A.  A Brief Overview of Virology

Viruses are very small organisms that can grow and replicate only inside of a living cell. The cell is required for the manufacture of the various components of the virus. These components are
DNA or RNA nucleic acids. These are the genes of the virus. Virus particles contain either DNA or RNA but not both, whereas cells, including bacterial cells, contain both DNA and RNA. The amount of DNA or RNA in a virus varies with different viruses. Small viruses, e.g., a parvovirus have about 3,000 nucleotide molecules strung in a single strand. Big viruses, e.g. a herpesvirus can have over 150,000 nucleotides in each of two complimentary nucleotide strands.

A protein (or proteins), which associate with the nucleic acids and provides some protection from nucleases and other environmental injury as the virus passes between cells. These proteins comprise the capsid of the virus.

Some viruses also have an outer protective layer of proteins in a lipid membrane. These envelope proteins can also assist in the attachment of virus particle to the surface of cells allowing for more efficient cell penetration.

Finally, many of the larger viruses contain additional proteins between the capsid and the envelope that can serve various functions. These proteins are commonly referred to as tegument proteins.

B. Virus Classification

Viruses can be classified in several of the following ways: i) Whether the vial particles contain RNA or DNA. Some RNA viruses convert by reverse transcription to a DNA copy after entering a cell. These viruses are called retroviruses. ii) Whether the nucleotides of the virus genome exist as a single strand (ss) or as a double stranded (ds) molecule. iii) For ss RNA viruses, the nucleotide sequence may directly code for the viral proteins (positive stranded viruses), or consist of the complimentary, or negative strand, that has to be transcribed to yield the protein coding sequence. iv) The virus genome may be linear or circular and may be in a single segment, or in multiple segments. Multiple segmented viruses can occupy the same particle, or be partitioned into two or even three particles (bi-partate and tri-partate viruses). v).The genome size, overall shape of the virus particles and whether an envelope is present, are also major criteria used for virus classification. vi) The kinds of cells that are susceptible to a particular virus and the clinical features of the diseases produced by a virus can also help categorize different viruses. Related viruses are successively grouped into species, genera, families and orders (White and Fenner Clinical Virology, Academic Press)

C. Virus Pathogenesis

All viruses have the potential to mediate cellular changes. They can do so by altering the normal metabolic balance within the cell through the over utilization of the cell's energy resources. While this can eventually lead to cell death, an earlier result can be the failure of the cell to perform all of its normal functions. A disruption of normal function can be especia and stealth virus infected animals, have shown clinical improvement with time. This finding may be related to a genetic instabiopsychiatric activities compared to similar damage to extra-neural tissues.

Continued metabolic drain on a cell can lead to a loss of crucial components. For example, since ATP is required by the mitochondria to maintain structural integrity and their capacity for lipid catabolism and oxidative phosphorylation, a lack of ATP can impair even the ability to manufacture this essential component. Mitochondria damaged cells can show foamy vacuolization, swelling and intercellular fusion. Accumulation of virus products can also lead to physical cell damage especially if the products tend to aggregate into insoluble debris. Certain viruses trigger a more active form of cell death, called apoptosis, characterized by shrinkage and condensation of cellular components. While herpesviruses, especially Herpes Simplex viruses, and human cytomegalovirus (HCMV), adenoviruses, influenza , measles, several enteroviruses and others viruses can be cytopathic for cultured cells, many of the known human viruses are essentially incapable of producing a readily discernable cytopathic effect (CPE) in cultures. For example, rubella, hepatitis A, B, C and D; human T lymphocytotrophic virus (HTLV), Borna and Hartaan viruses, are non-cytopathic. Moreover, primary clinical isolates of measles, mumps and polioviruses induce far less CPE on human when compared to animal cell lines. For many non-cytopathic viruses, the in vivo damage that occurs following infection is a consequence of immunological activation.

D. Virus Immunity

The immune system can both reduce and enhance the extent of virus damage. A major component of the immune system is mediated by circulating antibodies. Anti-viral antibodies can provide an effective blockade preventing viruses from gaining access to normally permissive cells. In particular, antibodies reactive with virus particles, can help prevent viruses passing from the blood into the brain. Protective virus reactive antibodies can be elicited by immunization with any materials containing the antigens expressed by the virus particles. The other major component of the immune system is mediated by lymphocytes and referred to as cellular immunity. This type of immunity can reduce virus load by destroying infected cells prior to the release of infectious virus particles and by eliciting an inflammatory response. Cellular immunity against virus antigens expressed on the surface of cells can also lead to immune damage of cells beyond that achieved by the virus itself. Moreover, virus modification of, and/or inappropriate expression of, various cellular components can trigger an immune response against these self-antigens leading to auto-immune mediated tissue damage.

E. "Stealth Adaptation" as an Escape Mechanism from Anti-Virus Cellular Immunity

While researchers had envisioned various ways in which viruses might avoid immune elimination, it had not previously been suggested that certain viruses might simply bypass the immune system by deleting and/or mutating critical genes involved in the cellular immune recognition of virus infected cells. The inventor has pioneered the concept of a "stealth adaptation" as a means of evading the cellular immune system. In addition to the substantial experimental evidence in support of the existence of stealth viruses, one can proceed along the following lines of reasoning. The cellular immune system is mediated by T lymphocytes. As elegantly proven in the clonal selection theory of immunology, each individual T lymphocyte carries receptors for a single antigenic specificity. A corollary of the clonal selection theory is that to be effectively recognized by a T lymphocyte, a virus infected cell must restrict the diversity of different virus antigens presented to the cellular immune system. Even with large complex viruses, relatively few viral components actually serve as effective targets for cellular immune defenses. For certain viruses, e.g. HCMV, experimental studies had indicated that deletion (or mutation) of genes coding for the major viral components recognized by the cellular immune system, would result in a defective, non-replicating, non-cytopathic, viral sequence. One could have, however, a potential building block towards the evolution of a cytopathic, non-immunogenic virus. Potentially, the downsized gene-deleted virus could form a synergy with a replicating non-cytopathic virus and/or incorporate certain cellular genes by recombination, to yield an atypically structured cytopathic virus. This concept has provided a background for a series of studies leading to the detection and characterization of what have been termed "stealth viruses."

F. Stealth Viruses

Stealth viruses can be defined as a molecularly heterogeneous grouping of atypically structured, cytopathic viruses, that cause persistent systemic infection, frequently associated with neuropsychiatric symptoms, in the absence of significant anti-viral cellular inflammation. Stealth viruses typically induce a vacuolating foamy CPE in a range of human and animal cell lines. The formation, progression, and/or host range of the CPE distinguish stealth viruses from traditional human cytopathic viruses, including herpesviruses, enteroviruses and adenoviruses. Additional distinctions from conventional viruses can be made on the basis of electron microscopy, serology and molecular-based studies. The definition of stealth viruses does not follow the approach outlined above of using restrictive criteria based on either the virus genomic sequence or the virus morphology. Rather the approach taken to initially define stealth viruses was based on the foamy vacuolating CPE and other in vitro growth characteristics and on the exclusion of other known viruses. As the research work progressed, several individual stealth virus isolates were characterized in terms of more conventional criteria, including their electron micrographic appearance, electrophoretic pattern of isolated DNA and RNA, partial sequencing and determination of probable origin. Even with the best characterized of these stealth virus isolates, however, the strict chemical and morphological classification schemes fail to account for the microheterogeneity and sub-genomic expression that is observed within a single isolate. Moreover, the precise chemical features of one isolate do not adequately encompass the broader concept of a diverse group of cytopathic viruses in which deletion and/or mutation of the major immunogenic components has occurred. Stealth adaptation is viewed as a mechanism to facilitate persistent infection by structurally loss of the normal capacity of conventional viruses to evoke an effective anti-viral cellular inflammatory response. Given these considerations, stealth viruses have not originated from a single source. It is likely that stealth adaptation can occur with all of the presently known human herpesviruses and many of the herpesviruses known to infect animals. Viral sources other than herpesviruses are not excluded, and, in fact, have been suggested in various studies. Indeed, as viruses downsize and simplify, their initial distinguishing characteristics tend to become less important compared to their common pathogenic capacity of overtaxing the metabolic resources of the cell.

G. Stealth Virus Detection

Tissue culture methods can provide a broad screening method for the detection of stealth viruses. An early observation was that the intensity of the CPE could be enhanced by frequently feeding the cultures. This led to the detection of virus inhibitory material in the supernatant of infrequently fed cultures. Low stringency molecular probing using the polymerase chain reaction (PCR) can also be used to detect abnormal DNA and RNA sequences in patients' samples and in virus cultures. Empirically, it was found that a primer set originally designed to amplify HTLV viruses could yield varying products with several stealth virus isolates. The amplified products could then be cloned and sequenced.

An additional method of stealth virus detection is provided by serology, using either the patient's own sera, or sera collected from various individuals apparently exposed to a stealth or to a related conventional virus. Specific antibodies can also be used to distinguish stealth viruses from common conventional viruses, such as HCMV. Finally, electron microscopy and animal transmission studies have proven useful for stealth virus detection and partial characterization.

H. Stealth Virus Disease Associations

As indicated above, stealth virus infection was initially described in association with disorders of brain function. Based on unequivocal culture and PCR based findings, it was proposed that stealth virus infection could potentially account for a wide diversity of clinically distinct neuropsychiatric illnesses. The diseases included well defined common psychiatric disorders, including schizophrenia and manic-depression, for which a virus etiology had been largely discounted. Neurological illnesses implicated by positive culture findings as being potentially connected to stealth viruses, included several patients labeled as having Alzheimer's disease, post infectious encephalopathy, otherwise unexplained coma. As if this list of illnesses was not large enough, the culture and PCR based assays clearly suggested an involvement of stealth viruses in at least some patients labeled as having the chronic fatigue syndrome (CFS).

I. Chronic Fatigue Syndrome

Attempts to define the chronic fatigue syndrome (CFS) as a clinical diagnostic entity have mainly failed because of the lack of a clear separation of what would be considered normal variability in human functional capacity, and what should be considered as a medical illness. At the other extreme, many patients with a clearly manifested severe fatiguing illness are inappropriately grouped along with individuals with only minimal impairment in their daily activities. Some severely affected CFS patients eventually met criteria for neurological, psychiatric and/or immunological disease classifications. The possible connection between CFS and these other illnesses is, however, often unrecognized by specialist clinicians and possibly actively discouraged by medical insurance providers. For understandable reasons, CFS patients are also hesitant to accept a psychiatric component to their illness.

The thesis was advanced that CFS is but one of many differing manifestations of a persistent stealth viral infection within the brain. Involvement of the brain in CFS patients is implied by the historical use of terms such as neurasthenia, myalgic encephalomyelitis, epidemic diencephalomyelitis and limbic encephalopathy. In more recent years, however, several investigators have argued that the disordered brain function is a secondary phenomenon, resulting, for example, from the overproduction of neuromodulatory cytokines from an activated immune system, that may be responding excessively to a multitude of normally tolerated ubiquitous microorganisms, such as Epstein Barr virus, human herpesvirus-6, Candida albicans, mycoplasma fermentans, chlamydia pneumoniae, etc. Recent attention has also be given to possible brain damage resulting from exposure to environmental neurotoxins, including the potential release into the circulation of neurotoxic bacterial products from a damaged gastrointestinal tract .

The shift away from a primary infectious process within the brain had occurred in spite of numerous epidemic outbreaks of CFS-like illnesses. Reasons for this neglect include the failure of established CFS investigators to isolate viruses from CFS patients, and by the lack of correlation of disease with conventional anti-viral serology. Published studies using the polymerase chain reaction (PCR) to test for evidence of retroviruses, enterovirus and mycoplasma infections, were also flawed by erroneous assumptions concerning the specificity of PCR assays when performed under low stringency conditions. The imposition of a restrictive clinical definition of CFS has especially hindered the capacity to validate any suggested new assay, since it required that only patients with fatigue should test positive. This demand has also obscured epidemiological studies for potential disease transmission within families or communities.

J. Fibromyalgia

Complaints of excess pain and stiffness generally throughout the body led to the suggestion of a low grade inflammatory condition involving the body's connective tissue. The condition was referred to as fibrositis and categorized along with various auto-immune connective tissue diseases, such as lupus erythematosus and rheumatoid arthritis. Inflammation could not be demonstrated on tissue biopsy. An alternative term, fibromyalgia was introduced by rheumatologists and several clinical criteria established to enable a positive diagnosis. Patients with fibromyalgia were commonly fatigued and the overlap with chronic fatigue syndrome became to be appreciated by many physicians. More importantly, once the focus shifted away from pain, the underlying neurocognitive deficits, insomnia, personality and mood changes, heightened sensitivity to various stimulants, and other features clearly established a common bond between fibromyalgia and chronic fatigue syndrome.

K. Gulf War Syndrome

As if clinicians had never experienced patients with chronic fatigue/fibromyalgia disorders, a new category of illness was introduced to account for the fatigue, aches and pain, neurocognitive deficits, insomnia, and other features listed above, that developed in large numbers of the deployed, and even non-deployed, troops involved in the Gulf War. From a non-existent illness, the condition became a "catch all" for miscellaneous illnesses of supposedly multiple origins. There was reluctance to accept any contribution to the illness from chemical exposure and a active resistance to openly speak of a possible infectious origin, for example from contamination of the gamma globulin preparations received by troops as a preventative measure against hepatitis infection. Money to investigate this syndrome was safely placed in the hands of clinicians who lacked both the means or scientific perspective to uncover novel infectious agents.

L. Autism, Attention Deficit and Behavioral Disorders in Children

One of the major tragedies that has impacted modern society is the increasing numbers of children struggling with less than optimal brain function. Clinical labels such as autism, attention deficit and hyperactivity syndrome, oppositional defiance, conduct disorders, seizure disorders, anorexia nervosa, etc. serve to lessen the awareness of the overall enormity of the problem. With a national average of nearly 10% of children partitioned into so called special educational programs, the complacency of foregoing a possible biological explanation for the deteriorating mental health of children should is inexcusable. Autism generally appears within the first 1-2 years of life. It is reflected in an impaired ability to emotionally or verbally communicate. Interestingly, some cases of autism appeared to follow closely upon the administration of a vaccine. At least some cases of autism have a structural basis suggestive of a neurodevelopmental disorder. Again very few studies had pursued a potential virus contribution to this illness or to less severe forms of disability variously called high functioning autism, attention deficit and hyperactivity disorders, etc. The incidence of autism has reportedly greatly increased in recent years.

M. Alzheimer's Disease and Other Dementing Illnesses

At the other end of the age spectrum, many elderly individuals have experienced major losses in cognitive abilities. In some, the disease runs the protracted course merging with "old age." For others, however, the mental deterioration can be rapid and, one would think, would demand an explanation other than he or she has Alzheimer's disease. It is not even as if one understood the disease named after Dr. Alzheimer, or after Dr. Parkinson, etc.

N. Other Commonplace Neurological and Psychiatric Disorders

Selecting out individually named syndromes and diseases, runs the risk of not seeing various brain illnesses as a spectrum of disorders that may well have a unifying explanation in terms of a viral infection of the brain. It over categorizes patients and establishes competitive boundaries between patient advocate groups. Naming a disease, whether multiple sclerosis, amylotrophic lateral sclerosis, chronic Lyme disease encephalopathy, schizophrenia or manic depression, projects a sense of completeness; that the diseases are understood; that the patients should not expect further inquiry into what has caused the disease, or how the disease can be distinguished from that of others bearing the same label.

O. Malignancy

A prediction of the finding that stealth viruses can apparently "capture, amplify and mutate" various cellular sequences, is that stealth virus infection could lead to cancer. While, clinical examples in support of this possibility have been previously recorded, the regular finding of stealth virus infection in patients with multiple myeloma, has added a sense of urgency to this issue. Multiple myeloma is a malignancy in which relatively differentiated B lymphocytes, with the appearance of plasma cells, accumulate throughout the bone marrow, and other tissues of the body. Less well differentiated lymphocyte malignancies appear as lymphomas. There have been several suggestions that multiple myeloma patients may be infected with a newly defined human herpesvirus 8. The data are more consistent with a stealth virus infection. Other prominent examples of positive stealth virus cultures include two adults with glioblastomas and several patients with salivary gland tumors. There is a need to examine patients with other types of cancers.

P. Family Illnesses

A potentially infectious origin of the above conditions has important implications regarding the safety of the Nation's blood supplies, import and export of biological products, occupational exposure, etc. While these issues are undoubtedly topics for high level economic discussion, they offer little comfort to the families faced with various illnesses occurring within multiple family members. The "stressed out" mother no longer able to care for her autistic son or demented husband. The "depressed" siblings, one anorexic and striving for perfection, the other with failing grades and emotionally blunted.

Q. Multi-System Illnesses

The specialization of medicine has focused attention on disorders that are essentially restricted to a single organ system. Multi-system diseases tend to fall outside the purview of most physicians and a balanced, comprehensive approach to their assessment is often lacking. Clinical review of many patients with stealth viral infections, initially labeled as having either CFS, fibromyalgia, Gulf War Syndrome, Lyme disease, schizophrenia or autism, has confirmed the importance of viewing each of these conditions as a systemic infection, rather than simply as an encephalopathy. The systemic nature of the illnesses adds further diversity to the clinical categorization of these patients, leaving many without proper evaluation of their overall illness.

R. Clinical Diversity in Stealth Virus Infected Patients

As discussed earlier, the spatial distribution of its various functions renders the brain uniquely susceptible to localized viral-induced cellular damage. Moreover, the brain exerts important controls on the functions of other organs through neural and hormonal pathways. An encephalopathy can readily explain several of the non-organ disorders that have been associated with chronic fatigue syndrome, such as neurally mediated hypotension, adrenal deficiency, irritable bowel syndrome, dysphagia, etc. An indirect effect through the brain does not account for many additional signs and symptoms seen in stealth viral infected patients. Summarizing from the patients described above and from other patients seen in recent years, the following symptoms can be attributed to the direct effects of a viral infection on other organs.

Liver and gastrointestinal tract: Slight elevations in liver function enzymes are potentially reflective of viral infection of liver tissue. The microvesicular histological changes seen on the liver biopsy of patient JL are similar to changes described in other CFS patients on whom liver biopsy has been performed. Liver dysfunction can reduce detoxification capacity and account for the enhanced susceptibility to various noxious xenobiotic and environmental chemicals that is a characteristic of many stealth viral infected patients, and a hallmark of multiple chemical sensitivity. Subclinical viral infection of the gastrointestinal tract can add to the demands of liver detoxification by helping the breach the normal barrier function of the gut to products of its normal bacterial flora and possibly even to normally resident pathogens such as Candida and mycoplasma. Pancreatic dysfunction, coupled with malabsorption of required nutrients can potentially explain the wasting syndrome that has been observed in several patients. Furthermore, the gut and liver may be impaired in their synthesis of various compounds required for normal cellular function.

Endocrine glands: Hypothalmic and pituitary dysfunction can be included within the spectrum of an encephalopathy. They can have secondary effects on normal menstruation, water balance, thyroid and adrenal function, etc. In addition, there is evidence of primary thyroid and adrenal dysfunction in some stealth viral infected patients. Direct viral infection can also evoke secondary auto-immunity adding to endocrine disease.

Genital organs: The pelvic pain that is occasionally encountered in patients is probably triggered by direct viral infection of the genital organs. This can present as prostate and testicular complaints in men and vulvar, low back and menstrual pain in women. An accompanying reduction in the pain threshold due to the viral encephalopathy can aggravate the pelvic pain.

Joints and renal involvement: While anti-viral cellular immune responses may be impaired, stealth viral infections can evoke circulating antibodies. Indeed, there may be overproduction of antibodies reactive with a range of viral and auto-antigens. Antigen antibody complexes can cause arthralgia and can also lead to long term kidney disease. They can also cause secondary vasculitis with widespread manifestations.

Skin, hair and muscle: Although not discussed specifically in this section, several cases of vitiligo have been studied for stealth viral infection with positive findings. A more common report by patients is the occasional outbreaks of small vesicular lesions suggestive of herpesviruses. Indeed, when tested, cytopathic viruses have been cultured from such lesions. Several herpesviruses are known to target hair follicles (e.g. Marek?s disease in chicken) and hair loss is not an uncommon complaint among CFS patients. Muscle wasting can be secondary to nerve dysfunction, but may also be a direct result of viral infection with or without an added auto-immune component.

Immune system: The persistence of a systemic viral infection can explain the various immunological changes seen in stealth viral infection. As with many of the other effects of stealth viral infections, these changes fluctuate over time and vary considerably between different patients. Ongoing immunity to a specific viral infection, can modulate responses to other antigenic stimuli and may account for enhanced allergic reactions and depressed delayed hypersensitivity responses seen in various patients.

S. Recovery from Stealth Virus Infections

In spite of an apparent lack of effective cellular immunity, many stealth virus infected patients, and stealth virus infected animals, have shown clinical improvement with time. This finding may be related to a genetic instability of the stealth virus. It may also be a reflection of the non-progressive nature of the CPE that is often observed in infrequently fed virus cultures. A potent inhibitor of stealth viruses can be found in supernatants of infrequently fed stealth virus cultures. While the nature of this inhibitor has yet to be biochemically defined, it offers one of the best prospect for the suppression of a stealth virus infection. Various non-immunological mechanisms of virus resistance are clearly operative in plants and may also be relevant to the findings of disease regression in stealth virus infections in humans and in animals. Various drugs might also selectively suppress virus replication and help to minimize cellular damage. These can include common anti-viral agents such as ganciclovir, acyclovir, foscarnet and interferon.

T. Prevention of Stealth Virus Infection

Immunization designed to elicit protective antibodies can potentially provide protection against an initial infection. The source of antigen can be from a stealth virus or from a conventional virus with antigens structurally related to those on a particular stealth virus isolate. The primary goal of immunization is to elicit antibodies that could provide a barrier to infection. Antibodies on a mucosal surface could prevent initial infection, while circulating antibodies could help prevent blood to brain transmission of virus. Stealth viruses comprise a molecularly and immunologically heterogeneous grouping of atypically structured viruses. Even if a person is infected with a stealth virus, it may still be appropriate to immunize against other types of stealth viruses. The production of antibodies as a result of immunization can be tested in neutralization assays. In this type of assay, one would expose stealth virus particles to the antibody and check for residual infectivity either in culture or in animals.

U. Therapy for an Existing Stealth Virus Infection

An early observation made during the in vitro culturing of stealth viruses was that the CPE often failed to progress and, in fact, often receded. Frequent feeding of the cultures helped counteract the lack of CPE progression. It was further noted that virus inhibitory materials were accumulating in infrequently fed cultures. The in vitro findings were also consistent with the lack of marked progression of the disease in many stealth virus infected patients. Thus given the apparent lack of effective cell mediated immunity, one needed to explain why the virus was not pursuing a relentless progression throughout all of the tissues of an infected individual. Various approaches can be suggested towards the effective therapy of stealth viruses, e.g., nutritional to offset the metabolic inbalance caused by the virus; symptomatic to alleviate many of the disease manifestations; and anti-viral. Of these approaches, the anti-viral therapy would appear to hold the greatest promise. While certain isolates of stealth viruses are partially suppressed by some of the clinically approved anti-viral drugs (e.g. ganciclovir, Acyclovir, interferon) virtually all of the stealth viruses display the tendency for their CPE to recede if they are not appropriately cultured. This finding implies a potential therapeutic use of the inhibitor that has been identified in stealth virus cultures. The inhibitor has been termed Epione after the wife of the Greek physician Ascepius. Although Epione has not been characterized biochemically, the inhibitory activity has been partially purified.

V. Restatement and Summary of Work That Led to the Detection and Characterization of Stealth Viruses.

Because many of the concepts underlying the nature of stealth viruses and their importance in human and animal diseases, have yet to be widely accepted, it may be worthwhile to restate some of the studies that led to their detection. Copies of published articles are also being provided.