Intradermal Immunization with Live Attenuated Sporozoites in a Mouse Model
Published: May 31, 2024
Abstract
Source: Mac-Daniel, L., et al. Myeloid Cell Isolation from Mouse Skin and Draining Lymph Node Following Intradermal Immunization with Live Attenuated Plasmodium Sporozoites. J. Vis. Exp. (2016).
This video demonstrates an assay for immunizing a mouse model with live attenuated sporozoites. The radiation-attenuated Plasmodium sporozoites are injected into the dorsal side of the mouse's ear. After injections at multiple locations on both ears, the mouse is allowed to recover.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Materials and Reagents Use female Anopheles stephensi mosquitoes (Sda500 strain) that feed on infected mice 3-5 days after emergence and rear as described previously. Use parasites Plasmodium berghei ANKA clone expressing the Green Fluorescent Protein (GFP) gene under the control of the const…
Representative Results
Figure 1: Intradermal Injection of Sporozoites into the Ear of Mouse. (A) Image showing intradermal injection of sporozoites in the ear pinna of an anesthetized mouse. The ventral side of the ear is stabilized with clear tape previously placed under a dissecting microscope. The needle is carefully inserted on the dorsal side of the ear, beneath the epidermis, wit…
Declarações
The authors have nothing to disclose.
Materials
| Ketamine: Imalgene® 1000 | Merial | – | – |
| Xylazine: Rompun® 2% | Bayer | – | – |
| NanoFil syringe + 35 gauge needle | World Precision Instruments | – | – |
| Omnican® 50 Insulin syringe 0,5 ml/50 I.U. | B. Braun Medical | 9151125 | – |
| MultiwellTM 6 well tissue culture plate – Flat Bottom | BD Falcon | 353046 | – |
| 70 µm cell strainer | BD Falcon | 352350 | – |
| 2 ml syringe | Terumo | SS-02S | – |
| BLUE MAXTM 15ml Polypropylene conical tube | BD Falcon | 352097 | – |
| BLUE MAXTM 50ml Polypropylene conical tube | BD Falcon | 352098 | – |
| 5ml Polystyrene Round-Bottom Tube with 35µm Cell-Strainer Cap | BD Falcon | 352235 | – |
| DPBS 1X Cacl2– and MgCl2-free | Life Technologies | 14190-094 | – |
| DMEM 1X + GlutaMAXTM | Life Technologies | 31966-021 | – |
| Deoxyribonuclease I from bovine pancreas, type IV | Sigma-Aldrich | D5025 | 50 µg/ml |
| Female C57BL/6JRj mice (7-week-old) | Janvier Laboratories | – | – |
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Intradermal Immunization with Live Attenuated Sporozoites in a Mouse Model. J. Vis. Exp. (Pending Publication), e22279, doi: (2024).