Sukrit Kapoor Here are a few suggestions:

• Include a mock infection control (cells + media only) to account for background cytokine production from handling or incubation.
• Optimize your MOI (multiplicity of infection) carefully—too high may cause excessive cell death, too low might not trigger a detectable response.
• Be sure to run biological replicates (from different donors or PBMC preps) to capture variability, and technical replicates to ensure consistency within each condition.
• Lastly, time points should be based on expected cytokine kinetics—early (e.g., 4–6h) and late (e.g., 24h) can capture both innate and adaptive responses.