pre-clinical studies the animal model selected to perform the preclinical studies. the protocol of...
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Pre-clinical studies
The animal model selected to perform the preclinical studies.
The protocol of vaccination
Evaluating the safety, immunogenicity and efficacy of the PFP vaccine.
The expect results.
Out line:
Guniea pig as Animal model?
• guinea pig model has been extensively used to test vaccine candidates
• This model is more susceptible to the disease than the mouse
• Has elements of pathology very similar to that seen in humans
Toxicity study • 4 group • 12 Hartley guinea pig per group (male/female)• ~ 300 gm weight Reproductive toxicity study• 2 groups• 6 female Hartley guinea pig per group• ~ 500 gm weight
Toxcicity study design Single dose toxicity Repeated dose toxicity Immunotoxicty Reproductive toxicity studies Safty
Vaccination and challenge Protocol
Immunization route: intradermaly( Same route as planned for clinical trial addministration)
Prime immunization:• One Dose BCG (102 CFU)
Boost(s):• Ag85 A-PPE44-RV2660 (PFP)• Volume: 100 μl formulated in IC-31 as adjuvant
Challenge: low-dose aerosol of M.tb (105 CFU)
groups BCG 1st 2nd dose
A √ placebo - IC31
B1 √ √ - 25 μg
B2 √ √ - 50 μg
C √ √ √ 25 μg
Experimental groups
0 9 12 16 19 29 w
BCG 1st 2nd challenge
0 9 13 16 26 w
BCG 1st challenge
• BCG at day 0• First boost 9 weeks later• Group C, received 2nd boost 3 week later• Challenge 4 weeks after final boost• Animals bled 1 day before challenge • 4 animals sacrificed at 1 hr, 3 & 13 weeks after challenge
groups BCG 1st dose
D1 √ placebo adjuvant
D2 √ √ 50 μg
Reproductive toxicity studies
0 1 8 10 w
BCG 1st
• BCG at day 0• Pregnancy 1 week later• Boost 8 week after BCG immunisation• Expected to deliver 2 weeks after boost
Evaluating the safety and efficacy
Evaluating the safetySurvivalchange in Feeding behaviorMeasuring change in body
weight
Maternal toxicity
• Food consumption, body weight, • Clinical observations, fertility index and gestational index
Histopathology analysis
Lung section Comparing the histopathology of the lungs
of animals in diffrent groups
EfficacyCounting the bacterial load [CFU] in lung and spleen
( homogenizing the lungs and plating on nutrient agar H711, and calculating the number of colonies)
• Humoral immunity: IgG level
• T cell population in lung and spleen
• lymphocyte proliferation assay from spleen
• No change in feeding behaviour
• No body weight loss
• Fewer and smaller lesion in lung comparing
to animals immunized with BCG
• lower CFU in lung and spleen, comparing to control
group immunized with BCG (>105)
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