17
Study 38.2: Experimental Design and Summary
| Date post: | 18-Jan-2016 |
| Category: |
Documents |
| Upload: | nancy-perry |
| View: | 224 times |
| Download: | 0 times |
Study 38.2: Experimental Design and Summary
Study 38.2: Experimental Design
Day 07B2-AAA I.V.
infusion50 mg/kg
Day -6Pre-bleed and rectal biopsies
12 hRectal biopsies
52 h7B2-AAA I.V.
infusion50 mg/kg
60 hRectal biopsies
Study 38.2: Details
• Monkey IDs: 5105, 5100, 5109, 5104• All monkeys SIV-negative and non-immunized prior to
study• Intravenous infusion of 7B2-AAA, a non-neutralizing
mAb against the gp41 immunodominant epitope; 50 mg/kg 2X at 0 and 52 hours. Timing was determined based on a previous pharmacokinetic study.
• Rectal biopsies obtained pre and then 8 or 12 hours post each infusion. RNA was isolated from the rectal mucosa for microarray analysis.
Study 38.2: Objectives
• Global microarray analysis to determine differential expression patterns following the infusion of 7B2-AAA.
• We are looking for the upregulation/downregulation of any genes of interest—no underlying hypothesis.
Cytokine kinetics from rectal wecks taken in NHP 38.2
Cytokine Plasma Rectal Weck
G-CSF GM-CSF
IFNg IL-10
IL-12/23(p40) X IL-13 IL-15 X IL-17 IL-18 X XIL-1b X XIL-1ra X XIL-2 IL-4 IL-5 IL-6 IL-8 X X
MCP-1 X XMIP-1a MIP-1b TGFa X XTNFa VEGF
sCD40L X
7B2 NHP PK study Cytokine Elevations
Day 0,
Pre-Bleed
Day 0, 1 hr
Day 0, 2 hr
Day 0, 4 hr
Day 0, 6 hr
Day 0, 8 hr
Day 0, 12
hr
24 hr 52 hr 53 hr 54 hr 56 hr 58 hr 60 hr 72 hr 96 hr 120 hr
0.0
5.0
10.0
15.0
20.0
25.0
30.0
35.0
40.0
0.0
100.0
200.0
300.0
400.0
500.0
600.0
700.0
gp41 ID epitope
IL-1b
Time Point
µg /
ml
pg /
ml
IL-1b
IL-1b Kinetics in Rectal Wecks
Day 0,
Pre-Blee
d
Day 0,
1 hr
Day 0, 2 hr
Day 0, 4 hr
Day 0, 6 hr
Day 0, 8 hr
Day 0, 12 hr
24 hr
52 hr
53 hr
54 hr
56 hr
58 hr
60 hr
72 hr
96 hr
120 hr
0.0
10.0
20.0
30.0
40.0
50.0
60.0
0.0
10.0
20.0
30.0
40.0
50.0
60.0
70.0
80.0
90.0
100.0
gp41 ID epitopeIL-18
Time Point
µg /
ml
pg /
ml
IL-18
IL-18 Kinetics in Rectal Wecks
Day 0,
Pre-Blee
d
Day 0, 1 hr
Day 0, 2 hr
Day 0, 4 hr
Day 0, 6 hr
Day 0, 8 hr
Day 0, 12
hr
24 hr
52 hr
53 hr
54 hr
56 hr
58 hr
60 hr
72 hr
96 hr
120 hr
0.0
5.0
10.0
15.0
20.0
25.0
30.0
35.0
40.0
0.0
20.0
40.0
60.0
80.0
100.0
120.0
140.0
gp41 ID epitope
IL-1ra
Time Point
µg /
ml
pg /
ml
IL-1ra
IL-1ra Kinetics in Rectal Wecks
Day 0,
Pre-Bleed
Day 0, 1 hr
Day 0, 2 hr
Day 0, 4 hr
Day 0, 6 hr
Day 0, 8 hr
Day 0, 12
hr
24 hr 52 hr 53 hr 54 hr 56 hr 58 hr 60 hr 72 hr 96 hr 120 hr
0.0
10.0
20.0
30.0
40.0
50.0
60.0
0.0
500.0
1000.0
1500.0
2000.0
2500.0
3000.0
3500.0
4000.0
4500.0
5000.0
gp41 ID epitope IL-8
Time Point
µg /
ml
pg /
ml
IL-8
IL-8 Kinetics in Rectal Wecks
Day 0,
Pre-Blee
d
Day 0, 1 hr
Day 0, 2 hr
Day 0, 4 hr
Day 0, 6 hr
Day 0, 8 hr
Day 0, 12
hr
24 hr
52 hr
53 hr
54 hr
56 hr
58 hr
60 hr
72 hr
96 hr
120 hr
0.0
10.0
20.0
30.0
40.0
50.0
60.0
0.0
10.0
20.0
30.0
40.0
50.0
60.0
70.0
80.0
gp41 ID epitope
TGFaTime Point
µg /
ml
pg /
ml
TGFa
TGFa Kinetics in Rectal Wecks
Day 0,
Pre-Bleed
Day 0, 1 hr
Day 0, 2 hr
Day 0, 4 hr
Day 0, 6 hr
Day 0, 8 hr
Day 0, 12
hr
24 hr 52 hr 53 hr 54 hr 56 hr 58 hr 60 hr 72 hr 96 hr 120 hr
0.0
10.0
20.0
30.0
40.0
50.0
60.0
0.0
50.0
100.0
150.0
200.0
250.0
300.0
350.0
gp41 ID epitope MCP-1Time Point
µg /
ml
pg /
ml
MCP-1
MCP-1 Kinetics in Rectal Wecks
Quality Control of ArraysNo Sample outliers found. Distributions of expression values consistent
Exploratory AnalysisUnsupervised analysis results in Samples grouping by Donor. Will provide a report (Part B), with analysis blocked by donor.
Differentical Gene ExpresionHour 60 most DEG, h12 looks more like preInfusion.h60 vs pre on the left, F test on right
Gene Set Enrichment analysis h12 vs preInfusionh12, TGF-beta, IL-22 pathways. 39 pathways enriched, non meet significance.
Gene Set Encichment h60 vs Preinfustion120 pathways enriched; predominantly signally pathways responding, top genes presented here. Please see table in report. Many cytokine and Chemokines have beeen affected. 3 pathways meet significance. Will see if this signal gets clearer with Analysis part B, bloc by donor
