Sign the petition

Durban Declaration Rebuttal

A rebuttal to the "Durban Declaration" published in Nature on July 6 2000.

Compiled by Robert Johnston¹, Matthew Irwin² and David Crowe³

Appendix B:What "HIV" Researchers Mean by "Isolation" and Cloning

Many scientists define isolation rather loosely as the detection of indirect phenomena in cell cultures - like reverse transcription and the presence of proteins like p24, rather than isolation of whole, infectious virus. But other scientists point out that this is not isolation - especially since none of these markers are unique to HIV and are quite common elsewhere. They state that there is no scientific reason to assume that detection of these phenomena is an indisputable indication of the presence of HIV. Reverse transcription for example, is found in uninfected cells, and in cells infected by one of the hundreds of harmless retroviruses that are also claimed to exist. They further state that one cannot determine which particular properties are unique to HIV unless one has first isolated an infectious, retroviral particle. Since a positive test result has such serious consequences, they conclude that the criteria for the isolation of HIV and for a diagnosis of HIV infection should be more stringent, not less.

Here is what has been allowed to pass for "HIV isolation":

"Although there may be several acceptable methodologic variations, the usual culture procedure involves recovering peripheral blood mononuclear cells (PBMC) by density gradient centrifugation and then cocultivating the PBMC or plasma with (PHA)-stimulated PBMC from HIV-seronegative donors in culture medium containing the lymphokine growth factor, interleukin-2. The cocultures are maintained for a period of up to 1 month and periodically replenished with fresh media and fresh PHA/phytohemagglutinin-stimulated PBMC to sustain viral replication. Although not as sensitive, immortalized cell lines established from neoplastic T lymphocytes have been used in place of mitogen-stimulated PBMC. At regular intervals (3 to 4 days), aliquots of supernatant fluid from the cocultures are tested for HIV p24 antigen. Isolation of virus is confirmed by the detection of p24 antigen in two to three samples collected consecutively from the same culture. The detection of retroviral reverse transcriptase also has been used as the indicator of a positive culture."(1)

Problems with this method of "isolation":

• · In order to say that the detection of p24 antigen indicates the presence of "HIV", first "HIV" has to be purified and then be broken down into its constituent proteins. This has not been done.(2,3,4,5) There is also plenty of evidence that p24 is not specific to "HIV".(6) The protein p24 occurs in high percentages of people who are negative on the antibody tests. (12,13) We are regularly reminded in the literature on "HIV" immunoassays that there is no gold standard for validation of the "HIV" tests. This is because the constituent proteins of "HIV" have not been determined.
• Everybody knows reverse transcriptase is not "HIV" specific. Immunochemistry text books will tell you "the RT-based assays offer the advantage that they can be adapted for studies on other retroviruses [….] and various animal retroviruses".
• Why can't "HIV" be isolated from fresh patient plasma even in cases where "viral load" numbers are through the roof?

The rules for isolation of a retrovirus were thoroughly discussed at the Pasteur Institute, Paris, in 1973, and are the logical minimum requirements for establishing the independent existence of an infectious "HIV".(7,8) They are:

1. Culture of putatively infected tissue.
2. Purification of specimens by density gradient ultracentrifugation.
3. Electron micrographs of particles exhibiting the morphological characteristics and dimensions (100-120nm) of retroviral particles at the sucrose (or percoll) density of 1.16 gm/ml and containing nothing else, not even particles of other morphologies or dimensions.
4. Proof that the particles contain reverse transcriptase.
5. Analysis of the particles' proteins and RNA and proof that these are unique.
6. Proof that 1-5 are a property only of putatively infected tissues and can not be induced in control cultures. These are identical cultures, that is, tissues obtained from matched, unhealthy subjects and cultured under identical conditions differing only in that they are not putatively infected with a retrovirus.
7. Proof that the particles are infectious, that is when PURE particles are introduced into an uninfected culture or animal, the identical particle is obtained as shown by repeating steps 1-5.

For more details on the "Pasteur Rules"
read Isolated Facts About HIV - A Reply

Have Montagnier, Gallo or any other HIV researcher ever achieved this?
Not even close. Kurt Vanquill describes the problems with what HIV researchers call "isolation":

"When Montagnier and Gallo detected reverse transcription activity in their cultures, they concluded that these T cells from AIDS patients were indeed infected with a retrovirus. Unfortunately, reverse transcription activity of normal cells also tends to be promoted by the very cellular conditions to which Gallo and Montagnier subjected their patients’ T cells. Therefore, detection of reverse transcription activity in the T cell cultures of AIDS patients was not proof at all that there was a retrovirus in those cultures.

"The second piece of evidence that Gallo and Montagnier offered in support of the notion that there was a retrovirus in the T cell cultures in their patients with AIDS was that they detected retroviral-like particles in these cell cultures. The important thing to remember is they didn’t identify retroviral-like particles in isolates, i.e. pure HIV, from these cultures. They simply pointed to particles in impure cell cultures and asserted that not only were they retroviruses, but they were a specific retrovirus, HIV.

"Now that really defies all scientific good sense because as even Gallo admits, retroviral-like particles that are actually cellular in origin are, in fact, ubiquitous in cultures, especially when cultures are subjected to the conditions that Gallo and Montagnier used in order to cultivate HIV. Therefore, the identification of these particles in impure cell cultures was not by any means proof positive that those particles were a retrovirus, much less a specific retrovirus, HIV.

"The third piece of evidence that Gallo and Montagnier offered in support of the notion that these T cells cultures from AIDS patients actually harbored a retrovirus was that they identified certain proteins in these cultures as HIV proteins. These HIV proteins were then incorporated into the antibody and Western Blot and used to test for HIV antibodies. Unfortunately, Gallo and Montagnier identified proteins in their cultures as HIV proteins simply because these proteins reacted with antibodies from AIDS patients, and not from non-AIDS patients. Unfortunately, because AIDS patients had a high level of circulating antibodies, much higher than in normal, healthy individuals, that meant that AIDS patients were likely to have antibody cross reactions with any particular given protein more frequently than non-AIDS patients. Therefore, the identification of certain proteins as HIV proteins, simply because they reacted with antibodies of AIDS patients and not non-AIDS patients was insufficient proof that these proteins were actually HIV proteins.

"Those three pieces of evidence--reverse transcription activity, the identification of retroviral-like particles in impure cell cultures, and the identification of HIV proteins simply on the basis of antibody reactions--were the only pieces of evidence Gallo and Montagnier had in support of their claims to have isolated a retrovirus from their patients’ cultures."(9)

Pictures of "HIV"

We have all seen pictures of "the virus". Is this proof that HIV exists?
Simply flashing the inside cover of Robert Gallo's Virus Hunting and claiming the blobs in the EM photos are proof that "HIV" has been isolated is like claiming UFO photos prove we have been visited by ETs. Our bodies express these sorts of particles all the time; they are known as Human Endogenous Retroviruses (HERVs). HERVs are very likely footprints of ancient germ-cell infections and encompass 1% of the human genome.(10) They are not considered pathogenic; some scientists speculate HERVs may confer protection from related exogenous retroviruses; others suspect their expression may be provoked by disease (oxidative) processes. Therefore, even accomplishing steps 1-3 of the "Pasteur rules" (which "HIV" researchers have yet to do) would still not prove an exogenous, infectious particle had been isolated.

Cloning "the virus"

"HIV" researchers often shift to talking about molecular cloning when they are cornered in the isolation argument. Val Turner addresses that red herring in his email debate with Robin Weiss:

"I do not accept isolation and purification as being different. The word "isolation" means to place apart or alone. (From Latin "insulatus", made into an island). "Purification" means to obtain something free from impurities. For a particle such as a virus, isolation and purification are the same thing. The act of placing an object apart or alone, isolation, is totally different from multiplying, transmitting, that is propagating that object. The fact that something can be multiplied or propagated through many generations of cultures neither means that it is any more or less isolated nor that you are any more aware of its identity. No object including a virus can be isolated by propagation. [...] To claim propagation of a unique retrovirus, HIV, one must first prove its existence which can be done only by isolating, purifying it. To date, nobody, not even you, has achieved this.

"By molecular cloning it is meant the production of identical copies of, for example, a DNA molecule, any DNA molecule, from an ancestral molecule by splicing it into a suitable cloning vehicle, for example a bacteriophage. The DNA molecules can be either of viral or cellular origin. Or they could be artificial. The molecules within the bacteriophages can, in their turn, be introduced in any cells. If you want to call these molecules an "infectious form" (I will not) then you must accept that any piece of DNA, no matter what its origin, is an "infectious form".

"Unquestionably, by molecular cloning one can obtain large amounts of "purified" DNA, but equally unquestionable is the fact that molecular cloning tells you nothing about the origin of a particular DNA molecule. To clone "the HIV genome" one must know beforehand that a particular DNA is the HIV genome. In other words, one must start with "HIV RNA" which can be obtained only by prior purification of particles proven to be retroviral particles. Since this has not been achieved, then what is the origin of the DNA fragment which is known as the "HIV genome" which you routinely clone in your lab? If you are using "material" similar to that used by Montagnier in 1983, in which there is no proof there were even retrovirus-like particles, then surely nobody, not even you, can claim this RNA (cDNA) is the genome of a retrovirus, not to mention a unique retrovirus, HIV."(11)

A similar argument stands to invalidate viral load tests.

For more evidence against HIV see:
   * HIV has not been isolated
   * First Pictures of "Pure HIV"

References:

1. Proffitt MR & Yen Lieberman B (1993, June). Laboratory diagnosis of HIV infection. Infectious Disease Clinics of North America 7(2).; 203-215.
2. Gluschankof et al. Cell Membrane Vesicles Are a Major Contaminant of Gradient-Enriched Human Immunodeficiency Virus Type-1 Preparations. Virology 1997; 230(1): 125-133
3. Bess et al. Microvesicles Are a Source of Contaminated Cellular Proteins Found in Purified HIV-1 Preparations. Virology 1997; 230(1): 134-144
4. Papadopulos-Eleopulos,E., Turner, V.F, Papadimitriou, J.M., Causer, D. The isolation of HIV: Has it really been achieved? The case against. Continuum (supplement) 4, no.3, pp.1-24, September/October 1996 http://www.virusmyth.com/aids/data/epreplypd.htm
5. Tahi, D. Did Luc Montagnier Discover HIV? Continuum vol.5, no2, winter 1997/8; http://www.virusmyth.com/aids/data/dtinterviewlm.htm
6. Papadopulos-Eleopulos et al. Is a Positive Western Blot Proof of HIV Infection? Bio/Technology Vol.11 June 1993 http://www.virusmyth.com/aids/data/epwbtest.htm
7. Toplin I. Tumor Virus Purification using Zonal Rotors. Spectra 1973; 4:225-235.
8. Sinoussi F, Mendiola L, Chermann JC. Purification and partial differentiation of the particles of murine sarcoma virus (M.MSV) according to their sedimentation rates in sucrose density gradients. Spectra 1973;4:237-243.
9. Null, G. AIDS Dissidents Speak Out http://www.garynull.com/documents/aids.htm
10. Lower et al. The viruses in all of us: Charracteristics and biological significance of human endogenous retrovirus sequences. Poc. Natl. Acad. Sci. USA; 93: pp 5177-5184
11. Email Correspondence Between Val Turner and Robin Weiss http://www.virusmyth.com/aids/data/vtcorweiss.htm
12. Papadopulos-Eleopulos, et al. (June 1993). Is a Positive Western Blot Proof of HIV Infection? Bio/Technology 11.
13. Ranki, A., Johansson, E. and Krohn, K. (1988). Interpretation of Antibodies Reacting Solely with Human Retroviral Core Proteins. NEJM 318:448-449.