DNA

DNA

Manufacturing residue.

According to CDC's excipient and media listing, this is only found in Hepatitis A. However, I must beg to differ. ANTYHING (virus/bacterium) replicated on animal tissue carries the animals DNA also (i.e. monkey kidney tissue for the polio vaccine, chick embryo tissue for the measles and influenza virus, aborted fetal tissue-as confirmed on the package insert)

DNA

<cell biology, molecular biology> Deoxyribonucleic acid.

The molecule that encodes genetic information in the nucleus of cells. It determines the structure, function and behaviour of the cell.

DNA is a double-stranded molecule held together by weak bonds between base pairs of nucleotides. The four nucleotides in DNA contain the bases: adenine (A), guanine (G), cytosine (C), and thymine (T).

In nature, base pairs form only between A and T and between G and C, thus the base sequence of each single strand can be deduced from that of its partner.

(10 Nov 1998)

Further deliberate DNA experimentation by US government:

http://www4.od.nih.gov/oba/rac/SAE_rpts/Mod0902s/Sep02_MODs.htm

SERIOUS, POSSIBLY ASSOCIATED AND UNEXPECTED ADVERSE EVENTS
REPORTED FOR HUMAN GENE TRANSFER PROTOCOLS
REPORTING PERIOD: 05/01/02 -- 08/01/02
RECOMBINANT DNA ADVISORY COMMITTEE MEETING
September 2002

Event # OBA Date Event Date Protocol # Event Description

9510-130

Administration of Neomycin Resistance Gene Marked EBV Specific Cytotoxic T Lymphocytes to Patients with Relapsed EBV-Positive Hodgkin Disease.

4446 05/24/2002 09/16/1997 Research participant had a Grade 3 cachexia. Deemed possibly related to gene transfer product by the investigator. No further details provided.

9902-287

Phase I Pilot Trial of Adenovirus p53 in Bronchioloalveolar Cell Lung Carcinoma (BAC) Administered by Bronchoalveolar Lavage. Sponsor: NCI-Cancer Therapy Evaluation Program (NCI-CTEP)

4508 07/18/2002 10/26/2000 Research participant developed arrhythmia two days after receiving gene transfer agent. As per investigator, arrhythmia possibly related to gene transfer agent. Referred to cardiologist who treated research participant and problem resolved.

9904-304

Pediatric Phase I Study of AdV/RSV-TK Followed by Ganciclovir for Retinoblastoma

4479 05/21/2002 04/12/2000 After receiving gene transfer agent by intraocular injection, research participant developed grade 1 irritation in the eye. Possibly related to injection, as per investigator.

4480 05/21/2002 04/20/2000 One week after event described in 4479, research participant developed a grade 2 irritation in the eye. Possibly related to the intraocular injection, as per the investigator.

4481 05/21/2002 04/20/2000 Concurrent with event 4480 research participant also developed a grade 1 conjunctival reaction. Possibly related to intraocular injection, as per the investigator.

4484 05/21/2002 08/24/2000 After receiving gene transfer agent by intraocular injection, research participant developed grade 1 irritation in the eye. Possibly related to injection, as per the investigator.

4485 05/21/2002 08/24/2000 Concurrent with event 4484 research participant also had a Grade 1 conjunctival response. Possibly related to injection, as per the investigator.

4486 05/21/2002 09/11/2000 Approximately 3 weeks after the events described in 4484 and 4485, research participant noted to have grade 2 irritation in the eye. Possibly related to intraocular injection, as per the investigator.

4487 05/21/2002 04/03/2001 After intraocular injection of the gene transfer product, research participant developed grade 1 lens opacity.

4488 05/21/2002 04/04/2001 Concurrent with event 4487, research participant also developed grade 1 eye irritation. Possibly related to intraocular gene transfer injection, as per investigator.

4495 05/21/2002 11/19/2001 Research participant had a Grade 1 corneal edema. Possibly related to intraocular gene transfer injection, as per the investigator.

0002-388

A Double-Blind, Randomized, Placebo-Controlled, Dose-Ranging, 26-Week Study to Assess the Safety and Efficacy of CI-1023 (AD_GVVEGF121.10) in Peripheral Arterial Disease Patients with Severe, Disabling Intermittent Claudication. Sponsor: Parke-Davis Pharmaceutical Research

4298 05/16/2002 05/01/2002 Follow-up from sponsor (for reports 4280, 4283, and 4291): As per sponsor, renal insufficiency most likely due to concomitant use of NSAIDs and ascites due to underlying malignancy.

4314 05/20/2002 05/01/2002 Follow-up report to 4280. 4283, 4291, and 4298: Repeat echocardiogram of heart showed near normal cardiac function and clot in ventricle no longer seen. Research participant underwent bypass graft surgery for correction of right lower leg ischemia. At this time, the subject's diagnoses are: acute cardiomyopathy, left ventricular clot, lower extremity ischemia necessitating bypass surgery, a stroke due to embolism originating from the left ventricular clot, ascites most likely due to ovarian cancer, and renal insufficiency due to NSAID use.

4318 05/23/2002 05/01/2002 Follow-up from sponsor (to reports 4280, 4283, 4291, 4298, and 4314). Following diagnoses not attributed to gene transfer agent by either the investigator or sponsor: renal insufficiency, left ventricular clot, acute cardiomyopathy, worsening ischemia of right leg, and ascites. In the investigator's opinion, possible association between study medication and ovarian cancer. In the sponsor's opinion, ovarian cancer probably related to other illnesses and not study drug.

4324 05/28/2002 05/01/2002 Follow-up from the investigator: Discharge summary containing summary of information already in reports 4280, 4283, 4291, 4298, 4314 and 4318. Etiology of adenocarcinoma cells in ascitic fluid still not identified but strongly suspected to be ovarian cancer.

0005-396

A Phase I, Open-Label, Dose-Escalating Study of the Safety, Tolerability, and Anti-Tumor Activity of a Single Intrahepatic Injection of a Genetically Modified Herpes Simplex Virus NV1020, in Subjects with Adenocarcinoma of the Colon with Metastasis to the Liver and the associated, long-term follow-up protocol: Long-Term Follow-Up of the Safety and Survival of subjects with Adenocarcinoma of the Colon with Metastasis to the Liver Who Enrolled in a Phase I Dose-Escalating Study Evaluating a Genetically Engineered Herpes Simplex Virus, NV 1020. Sponsor: NeuroVir Therapeutics, Inc.

4380 06/19/2002 06/12/2002 Twelve hours after injection of the study agent the GGT level was elevated to 809 which was greater than 20 times the upper limit of normal. There was no baseline GGT level given. Pre-injection AST=30, ALT=21. Twelve hours post-injection AST=60, ALT=34. Conjugated bilirubin and LDH levels were elevated, but coagulation parameters were just at the upper limit of normal. By 24 hours post-injection, the GGT, AST, and ALT levels were normal. The research

4512 07/22/2002 06/12/2002 Follow-up to Event 4380 from the PI. After reviewing the laboratory information, the PI changed the causality of the elevation of G-GTP from possibly related to "most likely related to viral administration."

0006-402

Phase I Study to Evaluate the Safety of Cellular Immunotherapy for Recurrent/Refractory Neuroblastoma Using Genetically-Modified Autologous CD8+ T Cell Clones.

4322 05/28/2002 05/23/2002 Research participant developed fever 2 hours after receiving infusion of transduced T-cells. Positive blood cultures times two. Due to the timing of the fever and positive blood cultures, investigator suspected that gene transfer product contaminated. Results of further testing pending. (Follow-up Note [event 4360]: All tests on gene transfer product showed no contamination. Cause of positive blood cultures no longer suspected to be gene transfer product, as per

0007-407

A Phase I Double-blind, Placebo-Controlled, Escalating Dose, Multi-center Study of Ad2/Hypoxia Inducible Factor (HIF)-1-alpha/VP16 Gene Transfer Administration by Intramyocardial Injection During Coronary Artery Bypass Grafting (CABG) Surgery in Patients with Areas of Viable and Underperfused Myocardium not Amenable to Bypass Grafting or

4502 07/12/2002 06/14/2002 The research participant was undergoing double coronary artery bypass surgery and receiving blinded study drug injections. Subsequent to injection number four, and the lifting of the heart to visualize the next injection site, the research participant went into spontaneous ventricular fibrillation. Following defibrillation and drug therapy, the participant converted into sinus rhythm and recovered. All remaining study injections were administered. The participant recovered without sequelae. The PI reported that the intensity of the event was moderate and that the event was unrelated to the study drug. The Sponsor determined that a relationship between the study drug and the event could not be excluded and changed the causality assessment to possibly related.

0011-435

Vaccination in Peripheral Stem Cell Transplant Setting for Multiple Myeloma: The Use of Autologous Tumor Cells with an Allogeneic GM-CSF Producing Bystander Cell Line. Sponsor: Cell Genesys, Inc.

4323 05/24/2002 05/07/2002 Prior to receiving 6th cycle of vaccines, research participant admitted for 4 days of dry cough, fever, chills and slight dyspnea on exertion. CT scan showed ground glass infiltrates in both lung fields. Workup done to find cause of this event, but all results negative. Research participant improving. Investigator believes that events secondary to gene transfer product, while corporate sponsor believes that more likely due to self-limited viral infection.

0105-472

Phase I/II Study of Vaccination with Irradiated Autologous Lung Tumor Cells Mixed with a GM-CSF Secreting Bystander Cell Line (Lung Bystander GVAX^�) in Advanced Non-Small Cell Lung Cancer. Sponsor: Cell Genesys, Inc.

4313 05/20/2002 05/01/2002 Research participant with advanced lung cancer admitted to hospital due to increasing shortness of breath, leg swelling, and right sided chest pain. CT scan showed mass in lung field, two masses in liver, and multiple enlarged lymph nodes. As per investigator, these events most likely due to progression of disease but "possibly" related to gene transfer

0107-481

An Open-Label, Phase Ib/II Study of the Safety, Tolerability and Efficacy of G207, a Genetically Engineered Herpes Simplex Type-1 Virus, Administered Intracerebrally to Subjects with Recurrent Malignant Glioma. Sponsor: MediGene, Inc.

4312 05/20/2002 04/27/2002 Follow-up for events 4278, 4279 and 4286: Clinical nurse coordinator spoke with research participant and presently doing well. Is due in for 28 day post-procedure visit in late May.

4371 06/18/2002 04/27/2002 Follow-up from the investigator: (see reports 4278, 4279, 4286): Final viral culture information: No growth on viral cultures. Cause of event probably related to gene transfer product, as per the investigator.

http://www2.niaid.nih.gov/newsroom/releases/rabies.htm

Laurie K. Doepel
(301) 402-1663
ldoepel@nih.gov

DNA Vaccine 100 Percent Effective Against Rabies in Monkeys

Scientists at the Rocky Mountain Laboratories (RML), part of the National Institute of Allergy and Infectious Diseases (NIAID), have developed a DNA vaccine against rabies that protected eight of eight vaccinated monkeys from the disease. It is the first DNA vaccine to show complete protection in nonhuman primates against a virus that attacks the central nervous system (CNS). Their report describing the successful experiment appears in the August 1998 issue of Nature Medicine.

"There’s no gray area in this experiment. That’s what’s so beautiful about it," comments lead author Donald L. Lodmell, Ph.D., an expert in NIAID’s Laboratory of Persistent Viral Diseases located in Hamilton, Mont. In addition to perfect protection afforded by the vaccine, anti-rabies antibodies elicited by the vaccine neutralized a global range of rabies viruses. These results suggest, says Dr. Lodmell, that the DNA vaccine could be effective anywhere in the world.

Each year, more than 40,000 people worldwide die from rabies. It is one of the oldest and most feared human diseases, first described in 2300 B.C. Symptoms include agitation, convulsions, paralysis and delirium. Without prompt treatment, rabies almost inevitably ends in death.

In the United States, few people die from rabies because of widespread immunization of domestic animals: since 1994, only eight deaths have been reported to the Centers for Disease Control and Prevention (CDC). However, the CDC estimates that another 30,000 to 40,000 people each year receive shots to fend off the disease after possible exposure. Rabid bats, raccoons, skunks or other wild animals are the primary sources of human infection in the United States.

Most deaths occur in developing countries where rabies is endemic and resources are inadequate to provide optimal post-exposure treatment. Such treatment, which consists of injections of rabies virus grown in human cells and then inactivated, and human anti-rabies serum, costs about $2,000. Cruder concoctions used in developing countries, derived in animal brains, often cause severe neurological side effects such as allergic encephalitis, which can lead to paralytic reactions as well as death.

"About three years ago," says Dr. Lodmell, "I became very interested in DNA vaccination, and thought it was a logical step for the rabies problem." DNA vaccines are inexpensive, stable and easy to make, and don’t need refrigeration, qualities that make feasible the possible widespread use in developing countries.

A postdoctoral fellow in the lab at that time, Nancy B. Ray, Ph.D., made the vaccine from DNA encoding the surface glycoprotein of the rabies virus. After getting excellent immune responses and protection using this vaccine in mice, they decided to move into primates. "The vaccine worked beyond our wildest dreams," says Dr. Lodmell.

They vaccinated eight monkeys with the DNA vaccine, two monkeys with a current human diploid cell vaccine (HDCV) and two control monkeys with the DNA vector alone. All animals received at least one booster shot at 190 days. In all but the two control animals, the researchers could measure high levels of anti-rabies antibodies. Neutralizing antibodies are known to be the primary source of protection for humans and animals.

Dr. Lodmell then had all the monkeys flown to Atlanta, where his collaborators tested the efficacy of the vaccine. At the CDC, Charles E. Rupprecht, D.V.M., Ph.D., chief of the rabies section, and his colleagues exposed the monkeys to lethal doses of rabies virus. By day 11, the two control monkeys had developed clinical signs of the disease. Yet six months after challenge, the investigators still could detect no evidence of rabies virus in the eight monkeys that received the DNA vaccine and the two that received the HDCV vaccine.

The only drawback of the DNA vaccine, says Dr. Lodmell, is that the antibody response cannot be detected before 30 days. Hence, as currently designed, the vaccine would not be suitable for post-exposure prevention of disease. However, he believes researchers will be able to overcome this problem in the future. On the other hand, DNA vaccines typically provide long-lasting immunity, so they could be used prophylactically to protect people at high risk, such as veterinarians and individuals who live in developing countries. Currently, Dr. Lodmell and his colleagues are assessing the durability of the antibody response following just one immunization to investigate the requirement for booster vaccinations, as well as other issues related to protection.

Reference:

DL Lodmell, NB Ray, MJ Parnell, LC Ewalt, CA Hanlon, JH Shaddock, DS Sanderlin and CE Rupprecht. DNA immunization protects nonhuman primates against rabies virus. Nature Medicine 4(8):949-52 (1998).