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

• US 5,460,945 A
• Filed: 05/20/1992
• Issued: 10/24/1995
• Est. Priority Date: 05/30/1991
• Status: Expired due to Term

• Current Assignee: Immune Disease Institute, Inc.
• Original Assignee: Center For Blood Research Incorporated
• Inventors: Lawrence, Michael, Springer, Timothy A.
• Primary Examiner(s): SAUNDERS, DAVID A
• Application Number:
  US07/887,444
• Time in Patent Office: 1,252 Days
• Field of Search: 422/69, 422/58, 427/2, 435/2, 435/7.23, 435/7.24, 435/7.8, 435/29, 435/30, 435/174, 435/176, 435/177, 435/287, 435/240.2, 436/503
• US Class Current: 435/7.24
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61M 1/3679 by absorption A61M1/3675 takes precedence
• CPC Class Codes: A61M 2202/0042 filtrate, i.e. the fluid passing through the filter
• CPC Class Codes: A61M 2202/0057 retained by adsorption
• CPC Class Codes: A61M 2202/0429 Red blood cells; Erythrocytes
• CPC Class Codes: A61M 2202/0439 White blood cells; Leucocytes
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism
• CPC Class Codes: G01N 33/56972 White blood cells

Abstract

The present invention provides in vitro models of the in vivo rolling and arrest of leukocytes along the endothelial cell wall, which are important steps in the migration of leukocytes out of the blood stream and into tissue, as part of the inflammatory response. The in vitro models of the invention are functional under physiologic flow conditions resulting in physiologic shear stresses. In a specific embodiment, for modelling leukocyte rolling, the apparatus of the invention comprises a solid phase surface with rolling mediator molecules present thereon. Such rolling mediators are, for example, selectins and selectin ligands which have binding partners expressed on leukocytes. In another specific embodiment, for modelling leukocyte rolling followed by adhesion/arrest, the apparatus of the invention comprises a solid phase surface with both rolling mediators and integrin binding partners present thereon. The apparatuses of the invention can be used for collecting, concentrating, purifying, and analyzing blood and blood components, in particular, leukocytes and subsets thereof. The invention further relates to methods for identifying inhibitors or, alternatively, promoters (agonists, functional components) of the inflammatory response. Therapeutic and diagnostic methods, pharmaceutical compositions and kits are also provided.

Claims

1. 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;

(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;

(c) a means associated with the inlet for introducing said sample into the housing; and

(d) a means associated with the outlet for withdrawing said sample from the housing, said apparatus being capable of permitting detection of leukocyte rolling on the solid phase surface.

40. The apparatus of claim 1, 4 or 30 in which said molecules are CD62.

41. The apparatus of claim 1, 4 or 30 in which said molecules are selected from the group consisting of CD62, ELAM-1, MECA-79 antigen, and any combination of the forgoing.

2. 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.

3. The solid phase of claim 2 wherein said surface also contains a plurality of one or more substantially purified integrin binding partners.

4. 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.

5. 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.

6. The solid phase of claim 5 in which said surface also contains a plurality of one or more substantially purified integrin binding partners.

7. The solid phase of claim 5 in which said molecules are CD62.

12. The solid phase of claim 8 or 6 in which said molecules are CD62, and the integrin binding partners are ICAM-1.

8. 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.

9. The solid phase of claim 8 in which the integrin binding partners are selected from the group consisting of ICAM-1, ICAM-2, VCAM-1, and any combination of the foregoing.

10. The solid phase of claim 8 in which the solid phase is plastic.

11. 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.

13. An apparatus comprising:

(a) a solid phase having (i) a plurality of one or more substantially purified molecules contained on a surface of the solid phase, said molecules being capable of mediating rolling of a leukocyte on said solid phase surface, and (ii) a plurality of one or more substantially purified integrin binding partners contained on said surface;

(b) inlet means for receiving a fluid sample and for permitting the sample to enter onto the surface of the solid phase; and

(c) outlet means for permitting the fluid sample after it has flowed across at least a portion of the surface of the solid phase to exit said surface.

14. The apparatus of claim 13 which further comprises a means associates with the outlet means, for removing fluid from the outlet means.

15. The apparatus of claim 13 which further comprises a means associated with the inlet means, for introducing the fluid sample into the inlet means, and a means associated with the outlet means, for removing the fluid sample from the outlet means.

16. The apparatus of claim 15 further comprising a housing containing said solid phase, inlet means, and outlet means.

17. The apparatus of claim 16 in which said molecules are CD62, and the integrin binding partners are ICAM-1.

18. The apparatus of claim 16 in which said molecules are selected from the group consisting of CD62, ELAM-1, MECA-79 antigen, and any combination of the foregoing;

and the integrin binding partners are selected from the group consisting of ICAM-1, ICAM-2, VCAM-1, and any combination of the foregoing.

19. The apparatus of claim 13 further comprising a housing containing said solid phase, inlet means, and outlet means.

20. The apparatus of claim 19 in which the housing provides a container for fluid flowing past the surface of the solid phase.

21. The apparatus of claim 20 in which physiologic shear stress can be achieved at the surface of the solid phase.

22. The apparatus of claim 21 in which the shear stress is in the range of from about 0.5 to about 4.0 dynes per square centimeter.

23. The apparatus of claim 22 in which said molecules are CD62, and the integrin binding partners are ICAM-1.

24. The apparatus of claim 19 further comprising a video camera or microscope operatively linked for viewing the surface of the solid phase.

25. The apparatus of claim 13 which further comprises a pump associated with the outlet means.

26. The apparatus of claim 13 in which said molecules are CD62.

27. The apparatus of claim 13 in which said molecules are CD62, and the integrin binding partners are ICAM-1.

28. The apparatus of claim 13 in which said molecules are selected from the group consisting of CD62, ELAM-1, and MECA-79 antigen, and any combination of the foregoing;

and the integrin binding partners are selected from the group consisting of ICAM-1, ICAM-2, VCAM-1, and any combination of the foregoing.

29. The apparatus of claim 13, 27, or 17 in which the solid phase is a planar phase comprising glass, and a planar lipid bilayer is present on said surface, said lipid bilayer containing said molecules and the integrin binding partners.

30. An apparatus comprising:

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

(b) inlet means for receiving a fluid sample and for permitting the sample to enter onto the surface of the solid phase; and

(c) outlet means for permitting the fluid sample after it has flowed across at least a portion of the surface of the solid phase to exit said surface;

(d) a means associated with the inlet means, for introducing the fluid sample into the inlet means;

(e) a means associated with the outlet means, for removing the fluid sample from the outlet means;

said apparatus being capable of permitting detection of leukocyte rolling on the solid phase surface.

31. The apparatus of claim 30 further comprising a housing containing said solid phase, inlet means, and outlet means.

32. The apparatus of claim 31 in which the housing provides a container for fluid flowing past the surface of the solid phase.

33. The apparatus of claim 32 in which physiologic shear stress can be achieved at the surface of the solid phase.

34. The apparatus of claim 33 in which the shear stress is in the range of from about 0.5 to about 4.0 dynes per square centimeter.

35. The apparatus of claim 34 in which said molecules are CD62.

36. The apparatus of claim 31 in which said molecules are CD62.

37. The apparatus of claim 36 or 35 in which the solid phase is a planar phase comprising glass, and a planar lipid bilayer is present on said surface, said lipid bilayer containing said molecules.

38. The apparatus of claim 31 in which said molecules are selected from the group consisting of CD62, ELAM-1, MECA-79 antigen, and any combination of the foregoing.

39. The apparatus of claim 30 in which the inlet means is an inlet tube or pipe, and the outlet means is an outlet tube or pipe.

42. An apparatus comprising:

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

(b) a base having (i) an inlet manifold, (ii) an outlet manifold, and (iii) a recessed lip on its lower surface;

(c) a deck positioned below the base, having (i) an entrance slot communicating with the inlet manifold, and (ii) an exit slot communicating with the outlet manifold;

(d) a longitudinal compressible gasket positioned within the recessed lip of the base, its top surface contacting the base, and having an aperture of a size and shape substantially congruent with the deck;

in which the planar solid phase is positioned on the bottom surface of the gasket, such that the solid phase surface with said molecules is communicating with the aperture of the gasket.

43. The apparatus of claim 42 further comprising a means for applying force whereby the gasket and solid phase are compressed together and their disengagement from the base is prevented.

44. The apparatus of claim 42 further comprising pump connected to the outlet manifold.

45. The apparatus of claim 42 in which the solid phase comprises glass;

and a planar lipid bilayer is present on said solid phase surface, said lipid bilayer containing said molecules.

46. The apparatus of claim 45 in which said molecules are CD62.

47. The apparatus of claim 42 in which a plurality of one or more substantially purified integrin binding partners are also contained on said surface of the solid phase.

48. The apparatus of claim 47 in which the solid phase comprises glass;

and a planar lipid bilayer is present on said solid phase surface, said lipid bilayer containing said molecules and the integrin binding partners.

49. The apparatus of claim 48 in which said molecules are CD62, and the integrin binding partners are ICAM-1.

50. The apparatus of claim 47 in which said molecules are CD62, and the integrin binding partners are ICAM-1.

51. The apparatus of claim 47 in which said molecules are selected from the group consisting of CD62, ELAM-1, MECA-79 antigen, and any combination of the foregoing;

and the integrin binding partners are selected from the group consisting of ICAM-1, ICAM-2, VCAM-1, and any combination of the foregoing.

52. The apparatus of claim 42 in which said molecules are CD62.

53. The apparatus of claim 42, in which said molecules are selected from the group consisting of CD62, ELAM-1, MECA-79 antigen, and any combination of the foregoing.

54. An apparatus comprising:

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

(b) inlet means for receiving a fluid sample and for permitting the sample to enter onto the surface of the solid phase;

(c) outlet means for permitting the fluid sample after it has flowed across at least a portion of the surface of the solid phase to exit said surface,

(d) a housing containing said solid phase, inlet means, and outlet means; and

(e) a video camera or microscope operatively linked for viewing the surface of the solid phase.

55. An apparatus comprising:

(a) a planar solid phase comprising glass;

(b) a planar lipid bilayer present on a surface of the solid phase, said lipid bilayer containing a plurality of one or more substantially purified molecules, said molecules being capable of mediating rolling of a leukocyte;

(c) inlet means for receiving a fluid sample and for permitting the sample to enter onto the lipid bilayer; and

(d) outlet means for permitting the fluid sample after it has flowed across at least a portion of the lipid bilayer to exit said surface.

56. 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.

57. The method according to claim 56 in which the relative movement between the solid phase and the leukocytes is such that physiologic shear stress is achieved at the surface of the solid phase.

58. The method according to claim 56 in which the leukocytes are recovered by recovering the solid phase and removing the leukocytes from the solid phase.

61. The method according to claim 56 or 59 in which said molecules are selected from the group consisting of CD62, ELAM-1, MECA-79 antigen, and any combination of the foregoing.

59. 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.

60. The method according to claim 59 in which the solid phase is comprised of glass fibers, said molecules are CD62, and a planar lipid bilayer containing CD62 is present on the surface of the solid phase.

62. 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.

63. The method according to claim 62 in which the relative movement between the solid phase and the leukocytes is 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.

64. The method according to claim 62 in which said molecules are selected from the group consisting of CD62, ELAM-1, MECA-79 antigen, and any combination of the foregoing;

and the integrin binding partners are selected from the group consisting of ICAM-1, ICAM-2, VCAM-1, and any combination of the foregoing.

65. The method according to claim 63 in which the solid phase is comprised of glass, said molecules are CD62, said integrin binding partners are ICAM-1, said chemoattractant is N-formyl methionyl leucyl phenylalanine, and a planar lipid bilayer containing CD62 and ICAM-1 is present on the surface of the solid phase.

66. 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.

67. The method according to claim 66 in which the relative movement between the solid phase and the leukocytes is such that physiologic shear stress is achieved at the surface of the solid phase.

70. The method according to claim 66 or 68 in which said molecules are selected from the group consisting of CD62, ELAM-1, MECA-79 antigen, and any combination of the foregoing.

68. 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.

69. The method according to claim 68 in which the solid phase is comprised of glass, said molecules are CD62, and a planar lipid bilayer containing CD62 is present on the surface of the solid phase.

71. 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.

72. The method according to claim 71 in which the relative movement between the solid phase and the leukocytes is such that physiologic shear stress is achieved at the surface of the solid phase.

73. The method according to claim 71 in which said molecules are selected from the group consisting of CD62, ELAM-1, MECA-79 antigen, and any combination of the foregoing;

and the integrin binding partners are selected from the group consisting of ICAM-1, ICAM-2, VCAM-1, and any combination of the foregoing.

74. A method for diagnosing a disease or disorder in a subject, comprising

(a) contacting a surface of a solid phase with a fluid sample derived from a subject and 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) measuring the number or percentage of the leukocytes or a subset thereof that roll on the surface of the solid phase;

in which an increase or decrease in said number or percentage relative to the number or percentage of leukocytes or the subset thereof that roll on the surface from an equivalent fluid sample from a healthy individual, indicates the presence of the disease or disorder in the subject.

75. The method according to claim 74 in which the relative movement between the solid phase and the leukocytes is such that physiologic shear stress is achieved at the surface of the solid phase.

76. The method according to claim 75 in which the solid phase is comprised of glass, said molecules are CD62, and a planar lipid bilayer containing CD62 is present on the surface of the solid phase.

77. The method according to claim 74 in which said molecules are selected from the group consisting of CD62, ELAM-1, MECA-79 antigen, and any combination of the foregoing.

78. The method according to claim 74 in which the disease or disorder is a leukemia and the number or percentage of rolling leukocytes or a subset thereof in the sample derived from the subject is increased.

79. The method according to claim 74 in which the disease or disorder is a neutropenia and the number or percentage of rolling leukocytes or a subset thereof in the sample derived from the subject is decreased.

80. A method for diagnosing a disease or disorder in a subject comprising

(a) contacting a surface of a solid phase with a fluid sample containing a chemoattractant and containing leukocytes, and said leukocytes obtained from a subject, 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) measuring the number or percentage of the leukocytes or a subset thereof that are arrested on the surface of the solid phase;

in which an increase or decrease in said number or percentage, relative to the number or percentage of leukocytes or the subset thereof that are arrested on the surface from an equivalent fluid sample containing leukocytes from a healthy individual, indicates the presence of the disease or disorder in the subject.

81. The method according to claim 80 in which the relative movement between the solid phase and the leukocytes is such that physiologic shear stress is achieved at the surface of the solid phase.

82. The method according to claim 80 in which said molecules are selected from the group consisting of CD62, ELAM-1, MECA-79 antigen, and any combination of the foregoing;

and the integrin binding partners are selected from the group consisting of ICAM-1, ICAM-2, VCAM-1, and any combination of the foregoing.

83. 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.

84. The method according to claim 83 in which the relative movement between the solid phase and the leukocytes is such that physiologic shear stress is achieved at the surface of the solid phase.

85. The method according to claim 84 in which said molecules are CD62.

86. The method according to claim 83 or 85 in which the solid phase is comprised of glass, and a planar lipid bilayer in which contains said molecules is present on the surface of the solid phase.

87. The method according to claim 84 in which said molecules are selected from the group consisting of CD62, ELAM-1, MECCA-79, and any combination of the foregoing.

88. 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 (i) leukocytes, (ii) a chemoattractant, and (iii) 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, thereby allowing a plurality of leukocytes in the sample 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) determining the number or percentage of arrested leukocytes, or a subset thereof, in which a decrease in such number or percentage relative to the number or percentage of leukocytes or the subset thereof which are arrested in the absence of the test compound indicates that the test compound is capable of inhibiting the inflammatory response.

89. The method according to claim 88 in which the relative movement between the solid phase and the leukocytes is such that physiologic shear stress is achieved at the surface of the solid phase.

90. The method according to claim 88 in which said molecules are CD62 and the integrin binding partners are ICAM-1.

91. The method according to claim 88 in which the solid phase is comprised of glass, and a planar lipid bilayer which contains said molecules is present on the surface of the solid phase.

92. 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.

94. The method according to claim 92 or 93 in which the relative movement between the solid phase and the leukocytes is such that physiologic shear stress is achieved at the surface of the solid phase.

93. 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 (i) leukocytes, (ii) a chemoattractant, and (iii) 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, thereby allowing a plurality of leukocytes in the sample 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) determining the number or percentage of arrested leukocytes or a subset thereof, in which a increase in such number or percentage relative to the number or percentage of leukocytes or the subset thereof which are arrested in the absence of the test compound indicates that the test compound is capable of promoting the inflammatory response.

95. 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.

96. The method according to claim 95 in which the solid phase is comprised of glass, and a planar lipid bilayer which contains the test molecules is present on the surface of the solid phase.

101. The method according to claim 95, 97, or 99 in which the relative movement between the solid phase and the leukocytes is such that physiologic shear stress is achieved at the surface of the solid phase.

97. 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.

98. The method according to claim 97 in which the solid phase is comprised of glass, said molecules are CD62, said integrin binding partners are ICAM-1, and a planar lipid bilayer containing CD62 and ICAM-1 is present on the surface of the solid phase.

99. 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.

100. The method according to claim 99 in which the solid phase is comprised of glass, said molecules are CD62, and a planar lipid bilayer containing CD62 and the test compounds is present on the surface of the solid phase.