heat stable bacillus spores as vaccines - usda ars stable bacillus spores as vaccines. ... var....

Download Heat Stable Bacillus Spores as Vaccines - USDA ARS  Stable Bacillus Spores as Vaccines. ... var. Natto. Japanese staple. BioPlus 2B. Spores of . ... Christian Hansen (Denmark) Bacillus Spores: Probiotics,

Post on 12-Apr-2018

212 views

Category:

Documents

0 download

Embed Size (px)

TRANSCRIPT

  • 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

Recommended

View more >