Anti-Ly6C Monoclonal Antibody Treatment
Objective: To assess the effects of anti-Ly6C monoclonal antibody treatment on inflammatory monocyte modulation, monocyte recruitment to the spinal cord, neuronal loss, and survival in a mouse model of ALS
Gather these items before starting the experiment. Check off items as you prepare.
Materials1
Not explicitly stated in provided text • Not mentioned • Not mentioned • Not mentioned
As an Amazon Associate, we earn from qualifying purchases. Product links help support this free resource.
Protocol Steps
Establish ALS mouse model baseline
Use mouse model of ALS to establish baseline conditions prior to disease onset and monitor disease progression
Note: Monitoring includes assessment of splenic Ly6C hi monocytes and microglia phenotype
View evidence from paper
“Prior to disease onset, splenic Ly6C hi monocytes expressed a polarized macrophage phenotype (M1 signature)”
Monitor monocyte activation and recruitment
Track inflammatory monocyte activation and their progressive recruitment to the spinal cord, correlating with neuronal loss
Note: Recruitment to spinal cord but not brain was monitored; resident microglia decrease was also assessed
View evidence from paper
“inflammatory monocytes were activated and that their progressive recruitment to the spinal cord, but not brain, correlated with neuronal loss”
Assess microglia chemotaxis signaling
Evaluate microglia expression of chemotaxis-associated molecules including CCL2 as disease onset approaches
Note: Microglia-derived signals lead to monocyte recruitment to CNS
View evidence from paper
“As disease onset neared, microglia expressed increased CCL2 and other chemotaxis-associated molecules, which led to the recruitment of monocytes to the CNS”
Administer anti-Ly6C monoclonal antibody treatment
Treat ALS mice with anti-Ly6C mAb to modulate inflammatory monocytes
Note: Treatment timing relative to disease progression not specified in provided text
View evidence from paper
“Treatment with anti-Ly6C mAb modulated the Ly6C hi monocyte cytokine profile, reduced monocyte recruitment to the spinal cord”
Assess treatment effects on monocyte cytokine profile
Evaluate changes in Ly6C hi monocyte cytokine profile following anti-Ly6C mAb treatment
Note: Cytokine profile modulation is a primary outcome of treatment
View evidence from paper
“Treatment with anti-Ly6C mAb modulated the Ly6C hi monocyte cytokine profile”
Quantify monocyte recruitment reduction
Measure reduction in monocyte recruitment to the spinal cord following treatment
Note: Spinal cord-specific recruitment is the focus, not brain recruitment
View evidence from paper
“reduced monocyte recruitment to the spinal cord, diminished neuronal loss, and extended survival”
Assess neuronal loss
Evaluate neuronal loss in treated versus untreated ALS mice
Note: Neuronal loss is correlated with monocyte recruitment
View evidence from paper
“diminished neuronal loss, and extended survival”
Monitor survival
Track survival duration in anti-Ly6C mAb treated mice compared to controls
Note: Extended survival is a primary outcome measure
View evidence from paper
“Treatment with anti-Ly6C mAb modulated the Ly6C hi monocyte cytokine profile, reduced monocyte recruitment to the spinal cord, diminished neuronal loss, and extended survival”