Biology Reference
In-Depth Information
1.2
Conclusions
EPO and derivatives of EPO devoid of erythropoietic activity (e.g.,
CEPO, asialoEPO) ameliorate the clinical course of EAE. Efficacy
in several models has been reported. Other than inhibiting
inflammation, EPO and derivatives inhibit demyelination and pro-
mote neuronal and axonal protection, suggesting that they might
actively induce nervous system repair. In a pilot clinical trial, EPO
has shown efficacy in patients with primary progressive MS, improv-
ing motor and cognitive functions.
In the following section, we report an example of effectiveness
of EPO and CEPO in ameliorating the clinical score of EAE (
21
).
We describe the protocol to induce EAE in C57BL/6 by immuni-
zation with MOG peptide 35-55 and the schedule and doses of
EPO and CEPO administration. Importantly, in this model both
EPO and CEPO work not only when administered before onset
but also when given according to a therapeutic schedule. Detailed
protocols to induce EAE in C57BL/6 and SJL mouse strains by
immunization with several neuroantigens, including MOG peptide
35-55, have been previously reported (
13, 25
).
2
Materials
1. Female C57BL/6 mice, 6-8 weeks old.
2. Recombinant human erythropoietin (rhEPO).
3. Carbamylated EPO (CEPO) and asialoEPO, prepared as
described (
20, 26
).
4. MOG peptide 35-55 (MEVGWYRSPFSRVVHLYRNGK).
5. Incomplete Freund's adjuvant.
6. Heat-inactivated
Mycobacterium tuberculosis
, strain H37Ra.
7. Pertussis toxin from
Bordetella pertussis
.
8. Pyrogen-free phosphate buffered saline (PBS).
9. Pyrogen-free saline solution.
10. Two 5 ml glass syringes connected by a 2-way luer lock
adapter.
11. 1 ml syringes with removable needle.
12. 21-gauge needles.
13. Two small glass beakers.
14. Magnetic stirrer.
15. Infrared lamp.
16. Mouse restrainer for intravenous injections.
17. Ice.
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