All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Induction of the adjuvant-induced arthritis (AA) mouse model
<ol>
	<li>Suspend complete Freund's adjuvant (CFA) by vortexing at least 5 s, then draw 100 &#181;L of suspension into an insulin injector. 
	NOTE: We recommend using a completely new CFA in experiments to ensure sterility.</li>
	<li>Anesthetize mice as described.</li>
	<li>Mark the chosen paw and inject 20 &#181;L of CFA into the ankle joint space. Inject 20 &#181;L of suspension into four periarticular spots on the chosen paw (80 &#181;L in total).</li>
	<li>Remove mice from the breathing unit and put the processed mice in a new chamber. Monitor mice to ensure that they are breathing until they regain the ability to move.</li>
	<li>Every 3 days, assess the joint diameter by measuring the ankle joint diameter using a pocket thickness gauge (Figure 1A).</li>
	<li>Every 3 days, assess arthritis severity by arthritis scoring criterion (Figure 1C): 0, normal, no evidence of erythema and swelling; 1, the mildest arthritis, erythema and mild swelling confined to the tarsals or ankle joint; 2, moderate arthritis, erythema and mild swelling extending from the ankle to the tarsals; 3, severe arthritis, erythema and moderate swelling extending from the ankle to metatarsal joints; 4, the most severe arthritis, erythema, and severe swelling encompass the ankle, foot, and digits, or ankylosis of the limb.</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>15 mL Conical Polypropylene Centrifuge Tube</td>
			<td>Falcon</td>
			<td>14-959-53A</td>
			<td></td>
		</tr>
		<tr>
			<td>23 G x 1 1/4&#34; Needle</td>
			<td>BD</td>
			<td>305120</td>
			<td></td>
		</tr>
		<tr>
			<td>26 G x 3/8&#34; Needle</td>
			<td>BD</td>
			<td>305110</td>
			<td></td>
		</tr>
		<tr>
			<td>30 G x 1/2&#34; Needle</td>
			<td>BD</td>
			<td>305106</td>
			<td></td>
		</tr>
		<tr>
			<td>Complete Freund&#39;s Adjuvant, 10 mg/ml</td>
			<td>sigma</td>
			<td>1002036152</td>
			<td></td>
		</tr>
		<tr>
			<td>Eppendorf Microtubes, 1.5 mL</td>
			<td>Sigma</td>
			<td>Z606340</td>
			<td></td>
		</tr>
		<tr>
			<td>Dial Thickness Gauge</td>
			<td>Mitutoyo</td>
			<td>7301</td>
			<td></td>
		</tr>
		<tr>
			<td>Shu Rui U40 Disposable Sterile Insulin Injection Needle 1 mL</td>
			<td>BD</td>
			<td>328421</td>
			<td></td>
		</tr>
	</tbody>
</table>

a technique to generate an adjuvant-induced arthritis mouse model

A Technique to Generate an Adjuvant-Induced Arthritis Mouse Model

Source: Lu, Q., et al. <a href='https://app.jove.com/v/60543'>Detecting Migration and Infiltration of Neutrophils in Mice.</a> J. Vis. Exp. (2020).
This video demonstrates a method to generate adjuvant-induced arthritis in a murine model. Complete Freund's adjuvant (CFA) is injected in the mouse ankle joint and periarticular areas. CFA activates the immune system, leading to neutrophil infiltration and exaggerated inflammation.

<img alt="Rheumatoid arthritis analysis; A: caliper measurement; B-D: joint diameter and score graphs, mouse paws." src="/files/ftp_upload/21709/21709_Figure1.jpg" data-altupdatedat="1753371239000" data-alt="Figure 1"> 
Figure 1: Representative results of the adjuvant-induced arthritis (AA) mouse model. (A) The ankle joint diameter was measured using a pocket thickness gauge. (B) Joint swelling was assessed based on the ankle joint dia...

Source: Lu, Q., et al. Detecting Migration and Infiltration of Neutrophils in Mice. J. Vis. Exp. (2020).
This video demonstrates a method to generate ...

To generate an adjuvant-induced arthritis mouse model, begin with an anesthetized mouse in the prone position.
Take a syringe containing complete Freund's adjuvant or CFA, a suspension of heat-killed Mycobacterium tuberculosis in oil. Inject the CFA into the ankle joint space &#8212; a synovial fluid-filled region between the lower leg bone and foot bone &#8212; followed by injection into the tissue surrounding the joint, inducing chronic inflammation.
The fluid-filled cavity is bordered by a synovial membrane, consisting of a lining layer enriched with macrophages and fibroblasts &#8212; functioning as an immunological barrier &#8212; and a sub-lining interstitial tissue.
The macrophages recognize the bacterial antigen and transform to a phagocytic phenotype, disrupting the barrier integrity. The macrophages also release chemoattractants and cytokines. The released chemoattractants mediate the recruitment of circulatory neutrophils to the infection site.
Recognition of the bacterial antigen activates neutrophils, resulting in degranulation. Released matrix metalloproteinases and proteases cause cartilage damage and sustained inflammation.
Macrophage-secreted cytokines also bind to receptors on the synovial fibroblasts. The cytokine binding and antigen recognition induce fibroblast activation.
Activated fibroblasts undergo abnormal proliferation, resulting in synovial hyperplasia &#8212; leading to excess synovial fluid production and joint swelling, characteristic of arthritis.

Suspend complete Freund's adjuvant by vortexing at least five seconds. Then, draw 100 microliters of suspension into an insulin injector.
After anesthetizing, mark the chosen paw and inject 20 microliters of complete Freund's adjuvant from the injector into four periarticular spots on the ankle joint space. Put the processed mice in a new chamber. Monitor the mice to ensure that they are breathing until they regain the ability to move. Every three days, use a pocket thickness gauge to measure the ankle joint diameter.
Also, assess arthritis severity by arthritis scoring criterion. Zero is normal. No evidence of erythema and swelling.
One is the mildest arthritis. Erythema and mild swelling confined to the tarsals or ankle joint. Two is the moderate arthritis. Erythema and mild swelling extending from the ankle to the tarsals. Three is the severe arthritis. Erythema and moderate swelling extending from the ankle to metatarsal joints. Four is the most severe arthritis. Erythema and severe swelling encompass the ankle, foot, and digits, or ankylosis of the limb.

a-technique-to-generate-an-adjuvant-induced-arthritis-mouse-model

Research

JoVE Encyclopedia of Experiments

Immunology

Immunopathology

Host-Pathogen Interaction Models

2.8K Views. To generate an adjuvant-induced arthritis mouse model, begin with an anesthetized mouse in the prone position. Take a syringe containing complete Freund's adjuvant or CFA, a suspension of heat-killed Mycobacterium tuberculosis in oil. Inject the CFA into the ankle joint space — a synovial fluid-filled region between the lower leg bone and foot bone — followed by injection into the tissue surrounding the joint, inducing chronic inflammation. The fluid-filled ...

Abstract

Protocol

<img title="figure-results-1" alt="Rheumatoid arthritis analysis; A: caliper measurement; B-D: joint diameter and score graphs, mouse paws." src="/files/ftp_upload/21709/21709_Figure1.jpg" data-altupdatedat="1753371239000" data-alt="Figure 1"> 
Figure 1: Representative results of the adjuvant-induced arthritis (AA) mouse model. (A) The ankle joint diameter was measured using a pocket thickness gauge. (B) Joint swelling was assessed based on the ankle joint dia...

A Technique to Generate an Adjuvant-Induced Arthritis Mouse Model

In This Article

Abstract

Protocol

Results

Disclosures

Materials

Reprints and Permissions

Tags

Related Articles

Name	Company	Catalog Number
15 mL Conical Polypropylene Centrifuge Tube	Falcon	14-959-53A
23 G x 1 1/4" Needle	BD	305120
26 G x 3/8" Needle	BD	305110
30 G x 1/2" Needle	BD	305106
Complete Freund's Adjuvant, 10 mg/ml	sigma	1002036152
Eppendorf Microtubes, 1.5 mL	Sigma	Z606340
Dial Thickness Gauge	Mitutoyo	7301
Shu Rui U40 Disposable Sterile Insulin Injection Needle 1 mL	BD	328421