Statement_of_Work.pdf

RFP Number: N02-RC-91014-56

STATEMENT OF WORK

Independently and not as an agent of the Government, the Contractor shall be required to furnish all the necessary services, qualified personnel, material, equipment, and facilities, not otherwise provided by the Government, as needed to perform the Statement of Work below:

Definitions

HIV Human immunodeficiency virus SIV Simian immunodeficiency virus HTLV-1/2 Human T-cell leukemia virus type I/II BSL2/BSL3 Biosafety level 2/Biosafety level 3 PBMC Peripheral blood mononuclear cells SHIV Chimeric simian/human immunodeficiency virus Ad4 Adenovirus type 4 Ad5 Adenovirus type 5 Ad7 Adenovirus type 7 DNA Deoxyribonucleic acid mRNA Messenger ribonucleic acid PCR Polymerase chain reaction RNAs Ribonucleic Acids

The Contractor shall produce assays, cell lines, peptides and cellular and or viral DNA and protein products for use by NCI investigators of the Vaccine Branch, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland. The Contractor shall also provide storage for these products. In performance of this work, the Contractor shall:

1. Prepare between 3 to 8 B or T cell lines from blood of non-human primates or humans expanded to 1010 viable cells for use by the Project Officer and other NCI Investigators for in vitro or in vivo studies. The Contractor shall deliver cell lines in accordance with SECTION F.2. DELIVERIES unless otherwise directed by the Project Officer.

For purposes of the proposal, the offeror shall assume that 5 T cell lines derived from rhesus macaques under study by the government will be expanded to 1010 viable cells and provided each year.

2. Conduct in vitro culture and detection of retroviruses from between 30 and 70 samples of peripheral blood and/or tissues, provided by the Government, of humans or animals, the latter including rodents, leporidiae and non-human primates. When requested, the Contractor will determine viral infectious units in cells or tissues by end point dilution.

Retroviruses shall include HIV, SIV, HTLV-1/2, and novel retroviruses of animals and man. To accurately assess low level infections, isolations from a single donor may be required from several tissue specimens using target cells with different susceptibilities.

This work shall be conducted in BSL2/BSL3 laboratories. The Contractor shall receive samples from the Government and/or Government contractors, as directed by the Project

Statement of Work Attachment #3 02/10/2009

Officer. The Contractor shall commence in vitro culture and detection within 1 hour of notification by the Project Officer of a sample being delivered to the Contractor. In the event the Contractor has not received the sample within 50 minutes of the Project Officer’s notification of the sample delivery, the Contractor shall immediately notify the Project Officer. The Contractor shall provide the result of culture and detection of retroviruses in accordance with SECTION F.2. DELIVERIES.

For purposes of the proposal, offerors shall assume that isolation of HTLV-1/2 will be requested from 50 samples each year.

3. Provide between 2 and 7 titered stocks of approximately 100 1 ml aliquots of field isolates for in vitro use from virus-infected humans and non-human primates at low passage number on human or non-human primate PBMC. Provide one to two primary isolate stocks of SIV or SHIV strains for use as in vivo challenge stocks, of sufficient titer for mucosal as well as intravenous administration. Provide between 100 and 400 1 ml aliquots of high-titered T-cell-line-adapted (TCLA) stocks of HIV or SIV for in vitro use.

This work shall be conducted in BSL2/BSL3 laboratories. The viral stocks shall be maintained in liquid nitrogen storage until requested by the Project Officer. The Contractor shall deliver viral stocks in accordance with SECTION F.2. DELIVERIES unless otherwise directed by the Project Officer.

For purposes of the proposal, offerors shall assume that 5 titered field isolate stocks consisting of 100 1 ml aliquots each, 1 TCLA HIV and 1 TCLA SIV stock of 100 aliquots each, and 1 challenge stock with a titer appropriate for intravenous or mucosal challenge and in quantities sufficient for challenging 100 macaques, will be provided each year.

4. Provide between 1 and 5 purified and titered stocks containing approximately 100 ml each with titers of approximately 104 to 105 TCID50/ml of attenuated poxvirus and vaccinia virus recombinants as requested by the Project Officer. The initial viral inocula will be provided by the Government. The stocks of recombinant viruses shall be stored until requested by the Project Officer. The Contractor shall deliver viral stocks in accordance with SECTION F.2. DELIVERIES unless otherwise directed by the Project Officer.

For purposes of the proposal, offerors shall assume that three (3) such purified stocks, approximately 100 ml each with titers of 104 to 105 TCID50/ml, will be produced each year.

5. Conduct in vitro immunologic assays to assess the serologic status of human or animal sample donors, to evaluate the prevalence of virus present within its host population, and to evaluate secretion of cytokines or chemokines. Serum or plasma samples will be tested for the presence of antibodies to human and other mammalian viral proteins or peptides and for the presence of cytokines or chemokines using such techniques as ELISA, Western blot, and antigen capture assays. Pepscan assays shall be conducted to determine specific antibody epitopes recognized. Neutralization of HIV and/or SIV isolates and microtiter neutralization assays for Ad4, Ad5, and Ad7 antibodies shall also be conducted. In performance of these tasks, the Contractor shall provide the following:

a) between 2500 and 7500 ELISA and between 50 and 150 Western blot assays to detect antibodies to either HIV, SIV, or HTLV-1/2 proteins,

b) Provide between 200 and 600 antigen capture assays for beta chemokines,

c) pepscan analysis of between 10 and 30 serum samples with anti-HIV envelope activity, and

d) between 100 and 300 adenovirus neutralization assays.

The Contractor shall, upon completion of assays, document the results achieved and report data in sufficient detail in accordance with SECTION F.2. DELIVERIES.

For purposes of the proposal, offerors shall assume 5000 ELISA and 100 Western blot assays to detect antibodies to either HIV, SIV, or HTLV-1/2 proteins, 400 antigen capture assays for beta chemokines, pepscan analysis of 20 serum samples with anti-HIV envelope activity, and 200 adenovirus neutralization assays shall be performed per year.

6. Provide between 300 and 700 purifications of lymphocytes from blood, lymph nodes or other tissues from animals or humans and provide fresh and viable cells to the Project Officer or other NCI Investigator. The Contractor shall deliver fresh viable cells in accordance with SECTION F.2. DELIVERIES unless otherwise directed by the Project Officer.

For purposes of the proposal offerors shall assume that 500 such purifications will be performed each year.

7. Provide purified, biochemically characterized and biologically active (where appropriate) native viral or cellular proteins from viruses, infected cells, or conditioned media and from bacterial or mammalian expression systems. Recombinant vectors expressing the proteins or peptides will be supplied by NIH and could include regulatory, enzymatic and structural proteins. Purified proteins shall meet a predetermined level of purity specified by the Project Officer. The Contractor shall also furnish data to the Project Officer confirming that the proteins are of the quantity and meet the specifications of purity and biological activity agreed to between the Project Officer and the Contractor before work was started on the product. In performance of this task the Contractor shall provide:

a) between 10 and 30 mg of both SIVmac251 and HIV native gp120,

b) between 5 and 15 mg of SIVE660 native gp120,

c) between 5 and 10 mg of SIV p27,

d) between 3 and 7 mg of SIV Nef,

e) between 5 and 15 mg of active HIV Tat, and

f) IgG purified from between 900 and 2700 ml of rhesus macaque plasma. The purified

IgG shall contain low endotoxin levels .

The Contractor shall provide proteins and data in accordance with ARTICLE F.2.

DELIVERIES.

For purposes of the proposal, offerors shall assume that 20 mg of both SIVmac251 and HIV native gp120, 10 mg of SIVE660 native gp120, 10 mg of SIV p27, 5 mg of SIV Nef, and 10 mg of active HIV tat will be purified and supplied in the first contract year. In addition, IgG containing low endotoxin levels will be purified from 1800 ml of rhesus macaque plasma and supplied in the first contract year.

8. Design, synthesize, and purify between 1 and 3 codon-optimized genes encoding cellular or viral proteins for eukaryotic or prokaryotic expression. For the individual genes produced the Contractor shall provide between 3 and 6 mg of product. Examples include HIV primary isolate envelopes (gp160), HIV or SIV regulatory genes, and HTLV-1/2 genes. Deliver the synthesized gene and/or the purified expressed protein to the Project Officer in accordance with ARTICLE F.2. DELIVERIES.

For purposes of the proposal, offerors shall assume that 5 mg of an HIV primary isolate gp160 and 4 mg of an HIV or SIV regulatory protein will be designed, synthesized and purified during the first year.

9. Synthesize and provide between 3 and 9 sets of overlapping peptides, of approximately 5 mg each, representing proteins from the HIV, SIV, HTLV-1/2, or adenovirus genomes.

These shall be 15-mers overlapping by 11 amino acids unless otherwise specified by the Project Officer. The produced peptides shall be delivered in accordance with ARTICLE

F.2. DELIVERIES.

For purposes of the proposal, offerors shall assume that 6 sets of overlapping peptides, 5 mg each peptide, representing 2 HIV or SIV gp160s, 2 HIV or SIV pol, one SIV gag, and one Ad5 fiber protein will be provided in the first year.

10. Prepare and provide between 3 and 9 pre-clinical grade plasmid DNA products containing minimal levels of endotoxin and encoding HIV or SIV genes for vaccine studies. The Contractor shall deliver the plasmid DNAs upon completion in accordance with SECTION F.2. DELIVERIES unless otherwise directed by the Project Officer.

For purposes of the proposal, offerors shall assume that 6 DNAs encoding 3 SIV envelopes, 2 SIV gag and 1 SIV pol, 150 mg each, will be provided during the first year.

11. Conduct between 3500 and 10500 real time PCR assays for SIV gag in plasma and between 100 and 300 for SIV gag in tissues. Purify and quantitate mRNA in plasma or tissue samples of animals or humans by real time PCR. Quantitate viral RNAs, including HIV, SIV, HTLV-1/2, and others as requested by the Project Officer. The real time PCR for viral RNAs shall be sensitive to 50 copies/ml plasma. The Contractor shall document in sufficient detail each step taken in performing the tasks delineated in this section and provide data collected in accordance with ARTICLE F.2. DELIVERABLES.

For purposes of the proposal, offerors shall assume that 7000 real time PCR assays for SIV gag in plasma and 200 for SIV gag in tissues shall be performed per year.

12. Conduct between 500 to 1500 assays to quantitate proviral DNA load by real time PCR for SIV or HTLV-1/2 from cells of non-human primates and leporidiae. The Contractor shall provide the results of the proviral DNA assays in a report in accordance with

ARTICLE F.2. DELIVERABLES.

For purposes of the proposal, offerors shall assume that 1000 such assays will be conducted each year.

13. Provide between 2 and 6 characterized monoclonal and polyclonal antibodies to proteins of interest for use in affinity purifications, immunologic characterizations, and for VB use. Antibodies shall be well characterized for titer, for specific recognition of the protein antigen, and for blockage of the biological function of the protein where appropriate. The produced antibodies shall be delivered in accordance with ARTICLE F.2.

DELIVERABLES.

For purposes of the proposal, offerors shall assume that 4 polyclonal antibodies will be prepared, characterized and supplied per year.

14. Provide long-term storage for viable cell lines, titered field isolates, viral challenge stocks and viable cell specimens from inoculated and immunized animals as directed by the Project Officer. The Contractor shall maintain a backup system capable of maintaining materials stored at the required temperatures. The storage capacities and temperature requirements are:

a) 50 cubic feet of minus 180 degree Celsius storage maintained by liquid nitrogen,

b) 375 cubic feet of minus 80 degrees Celsius storage, and

c) 150 cubic feet of minus 20 degrees Celsius storage.

15. Locate, purchase, and maintain standard laboratory animals for conducting the activities stated in this contract requiring such animals. These animals shall include between 30 and 90 rabbits and between 10 and 30 mice. The Contractor shall house these animals under BSL2 conditions and/or provide special microinsolator cages if requested by the Project Officer or required by federal, state or local Safety and Health regulations.

For purpose of the proposal, the offeror shall assume that 60 rabbits and 20 mice will be required for conducting studies per year.

16. The Contractor shall arrange for shipment and delivery of specimens Monday through

Friday, except Government holiday, between the Contractor’s site and the laboratories on the NIH campus (9000 Rockville Pike, Bethesda, MD 20892-1360). Arrival times of specimen will be specified by the Project Officer or other Investigator and will be approximately between 8:00 AM and 4:30 PM local time. The Contractor shall package biological specimens according to U.S. Department of Transportation regulations.