Novel usage of soluble P-lectin and anthrax lethal toxin
A soluble, selectin technology, used in medical preparations containing active ingredients, resistance to vector-borne diseases, blood diseases, etc. The effect of promoting blood coagulation, good effect
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example 1
[0044] Example 1 Plasma agglutination test of hemorrhagic dengue fever and anthrax toxin
[0045] In the plasma agglutination test, the blood of mouse C57BL / 6J was centrifuged at 1,500×g for 25 minutes to obtain “platelet-poor plasma (PPP)” of the mouse. The resulting plasma was added to an equal volume of 20 millimolar calcium chloride (CaCl 2 ), at 800rpm and 37°C, the changes in absorbance due to agglutination were recorded with an aggregometer (aggregometer: model 600B, Ion-Trace, Stouffville, Canada) and the plasma clotting time was measured.
[0046] In this example, the following reagents were used for plasma agglutination test: recombinant P-selectin-Ig Fc protein (purchased from R & D Systems Inc., Minneapolis, Minnesota, USA) as the source of P-selectin. The dengue virus protein uses the first non-structural protein of dengue virus, which is derived from the first non-structural protein gene ( Chang et al., 2002, J Infect Dis 186, 743-51). The anthrax toxin uses r...
example 2
[0049] Example 2 Mice protection test of anthrax toxin and hemorrhagic snake venom
[0050] In the protective power test of P-selectin on mice, the mice were first administered with recombinant P-selectin-Ig protein. Four hours after P-selectin was added with neutralizing monoclonal antibody or human immunoglobulin IgG (both at a dose of 1.2 g / g), anthrax toxin and hemorrhagic snake venom were administered intravenously. Then the death time of the mice was observed. The sources of recombinant P-selectin-Ig protein and anthrax toxin are shown in Example 1. The hemorrhagic snake venoms used the venoms of western diamondback rattlesnake (Crotalus atrox) and eastern diamondback rattlesnake (Crotalus adamanteus) (purchased from Sigma, St. Louis, Missouri, USA).
[0051] Such as figure 2 As shown in A, after administering the venom of western diamondback rattlesnake (Crotalus atrox) and eastern diamondback rattlesnake (Crotalus adamanteus), all six mice (C57BL / 6J) died within 1 ...
example 3
[0053] Mice protection test of example 3 bacteremia
[0054] The reagents and methods of the mouse protection test are shown in Example 2. The mouse bacteremia line will be 1 x 10 8 The number of bacteria per gram of Escherichia coli was injected into the plasma of mice (C57BL / 6J) to simulate bacteremia. As shown in Table 1, all 6 mice injected with the E. coli solution died within 24 hours. However, mice given prior recombinant soluble P-selectin survived an average of 2 days longer. Therefore, soluble P-selectin does have a protective effect on bacteremic mice.
[0055] Mice Handling
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