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LAMP-DNA Vaccines for Japanese Red Cedar Allergy
Conclusions and Future Directions
• ITI’s JRC2-LAMP-VAX met safety and immunological endpoints in Phase I clinical trial of 24 subjects.
• In mouse model for JRC allergy, we demonstrate that JRC2-LAMP VAX functions through inducing Th1 immune
response and suppressing allergic Th2 response.
•J
RC2-LAMP VAX elicits antigen-specific CD4+, but not CD8+, T cell activation, which promotes the protective IgG2a antibody
production & suppresses allergic IgE response.
•R
egulatory T cells might not be involved in the mechanism of action.
•D
NA vaccination through the needle-free Bioject delivery is effective, particularly in the initial stage of the vaccination regimen.
• ITI is a private company and welcomes collaboration in the fields of cancers and allergic and infectious diseases for human
and animal health.
JRC2-LAMP-VAX INDUCES A PREDOMINANTLY TH1 RESPONSE
JRC2-LAMP-VAX MODEL AND HYPOTHESIS
Inflammatory cell
Th2 Memory
POLLEN
Th2 cell
IL-4
B cell
IgE
+LAMP
LAMP-vax
Interferon-gamma
Killer T cell
Th1 cell
IgG
Th1 Memory
IL-12
Interferon-gamma
B cell
Dendritic cell
Without LAMP, allergens are processed through the
Th2 pathway resulting in an inflammatory response.
LAMP drives an MCH-11 mediated Th1 response resulting
in a antigenic immune response.
• Lysosomal Associated Membrane Protein 1 (LAMP1), a
lysosome residential protein, is utilized in the ITI’s DNA
vaccine platform to direct DNA products toward the
lysosomal/endosomal compartments in antigen
presenting cells.
Aim 1: To evaluate the effects of JRC2-LAMP VAX in a prophylactic
protocol in a mice model.
Aim 2: To compare the efficacy of ID needle-free Biojector, intramuscular
(IM) and intradermal (ID) injections.
Method: 50µg CryJ1-LAMP and 50µg CryJ2-LAMP DNA plasmids were
administrated to BALB/c mice (n=7) as indicated. 50µg Control vector
was injected intramuscularly (n=7). 5µg recombinant CryJ1 and CryJ2
proteins were given as indicated. Mice were bled at indicated times.
Serum anti-CryJ1 or -CryJ2 IgG1 (a Th2 type antibody) and IgG2a
(a Th1 type antibody) titers were assayed by endpoint ELISA method.
Results and Conclusions: JRC2-LAMP VAX delivered through Biojector,
or IM and ID injections elicited strong IgG2a antibodies against both
CryJ1 and CryJ2, suggesting a predominant Th1 response is induced.
The efficacy of Biojector delivery in induction of immunity is comparable
to, or even better than the classic ID and IM routes, particularly in the
initial stages of DNA vaccination.
• Without LAMP, antigens encoded by DNA vaccines
are either secreted or degraded in the proteasomes,
eventually leading activation of CD8+ T cells.
• We propose that LAMP directs the processing of
antigens in the lysosomes/endosomes and the
subsequent presentation by MHC class II molecules,
which activates CD4+ T cells.
www.immunomix.com For more information email: [email protected]
LAMP-DNA Vaccines for Japanese Red Cedar Allergy
CRYJ SPECIFIC CD4+ T CELLS MEDIATE PROTECTIVE TH1 RESPONSE
1 3 4 5 10 CD4+ T cell transfer 14 Bleed Weeks 24 19 Cry J1/Cry J2/Alum Anti-CryJ IgE responses
An3-­‐CryJ IgE responses Cry J1/Cry J2/PBS An3-­‐CryJ1 IgE Anti-CryJ
IgG
AnA-­‐CryJ IgG responses
responses Two Sites One Site AnA-­‐CryJ1 IgG Titers 10000000 10000000 1000000 1000000 100000 100000 10000 10000 IgG1 100000 10000 1000 1000 100 100 100 21 35 70 98 133 167 Two Sites Control Vector 10000000 1000000 1000000 1000000 100000 100000 100000 10000 10000 10000 1000 1000 1000 100 100 21 35 70 98 133 167 Days An3-­‐CryJ2 IgE two sites one site control 0.4 1.2 1 0.8 0.3 0.6 0.2 0.4 0.1 0.2 0 0 70 98 133 Days 167 35 70 98 133 Days 167 Days One Site 10000000 0.5 35 21 35 70 98 133 167 Days Days 10000000 IgG2a 1000000 1000 21 35 70 98 133 167 AnA-­‐CryJ2 IgG Titers Control Vector 10000000 OD450 (1:20 dilu3on) 0 IgG1 IgG2a Days Aim 2: To study whether administration of both the CryJ1 and CryJ2
plasmids in one site has the same efficacy as separate sites.
Generation of CryJ specific T cells: CryJ1-LAMP and CryJ2-LAMP
DNA plasmids were delivered by Biojector either on one site (mixed)
or on two sites (separated) at the flank of mice. Two weeks after the
3rd dose, splenic CD4+ T cells were isolated and then transferred into
naïve BALB/c mice (n=5).
100 21 35 70 98 133 167 Aim 1: To investigate whether T cells are responsible for the effects of
DNA vaccination.
21 35 70 98 133 167 Days Conclusions: Transfer of CD4+ T cells from vaccinated mice is
sufficient to induce protective immune responses in recipient mice.
The two plasmids, whether given in either one or two injections,
exhibited excellent efficacy in inducing Th1 type immunity.
CLINICAL STUDIES OF JRC2-LAMP-VAX SUMMARY OF PH. I RESULTS IN 24 SUBJECTS
Efficacy & Safety Goals for JRC2-­‐LAMP-­‐VAX Phase 1A Phase 1B No anaphylactic / allergic responses
Achieved Achieved No anti LAMP antibody generation
Achieved Achieved No severe adverse reactions
Achieved Achieved Skin test negative patients remain negative
8 of 8 subjects Achieved IgE levels stable or decreasing
Achieved Maintained 14 of 16 subjects 100% conversion Conversion skin test from positive to negative 15 of 16 subjects 100% conversion SAFETY:
IMMUNOLOGICAL ACTIVITY:
Elimination of JRC/Mtn C/CryJ2 skin test
reactivity
*Study results above based on Phase IA Study in 24 subjects
receiving 4 doses, 1x every 2 weeks, including 8 JRC non-allergics
receiving 4mg, 8 JRC allergics receiving 4mg, and another 8 JRC
allergics receiving a 2mg dose 1x every 2 weeks. Further studies
required before any safety and potential efficacy claims can be
substantiated
*Adverse events were observed, which were classified by the clinical
trial site medical director as primarily associated with the injection
of the vaccine and included injection site soreness, redness and
minor pain, which in the majority of incidences was observed to be
transitory.
Current Status: Phase IA and 1B in US complete, moving towards Phase I & II in Japan
www.immunomix.com For more information email: [email protected]