Influenza genetics appears bedazzling to us lay people but looking for understanding is best found in the historic research on “bacteriophage”, or simply called “phage” research. Phages are described as the viruses that infect bacteria, although use of the word “infect” needs qualification. All complex living things are composites of cellular material including bacteria and viruses. The “Origin of Life” theorists claim that human beings are composed of 90% microbes. Evolutionary biologists are recognizing that viruses are essential biologically derived “chemicals” that further the processes of life. Living organisms synthesize their own viruses to perform complementary activity as part of their natural adaptation to the environment. Using the word “phage” instead of the word “virus”, the phages of bacteria “contribute to bacterial homeostasis in nature, keeping bacteria under control”. They “do not leave an ecological footprint — phages are comprised of and disintegrate into amino acids and nucleic acids, and are normal commensals of humans and animals.” Phages and viruses also arise from the very same acids and other available material in the environment. The shifting “transitional” forms are called “polymorphisms”. In the 19th century, Antoine Bechamp and Claude Bernard pioneered the study of polymorphisms and came into conflict with the “monomorphism” of Louis Pasteur from whom we get the basic “Germ Theory”. Pasteur, apparently, received his funding and support from Gustave de Rothschild, building state of the art laboratories in Paris and elsewhere. The commercial interests at the time were focused on the processes of fermentation, or harnessing bacteria for industrial use in food and beverage production.
A self-taught bacteriologist named Felix d’Herelle dubbed the “phage” in 1917 with a Greek name, meaning roughly “eater”, and became the most eminent of proponents for phage therapy. He believed they were organisms, as early virologists believed in the living status of virus. –”D’Herelle’s theory that the material is a living virus parasite of bacteria has not been proved. On the contrary, the facts appear to indicate that the material is inanimate, possibly an enzyme” — [ref. below].
Felix d’Herelle traveled the world as a contract bacteriologist to national governments, setting up fermentation operations in Canada, Guatemala, Mexico, Argentina, and France. In 1925 d’Herelle was hired as a ‘health officer’ of the League of Nations to monitor infectious diseases on board ships passing through the Suez Canal (Suez was the one of the most strategic shipping channels on the globe). He lectured and taught in the United States at Stanford and Yale, and in 1933 he left the US and went to the USSR to found the Bacteriophage Institute in Tbilisi. He appears to have been a dedicated communist throughout his life and did not return to the US, although he maintained his activities including his businesses in France and Russia.
D’Herelle’s legacy in the US was organizing “one of the most extensive trials of phage therapy” called the Bacteriophage Inquiry, a project spanning the years 1927 to 1936, involving a lot of human experimentation. After him, phage work was taken up by Max Delbruck, Salvador Luria and Alfred Hershey. A “phage group” was formed, involving the institutions primarily of Cold Spring Harbor Lab (Long Island, NY), California Institute of Technology (Caltech, Pasadena CA) and Vanderbilt University (Nashville, TN). Summer sessions at Cold Spring Harbor on phage research became a yearly feature of genetic research. It is documented that over 800 scientific papers were published on phage research from its beginnings up through 1956. Influenza researchers avidly used phages to study human, swine and avian influenza viruses. One of the most prominent flu researchers, George K. Hirst, who worked under the auspices of the US Army as well as private institutions, notes that phage-influenza researches were “dropped” after this time. Where did it go? What were they learning? I’ll be back with answers to those questions and more.
In 1959, a textbook, “Bacteriophage”, was published from the career work of Mark Hannock Adams at Cold Spring Harbor Lab
who died suddenly in 1956 of an acute infection at age 44. Far from disappearing in the late 50s, the phage research had become complicated and difficult to access. Max Delbruck wrote in the introduction to Bacteriophage
that “the fact is that the literature on this subject has grown so complex as to deter everybody, except Mark Adams.
” Adams’ untimely death led to the revision and completion of his work by a team of 15 scientists, overseen by executive editor Alfred Day Hershey
, who later directed the Carnegie Institute. It is their version of Adams’ book linked here: www.archive.org/stream/bacteriophages00adam/bacteriophages00adam_djvu.txt
A tantalizing passage that holds a clue to the Spanish Flu suggests that “In 1917, Felix d’Herelle…published his independent discovery…and in 1921 published his classic book…stimulated by the announcement that hog cholera [swine flu] was due to the synergistic action of a bacterium and a virus…” This scenario is very near to a report claimed by Henry Makow today that a captured Nazi told of a “double-blow” bug made into a bioweapon that caused the Spanish Flu (1918-1919). Other hints in the text continue: “There is no doubt that numerous early bacteriologists saw and described signs of phage action…however, no intensive investigation of these phenomena was undertaken prior to the appearance of a brief but provocative paper by F.W. Twort (1915)…Twort considered various hypotheses to explain this phenomena [of synergy]; among others that it was a filterable virus analogous to the virus pathogens of animals and plants…yet the paper remained unnoticed by scientists..”
Delbruck is one of a host of generalists on the subject pushing not just the public, but the science community away from the idea that the value of phage research was embraced, offering disparagment and dismissal in place of acceptance. Were this really true, the breakthrough of ‘lysogenic’ discoveries in the 1920s would have been unlikely. Joshua Lederberg reports in a 1951 paper [see the ‘Mutation’ post on the blog] in the section called “Transformations” that “the history of ‘type transformation’ is often regarded as dating from Griffith’s classical experiment on pneumococcal types in 1928 but this work is andedated by a confusing array of studies which date at least as far back as the controversy over the etiology of typhus fever….Griffith’s demonstration was..reproducible by other workers and remains a proper starting point for modern discussions of transformation. [‘transformation’ describes the inclusion of transferred genetic material, changing the properties of a cell]…there are a number of more recent reports [as of the 1950s] of transformations in various organisms. These include E. coli, Hemophilus influenzae, Shigella paradysenteriae, Alkaligenes radiobacter and staphylococcus.”
Among the possibilities I’ve considered of an intentional bioweapon seeded at select locations in 1918 is the potential of poliovirus as an analog of influenza. In the polio epidemics that preceeded the Spanish Flu, a cause of death was overwhelmingly recorded as bacterial atypical pneumonia, the same diagnosis that accompanied a huge proportion of deaths in the 1918 pandemic. Intestinal bacteria were commonly found in the lungs, and as Frank Macfarlane Burnet observed in 1931, the Pfeiffer’s bacillus (hemophilus influenzae) appeared in the respiratory tract two to three days after a flu infection had begun (first recorded in the 1889-93 “Russian Flu” pandemic). Simon Flexner, director of RIMR (Rockefeller) was convinced that polio was spread exactly in the manner of influenza, holding this conviction against the opinions of his peers and against the evidence of experimentation . Polio appeared noncontagious. It was Flexner’s work in 1896, published in the JEM, that noted the presence of gut bacteria in the lungs of pneumonia victims. The work involved subjecting bacteria to toxins and ‘treatments’ that caused increased virulence in the organisms. Shortly after, incidentally, he went to the Philippines as a journeyman bacteriologist and discovered a new strain of Shigella (shigella flexneri), the most reliably virulent of the shiga toxin bacteria.
But if indeed a bioweapon was released in 1918 to account for the devastating effects of the ‘black influenza’, the science community was no more able to find it in the following decades. That is until 1933 and the isolation of human influenza virus
, as this British version would have you know. www.nimr.mrc.ac.uk/MillHillEssays/2000/influenza.htm
. In this account of the discovery of influenza, researchers at ”Mill Hill” near London accidently infected their lab ferrets with their own flu. At the time they were experimenting with ‘distemper’, developing a vaccine for the flu-like illness in dogs. According to this website, influenza was unusually harsh among the British people in the winter of 1933. Interestingly, polio outbreaks had spiked dramatically throughout the year before, especially in the U.S. Distemper appears like this in dogs: “Infected animals have high temperatures and may vomit and have diarrhea. There is a discharge from the nose and eyes and the disease is spread when virus in droplets of discharge is inhaled. The animals cough and may have pneumonia. About half of them have fits and tremble uncontrollably, even when they are alseep. Sometimes their hind legs are paralyzed. If they survive, distemper can also cause long-term problems..[like]“old dog encephalitis
“, a sort of canine senile dementia
“. This webpage continues the polio correlation, “It’s interesting that the use of this proceedure [in distemper] of vaccinating with inactivated virus vaccine and then with infectious virus is now recommended for vaccination against poliomyelitis
The London researchers, 3 who were given the credit of discovery, represent a team working for the National Institute of Medical Research’s “World Influenza Centre”, a study establishment created after the 1918 pandemic. Animal viruses were the subjects. Frank M. Burnet was there on a Rockefeller grant along with Sir Patrick Laidlaw, Sir Christopher Andrewes, Wilson Smith, W.I. Elford and J.E. Barnard. The first human influenza virus has the distinction of being labeled “WS”, for Wilson Smith whose throat-washings provided the culture. Frank Burnet, perhaps the foremost influenza researcher in history, is notorious for advising his native Australian government to develop “intestinal agent” bioweapons in 1947, the same year he contributed key research to polio investigators. After 1933, however, Burnet returned to his home in Melbourne to co-direct the Walter and Eliza Hall Institute where later , as the director “he decided that the future activities of the Institute should be concentrated on animal virology, especially influenza virus…”
17) Bacteriophage I
18) Bacteriophage II
19) Bacteriophage III
Understanding modern influenza is a matter of knowing the work of Frank Macfarlane Burnet, generalized in this document:
“In his autobiography, Burnet notes that he was ‘by temperament an ecologist, a naturalist’…[with] a serious interest in the major problems..of war and overpopulation…the long-term problems of man as a mammal”. In addition to chairing a plethora of official committees on public-health and national defense over his career, Burnet spent the later years of his life churning out his views in print. “His last two books..examined human aggression as the expression of the genetic make-up of man, selected-for during his long evolution as a hunter-gatherer, but totally inappropriate for civilized life. [He wrote a]paper that dealt explicitly with the concept that the lifespan of man and other mammals was genetically determined….He accepted the biological necessity for death and was impatient with proposals designed to prolong the human lifespan. However, he saw “…wide scope for research on the best means of minimizing the depression and misery of pre-death…”
Burnet’s suggestion to his government for biowarfare agents is addressed here:
Born in 1899 Australia, to bank manager Frank Burnet and Hadassah Pollack (Mackay)Burnet, Frank M. Burnet grew up as a shy loner who took especial interest in collecting specimens of flora and fauna. He graduated from university in 1922, got a job working at the Melbourne Hospital and the Walter and Eliza Hall Institute as the pathology registrar. “In 1924, shortly after beginning work at the Institute, he had acquired a copy of Felix d’Herelle’s first book on bacteriophage. His fascination with this subject…was to dominate Burnet’s research for the next decade.” Over that decade, half of his years were spent in London where Rockefeller funding supported his collaborations at the British National Institute for Medical Research (NIMR). When the “first human influenza virus” (labeled WS) was discovered and mutated for experimental purposes (becoming the WSN or ‘NWS’ strain, as Burnet referred to it), he returned to Australia and launched into an extensive phase of viral research focusing on influenza among others and the “lysogenic” (gene transfer) properties of virus. Burnet favored the use of chick embryos in which to colonize bacteria, phage, and virus, and developed the “plaque formation” techniques (called ‘pock-counting’ in Aussie) which occurred in the “chorioallantoic membrane” which is still a basis of generating viral production for influenza vaccines [i.e. grown in eggs].
Burnet had a strong preference for “avianized” viral production through the method of egg-passage over the standard “mouse brain” passage that produced the original (O type) experimental mutant, the WSN influenza A strain, written as A/WSN/33 (influenza ‘A’–slash– ‘W’ilson ‘S’mith ‘N’ for altered neuraminidase–slash– isolated in 1933). By the time that the second World War had begun “Burnet decided that his war effort should be the development of a method of immunization against influenza…influenza virus became the major focus of his work..until 1957 when he made an historic shift to immunology. [Over] the period 1942 to 1959 Burnet’s name was attached to some 114 papers on influenza virus…almost every other independent worker in the Hall Institute at that time..was working on influenza virus, the volume of investigations on this topic in which he was involved as an advisor was perhaps 3 times greater than this…however, it should be remembered that all of this work was strongly influenced by Burnet’s perceptions and often by his advice.”
Burnet’s experimental achievement was to define the mutability of influenza genes through recombination, a process called reassortment, and show that the influenza genome was “fragmented”. He “demonstrated that reciprocal recombination occurred between two different strains of influenza A virus in first-cycle viral multiplication….back-cross experiments were also positive [for mutant recombinants] and provided suggestive evidence of ‘heterozygotes’….[he] obtained recombinants with a wide range of virulence for the mouse lung, a result which led him to postulate the possibility that the genome of influenza virus ‘may fracture and the fragments themselves replicate independently.” Over the next few years Burnet produced an “incomplete virus which he showed could contribute genetic information in recombination experiments and the reactivation of inactivated influenza virus, which he interpreted correctly as being due to genetic recombination. He also…probed further into the genetic control of viral virulence”.
The central mystery of influenza A viruses, for the researchers, was their ability to reassort and alter their characteristics of virulence. They were, in a way, ’super–phage’; reconfiguring what was then known about the behavior of phages. Early investigators of the 1920s and 30s had a hard time accepting the evidence. American George K. Hirst, of the Rockefeller Institute and the Armed Forces Epidemiological Board, described in a 1961 speech to an academic audience that “It was surprising to hear just a short time after the birth of bacterial genetics that genetic recombination takes place between two phage particles invading the same cell [co-infection]. Shortly thereafter, Burnet described influenza recombination. Phage genetics turned bacterial virology into one of the central sciences of modern biology”.
Hirst continues, “influenza..gives readily demonstrable recombination, even in crude systems, but it has been very difficult to develop quantitative [virus multiplication] methods….an essential step for good quantitation for viruses is a plaque-forming system, and it seemed unfortunate that most strains of influenza A either do not kill the cells in monolayers in such a way that plaques can be detected or else the efficiency of plaque formation is so low as to make the technique virtually unusable.
“Most strains of influenza are rather strictly pneumotropic [replicating in lung tissue] in the mouse, but a mutant of the original human influenza strain was isolated many years ago which has the capacity of multiplying in the endothelial cells of the mouse and as a consequence it can multiply in the brain where it is lethal. This strain, (WSN) inaccurately described as being neurotropic, also produces excellent sharply outlined plaques on chick fibroblast monolayers and does so with a very high efficiency. As noted before, influenza strains in general either produce poor plaques at low efficiency on this medium or fail to produce plaques at all.”
A Cold Spring Harbor ‘book’, linked here on the blog in the ”Mutation” post, generalizes the problem for experimentation with the slowness of natural mutations in microbes (one in one million or one billion replicants) and was overcome in the labs by providing an environment of constant adaptive pressure. ‘high yielding’ microbe forms by survivor-selection had already paid off big commercially in the production of antibiotics. Mutants were outstanding performers. Frank Burnet “also noted that with some strains of influenza virus that multiplied to high titre, the embryo was unaffected and hatched normally, however no antibody to influenza virus was produced”. For Frank Macfarlane Burnet, this discovery made with flu mutants led to his greatest scientific achievement: the elucidation of “acquired immunological tolerance”, what he called the “clonal selection theory” and what we would call the recognition of ’self’ and ‘not self’ by the immune system. What these flu scientists had discovered were the principles of successful gene transfer.
In 1940, with the world at war, Burnet ran a series of experiments to develop a flu vaccine that were repeated in kind in 1943 by the U.S. Army doctors; Jonas Salk and Thomas Francis Jr., et.al. The Aussies used ‘volunteers’ and the Americans used the institutionalized wards of state hospitals, also recorded as ‘volunteers’. Gordon N. Meikeljohn, distinguished flu researcher who took up the writing of Army medical history after Dr. Francis in 1945, had little to say about the experiments of the “Commission on Influenza” except “Performed today, experiments of this type would be considered highly unethical”. What the experiments revealed, is that it’s “clear that immunity was far from solid….The Surgeon General of the Army at that time was especially concerned that an attenuated live virus vaccine might revert to virulent virus”. It was sufficiently known by this time that ‘killed’ and ‘fractioned’ virus was just as likely to ‘revert’. Thomas Francis was the director of the Commission on Influenza, one of the first people selected to join the Army Epidemiological Board (AEB, changed to AFEB) at inception, and he remained the director until 1957. Concurrent with his military duties, Thomas (and Salk) produced, field tested, evaluated, and announced to the public the news of polio vaccine. As Burnet had discovered, experimental poliovirus was nearly as efficient in productivity and recombination as influenza. After the war, in 1947, Albert Sabin proved that poliovirus was the cause of ‘grippe’, merely the French word for the Italian ‘influenza’.
Army flu history (in part, of many parts):
In 1951, Richard E. Shope of the Rockefeller and AFEB, took the lead of the ‘Commission on Epidemiological Survey’, the second of the commissions formed after the Commission on Influenza. The Survey team with Shope as its chair, were the nominal directors of Fort Detrick’s bioweapons program over the most intensely secretive years of the Cold War and nuclear testing. Shope was a ‘hog cholera’ expert who held his post at Ft. Detrick until 1959. Shope came up with a theory of the swine–flu cycle that involves lung worms being swallowed by their hosts and ‘recycled’ back into the environment…I suggest you read Burnet.
There is a Continuum that has become visible to my eyes since delving into modern disease history, looking roughly like this: POLIO to INFLUENZA to CANCER to AIDS…and on it goes, cycling and diversifying. Certain genetic ‘agents’ have proven to be better ‘modifiers’ of the public body than others, because, as this website explains, “Cellular viability and propagation require that genomes be reasonably stable. Cancer is an example of a genetic disease often associated with and in some cases perhaps, engendered by the loss of genomic stability….DNA motifs that are predisposed to genetic change have been referred to as at-risk motifs….structural features prone to strand breakage would be expected to be particularly active in homologous recombination….A long inverted repeat (LIR) is an example of an at-risk motif….”
read about how viruses can be used to induce LIR replication alterations….
In the words of Dr. Stefan Lanka, who holds that genetic material is not stable, “the first genetic material of life was RNA, and only later in evolution did DNA come into existence. Every one of our genomes..is the product of so-called reverse transcription: RNA transcribed into DNA…Later, I learned that reverse transcription is a repair mechanism for chromosomal DNA.”
….for your consideration.