heat stable bacillus spores as vaccines - usda ars stable bacillus spores as vaccines. ... var....
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Heat Stable Bacillus Spores as Vaccines
Professor Simon CuttingSporeGen Ltd.School of Biological SciencesRoyal Holloway University of LondonEgham, SurreyTW20 0EX, UKs.cutting@rhul.ac.uk
Ethical Non-pathogenic Storage Robust (heat-stable), suitable for 3rd World Cost Straightforward production Route Oral edible vaccine. Intra-nasal (killed spores)
Licensed OTC drug (Bacillus clausii)100m doses sold/yr in Italy by Sanofi-Aventis
Bacillus spores
The Concept of Spore Vaccines
Stomach
Spores survive intact
vegetative Bacillus killed
Small Intestine
Nutrient rich
Spore germination &
dissemination to GALTColon
Re-sporulation
GIT
Excretion of sporesinto the soil
Symbiotic interactions with plants
Consumption ofplants and spores
spores
vegetative cells
Bacillus as a gut commensal
NattoB. subtilis var. Natto
Japanese staple
BioPlus 2BSpores of B. subtilis & B.
licheniformisAnimal feed
Christian Hansen (Denmark)
Bacillus Spores: Probiotics, Functional Foods and Feed Supplements
Main Street Gourmet (USA)obtained from Isabellas Healthy Bakery
ActivateProbiotic muffins impregnated with spores
1.E+00
1.E+01
1.E+02
1.E+03
1.E+04
1.E+05
1.E+06
1.E+07
1.E+08
1.E+09
1.E+10
1 2 3 4 5 6 7 8
HU58
Protexin
Control
Before cooking
0 7 14 28 42 56 84
Days
Spor
es co
unt /
gra
m
Survival of spores incorporated in biscuits (McVities) after 84 days
These are background countsAnd are Bacillus spores present in your biscuit mix. The biscuit mix is not sterile and data shows that cereals, wheat etc have a basal level of spore counts.
Starting CFUHU58 Tube 1 + 1kg mix = 6.57 X 108 spores/gPXN21 Tube 1 + 1kg mix = 1.33 X 109 spores/g
235oC 8 min.
+Killed Spore
Killed Spore
Antigen(protein or virus)
Vaccinate
Expression on Spore CoatRecombinant (GMO)
Adsorption onto the Spore CoatNon-recombinant/non-GMO
Vaccinate
Spore gene X + Antigen gene Y
X Y
rec spore
SporeVax
Strengthening Animal Production and Health through the Immune Response
The European research project SAPHIR aims to develop innovative, safe , affordable and effective vaccine strategies effective against endemic pathogens responsible for high economic losses in livestock.
SporeGen is collaborating on the development of spores for 2nd generation oral/feed vaccines for Necrotic Enteritis
Shrimp Aquaculture(>$11bn/annum)
Penaeus monodonBlack Tiger Shrimp
Massive reduction in production to disease
Litopenaeus vannameiWhiteleg shrimp
Originally from Central/South AmericaIntroduced to Asia to replace P.
monodonBut stock now suffering epidemics of
disease
Shrimp Pathogens
About 30 viral diseases of shrimps White Spot Syndrome Virus (WSSV) is universally considered the worst
Expression on Spore CoatRecombinant (GMO)
Vaccinate
Spore gene X + VP28/VP26
X Y
rec spore
Spores display VP28 and VP26capsid proteins
Shrimps that eat the vaccine in feed pellets are protected against White Spot Syndrome Virus
Mortality rate (%)
Days post challenge
Protection trial: feeding during 14 days, then challenge
SPR at day 14 by CotB-VP28 at 1 x 10*9 cfu/g = (1-19.23/100)*100% = 81% SPR at day 14 by CotB-VP28 at 1 x 10*11 cfu/g = (1-10/100)*100% = 90%
0102030405060708090
100
1 2 3 4 5 6 7 8 9 10 11 12 13 14
UnchallengedUntreatedPY79 (1 x 10*9 cfu/g)CotB-VP28 (1 x 10*9 cfu/g)CotB-VP28H (1 x 10*11 cfu/g)
Does it work?
Phase 1/2a clinical trialRecruiting nowin Germany
www.cdvax.org
SporeGen (www.sporegen.com)
Operating in UK and Vietnam
SporeVax for development of human and animal vaccines
Spore mediated delivery of heat-stable feed enzymes (phytase, xylanase etc)
Clostridium difficilevaccinesnovel treatmentsdiagnostics
Helicobacter pyloritreatments
Animal feed productsHanoi, Vietnamhttp://biospring.com.vn
+ SGF + SIF
72 kDa
72 kDa
1 2 3 4
41 kDa
A Cytoplasmic expression (rrn0-GST-Cpa247-370)
B Spore coat expression (CotB-GST-Cpa247-370)
58 kDa
41 kDa
58 kDa
41 kDa
15 kDa
15 kDa
C. perfringens
A (Oral) B (Nasal)
Cpa (Clostridium perfringens) toxin A expressed on spores
3.6 g/dose; 2 X 1010 spores 0.15 g/dose; 2 X 109 spores
Seroconversion, IgG
Chart3
4040.214042.340.3023.216338907905.360206561822.97873957
406641.3112.461.2019.69300576513.400409163717.369686654413.6303805758
37.97354.9295.641222.954745831720.758675798123.4416478273119.4987208581206.7500835338
4040402911.74937.4004.0424422064985.13130851562556.0593300985
Nave
PY79
GST-
PY79+ GST-
HT251
Days post primary vaccination
End-point titer
test IgG, IgA
0.2930.0340.2420.080.1430.1030.3170.016
0.2080.2170.2720.170.0260.1050.1940.286
0.240.210.2750.2530.1620.1530.1830.198
0.140.1640.2380.2410.0430.0820.1270.197
0.2450.2070.2730.2560.1620.1710.2110.274
0.1930.1760.2430.2440.0310.0620.1240.197
0.2350.2070.2710.2590.1630.1660.2060.243
0.2150.1970.2680.2650.1580.190.2040.22
0.4740.3570.3540.4010.02
0.480.4060.4530.5060.023
0.5020.4760.5050.5070.468
0.5190.5020.5030.5060.518
0.3020.0490.1720.1230.019
0.4080.2050.2620.2890.025
0.4370.3430.3340.4250.435
0.4950.4680.4310.4640.181
17-Sep
IgA_266_Nnasallung-IgA
12-80.6050.3950.2910.1880.1480.1070.0570.0630.060.060.0770.077Blank
12-70.5910.4710.3630.2230.1710.1260.8630.8680.7390.330.4710.344Pos
12-60.7640.5820.4170.2620.1740.1250.260.2050.1360.0830.1080.101Py-4
12-50.6980.5480.4180.2850.1980.1290.2620.1730.1340.080.1050.095py-3
12-40.9420.9380.8480.6890.4810.30.2180.1490.1220.070.0980.086py-2
12-30.4020.350.240.1680.1180.0920.2660.1770.1290.070.0970.098py-1
12-20.3450.3010.20.1310.1010.0890.1990.1260.1170.0720.1080.102naive3
12-10.6820.4630.3160.2220.1440.1230.2260.1250.120.0780.1110.1naive2
IgG-266-NnavePy12-1
0.2550.1870.1610.1060.1820.1310.1660.1870.1560.140.1771.131
0.1480.090.0830.0640.1020.0720.1020.1080.0940.0840.1020.999
0.0960.0620.0580.0480.0760.0490.0750.0750.0740.0580.0670.896
0.0940.0470.0460.0380.0640.0470.0780.0550.0610.0480.0540.664
0.0760.0430.0390.0360.050.0370.0460.0440.0490.0410.0420.449
0.0810.0410.0360.0330.0420.0360.0410.0380.0430.0290.040.279
0.0750.0420.0360.0330.0390.0360.0370.0350.0370.0340.0370.173
ip-pos1.4761.4431.3371.2021.1341.0710.8980.7170.0610.0540.070.088
1/4001/51200
18-Sep
IgA_266_Nnasallung-IgA
12-80.7610.5010.3460.2240.1780.140.1180.1250.1250.1250.1190.127Blank
12-70.6910.4710.3420.2370.2230.1421.1231.0680.9730.8030.5750.418Pos
12-60.7320.6060.4150.2960.1960.1560.270.2210.1810.1490.1280.134Py-4
12-50.8120.6690.4790.360.2230.1690.2230.2040.1680.1480.1240.112py-2
12-41.0811.0340.9210.7720.4280.3730.2280.1970.1580.1380.1130.113py-2
12-30.4510.3470.2420.1850.1010.1140.3260.2490.2070.1720.1460.139py-1
12-20.3920.2630.1920.1470.0590.1020.3170.2620.2170.1740.1510.131py-3
12-10.8790.5030.3210.2320.1040.1460.7960.6120.3920.2640.2180.196naive2
IgG-266-Npy12-1-paper12-1-serum
0.2050.1560.1910.2230.2020.250.330.8740.210.1710.1580.8921/40
0.1410.0750.0980.1220.1330.2030.2610.7760.1340.0830.0930.8181/80
0.1010.0550.0650.0890.0950.1250.1630.730.0950.0610.0630.6891/160
0.0730.0430.0490.0670.0650.0810.1110.5710.0720.0450.0430.5171/320
0.0540.0360.0380.0490.0490.0630.0710.430.0490.0370.0360.3621/640
0.0460.030.0310.0410.0420.050.0530.2870.0410.0330.0320.2251/1280
0.0370.0290.0290.0320.0350.0330.0420.1460.0340.0260.0280.146
IP-pos0.9690.9910.8450.8080.7230.5630.4910.2650.0440.0440.0490.061
1/800102400
neutralisation
NaCL
11-SepHT251IPnasalORAL
0.3540.2980.310.2010.30.2280.180.1180.3370.2710.2050.0861
0.3750.3320.3030.2120.1310.0170.0170.020.3480.3250.3050.3142
0.2990.3320.2750.2140.4570.4410.3760.2490.0130.0160.020.0283
0.2650.1890.1560.0830.3180.2430.1680.10.3840.3070.30.2654
ip-Pos0.140.0390.0170.0150.2870.2670.2260.1790.310.2720.2590.2625
0.2950.2490.2520.1910.2650.2270.190.1520.3350.2950.2820.2396
0.3240.2910.2920.260.3110.2940.2750.2380.0210.0130.0170.0227
0.3020.2940.4290.0460.270.1550.0230.0310.2990.2180.1180.0318
toxintriton-XNaCL
HT266IPnasalORAL
0.3450.3220.2730.2410.2910.2570.1780.0790.4510.3510.2850.3021
0.3230.2870.2580.2760.3480.290.2670.2290.3860.3360.2930.2792
0.3420.3080.2710.2630.3010.2830.2420.2160.3620.3130.2650.2583
0.3020.2880.2650.2350.3010.2730.2210.1940.3640.3340.2890.264
0.3470.3420.310.2850.2850.2350.1610.0710.3440.3090.2910.2835
0.3520.3290.3110.3140.310.2930.2610.2650.3590.3170.2890.2886
ip-Pos0.2030.0940.0250.0190.2980.2620.2270.1780.3430.2780.2210.1367
0.2150.3380.4140.0180.3930.290.2750.2830.380.3580.3450.3128
toxintriton-XNaCL
HT230ORALnasalIPtriton-Xtoxin
80.3180.3530.3670.3950.2090.2790.3190.3190.3820.4450.5180.379
70.2980.3050.3210.3560.3070.3040.3360.3590.5290.3220.3640.383
60.2160.3110.3090.3340.2330.2510.2650.2880.5360.2950.3230.385
50.4770.3790.2010.2850.1670.1950.2470.2650.530.0260