a fluid sample containing leukocytes, said surface containing a plurality of one or more substantially purified molecules, said molecules being capable of mediating rolling of a leukocyte on said solid phase surface, in which the contacting is under conditions that provide relative movement between the solid phase and the leukocytes in the sample such that bulk flow is induced and shear stresses at the surface of the solid phase are in the range of from about 0.5 to about 4.0 dynes per square centimeter, thereby allowing a plurality of leukocytes in the sample which express a binding partner to said molecules to roll on the surface of the solid phase;
• and
• (b) recovering the leukocytes that roll on the surface.

A method for collecting or purifying leukocytes comprising:

• (a) contacting a surface of a solid phase with a fluid sample containing a chemoattractant and containing leukocytes, said surface containing a plurality of one or more substantially purified molecules and a plurality of one or more substantially purified integrin binding partners, said molecules being capable of mediating rolling of a leukocyte on said solid phase surface, in which said contacting is under conditions that provide relative movement between the solid phase and the leukocytes in the sample, thereby allowing a plurality of leukocytes which express (i) a binding partner to said molecules, (ii) a receptor for the chemoattractant, and (iii) an integrin recognizing the integrin binding partners, to become arrested on the surface of the solid phase;
• and
• (b) recovering the arrested leukocytes from the solid phase.

A method for analysis of a fluid sample containing leukocytes, comprising

• (a) contacting a surface of a solid phase with a fluid sample containing leukocytes, said surface containing a plurality of one or more substantially purified molecules, said molecules being capable of mediating rolling of a leukocyte on said solid phase surface, in which said contacting is under conditions that provide relative movement between the solid phase and the leukocytes in the sample, thereby allowing a plurality of leukocytes which express a binding partner to said molecules to roll along the surface of the solid phase;
• and
• (b) detecting or quantifying the leukocytes that roll along the surface.

A method for analysis of a fluid sample containing leukocytes, comprising:

• (a) contacting a surface of a solid phase with a fluid sample containing leukocytes, said solid phase containing a plurality of one or more substantially purified molecules, said molecules being capable of mediating rolling of a leukocyte on said solid phase surface, in which said contacting is under conditions that provide relative movement between the solid phase and the leukocytes in the sample such that bulk flow is induced and shear stresses at the surface of the solid phase are in the range of from about 0.5 to about 4.0 dynes per square centimeter, thereby allowing a plurality of leukocytes which express a binding partner to said molecules to roll along the surface of the solid phase;
• and
• (b) detecting or quantifying the leukocytes that roll along the surface.

A method for analysis of a fluid sample containing leukocytes, comprising:

• (a) contacting a surface of a solid phase with a fluid sample containing a chemoattractant and containing leukocytes, said surface containing a plurality of one or more substantially purified molecules and a plurality of one or more substantially purified integrin binding partners, said molecules being capable of mediating rolling of a leukocyte on said solid phase surface, in which said contacting is under conditions that provide relative movement between the solid phase and the leukocytes in the sample, thereby allowing a plurality of leukocytes which express (i) a binding partner to said molecules, (ii) a receptor for the chemoattractant, and (iii) an integrin recognizing the integrin binding partners, to become arrested on the surface of the solid phase;
• and
• (b) detecting or quantifying the leukocytes arrested on the solid phase.

A method for identifying a candidate compound capable of inhibiting a component of the inflammatory response, comprising

• (a) contacting a surface of a solid phase with a fluid sample containing leukocytes and containing a test compound, said surface containing a plurality of one or more substantially purified molecules, said molecules being capable of mediating rolling of a leukocyte on said solid phase surface, in which said contacting is under conditions that provide relative movement between the solid phase and the leukocytes in the sample, thereby allowing a plurality of leukocytes which express a binding partner to said molecules to roll on the surface of the solid phase;
• and
• (b) determining the number or percentage of leukocytes or a subset thereof rolling on the surface of the solid phase, in which a decrease in said number or percentage relative to the number or percentage of leukocytes or the subset thereof which roll in the absence of the test compound indicates that the test compound is capable of inhibiting the inflammatory response.

A method for identifying a candidate compound capable of promoting a component of the inflammatory response, comprising:

• (a) contacting a surface of a solid phase with a fluid sample containing leukocytes and containing a test compound, said surface containing a plurality of one or more substantially purified molecules, said molecules being capable of mediating rolling of a leukocyte on said solid phase surface, in which said contacting is under conditions that provide relative movement between the solid phase and the leukocytes in the sample, thereby allowing a plurality of leukocytes which express a binding partner to said molecules to roll on the surface of the solid phase;
• and
• (b) determining the number or percentage of leukocytes or a subset thereof rolling on the surface of the solid phase, in which an increase in said number or percentage relative to the number or percentage of leukocytes or the subset thereof which roll in the absence of the test compound indicates that the test compound is capable of promoting the inflammatory response.

A method for identifying a molecule capable of mediating rolling of a leukocyte on a solid phase surface containing said molecule, comprising:

• (a) contacting a surface of a solid phase with a fluid sample containing leukocytes, said surface containing a plurality of a substantially purified test molecule, in which said contacting is under conditions that provide relative movement between the solid phase and the leukocytes in the sample;
• and
• (b) detecting any leukocytes that roll on the surface, in which rolling of leukocytes on the surface indicates that the test molecule is capable of mediating rolling of a leukocyte on a solid phase surface containing the test molecule.

A method for identifying a leukocyte chemoattractant, comprising:

• (a) contacting a surface of a solid phase with a fluid sample containing leukocytes and containing a test compound, said surface containing a plurality of one or more substantially purified molecules and a plurality of one or more substantially purified integrin binding partners, said molecules being capable of mediating rolling of a leukocyte on said solid phase surface, in which said contacting is under conditions that provide relative movement between the solid phase and the leukocytes in the sample;
• and
• (b) detecting any leukocytes that are arrested on the surface, in which arrest of leukocytes on the surface indicates that the test compound is a leukocyte chemoattractant.

A method for identifying an integrin binding partner, comprising:

• (a) contacting a surface of a solid phase with a fluid sample containing a chemoattractant and containing leukocytes, said surface containing a plurality of one or more substantially purified molecules and a plurality of a substantially purified test compound, said molecules being capable of mediating rolling of a leukocyte on said solid phase surface, in which said contacting is under conditions that provide relative movement between the solid phase and the leukocytes in the sample;
• and
• (b) detecting any leukocytes that are arrested on the surface, in which arrest of leukocytes on the surface indicates that the test compound is an integrin binding partner.