From Steven B. Harris, M.D. Here are some more complete remarks on the long paper _HIV, Reality or Artifact_, the product of a purported virologist, one "Stefan Lanka, Ph.D." and recently posted to this forum by John Lauritsen. Excerpts from Lanka's paper are below, and my comments follow. Lanka >>"In going back to the origins of HIV virology and telling the HIV story, a view will be presented which will make clear that HIV itself, the very object of this Manhattan Project of modern medicine, AIDS research, does not exist,"<< Comment: Strangely enough, Lanka does not explain what we're seeing in all those electron micrographs. Lentiviruses (the viral family to which HIV belongs) are not just blobs, but are highly complex and stylized structures which are difficult to missinterpret when seen. They are spherical membrane-covered viruses of about 100 nm diameter, with glycoprotein knobs which easily shear off. When they bud from cells (a picture one can see in any AIDS text), the nuclear material in them forms a crescent shaped structure around the outer limb of the bud, as it does in the HTLV viruses. Unlike HTLV viruses, however, mature lentiviruses (with the exception of non-lymphotrophic lentiviruses like visna/OLV/MVV) have an eccentric nucleoid containing RNA, which lies at one end of a truncated-cone shaped viral core, much like type D retroviruses. At either side of the core are dense lateral bodies, which D retroviruses lack (the D types bud differently also). For a primer on lentivirus structure, contrasting it the other retrovirus structures, I recommend AIDS 5:617-638, 1991. Lymphotropic lentiviruses are visually unique, and readily identifiable. These visually identical lymphocyte-infecting lentivirus retroviruses EIAV, BLV, FIV, and SIV, infect cows, horses, cats, and monkeys respectively, and these infections have all been done experimentally. The results for the last three viruses are immunocompromise, CD4 cell loss, lymph tumors, opportunistic infection, brain infection, wasting, and death. A review of these studies (plus pictures of the viruses) can be seen in my article in this quarter's SKEPTIC magazine, on your newsstand now (SKEPTIC vol 3. no. 2, 1995). HIV-1 and HIV-2, the two viruses isolated from human AIDS patients, are identical in appearance to the other viruses in this class. They share protein antigens, and they share genetic organization. They are infectious, and cell-free preparations of them are capable of killing cells in placque assay, which is a time-honored standard for presence of a virus. They have been molecularly cloned, and the clone DNA, when added to cell culture, produces viruses of identical appearance, which can again be separated from cells, and which show the typical morphologic characteristics of lentiviruses (Virology 189:695- 714, 1992), as well as their typical proteins and enzyme activities. It is even possible to grow this virus in the prepense of an inhibitor of an enzyme which cleaves proteins for the core of this virus, and show that the resulting viruses which are produced from cells have defective-looking cores which mark them clearly as abnormal lentiviruses (AIDS Res. Hum. Retro. 10:735-743, 1994). Such studies are now routine. Dr. Lanka states: >>"Less well-known is the existence of other particles which look like viruses but aren't, and are nonchalantly referred to as "virus-like" particles. Such partic- les are far from rare, found, for example, always in placentas, and very frequently in the artificial environment of laboratory cell cultures. They have served to muddy the waters considerably as far as AIDS research is concerned, because particles just like these have been called HIV," << But here, it is Dr. Lanka who is muddying the waters. There are existent studies in which less than perfect-looking particles have been called HIV, and assumed to be HIV, but some of these studies have no doubt also seen cellular debris, or other retroviruses which can be mistaken for HIV (or other lentivirus- es) if not all lentivirus identifying features are present (for instance, see the overcited Hum. Path. 19:545-549, 1988 for some particles which could be anything). However, there are quite enough studies in which all the lentivirus characteristics noted above ARE present in EM photographs, and morphology is considera- bly more clear. In these studies, the viruses in the electron micrographs are clearly lentiviruses: for example either EIAV, FIV, SIV, or HIV-1, or HIV-2. Lanka goes on to expose an "error" of the past: >> "An error of the past: cancer caused by viruses. It was believed that the new enzyme was a marker for a virus, because the cells in which it was detected, and which were used to study cancer,7 were thought to have become cancerous through being infected by a virus. New to the idea of cancer viruses8 was that nucleic acid, when in the form of RNA could be converted into DNA by the enzyme, thus providing a mechanism for viral nucleic acid to be inserted anywhere in the chromosome of the cells.9 These "new" viruses became known as retroviruses.10 The insertion of certain retroviral genes was thought to trigger cancer. The idea that these postulated viruses caused cancer quickly became "hot news" the world over, but did not survive investigation11 and other explanations were sought.12 The theory did not predict or explain the dramatic increase in cancer cases, cancer could not be shown to be transmissible, nor could it suggest any remedy in the form of a vaccine.13"<< Comment: This is, in general, untrue. Not all cancers were blamed on viruses. Many viruses, such as FLeV, FIV, SIV, and even DNA viruses with reverse transcriptase activity, like hepadnaviruses in woodchucks (Gastroenterol Jpn 25 Suppl 2:38-42, 1990) were proven experimentally to cause cancer in lab animals J Acquir Immune Defic Syndr 4:547-557,1991 and even in random source animals (pet store animals, such as cats) brought to the lab and given virus (J. Vet Med Sci 55:387-94, 1993; J Infect Dis 170:543-52,1994). The idea that retroviruses other than the Rous sarcoma virus could not cause cancer is an error; the reverse has been proven in multiple experiments, including experiments with lentiviruses. Lanka goes on: >> The isolation and purification of a real virus is a straightforward matter, because unlike cells, viruses of one species are always of the same size and shape, and can be readily separated from other cell components by standard techn- iques. A control experiment is to try an isolation with putative non-infected material in exactly the same way as the supposedly infected material. Nothing should be isolated in this case. To identify a virus definitively, a first and simple step is to photograph isolated particles of it in an electron microscope, and they must look like the viral particles observed in cells, body fluids or cell cultures to distinguish them from other cellular particles which look like viruses, but are not. Proteins making up the viral coat must then be separated from each other and photographed. This produces a pattern which is characteristic of the species of virus. A similar separation and identification procedure must be gone through for the DNA or RNA of the virus. Only after the viral proteins and nucleic acid components have been properly identified, is it legitimate to speak of a new virus.<< Comment: All this has been done for HIV. Lanka's problem is that he has not spent enough time in the library. He says: Lanka >>Such evidence has up till now never been produced for HIV. No photograph of an isolated HIV particle has ever been published<< Comment: see Virology 189:695-714 (1992), particularly p. 700 where some nice photographs of isolated HIV particles are published, with all lentiviral morphology clearly seen. Lanka >>nor of any of its proteins or nucleic acids. Comment: This is not clear: he wants photographs of proteins and nucleic acids? For proteins from the virus to be separated and photographed as spots on gels is easy, and these do produce characteristic patterns for HIV. For instance, Cohen et al (J. Virology 64:3097-3099, 1990) in the course of looking for a vpr gene protein in HIV, separated HIV particles and banded them on a sucrose density gradient, where reverse transcriptase activity was detected in just the fractions expected for a retrovirus. These same fractions (but not others) gave the characteristic multiple protein bands of HIV on radiolabeled SDS-PAGE electrophoresis, as well as the vpr protein product. Here are all the components of the virus, banding together in packages in a density gradient in a centrifuge, just where HIV or any retrovirus should band. This repeats the classic Barre-Sinoussi work. Yuan et al (AIDS Research and Human Retroviruses 6:1265- 1271, 1990) repeats and extends this same work: it is real. For an interesting variation see Ratner, et al., (AIDS Res and Hum Retrovir 7:287-294, 1991) in which various clones of HIV with gene deletions for gp envelope proteins are shown to all produce p24 activity which segregates at the proper sucrose density of 1.12-1.14. This p24 protein at this band is protected by trypsin by something, and that something is obviously a lipid membrane, which is disrupted by Triton detergent. Here again are lipid covered p-24 protein particles which have the proper lipid density and are infectious (this paper also contains EMs of lentiviruses budding from cells when the clones are added to them). Lanka >> No control experiments as mentioned above have been published to date.<< Comment: That depends on what one wants for a control. There are many papers in which it is shown that when replication is interfered with in HIV, the virus product in culture is either missing or defective. Again, see AIDS Res and Hum Retroviruses 10:735-743, 1994. Lanka >> What has been shown are photographs of virus-like particles in cell cultures, but none of isolated viruses, let alone of a structure within the human body having the shape ascribed to HIV.<< Comment: First, I have quoted a paper above in which the virus is totally isolated from cells in culture (Virology 189:695-714, 1992). It's not clear why totally isolated viruses should be needed here-- so long as the virus is not connected to the cell, what more isolation is needed? It is known that HIV can be totally isolated from cells (see Virology paper above), so why do this every time, when the separation is known to damage some virions and sometimes give worse quality photos? (although the paper cited above manages to have isolated virions with excellent photos, it isn't easy). Once again, these are not "virus-like" particles in cell cultures, these are particles with as clear morphologic characteristics as are needed to identify them as lentiviruses. I know of no really good photos of HIV isolated directly from humans, but the virus is present in low concentrations outside cells in humans, and contamination is great. What is wrong with culturing it first? Lanka >> What the whole world has seen are models representing HIV with dish aerials, said to be receptors with which the virus attaches itself to cells. << Comment: These glycoprotein knobs can be seen directly in electron micrographs of some isolated viruses in Virology 189:700, 1992. What more is needed? If Dr. Lanka needs a good primer (as it appears he does) on lentivirus structure, as opposed to the other retrovirus structures, I again recommend AIDS 5:617-638, 1991. The evidence that these gp-120 "knobs" attach HIV to cells is massive, and it is ridiculous that Lanka refers to it as "said to be." One can construct HIV with the gene for these knobs deleted, and show that HIV is produced (as seen by its isolation on sucrose gradient) but not infective. However, if another plasmid for this protein is added to cells making HIV, the HIV produced has the knob attachment protein available and IS infective. One can even add attachment protein from a mouse virus, and produce HIV which will infect mouse cells. Infection itself can be shown by making the virus carry yet another gene for antibiotic resistance, to allow make cells it infects to survive in special media. For an elegant set of such experiments done with HIV, this "nonexistent" virus, I suggest the Ratner paper above, or looking up Page et al, J. Virol. 64:5270-5276, 1990. Lanka >> This is the crux of the problem facing all HIV (AIDS) tests. The inability to isolate a viral entity, and to obtain proteins from it which are free from proteins derived from the cells in which the alleged virus is grown, reduces the evidence for the existence of HIV using antibodies to arguing in circles..... It is consequently quite illogical to claim that a positive test results from prior contact with the virus....It is of no help that nowadays "second" and "third" generation tests exist using synthetic proteins which give greater consistency and comparability, because only by an unscientific stretch of the imagination are they viral proteins!<< Comment: These are the same proteins which are coded for by the viral genome, and which copurify with the infectious virus on sucrose gradient, as noted above. Why does it take a stretch of imagination to imagine that they are viral proteins? Lanka: >> The dilemma cannot be stated more poignantly than by quoting from the leaflet accompanying one such test kit: The test for the existence of antibodies against AIDS-associated virus is not diagnostic for AIDS and AIDS-like diseases.<< What's the problem with that? Not everybody with HIV has AIDS. This is elementary. Lanka >> Negative test results do not exclude the possibility of contact or infection with the AIDS-associated virus.<< No, since it does take from 2 to 6 months for antibodies to show up. So? Lanka >> Positive test results do not prove that someone has an AIDS or pre-AIDS disease status nor that he will acquire it.20 << Again, since all people with HIV don't get AIDS (maybe 90% do). Again, so what? Lanka >> Quite! << Quite what? Here one has a test of good prognostic value. It's so good that insurance companies are not allowed to use it. If it showed nothing useful about a person's future, insurance companies would want to save their money. Lanka >> The direct proof of HIV Some HIV researchers have tried to circumvent the problem by pointing to something called "direct" evidence for the virus. All that this meant, though, was arbitrarily selecting a protein of a certain size which happened to coincide with that shown in HIV models. The delusion of such "evidence" was illustrated when the protein later turned out to be of human origin!21 << Comment: Neither gp-160 nor the viral reverse transcriptase of HIV (which is magnesium dependent, unlike any in your cells) are of "human origin" You will find none in any human cell uninfected with HIV. Both are coded in the genome of HIV. ------------------ There follows a sorry theory in which Lanka theorizes how the genome of HIV, now fully sequenced and cloned (and found to be very much like previously known lentiviruses), was supposedly "manufactured." Lanka >> The real explanation of what happens is as follows. In the mixture of cell cultures and stressed human cells, RNA and reverse transcriptase come to be produced in large amounts, because the cells have been specially selected and treated to do this.<< This is baloney. Cells produce no magnesium dependent reverse transcriptase, no matter how you stress them. There follows a fantasy that the HIV genome, so similar in structure to the 9-gene genome of other lentiviruses, was produced by mixing up and producing random cell DNA, and fishing out the "right" retroviral like sequences with HTLV sequence probes; Lanka >> Since no DNA from HIV existed to hybridize with the prepared DNA, Gallo and Montagnier simply used stretches of DNA from what they said was specific to HTLV-I, a retrovirus Gallo had earlier claimed to have discovered, and which they deemed suitable for this purpose. The DNA detected in this way was replicated and certain stretches of it cloned and declared to be the DNA of HTLV-III (later to be called HIV). << Comment: A nice scenario, except for a couple of small problems. HIV looks like a lentivirus, it doesn't look like HTLV visually at all. Furthermore, HIV has no antigenic similarities to HTLV-I, something that caused Gallo real problems until he got a sample of Montagnier's LAV virus (the real HIV). Instead, HIV antisera cross reacts to EIAV (equine infectious anemia virus) which really is a lentivirus (one that infects horse macrophages, and gives the animals lymphadenopathy). So, in order to get a lentivirus from DNA already in human cells, we must posit, at the very least, that Montagnier used EIAV probes, and managed to "make" a human lentivirus (which now kills and buds from human cells!) out of normal cell DNA, fishing it out with horse virus DNA probes (!). All Nobel prize winning work, creating a new lentivirus for humans, when there had never been one-- and all supposedly done by accident. Lanka >> To summarize, the purpose of the exercise is to grow HIV, but it actually produces a mixture of different lengths of DNA, contrary to theory which says they should all be identical, and no virus at all.<< Comment: No, it actually produces fine looking viruses-- lentiviruses in fact, in good form. Again see all the papers I quote above, particularly the Virology 189:695-714, 1992. Lanka >> It is then claimed that the "correct" DNA has been prepared by finding certain strands in this heterogeneous mix by hybridizing them with an HTLV-I DNA probe whose sequence is known and defined to be similar to HIV. However, non-hybridizing strands of DNA should not be there at all, and the fact that they are, proves that just a rag-bag of endogenous DNA from the pool of repetitive elements has been prepared.<< Comment: Wrong. If this were so, DNA PCR hybridization would be able to "get" HIV DNA out of any human cell. But there have been many studies of HIV infected people, along with HIV-negative controls, and the PCR or viral culture is negative on the controls. No "HIV" DNA is there. See for instance N Engl J Med 326:1385-1391, 1992, N Engl J Med 321:1621-5, 1989, AIDS 6:373- 377, 1992, AIDS 8:895-900, 1994. Lanka >> One cannot help asking why no-one had not long ago spotted the flaw in the techniques employed by the Gallo and Montagnier groups.<< Comment: Actually, one cannot help asking who nobody has spotted the flaw in Lanka's argument, which suggests that "HIV DNA" must be in every normal cell, since HIV as a separate entity does not exist, and therefore no DNA PCR test can ever be "negative" on anyone. Yet these tests routinely find no HIV DNA in HIV negative people. Lanka >> Since "HIV" has been shown to be a laboratory artefact it must be assumed that, when not just cross-reacting with other known antibodies, the "AIDS" test detects antibodies against proteins produced in the procedure itself. They must be of human origin because the cells used originated from leukaemic patients. Test positivity, logically, results from immunological contact with them. However, since positivity actually correlates with otherwise unrelated factors such as rheumatism and sun bathing, no specificity can be ascribed to the test.29 << Comment: Then why does a positive test give you a 50% chance of dying of immunodeficiency in 10 years? That's a harsh punishment from rheumatology and sunbathing. And why do positive antibody tests correlate so well (both positively and negatively) with viral culture tests (which measure a protein) or with direct PCR (which measures DNA). If there's no virus to explain all this, how does Lanka explain it? Remember, one must to explain not only why people who test positive for "HIV" antibodies almost always test positive HIV by for culture and PCR (which don't test for antibodies), but also why people who test negative for antibodies test *negative* by culture and PCR (see last set of 4 papers quoted). Lanka: >> Whether antibody positivity really correlates with disease as is commonly supposed, remains to be determined by a critical re-evaluation of the data.<< Comment: Is the claim made that it was done wrong the first time? Showing how it was done wrong (i.e., the burden of proof) is on Lanka. HIV is the only variable which independently correlates with AIDS risk in all AIDS groups. None other has been found. Lanka: >> AIDS research is therefore back at square one and not at Basic Science as suggested elsewhere.30 The main players have since 1993 begun to slink off, arguing that the virus having mutated so much is now no longer detectable.<< Comment: Utter nonsense. References? Lanka: >> Whatever happens, the use of AZT and other "anti-- virals" which are supposed to target HIV replication, but actually kill cells indiscriminately (and ultimately the whole body), must be stopped immediately. It is especially distressing to note that AZT and its analogues preferentially attack those cells which divide most rapidly, namely, cells in the intestines causing diarrhoea and malabsorption of food, and in bone marrow, ironically, the primary production site for cells of the immune system.36 << Comment: There is no evidence that these drugs produce deficits in the cell-mediated immune system which are important clinically. There is no evidence that they produce AIDS (as Lanka may be trying to suggest here) and a great deal of evidence they don't (the Concorde trials). Lanka >> The most important and delicate task is to convince antibody test positives that their result is not a death sentence,<< Comment: Nobody said it was. At least 10% of newly HIV infected people today will still be well and healthy in 15 years, even if medicine does not improve, and only 75% will be dead. For hemophiliacs, that may be as low as 50%. And, of course, things will not get that bad, because medical treatment will improve drastically in the next 15 years. So there is real hope. Lanka: >> To be generally supportive of them, to assuage their anxiety, and to help them understand that with appropriate treatment of any specific disease, they have a good chance to retain or regain their health.<< Comment: That depends entirely upon the state of the destruction of their immune system. It is unrealistic to tell someone with no CD4 cells and 100 CD8 cells, for example, that they have a "good chance" to regain their health. Some people don't like to be lied to, by their doctors. Lanka >> The large number of long-term positives, whose condition cannot be explained by conventional AIDS theory,<< Comment: There are not a "large number" of long term positives-- it is a small percent (10%). And "theory" has nothing to say about their existence either way. That is simply a cheap shot by Lanka at the establishment. Lanka >> as well as the phenomenon of sero-reversion (return to negative test status), provide eloquent testimony to this. HIV/AIDS researchers and health officials are herewith called upon to debate the whole subject of HIV/AIDS openly and humanely, and to recognize the mistake of assuming that immune deficiency was acquired by an infectious agent. << Comment: Sure we will. Just as soon as Dr. Lanka can tell us how it is that when people got AIDS after a blood transfusion, their donors were found to more likely to be high-risk gay men, than were donors for people who got the identical amount of blood from the same bank, but didn't get sick. When the *lifestyle* of a stranger donor statistically predicts disease (p < .01) in the transfusion recipient, perhaps Dr. Lanka can explain this without resort to an infections agent? Please read New England Journal of Medicine 310:69-75, 1984. It's as good an article now as it was then, and as frightening. Steve Harris, M.D. 71450,1773@compuserve.com