Claims
- 1. A method for in vitro detection of C5b-9-mediated activation or injury of platelets intended for transfusion comprising
- exposing the donor platelets in a solution suitable for transfusion, prior to transfusion, to blood serum or plasma from the intended recipient containing active complement which may activate the platelets,
- adding to the platelets a fluorescent potentiometric dye indicator, wherein the fluorescence emission intensity changes in response to changes in the cell plasma membrane potential, and
- measuring the resulting change in fluorescence emission intensity of the potentiometric dye indicator relative to the fluorescence emission intensity of a fluorescent potentiometric dye indicator added to control platelets exposed to autologous human plasma or serum to determine the amount of C5b-9 activation of the platelets by complement.
- 2. The method of claim 1 wherein the platelets to be transfused have been treated prior to transfusion with a C5b-9 inactivator having the ability to inhibit C5b-9 mediated platelet or endothelial cell C5b-9 activation and cytolysis selected from the group consisting of an 18 kDa C5b-9 inhibitory protein on erythrocyte membranes, peptide fragments thereof having C5b-9 inhibitory activity, monoclonal antibodies to C7 that block membrane binding of the C5b-9, monoclonal antibodies to C9 that block C9 polymerization and insertion into the membrane, monoclonal antibodies that block C9 binding to C5b-9, and anti-idiotypic antibodies which inhibit the function of the cell surface or membrane bound molecules in inhibiting C5b-9 activity, wherein the molecular weights are determined by SDS-PAGE under non-reducing conditions, and the inhibitor proteins are of the same origin as the complement proteins to be inhibited.
- 3. The method of claim 1 wherein the dye is a carbocyanine.
- 4. The method of claim 1 wherein the fluorescence of the platelets after addition of the dye is measured by fluorescence spectrophotometry.
- 5. The method of claim 1 wherein the fluorescence of the platelets after addition of the dye is measured by fluorescence flow cytometry.
- 6. The method of claim 1 wherein the potentiometric dye indicator is a cyanine dye.
- 7. The method of claim 6 wherein the potentiometric dye indicator is diS-C3(5).
- 8. The method of claim 1 wherein the platelets are in a blood banking unit and are exposed to complement from the intended recipient.
Parent Case Info
This application is a Divisional of U.S. Ser. No. 08/465,548 filed Jun. 5, 1996, now U.S. Pat. No. 5,660,825, by Peter J. Sims and Therese Wiedmer entitled "Inhibition of Complement Mediated Inflammatory Response," which is a divisional of U.S. Ser. No. 08/243,540 filed May 16, 1994, now U.S. Pat. No. 5,550,108, which is a continuation of U.S. Ser. No. 07/813,432 filed Dec. 24, 1991, now abandoned, which is a divisional of U.S. Ser. No. 07/365,199 filed Jun. 12, 1989, now U.S. Pat. No. 5,135,916.
Government Interests
The U.S. Government has rights in this invention by virtue of certain government grants.
US Referenced Citations (1)
Number |
Name |
Date |
Kind |
4762701 |
Horan et al. |
Aug 1988 |
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Non-Patent Literature Citations (2)
Entry |
Sims et al J. Biol. Chem. vol. 263 p. 18105, Dec. 1988. |
Sims et al, Biochemistry vol. 13 p. 3315, 1974. |
Divisions (3)
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Number |
Date |
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Parent |
465548 |
Jun 1996 |
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Parent |
243540 |
May 1994 |
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Parent |
365199 |
Jun 1989 |
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Continuations (1)
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813432 |
Dec 1991 |
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