The request remains the same and is still pure and simple 12 June 2003
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Eleni Papadopulos-Eleopulos,
Biophysicist
Department of Medical Physics, Royal Perth Hospital, Western Australia, 6001,
Valendar F Turner, John Papadimitriou, Barry Page, David Causer, Helman Alfonso

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Re: The request remains the same and is still pure and simple

The request remains the same and is still pure and simple.

Brian Foley’s rapid response “Re: Re: Re: A modest proposal” (5 June 2003) raises several questions. In this reponse we would like to address only one of them. In responding to Jamie Mills’ question: “Did either Bess et al, or Gluschankof et al, approach 80% or even 98% purified virus, or what looks like purified virus?”, Foley wrote: “And the answer is YES. In the Bess paper, the HIV-1 isolate MN, clone 4 virus grown of CEM-SS cells produced a preparation that was more than 99% pure.” Our question is: “Where in the Bess paper is there proof that the “the HIV-1 isolate MN, clone 4 virus grown of CEM-SS cells produced a preparation that was more than 99% pure.”?”

From our reading of the Bess paper we found that the only reference for a quantitative estimation of purity is as follows: “As an index of relative purity, the ratio of viral p24CA concentration to the sum of the HLA DR and â2-M concentrations was calculated for each preparation. The HIV-1(B)/H9 Clone 4 preparation was significantly MORE PURE IN THIS SENSE sense than the two prototypical HIV-1 producing cell lines, HIV-1(B)/H9 and HIV-1(MN)/H9. However, much better ratios were found for the HIV-1(MN) Clone 4/CEM-SS and HIV-1(MN) Clone 4/Daudi.”(1) (emphasis ours).

For this ratio to be used even as an “index of relative purity”, evidence must exist which proves at least the following: i. The p24 protein is an HIV protein. No such evidence exists. According to Montagnier: “analysis of the proteins of the virus demands mass production and purification. It is necessary to do that.”(2) In 1983 Montagnier claimed to have purified HIV and thus to have shown the existence of the p24 HIV protein. This claim has been widely accepted by many people including Brian Foley. However, in an interview he gave to the French journalist Djamel Tahi, after repeated questioning, Montagnier gave the astonishing reply, namely, that in what they called “purified virus” they did not have even particles with “the morphology typical of retroviruses”. (2) This means that there is no proof that p24 is even a retroviral protein. ii. That the ratio between the different cellular proteins is constant. No such evidence was presented. In fact, according to Bess et al: “Previous reports have indicated that infection of T-cell lines can up- regulate, down-regulate, or have no effect on the cell surface expression of various cellular proteins”.(1) In addition, the CEM-SS cells contains low amounts of â2-M proteins and “Implicitly, microvesicles purified from cell lines such as CEM-SS and MOLT-3 that do not produce HLA-DR did not contain any HLA-DR”. (1) This fact alone will result in a high ratio even if the purity is extremely low.

This means that the purity of the viral preparation cannot be determined from the “ratio of viral p24CA concentration to the sum of the HLA DR and â2-M concentrations” as Brian Foley seems to have done.

The only way to estimate purity is to perform a careful examination of the electron micrographs. Perhaps we need to remind Dr. Foley how to judge such purity. One studies a given, single object in the EM and then compares it with all those immediately surrounding it. Then this procedure is repeated for every other object in the EM. If the overwhelming objects are identical then we can deduce the material is virtually pure. Applying this procedure to Bess’ EM it is child’s play to recognise that whatever Fig 3 in the Bess paper represents, it is not pure. That is, neither the purified HIV-1(MN) or the purified HIV-1(MN) Clone 4 electron micrographs show an HIV-1 purity of either 99% or in fact even 80%. Furthermore, none of the particles arrowed as being HIV have the morphological characteristics attributed to HIV or even to retroviruses. There is no correlation even in their diameters. Retroviruses have a diameter of 100-120 nm while the diameters of the particles in the two electron micrographs assumed to be HIV have an average diameter of 234 nM with none of these particles having a diameter of less than 160 nM. By just this criterion alone it is impossible to claim this material contains even a retrovirus, pure or impure.

In his rapid response “Re: A simple request from the Perth Group” (6 June 2003), Brian Foley wrote: “The Perth group claims that “…under suitable culture conditions, the very act of transfection may result in retroviral expression including the production of retroviral particles. …” This is not a claim. This is a fact.

Brian Foley also wrote: “Lentiviruses are easily differentiated from other retroviruses both at the morphological level…and at the molecular level…”. If HIV is a lentivirus and lentiviruses can be so easily differentiated from other retroviruses then why, on the basis of morphology, did Montagnier describe HIV firstly as a typical type-C particle then a type-D particle and ultimately as a lentivirus? Similarly, why did Gallo for many years insist HIV belongs to the same family as HTLV-I (an oncovirus)? (3) To differentiate HIV from other retroviruses at the molecular level, firstly HIV must be purified and its genomes sequenced. Since HIV has not been purified it follows the HIV genome could not have been characterised.

Brian Foley further wrote: “…using molecular clones to observe viral morphology…the Perth group is surely aware that molecular clones have also been used to tease apart the individual regions of the HIV and SIV genomes that contribute to pathology.” Retroviruses are not simply molecules but particles whose constituents include lipids, proteins and RNA. Their morphology cannot be determined by using molecular clones which are only a string of nucleic acids. How can we be aware of “individual regions of the HIV” genome “that contribute to pathology” when as we have pointed out in a previous rapid response “A simple request from the Perth group” (5 June 2003) that the “HIV genome” “originated from material that either predominantly contained cellular fragments and thus cellular RNA and DNA and a few particles which have some but not all of the morphology attributed to either HIV or other retroviral particles; or consist of material which contain no particles having any of the morphology attributed to retroviruses.”?

Again, we repeat our request, pure and simple: “Where are the experiments which prove HIV isolation and thus the existence of the HIV genome, sexual transmission and antibody specificity?”

References

(1) Bess JW Jr, Gorelick RJ, Bosche WJ, Henderson LE, Arthur LO. Microvesicles are a source of contaminating cellular proteins found in purified HIV-1 preparations. Virology. 1997 Mar 31;230:134-44.

(2) Tahi D. Did Luc Montagnier discover HIV? Text of video interview with Professor Luc Montagnier at the Pasteur Institute July 18th 1997. Continuum 1998 5:30-34.

(3) Papadopulos-Eleopulos E, Turner VF, Papadimitriou JM. (1993). Is a positive Western blot proof of HIV infection? Bio/Technology 11:696-707.

Competing interests:   None declared