Anthrax Vaccine: Model of a Response to the Biologic Warfare Threat

Meryl Nass, MD

Infectious Disease Clinics of North America, Volume 13, Number 1

March 1999

Anthrax in nature is a usually fatal zoonotic disease that was a scourge of livestock until vaccines were developed in the 1880s. Animals acquire the disease from consuming contaminated soil in which pre-existing anthrax spores are likely to have germinated, then resporulated under appropriate soil and weather conditions, increasing their concentration in soil to infectious levels. [60] [91] Human disease results from exposure to contaminated animal products. Cutaneous disease is most common. The mortality is neglible if treated, and has been 15% due to septic complications when untreated. Gastrointestinal and meningeal cases are rarely seen. Inhalation anthrax (woolsorters’ disease) results from inhaling spores, most often in poorly ventilated areas. It is about 90% fatal. Antibiotics and standard interventions begun after symptoms develop rarely prevent a fatal outcome. The human infectious dose is unknown, but is estimated to be between 100,000 and 100,000,000 spores. [54] [78] In animals the infectious dose is highly strain dependent, and this is likely to be true in humans as well. The high infectious dose probably accounts for the rarity of human cases.

Animal Vaccines

Pasteur, Toussaint, and Greenfield developed the first animal anthrax vaccines about 1880. [27] [87] Sterne developed an attenuated live animal vaccine in 1935 that is still used, and derivatives of this strain account for almost all vaccines used in the world today. [87] The most significant problem with the Sterne vaccine is that it retains some virulence. Goats, llamas, and occasionally other animals may die following vaccination. [22] [40] [41] [43] [89] [95] This vaccine, along with improvements in animal husbandry and industrial hygiene, has made anthrax an almost negligible problem in the developed world. There are rare animal outbreaks in the United States, and less than one human case reported per year. [97]

The Sterne vaccine strain lacks the plasmid pX02, which codes for the d-glutamic acid polypeptide capsule. The capsule inhibits phagocytosis and opsonization. The Sterne strain does retain the toxin plasmid pX01, which codes for the three toxin proteins: (1) protective antigen (PA); (2) lethal factor (LF), a zinc metalloprotease that inhibits mitogen-activated protein (MAP) kinase-kinase [19] ; and (3) edema factor (EF), a calmodulin-dependent adenylate cyclase that generates cyclic adenosine monophosphate in the cytoplasm of eukaryote cells. [22] [55] PA is an 82-kD protein that binds to receptors present on most mammalian cells. It is then cleaved by a cell surface protease to a 63-kD fragment, exposing a site that binds competitively to either EF or LF. The PA-LF or PA-EF complex then enters cells. [51] [52]

Human Vaccines

Japan developed anthrax as a biologic weapon in the 1930s, [98] and the United States and Great Britain followed in the 1940s. [5] [6] [28] Little is known about the actual use of anthrax in biologic warfare (BW), although it is reported to have been used by Japan against China in World War II, [98] against livestock belonging to blacks during the Rhodesian civil war in 1978, [60] and by Germany against pack animals in World War I. [10]

Human vaccines were developed in the Soviet Union by 1940 [1] [80] and in the United States and Great Britain in the 1950s. [88] The current US vaccine was formulated in the 1960s and licensed in 1970, 2 years before efficacy data were required for licensing. [3] [88] Russia and China use live attenuated strains for their human vaccines. The Chinese and Russian vaccines may be given by aerosol, scarification, or subcutaneous injection. [80] [81] The Russian vaccine was manufactured at the George Eliava Institute of Bacteriophage, Microbiology and Virology in Tblisi, Georgia, until 1991 (Nina Chanishvili, PhD, personal communication, June 1998). Efficacy of the live Russian vaccine is reported to be greater than that of the killed US or British vaccines. [31] [53] [80] [90]

The US and British vaccines are filtrates from two different anthrax strains, each lacking the capsule plasmid. They are composed chiefly of PA. The British vaccine consists of alum-precipitated toxin proteins and has larger amounts of EF and LF than the US vaccine. [88] The US vaccine uses aluminum hydroxide (alhydrogel) to adsorb PA, and to serve as an adjuvant that is believed to stimulate humoral but not cell-mediated immunity. [95] According to Hambleton and Turnbull “,[S]uch vaccines can produce some protective activity in experimental animals and may be effective in humans.” [31]

The US vaccine is termed MDPH-PA (produced until February 1998 by the Michigan Department of Public Health, at the Michigan Biologic Products Institute [MBPI]), under contract to the Department of Defense [DOD]) or MDPH-AVA (anthrax vaccine adsorbed). The facility was sold in September 1998 and has been renamed BioPort.

The vaccine consists of a culture filtrate from the toxigenic, nonencapsulated strain of Bacillus anthracis V770-NP1-R. [74] It was administered to only several thousand people until 1990. One textbook states that there have been no controlled clinical trials in humans of the efficacy of the currently licensed US vaccine [7] and no published studies of its safety exist.

Three problems with this vaccine have stimulated interest in an improved human anthrax vaccine [17] [31] [33] [36] [37] [38] [39] [40] [41] [42] [43] [56] : (1) the immunization schedule involves six initial doses over 18 months followed by yearly boosters; (2) immunity is not protective against all natural anthrax strains in guinea pigs; and (3) there is a high incidence of local reactions (30% according to the package insert). The vaccine is an undefined mix of bacterial products. [7] [33] [47] Furthermore, the potency of both the UK and MDPH-PA vaccines is found to vary significantly between lots. [37] [40] [68]

Therefore, attempts have been ongoing since the early 1980s to develop an improved human vaccine. It is proposed that better vaccines should generate cell-mediated as well as humoral immunity, inhibit spore germination, and possibly other factors, and be well-defined chemically. Although virulence factors other than the toxin proteins and capsule have been identified, their roles are only beginning to be defined. [85] These include a type 1 DNA topoisomerase coded for by pX01, [24] and chromosomally encoded factors including extracellular proteases. [82] [83] [86] Vaccine development has been hampered by limited understanding of anthrax pathogenicity and lack of knowledge of epitopes that contribute to the improved immunity conferred by live vaccines. [40] [43] [81] [84] [86]

In the United States, two approaches toward an improved vaccine have been taken. [22] First, a chemically pure PA vaccine has been sought. [40] [42] [90] One candidate has been derived from a recombinant anthrax strain that lacks the capsule, LF and EF, achieving 98% purity and retaining PA’s biologic activity. [21] Whether it stimulates adequate immunity is not yet known. In the second approach, a live vaccine that is safe, produces PA but also contains other immunogenic epitopes, and presents the antigens more effectively than a chemical vaccine is being sought. [42] Although a variety of live vaccine candidates have been tested, none yet has been found to be ideal. [17] [20] [40] [41] [95]

The only human field trial of an anthrax vaccine in the West was conducted by Brachman et al [8] in goat hair mills in New England in the late 1950s. During the 4-year study period there were 26 cases of anthrax, of which 21 were cutaneous and 5 were due to inhalation. One case of cutaneous anthrax occurred in a recipient of the complete injection series, and two in recipients of the partial vaccine series. The five inhalation cases were too few to draw any conclusion about vaccine efficacy with regard to inhaled organisms. [8]

Since then, the vaccine formulation has been changed, and the current MDPH-PA vaccine contains an estimated four to six times more PA than the earlier vaccine. [37] The in vitro assessment of vaccine-induced immunity in humans is limited, however, because no reliable chemical correlates of immunity have been identified. [38] Although PA appears to be a primary and necessary component of all anthrax vaccines, the serum levels of anti-PA antibodies (by enzyme-linked immunosorbent assay) are not predictive of immunity in animals, when compared with the results of an anthrax spore challenge. [11] [37] [39] [40] [43] [87] [88] [90] The reason for this is obscure. Thus, vaccine efficacy and potency cannot be assessed reliably without resorting to anthrax challenge studies, so one is left with the conundrum of how these results translate to humans. [31] [38]

The Russians use an anthraxin skin test (similar to the purified protein derivative) to detect immunity. [79] The test reliably identifies recent or remote anthrax infection, and even after many years 73% of those with prior infections remain positive. Following immunization, however, skin test sensitivity wanes rapidly with only 34% positive at 1 year. Whether immunity fades as rapidly is unclear. [81]

Vaccine Efficacy

Both the US and British vaccines have been extensively tested for efficacy in animal challenge studies. Table 1 provides data from six studies of anthrax vaccines, showing survival rates following parenteral anthrax challenge of vaccinated guinea pigs. Table 2 does the same for studies in mice. Table 3 reviews the data from aerosol challenge studies in guinea pigs. Although there are large variations in design of the different studies, in the infectious doses used, and in the results, it is obvious that protection conferred by the MDPH vaccine to vaccine-sensitive Vollum anthrax strains is modest in guinea pigs, but that protection is unsatisfactory for other vaccine-resistant anthrax strains as well as for all anthrax strains in mice. Protection is only slightly better with the UK human vaccine. [11] According to Jones et al, “[F]urther work on aerosol infection conducted at CAMR (Porton Down) in 1991 (unpublished data) confirmed that the current UK vaccine affords poor protection to guinea pigs challenged with an aerosol of spores of the Ames strain.” [47]

Different animal species vary greatly in their resistance to infection by anthrax, as well as in their sensitivity to intravenous challenge with lethal toxin, composed of LF plus PA. [40] [94] Because it is not known how closely the responses of experimental animals parallel the human response, it is also not known how the animal studies of vaccine efficacy can be extrapolated to humans. [31]

As noted previously, a long history of research using guinea pigs and mice suggests that the US and UK vaccines confer moderate protection against the types of exposures being faced in agriculture and industry, but may confer less protection against strains selected for virulence or vaccine resistance. Two recent reports from Fort Detrick of monkeys immunized with MDPH-PA and exposed to up to 900 times the LD50 of aerosolized Ames spores, however, gave quite different results. [38] [68] Nearly every immunized monkey survived, and all the controls died. The findings were presented to the International Workshop on Anthrax in 1995, but they have not been published elsewhere. [61] [62] In earlier monkey studies done by the same group, anthrax vaccine administered to monkeys on days 1 and 15 after an aerosol challenge led to survival no better than unvaccinated controls. [26]

Exposure Issues Specific to Biological Warfare (BW)

The ways vaccines will be required to function to mitigate the effects of a BW attack may be different in some ways than their use against natural pathogens. BW exposures could be characterized by the following:

  1. Exposure will probably be via aerosol, although subsequently there may be an increased risk of exposure from soil. [60]
  2. Especially virulent strains are likely to be used. [73] There may be a mixture of strains, as suggested by the work of Jackson et al, [44] who strain-typed specimens from the Sverdlovsk outbreak.
  3. More than one type of disease organism may be used simultaneously or in tandem. For example, a pathogen that attacks mucous membranes may be used to enable easier penetration of the host by a second pathogen. This reduces the infectious dose, increasing the attack rate.
  4. Strains are likely to be selected for antibiotic resistance and vaccine resistance, or genetically engineered to maximize these properties and perhaps add unusual virulence factors. [48] [73]
  5. Recombinant anthrax strains may not require PA to introduce LF and EF into cells, as suggested by work done by Pomerantsev, et al. [9] [70]
  6. Very high concentrations of microorganisms are likely to be faced, which may overcome vaccine protection. [34] [46]
  7. Exposure may be detectable early, in advance of symptomatic illness, using sensors and intelligence information.
  8. Novel syndromes may occur.
  9. Herd immunity has allowed existing vaccines to keep infectious diseases out of immunized populations, although immunity within a population is significantly less than 100%. The advantage conferred by herd immunity will not be experienced in a BW setting; therefore, successful vaccines will need to approach 100% efficacy.

Consequently, for prophylaxis of BW, an anthrax vaccine should be effective against aerosol exposure, high doses, the most virulent strains, and have extremely high efficacy. Ideally, it should inhibit spore germination, to suppress the disease at an earlier stage than the current vaccine, [87] as well as neutralize other virulence factors. If it is solely directed against PA, it would probably provide no protection against a recombinant anthrax strain lacking PA. Postexposure therapies would also be highly desirable.

Adjuvant Studies

Experiments beginning in the 1980s showed that the addition of certain adjuvants, either killed or attenuated bacteria, or novel adjuvant formulations, greatly improved the efficacy in terms of survival rates of the US and British anthrax vaccines in animal studies. [31] [42] [43] [87] [88] [90] [94] Adjuvant use led to more rapid development of immunity than the standard vaccines, so fewer doses were needed. [88]

Tables 4 and 5 show the effects of boosting the British and US vaccines with these adjuvants. To summarize the results, guinea pigs fared much better with the novel adjuvants than without them. Mice could not be adequately protected by MDPH-PA alone; CBA/J mice benefited somewhat from adjuvants, whereas A/J and Balb/c [17] did not. [41] [43] [95]

Boosting Vaccine Effects in Humans

Just prior to the Gulf War, amid fears of BW use, and with limited available supplies of anthrax vaccine, the British immunized their troops against various infectious diseases including anthrax, and included a killed Bordetella pertussis vaccine preparation as an adjuvant for the anthrax vaccine. [14] [57] [65]

The basis for this appears to have been research carried out on animals by Turnbull et al who wrote: “the human chemical vaccines as constituted have limited protective activities, lower than that induced by live spore vaccines available for use in the USSR … however, non-specific killed microbial additives, such as Freund’s Complete Adjuvant, Bordetella pertussis (as in the human vaccine) or Corynebacterium ovis can enhance the protective action of PA in the chemical vaccines to levels equivalent to or exceeding those of the live spore vaccines.” [90]

Butler [14] felt that use of the pertussis vaccine in this way was highly experimental, relying on preliminary results from Ministry of Defense-sponsored research at the Centre for Applied Microbiology Research at Porton Down, but was done to get troops out to the Gulf quickly. Questions have since arisen regarding possible adverse effects from this anthrax-pertussis combination in humans. It was noted in the animal trials that a group of guinea pigs that received three doses of a PA preparation plus B. pertussis and one dose of Freund’s complete adjuvant, showed signs of aging (hair loss, anorexia, and wasting in a proportion of the animals). [90] The researchers therefore challenged the animals with anthrax spores 4 months earlier than originally planned.

Might these vaccines contribute to immune system disease? Rook and Zumla [77] have proposed that Gulf War illnesses may be caused by a shift in cytokine balance from Th1 to Th2. This can be induced by use of multiple vaccinations, particularly pertussis, and possibly by exposure to carbamate and organophosphate insecticides as well, which inhibit Th1 function. [77] The British Ministry of Defense is now sponsoring a trial on the effects of the combination of vaccines and other treatments given to troops. [14]

In the United States, the most effective adjuvant formulations used with PA in animals were Detox, Triple mix (also known as Tri-Mix); and monophosphoryl lipid A (MPL) (all produced by Ribi ImmunoChem Research Hamilton, Montana). MPL was mixed with a squalene-lecithin-tween 80 emulsion. [39] [43] [88] Tables 4 and 5 show a comparison of efficacy with and without these adjuvants. Detox contains the cell wall skeleton of either Mycobacterium phlei or bovis, which includes muramyl dipeptide (MDP), and MPL (in a 10:1 ratio) in a base of 2% squalene. Triple mix contains the same ingredients with the addition of trehalose dimycolate, all in equal proportions, also in a 2% squalene base. These materials are immunogenic, but they are not harmless. Significant side effects, including uveitis, and adjuvant arthritis, have been described. [13] In one experiment where guinea pigs were given PA plus threonyl-MDP in an emulsion vehicle, five died unexpectedly 1 to 5 days postvaccination. [39]

Only aluminum-based adjuvants are licensed for human use in the United States, and military spokepersons have denied that unlicensed adjuvants or anthrax vaccines were given to US soldiers at the time of the Gulf War. [76] Asa and Garry, however, describe having found antisqualene antibodies in the blood of hundreds of patients reporting symptoms of Gulf War illness, and in rare controls (Robert Garry, PhD, personal communication, April 1998). This observation has reopened the issue of whether the US military, like the British Ministry of Defense, may have adjuvanted its anthrax vaccine in a novel way, in this case presumably using squalene-containing adjuvants. [62] Although efficacy may be improved by adding such adjuvants to a human anthrax vaccine, safety questions must be resolved before any large scale use in humans is attempted. [43]

Other Approaches to Anthrax Prophylaxis: Postexposure Therapies

In contrast to the limited efficacy of vaccines, antimicrobial treatment begun either shortly before or shortly after exposure to anthrax (though prior to development of clinical illness) has led to significant long-term survival. [26] [46] It promises to be very useful therapy, as long as the anthrax challenge strain is not antibiotic resistant. Doxycycline and ciprofloxacin have been the best performers of the drugs tested. [26] [46] A combination of postexposure vaccination plus antibiotics may be more efficacious than antibiotics alone. The Russians have created an antimicrobial-resistant vaccine strain for which they can use postexposure, live spore vaccination, and antibiotic therapy simultaneously. [69] [84]

One problem found with use of postexposure therapy is that a significant number of residual spores can be seen in the lungs of experimental animals after 3 or more weeks of antibiotic therapy, and may germinate following cessation of antimicrobial therapy, leading to death. [26] [46] [47] [84] It has been reported that the Russian live vaccine inhibits spore germination, whereas the chemical vaccines do not. [85] Whether administering a live vaccine after anthrax exposure, simultaneously with antibiotics, improves survival remains to be seen.

The use of antisera in the treatment of anthrax goes back to the preantibiotic era. Multiple case reports as well as experimental studies have documented some benefit of this approach, particularly because other treatments had little impact on systemic anthrax infections. [30] [32] [50] [55] [75] [92] [95] [99] In China, the use of a polyclonal antiserum, in addition to antibiotics (obtained after equine hyperimmunization with a mix of avirulent strains) is standard therapy for anthrax infection. [18] The development of monoclonal or polyclonal antianthrax antibodies and antisera for clinical use, however, has lagged in the West.

Anthrax Vaccine Manufacture in the United States

The sole US supplier of human anthrax vaccine is now Bioport– formerly MBPI and MDPH. Persistent quality control problems have plagued these vaccine manufacturers, as noted by a series of Food and Drug Administration (FDA) inspection reports over the past 5 years. [23] In 1997, the FDA threatened to revoke MBPIs license, citing an inadequate response to previous concerns raised by the FDA. [101] The FDA report of the most recent February 1998 inspection contains 11 pages of quality control failures in anthrax vaccine manufacture, including grossly inadequate standardization and testing, placing lots that failed testing into use, and use of vaccine from lots in which contaminants were found in some vials, without further testing of the remaining lot. [23]

In December 1997, the US DOD announced a program to vaccinate all 2.4 million US active duty and reserve servicemembers with MDPH-PA as prophylaxis against a BW attack, and vaccinations began in March 1998. Canada and Great Britain also began vaccinating some troops. Suggestions for using anthrax vaccine in civilians have been made as well, although at the present time the current anthrax vaccine is not stockpiled for civilian use. [58]

The first shipment of anthrax vaccines to the Gulf was recalled because it had apparently frozen during shipment. The anthrax vaccine lot then used for troop immunization by DOD beginning in March 1998 was FAV 020. This lot had been identified in the February 1998 FDA inspection report as having expired, been retested, and returned to use. [23] The report noted that the lot was mislabeled, so that it appeared to remain within the original expiration period.

Although lots received potency testing in order to return them to use following expiration, no testing was done for the presence of degradants, and no testing for stability was done prior to 1997. [23] One issue that does not appear to have been considered is that of adjuvant stability. Newman and Powell [64] state that adjuvants, such as alum, are prone to physical denaturation and any degradation that alters the protein-binding capacity can decrease the adjuvant activity. The reverse situation, where protein cannot undergo normal desorption in vivo, can also be a problem in aged alum formulations. Alhydrogel has a recommended 2-year shelf-life [93] but some lots of vaccine have been stored for at least 6 years. [23]

The MBPI anthrax line was shut down in February 1998 for refurbishing at DOD expense. The DOD intends to utilize its stockpile of 7 million doses (a number of which were cited for problems in the FDA report) while the plant is being improved. The sale of the MBPI by the state of Michigan to BioPort Corp, a firm directed by former chairman of the Joint Chiefs of Staff Admiral Crowe, was announced in July 1998. [59]

When the Department of Defense Drives the Vaccination Process

Ethical concerns arise when a medical treatment is used primarily or exclusively by the DOD. In the case of the anthrax vaccine, several issues have emerged:

  1. Annas [2] states that military regulations hold that, although a serviceperson must accept standard medical treatment or face court-martial, soldiers have no obligation to accept interventions that are not generally recognized by the medical profession as standard procedures. Because anthrax vaccination is not a standard civilian treatment, there may be no legal justification for forcing servicemembers to be vaccinated. Despite this, the current anthrax vaccination program is mandatory, and reprisals have been used against service personnel who have refused vaccination. [4] [29]
  2. After reviewing the history of the troubled MBPI, it might be suspected that FDA oversight of the manufacturing process was influenced by DOD concerns, because numerous infractions of federal regulations were allowed to persist. [23] [101]
  3. Military recordkeeping has not complied with civilian guidelines. The records of anthrax vaccinations administered at the time of the Gulf War apparently cannot be found, and the injections were not entered into the personal medical records of servicemembers. [15] [16] [71]
  4. Postmarketing surveillance must be performed to detect rare, adverse events associated with vaccination and to monitor the safety of practices such as simultaneous vaccination. [67] Postmarketing surveillance did not take place after the first large-scale use of MDPH-PA vaccine, at the time of the Gulf War, and evaluation of long-term safety of the anthrax vaccine administered to Gulf troops has not yet taken place. The safety profile of the vaccine therefore remains unknown, and questions persist regarding its role in the development of Gulf War illnesses. [15] [62]

According to the head of the FDAs Center for Biologics Evaluation and Research, data for clinical studies conducted on the long-term health effects of taking the anthrax vaccine have not been submitted to the FDA. [102] Yet, the DODs independent reviewer of the anthrax vaccination program reported, in answer to a question about the need for medical surveillance of vaccinees, that “anthrax vaccine is an FDA-licensed vaccine and no followup is required.” [12]

Expansion of Military Vaccinations

The DOD has proposed a large vaccine initiative, termed the Joint Vaccine Acquisition Program, for which initial funding of $321 million was approved in November 1997. [25] [45] [66] The goals of the project are to develop about 17 vaccines directed at BW threat agents, take them through the FDA licensing process, and arrange for manufacturing and administration to military personnel. The vaccinations against anthrax may be considered the prologue to this program.

As the Joint Vaccine Acquisition Program moves forward, the DOD is to fund and control all steps in the vaccine process, from initial research and development to manufacturing and administering the vaccines. If history is a guide, assessment of efficacy and safety, stringent manufacturing controls, and normal FDA oversight may be compromised. If the vaccines are licensed, as proposed, no informed consent need be obtained and vaccinations will probably be mandatory.

The DOD is assuming greater authority over the medical interventions given to troops at the same time that it has failed to follow agreed upon procedures for the use of experimental drugs and vaccines. This was noted both during the Gulf War and later in Bosnia. According to the October 1997 Presidential Advisory Committee on Gulf War Veterans Illnesses Special Report, “As determined by FDA, DOD’s use of tick-borne encephalitis vaccine during Operations Joint Endeavor/Joint Guard has violated federal regulations pertaining to investigational products on several accounts, including: recordkeeping failures; failure to monitor fully the study’s progress; failure to ensure the protocol was followed so safety and efficacy can be assessed; promotion of safety and efficacy for the investigational product; and failure to obtain Institutional Review Board approval of informed consent documents. FDA also expressed uncertainty about whether there had been a violation of Army recordkeeping and documentation requirements, which mandate that servicemembers’ permanent records accurately reflect TBE immunizations.” [72]

Should DOD be given carte blanche over the use of vaccines and therapeutics, when DOD will control all facets of production of those same materials, and the normal checks and balances of our proprietary system may be missing or unenforced? The ethics and legal implications of this situation have yet to be examined publicly.

Are Vaccines a Good Strategy to Counter the Biological Warefare Threat?

Vaccination may be a good idea in the short-term, particularly if national intelligence indicates that potential perpetrators are limited to microorganisms for which effective vaccines are available. Naturally occurring strains of anthrax, however, routinely overwhelm MDPH-PA vaccination in mice and guinea pigs. Whether this will carry over to humans has not been established. The ability of new adjuvants or new immunogens to solve this problem safely is uncertain.

In the longer term, vaccines appear to be less likely successfully to defend against BW threats. Given advances in biotechnology, and allegations that recombinant strains of anthrax and other microorganisms with both greater virulence than the native strains, and with added resistance to vaccines and antibiotics already exist, [73] it is unlikely that the present generation of vaccines will be protective.

Furthermore, it can take years to develop, test, license, manufacture, administer, and induce immunity using vaccines. Once one has created or obtained novel organisms, however, it takes only days or weeks to produce weaponizable quantities. Because of the necessity to perform human testing of vaccines and therapeutics, and make longitudinal observations prior to large-scale use, [3] [13] the development of medical countermeasures will necessarily lag far behind the creation of new bioweapons. The fact that we have no better vaccine to protect troops against anthrax 7 years after the end of the Gulf War puts this sharply into focus.

Development of biologic weapons has historically involved multiple microorganisms or toxins. [28] [100] If our troops are vaccinated against anthrax, an enemy may simply choose to use a different microorganism or toxin for which no vaccine is available. Although there is probably a role for vaccines, they should not be expected to provide a robust BW defense.

Conclusions: Thoughts on the Medical Response to Biological Warfare

Therapeutic Strategies
The present human anthrax vaccine probably provides only limited protection for troops facing a BW attack by anthrax. Postexposure antibiotics are likely to improve the outcome for victims following exposure, if the exposure is identified prior to the onset of illness, and the anthrax strain used is not antibiotic resistant. Monoclonal antibodies and antisera may well have a role in the treatment of anthrax: additional research should be done in this area. Defense planners should consider developing a library of monoclonal or polyclonal antibodies and vaccines to virulence factors of existing microorganisms. This will produce therapies that may be effective for novel organisms, when the recombinant organisms make use of known virulence factors.

Problems with Military Vaccines
If the DOD controls all steps in the vaccine development and production process, and is the employer of both physicians administering vaccines and servicemembers receiving vaccines, some or all of the following may result:

  1. Ethical conflicts of interest for those engaged in the process
  2. Insufficient testing of products or combinations
  3. Inadequate quality control of production
  4. Enforced administration of medical treatments or procedures that are not standard practice for civilians
  5. Inadequate record keeping
  6. Lack of proper surveillance for side effects following use.

There will be loss of the checks and balances implicit in the civilian medical system. Effective oversight of DOD’s health-related activities, such as vaccination, is therefore imperative.

Physical Protection
Protection for troops is challenging in a bioweapons attack; for civilians, it may be impossible. Medical interventions that protect troops will be accompanied by other measures, such as ready availability of barrier masks and suits, and use of air samplers and biodetectors. Although detectors are still rudimentary, they hold the promise of early bioagent detection in the future. These measures are not routinely achievable for the civilian population. Yet, is a perpetrator more likely to use bioweapons on vaccinated troops, or on unprotected civilians?

Primary Protection
BW is now, and will continue to be, difficult to deal with medically. There may well be no effective medical response to weapons that already exist, as well as to those that may be created. Therefore, the utmost efforts at primary prevention are demanded. Creation of and adherence to an international biologic weapons treaty regime that contains the most rigorous verification measures possible, including surprise inspections and stiff penalties for possession or use of biologic weapons, is needed. [48] Improvements in global infectious disease surveillance and communication of outbreaks should be made.

Although measures of this kind may not be completely effective at preventing biowarfare incidents, it is generally agreed that a strong BW treaty would still have significant positive effects. The possibility of being inspected without warning would deter many bioweapons programs. UN inspections in Iraq have clearly established the usefulness of such strategies at uncovering BW programs.

Before a multibillion dollar commitment is made to use vaccines as the primary strategy for mitigation of BW, a careful evaluation of their benefits and costs should be made. Strategies for prevention of BW should be moved to their rightful place at the forefront of the biologic weapons debate.

References

1. Anaisimova TI, Pimenov TV, Kozhukhov VV, et al: Development of method for preparation and maintenance of the anthrax strain STI-1 and test strain Zenkovsky. Salisbury Medical Bulletin 87(suppl):122, 1996

2. Annas G: Changing the consent rules for Desert Storm. N Engl J Med 326:770, 1992 Citation

3. Anthony BF, Sutton A: The role of the FDA in vaccine testing and licensure. In Lelvine MM, Woodrow GC, Kaper JB, et al (eds): New Generation Vaccines, ed 2. New York, Marcel Dekker, 1997, p 1185

4. Anthrax vaccine is refused. Washington Post. April 9, 1998, A5

5. Bernstein BJ: The birth of the US biological warfare program. Sci Am 256:116, 1987 Citation

6. Bernstein BJ: Churchill’s secret biological weapons. Bull At Sci 43:46, 1987

7. Brachman PS, Friedlander AM: Anthrax. In Plotkin SA, Mortimer EA (eds): Vaccines, ed 2. Philadelphia, WB Saunders, 1994, p 729

8. Brachman PS, Gold H, Plotkin SA, et al: Field evaluation of a human anthrax vaccine. Am J Public Health 52:632, 1962

9. Broad W: Gene-engineered anthrax: Is it a weapon? New York Times, Feb 14, 1998

10. Broad W: Norway’s 1918 lump of sugar yields clues on anthrax in war. New York Times June 25, 1998, A11

11. Broster MG, Hibbs SE: Protective efficacy of anthrax vaccines against aerosol challenge. Salisbury Medical Bulletin 68(Suppl ):91, 1990

12. Burrow GN: Letter to Rudy de Leon, Undersecretary for Defense. 2/19/98 http://www.defenselink.mil/other_info/burrows.html

13. Bussiere JL, McCormick GC, Green JD: Preclinical safety assessment considerations. Pharm Biotechnol 6:61, 1995 Citation

14. Butler D: Admission on Gulf War vaccines spurs debate on medical records. Nature 390:3, 1997 Citation

15. Committee on Government Reform and Oversight: Gulf War veterans’ illnesses: VA, DOD continue to resist strong evidence linking toxic causes to chronic health effects. HR 105–388. Washington, DC, US Government Printing Office, 1997

16. Committee on Veterans’ Affairs, Staff Report, US Senate: Is military research hazardous to veterans’ health? Lessons spanning half a century. S. Prt. 103-97, December 8, 1994

17. Coulson NM, Fulop M, Titball RW: Bacillus anthracis protective antigen, expressed in Salmonella typhimurium SL 3261, affords protection against anthrax spore challenge. Vaccine 12:1395, 1994 Abstract

18. Dong SL: Progress in the control and research of anthrax in China. Salisbury Medical Bulletin 68(suppl):104, 1990

19. Duesbury NS, Webb CP, Leppla SH, et al: Proteolytic inactivation of MAP-kinase-kinase by anthrax lethal factor. Science 280:734, 1998 Abstract

20. Ezzell JW, Abshire TG: Immunological analysis of cell-associated antigens of Bacillus anthracis. Infect Immun 56:349, 1988 Abstract

21. Farchaus JW, Ribot WJ, Jendrek S, et al: Fermentation, purification, and characterization of protective antigen from a recombinant, avirulent strain of Bacillus anthracis. Appl Environ Microbiol 64:982, 1998 Abstract

22. Farrar WE: Virulence and vaccines. Ann Intern Med 121:379, 1994 Citation

23. Food and Drug Administration: Report of the Inspection of Michigan Biologic Products Institute (Form FDA 483). February 20, 1998

24. Fouet A, Sirard J-C, Mock M: Virulence gene determinants. Salisbury Medical Bulletin 87(suppl):84, 1996

25. Fox J: ASM News. May, 1998, p 255

26. Friedlander AM, Welkos SL, Pitt MLM, et al: Postexposure prophylaxis against experimental inhalation anthrax. J Infect Dis 167:1239, 1993 Abstract

27. Geison GL: The private science of Louis Pasteur. Princeton, NJ, Princeton University Press, 1995

28. Geissler E: Biological and Toxin Weapons Today. Stockholm International Peace Research Institute. Oxford, Oxford University Press, 1986

29. Ginburg Y: Sailors refuse vaccine. Navy Times April 20, 1998

30. Gold H: Studies on anthrax. J Lab Clin Med 21:134, 1935

31. Hambleton P, Turnbull PCB: Anthrax vaccine development: A continuing story. Adv Biotechnol Processes 13:105, 1990 Citation

32. Heeren RH: Anthrax in Louisiana. New Orleans Medical and Surgical Journal 99:545, 1947

33. Holmes RK: Anthrax. In Fauci AS, Braunwald E, Isselbacher KJ, et al (eds): Harrison’s Principles of Internal Medicine, ed 14. New York, McGraw-Hill, 1998, p 897

34. Huxsoll DL, Patrick WC, Parrott CD: Veterinary services in biological disasters. J Am Vet Med Assn 190:714, 1987

35. Ivins BE: Anthrax vaccines — how stable is the potency? Presented at the ASM 98th General Meeting, Atlanta, May 18, 1998

36. Ivins BE, Ezzell JW, Jemski J, et al: Immunization studies with attenuated strains of Bacillus anthracis. Infect Immun 52:454, 1986 Abstract

37. Ivins BE, Fellows PF, Nelson GO: Efficacy of a standard human anthrax vaccine against Bacillus anthracis spore challenge in guinea pigs. Vaccine 12:872, 1994 Abstract

38. Ivins BE, Fellows PF, Pitt MLM: Efficacy of a standard human anthrax vaccine against Bacillus anthracis aerosol spore challenge in rhesus monkeys. Salisbury Medical Bulletin 87(suppl):125, 1996

39. Ivins BE, Fellows P, Pitt L, et al: Experimental anthrax vaccines: Efficacy of adjuvants combined with protective antigen against an aerosol Bacillus anthracis spore challenge in guinea pigs. Vaccine 13:1779, 1995 Abstract

40. Ivins BE, Welkos SL: Recent advances in the development of an improved, human anthrax vaccine. Eur J Epidemiol 4:12, 1988 Abstract

41. Ivins BE, Welkos SL, Knudson GB, et al: Immunization against anthrax with aromatic compound-dependent (Aro-) mutants of Bacillus anthracis and with recombinant strains of Bacillus subtilis that produce anthrax protective antigen. Infect Immun 58:303, 1990 Abstract

42. Ivins BE, Welkos SL, Little SF, et al: Cloned protective activity and progress in development of improved anthrax vaccines. Salisbury Medical Bulletin, 68 (suppl):86, 1990

43. Ivins BE, Welkos SL, Little SF, et al: Immunization against anthrax with Bacillus anthracis protective antigen combined with adjuvants. Infect Immun 60:662, 1992 Abstract

44. Jackson P, Hugh-Jones ME, Adair DM, et al: PCR analysis of tissue samples from the 1979 Sverdlovsk anthrax victims: The presence of multiple Bacillus anthracis strains in different victims. Proc Natl Acad Sci U S A 95:1224, 1998 Abstract

45. Joint Vaccine Acquisition Program: Final Programmatic Environmental Assessment. Joint Vaccine Acquisition Program Project Management Office, Department of the Army, August 1997

46. Jones MN, Beedham RJ, Turnbull PCB, et al: Antibiotic prophylaxis for inhalation anthrax. Salisbury Medical Bulletin 87(suppl):127, 1996

47. Jones MN, Beedham RJ, Turnbull PCB, et al: Efficacy of the UK human anthrax vaccine in guinea pigs against aerosolized spores of Bacillus anthracis. Salisbury Medical Bulletin 87(suppl ):123, 1996

48. Kadlec RP, Zelicoff AP, Vrtis AM: Biological weapons: Control, prospects and implications for the future. JAMA 278:351, 1997 Abstract

49. Kaufman A: Anthrax Vaccine Safety and Efficacy. ProMED-Mail, April 16, 1998. http://www.healthnet.org/programs/promed-hma/9804/msg00104.html

50. Knudsen GB: Treatment of anthrax in man: History and current concepts. Mil Med 151:71, 1986 Citation

51. Leppla SH, Friedlander AF, Singh Y, et al: A model for anthrax toxic action at the cellular level. Salisbury Medical Bulletin 68(suppl):41, 1990

52. Leppla SH, Klimpel KR, Singh Y, et al: Interaction of anthrax toxin with mammalian cells. Salisbury Medical Bulletin 87(suppl):91, 1996

53. Lesnyak OT, Saltykov RA: Comparative assessment of anthrax vaccine strains. Zh Mikrobiol Epidemiol Immunobiol 47:32, 1970

54. Lincoln RE, Walker JS, Klein F, et al: Value of field data for extrapolation in anthrax. Fed Proc 26:1558, 1967 Citation

55. Little SF, Ivins BE, Fellows PF, et al: Passive protection by polyclonal antibodies against Bacillus anthracis infection in guinea pigs. Infect Immun 65:5171, 1997 Abstract

56. Little SF, Knudsen GB: Comparative efficacy of Bacillus anthracis live spore vaccine and protective antigen vaccine against anthrax in the guinea pig. Infect Immun 52:509, 1986 Abstract

57. Miller C: Reserve troops suffering from syndrome. PA News (UK) September 24, 1997

58. Miller J: Clinton seeks additional $300 million to fight bioterrorism. New York Times June 9, 1998, A16

59. Miller J: Company led by top admiral buys Michigan vaccine lab. New York Times July 8, 1998

60. Nass M: Anthrax epizootic in Zimbabwe, 1978-1980: Due to deliberate spread? Physicians for Social Responsibility Quarterly 2:198, 1992

61. Nass M: Anthrax vaccine and the prevention of biological warfare? ASA Newsletter #65:1, 20, 1998

62. Nass M: Anthrax Vaccine Safety and Efficacy: Response to Army Surgeon General Ronald Blanck’s posting. Pro-MED Mail, May 7, 1998. http://www.healthnet.org/programs/promed-hma/9805/msg00044.html

63. Nass M: The labyrinth of biological defense. PSR Quarterly 1:24, 1991

64. Newman MJ, Powell MF: Immunological and formulation design considerations for subunit vaccines. Pharm Biotechnol 6:1, 1995 Citation

65. Norton-Taylor R: MOD ignored warning on Gulf drugs. The Guardian (UK) October 29, 1997

66. OraVax Joins DynPort in Department of Defense Contract to Develop FDA-Licensed Vaccines Against Potential Biological Warfare Agents. Cambridge, Mass., May 04/98 PRNewswire/ — OraVax

67. Orenstein WA, Hinman AR, Bart KJ, et al: Immunization. In Mandell GL, Bennett JE, Dolin R, (eds): Principles and Practice of Infectious Diseases, ed 4. Churchill Livingstone, NY, 1995, p 2770

68. Pitt MLM, Ivins BE, Estep JE, et al: Comparison of the efficacy of purified protective antigen and MDPH to protect non-human primates from inhalation anthrax. Salisbury Medical Bulletin 87(suppl):130, 1996

69. Pomerantsev AP, Mockov YUV, Marinin LI, et al: Anthrax prophylaxis by antibiotic resistant strain STI–AR in combination with urgent antibiotic therapy. Salisbury Medical Bulletin 87(suppl ):131, 1996

70. Pomerantsev AP, Staritsin NA, Mockov YV, Marinin LI: Expression of cereolysin AB genes in Bacillus anthracis vaccine strain ensures protection against experimental hemolytic anthrax infection. Vaccine 15:1846, 1997 Abstract

71. Presidential Advisory Committee on Gulf War Veterans’ Illnesses. Interim Report. US Govt Printing Office, Wash., DC, February, 1996

72. Presidential Advisory Committee on Gulf War Veterans’ Illnesses. Special Report. US Govt Printing Office, Wash., DC, October 31, 1997

73. Preston R: The Bioweaponeers. New Yorker, March 9, 1998, p 52

74. Puziss M, Manning LC, Lynch JW, et al: Large-scale production of protective antigen of Bacillus anthracis in anaerobic cultures. Appl Microbiol 11:330, 1963

75. Regan JC: The local and general serum treatment of cutaneous anthrax. JAMA 77:1944, 1921

76. Rodriguez PM: Anti-HIV mix is found in blood of Gulf War veterans. Washington Times, August 24, 1997

77. Rook G, Zumla A: Gulf War syndrome: Is it due to a systemic shift in cytokine balance towards a Th2 profile? Lancet 349:1831, 1997 Abstract

78. Schlingman AS, Devlin HB, Wright GC: Immunizing activity of alum-precipitated protective antigen of Bacillus anthracis in cattle, sheep and swine. Am J Vet Res 17:256, 1956

79. Shlyakov E: Anthraxin–a skin test for early and retrospective diagnosis of anthrax and anthrax vaccination assessment. Salisbury Medical Bulletin 87(suppl):109, 1996

80. Shlyakov EN, Rubinstein E: Human live anthrax vaccine in the former USSR. Vaccine 12:727, 1994 Abstract

81. Shlyakov E, Rubinstein E, Novikov I: Anthrax post-vaccinal cell-mediated immunity in humans: Kinetics pattern. Vaccine 15:631, 1997 Abstract

82. Stepanov AS, Klimpel KR, Leppla SH: Extracellular proteases in Bacillus anthracis. Salisbury Medical Bulletin 87(suppl ):87, 1996

83. Stepanov AS, Leppla SH: Macrophages are killed by the plasmid and chromosomally encoded factors synthesized by Bacillus anthracis inside and outside the host cell. Salisbury Medical Bulletin 87(suppl):89, 1996

84. Stepanov AV, Marinin LI, Pomerantsev AP, et al: Development of novel vaccines against anthrax in man. J Biotech 44:155, 1996

85. Stepanov AS, Marinin LI, Staritsyn NA, et al: Molecular mechanisms underlying Bacillus anthracis infection at early stages and search for novel vaccines. Vestn Ross Akad Med Nauk 6:16, 1997 Abstract

86. Stepanov AS, Mikshis NI, Bolotnikova MF: Role of chromosomally-encoded factors in virulence of Bacillus anthracis for mice and guinea pigs. Salisbury Medical Bulletin 87(suppl):86, 1996

87. Turnbull PCB, Leppla SH, Broster MG, et al: Antibodies to anthrax toxin in humans and guinea pigs and their relevance to protective immunity. Med Microbiol Immunol 177:293, 1988 Abstract

88. Turnbull PCB: Anthrax vaccines: Past, present and future. Vaccine 9:533, 1991 Abstract

89. Turnbull PCB, Broster MG, Carman JA, et al: Development of antibodies to protective antigen and lethal factor components of anthrax toxin in humans and guinea pigs and their relevance to protective immunity. Infect Immun 52:356, 1986 Abstract

90. Turnbull PCB, Quinn CP, Hewson P, et al: Protection conferred by microbially supplemented UK and purified PA vaccines. Salisbury Medical Bulletin 68(suppl):89, 1990

91. Van Ness GB: Ecology of anthrax. Science 172:1303, 1972

92. Vick JA, Lincoln RE, Klein F, et al: Neurological and physiological responses of the primate to anthrax toxin. J Infect Dis 118:85, 1968 Citation

93. Vogel FR, Powell MF: Compendium of vaccine adjuvants and excipients. In Powell MF, Newman MJ (eds): Vaccine Design: The Subunit and Adjuvant Approach. New York, Plenum Press, 1995, p 1464

94. Welkos S, Becker D, Friedlander AF, et al: Pathogenesis and host resistance to Bacillus anthracis: A mouse model. Salisbury Medical Bulletin 68(suppl):49, 1990

95. Welkos SL, Friedlander AM: Comparative safety and efficacy against Bacillus anthracis of protective antigen and live vaccines in mice. Microb Pathog 5:127, 1988 Abstract

96. Welkos SL, Friedlander AM: Pathogenesis and genetic control of resistance to the Sterne strain of Bacillus anthracis. Microb Pathog 4:53, 1988 Abstract

97. Whitford HW: Incidence of anthrax in the USA: 1945-1988. Salisbury Medical Bulletin 68(suppl):5, 1990

98. Williams P, Wallace D: Unit 731: Japan’s Secret Biological Warfare in World War II. New York, The Free Press, 1989

99. Williamson ED, Percival DA, Frith NJ, et al: Cell-mediated immune response to the toxins of anthrax. Salisbury Medical Bulletin 68(suppl):92, 1990

100. Zilinskas RA: Iraq’s biological weapons. JAMA 278:418, 1997 Abstract

101. Zoon KC: Letter from the Director of the FDA Center for Biologics Evaluation and Research to Robert Myers, Responsible Head, Michigan Biologic Products Institute. Available: www.dallasnw.quik.com/cyberella/Anthrax/Zoon4_98.htm March 11, 1997

102. Zoon KC: Letter to Patrick Eddington. FDA via FOIA. Available: www.dallasnw.quik. com/cyberella/Anthrax/Mar97.htm April 28, 1998

_____________________

Meryl Nass, M.D. is board certified in Internal Medicine, and practices at Mount Desert Island Hospital in Bar Harbor, Maine. She cares for hospitalized patients, and has a clinic where she treat patients with Fibromyalgia, Chronic Fatigue Syndrome, Gulf War Syndrome, Multiple Chemical Sensitivity and related disorders.

She has studied biological warfare and bioterrorism since 1989. In 1992 Dr. Nass identified the use of anthrax as a biological weapon in Rhodesia (now Zimbabwe) between 1978 and 1980, during a civil war between the white minority and black majority. Only areas populated by blacks were affected. As Rhodesia was then an apartheid nation, it was possible to confine the effects of the epidemic to areas where only black people lived and worked. The epidemic was part of a strategy of low intensity warfare, designed to wreak economic havoc by killing cattle needed for ploughing. At least 182 people were also killed.

Dr. Nass has worked to prevent biological warfare by creating ways of investigating suspect epidemics. A report she coauthored was presented to the 1996 international Biological Weapons Convention Review Conference in Geneva.

In 1998 Dr. Nass found that the anthrax vaccine being given to hundreds of thousands of military service-members was making many of them sick, and has since helped publicize the reactions and treated many affected individuals. She has pointed out the serious flaws in relying on vaccines as a panacea for the threat of biological warfare, as they are a medical Maginot Line. This was pointed out by the Congressional Committee on Government Reform in its 2000 report, Unproven Force Protection.

Dr. Nass volunteers to answer medical questions related to biodefense vaccines, host the website anthraxvaccine.org, advocate for better vaccines and drugs for bioterrorism, and continues to write academic and popular articles on anthrax, bioterrorism and biodefense vaccines in her spare time. Dr. Nass has provided five testimonies at the request of four Congressional committees on these topics since 1999.