EPIVAX, INC.
The goal of this new R21 proposal is to develop a pro-inflammatory and non-tolerogenic HIV vaccine delivery system based on the dendritic cell targeting anti-DEC-205 antibody. The success of anti-DEC-205 as a stimulator of strong inflammatory immune responses depends on co-administration of non-specific dendritic cell maturation factors. In their absence, anti-DEC-205 induces antigen-specific tolerance rather than immunity. We hypothesize that regulatory T-cell epitopes contained in anti-DEC-205 promote a tolerogenic reaction that is only overcome through the co-administration of non-specific immuno-stimulators. This hypothesis is based on our discovery of a set of natural regulatory T-cell epitopes derived from human immunoglobulins that induce tolerance by stimulating regulatory T cells. We have verified experimentally that these epitopes cause antigen-specific expansion of regulatory T cells and suppress inflammatory immune responses. We propose to develop a modified pro-inflammatory and non-tolerogenic anti-DEC-205 antibody. We expect that modification of regulatory T-cell epitopes will significantly diminish tolerogenicity, enabling use of anti-DEC-205 as a stand-alone HIV antigen delivery system that obviates the dangers associated with non-specific activation of the immune system. We will de-tolerize anti-DEC-205 by epitope modification using the process we developed to reduce immunogenicity of protein therapeutics. We will substitute key amino acids in the regulatory T-cell epitopes with those that are experimentally shown to interfere with MHC binding. We will then (1) produce a set of antibody variants recombinantly conjugated to HIV Gag, (2) identify de-tolerizing mutations that do not interfere with dendritic-cell targeting, and (3) and evaluate variants for reduced tolerogenicity, as well as for enhanced Gag immunogenicity. Finally, we will produce and characterize the immunogenicity of a de-tolerized anti-DEC-205-based HIV vaccine composed of immunogenic consensus sequences. PUBLIC HEALTH RELEVANCE: This project will develop an improved delivery vehicle for HIV vaccine components. Immune-dampening portions of the delivery vehicle will be silenced so that potent and effective immune responses can be raised against HIV.
Administrative Outcome: One new hire of a Senior Research Associate.
Scientific Outcome: An optimized áDEC-205 DC-targeting vaccine delivery vehicle combined with a multi-epitope string containing conserved and immunogenic HIV sequences.
| AWARD OVERVIEW |
| Award Number |
3R21AI078800-02S1 |
Funding Agency |
Department of Health and Human Services |
| Total Award Amount |
$215,016 |
Project Location - City |
Providence |
| Award Date |
09/17/2009 |
Project Location - State |
RI |
| Project Status |
Completed |
Project Location - Zip |
02903-4118
|
| Jobs Reported |
1.00 |
Congressional District |
02 |
| Project Location - Country |
US |
|
|
Recipient Information
(Grants)
| Recipient Information (Grants) |
|
Recipient Name
|
EPIVAX, INC. |
| Recipient DUNS Number |
135531015
|
| Recipient Address |
146 CLIFFORD ST |
| Recipient City |
PROVIDENCE |
| Recipient State |
Rhode Island |
| Recipient Zip |
02903-4163 |
| Recipient Congressional District |
02 |
| Recipient Country |
USA |
Required to Report Top 5
Highly Compensated Officials |
No |
Projects and Jobs Information
| Projects and Jobs Information |
| Project Title |
Optimization of HIV vaccine subunit delivery |
| Project Status |
Completed |
| Final Project Report Submitted |
Yes |
| Project Activities Description |
Testing Laboratories |
| Quarterly Activities/Project Description |
We've previously reported progress on both hiring and scientific fronts. To fill the 'new hire' position in a timely manner, a temp-to-hire highly-skilled scientist with the proficiency to complete our research proposal was hired and continues in this position. She remains a valuable asset, bringing expertise, skills and knowledge in an important area of our laboratory which we intend to retain.
Prior reports described the finding of a small sequence error of the wildtype (WT) antibody. We proceeded with testing of binding to dentritic cells and found strong binding of the original WT unrepaired antibody, the WT fusion we created and most of the variant antibodies. A small preliminary murine experiment to look at uptake of antibodies in vivo showed no significant results.
We have decided to advance our ARRA supplement proposal by creating variant fusions, applying multiple modifications (versus one) to our WT fusion antibody to produce a stronger non-tolerizing effect. GeneArt supplied us with the WT antiDEC205:39epi construct subcloned into pVAX-1 in the second quarter of 2010.
In this quarter, we have established a HEK293F cell line to be transfected with these constructs in order to produce protein. A modified antiDEC205 antibody construct has been synthesized that contains two modifications, one in the heavy chain and one in the light chain, which we expect to enhance immunogenicity. We have also swapped this modified antiDEC205 with the WT antiDEC205 in the WT 39epi fusion construct.
Our current efforts are focused on producing purified WT antiDEC205:39epi and modified antiDEC205:39epi proteins. Transfection conditions have been optimized by transfecting a pBud-GFP expression vector into HEK293F cells. Using these conditions, we see efficient transfection of all DEC205 constructs, but poor secretion into the supernatants. Alternative strategies for enhancing secretion or extracting intact DEC205 variants from these cultures are being explored. |
| Jobs Created |
1.00 |
| Description of Jobs Created |
A full-time Senior Research Associate position has been created due to this ARRA funded award. We were able to offer a temp-to-hire researcher a permanent full-time position as Senior Research Associate whose primary work encompasses the research aims of this proposal.
This position has greatly impacted our workforce as it filled a gap in our laboratory staffing. We were in need of an experienced molecular biologist to assume responsibility of molecular work for this project and several internally funded projects. We hired a temporary worker to carry out the workload because we were not able to offer a full-time position during that time. Originally, we had planned on hiring a graduate student to perform all of the proposed research for this ARRA award. The identified graduate student decided to take time off from her studies and left our company shortly after the award was funded. We saw an opportunity to use the funding to offer a full-time appointment to our temporary researcher who immediately accepted and assumed the role of Senior Research Associate. She continues to work full-time, at EpiVax on internally funded projects. |
Purchaser Information
(Grants)
| Purchaser Information |
| Contracting Office ID |
Not Reported |
| Contracting Office Name |
Not Available |
| Contracting Office Region |
Not Available |
| TAS Major Program |
75-0900 |
| Award Information |
| Award Date |
09/17/2009 |
| Award Number |
3R21AI078800-02S1 |
| Order Number |
|
| Award Type |
Grants |
| Funding Agency ID |
75 |
| Funding Agency Name |
Department of Health and Human Services |
| Funding Office Name |
Not Available |
| Awarding Agency ID |
75 |
| Awarding Agency Name |
Department of Health and Human Services |
| Amount of Award |
$215,016 |
| Funds Invoiced/Received |
$215,016 |
| Expenditure Amount |
$215,016 |
| Infrastructure Expenditure Amount |
$0 |
| Infrastructure Purpose and Rationale |
Not Reported |
| Infrastructure Point of Contact Name |
Not Reported |
| Infrastructure Point of Contact Email |
Not Reported |
| Infrastructure Point of Contact Phone |
Not Reported |
| Infrastructure Point of Contact Address |
Not Reported |
| Infrastructure Point of Contact City |
Not Reported |
| Infrastructure Point of Contact State |
Not Reported |
| Infrastructure Point of Contact Zip |
Not Reported |
Product or Service Information
(Grants)
| Product or Service Information |
| Primary Activity Code |
541380 |
| Activity Description |
Testing Laboratories |
| Sub-Awards Information |
| Sub-awards to Organizations |
1 |
| Sub-award Amounts to Organizations |
$43,574 |
| Sub-Awards to Individuals |
1 |
| Sub-Award Amounts to Individuals |
$43,574 |
| Number of Sub-awards less than $25,000/award |
0 |
| Amount of Sub-awards less than $25,000/award |
$0 |
| Number of payments to vendors greater than $25,000 |
0 |
| Total Amount of payments to vendors greater than $25,000/award |
$0 |
| Number of payments to vendors less than $25,000/award |
0 |
| Total Amount of payments to vendors less than $25,000/award |
$0 |
Sub-award 741626779398 - SAINT LOUIS UNIVERSITY
| Sub-Award Amount |
$43,574 |
| Sub-Award Date |
09/17/2009 |
| Sub-Awards Disbursed |
$39,569.43 |
| Project Location - City |
St. Louis |
| Project Location - State |
MO |
| Project Location - Zip Code |
63103-2006 |
| Project Location - Congressional District |
01 |
| Sub-Recipient DUNS Number |
050220722
|
| Sub-Recipient Address |
221 N GRAND BLVD |
| Sub-Recipient City |
SAINT LOUIS |
| Sub-Recipient State |
Missouri |
| Sub-Recipient Zip Code |
63103-2006 |
| Sub-Recipient Congressional District |
01 |
Required To Report Top 5
Highly Compensated Officials |
No |
| Location Information |
| Latitude, Longitude |
41º 49' 5",
-71º 24' 42" |
| Congressional District |
02 |
| Address 1 |
|
| Address 2 |
|
| City |
Providence |
| County |
Providence |
| State |
RI |
| Zip |
02903-4118 |
|
 |