Wistar Institute, Dr. Elaine DeFreitas, and the Cheney-Bell-DeFreitas Work: Startling Revelations from Wistar's World Patent and Serious Reasons for Concern Now Revealed!
By Alan Cocchetto

 As many of you can remember, Dr. Elaine DeFreitas, Dr. Paul Cheney, Dr. 
David Bell, and others published the work done at Wistar, in the Proceedings of the
National Academy of Science in April 1991.  This created quite the excitement 
and stir as information was released by personal interviews that even made the 
cover of the CFIDS Chronicle.  
     It would not be surprising if many of the researchers involved with 
Wistar scientists were unaware of a world patent that was subsequently issued in 
April 1992, one year after the PNAS(Proceedings of the National Academy of Science)
article!  I myself was quite surprised since the contents of this patent have 
major implications due to the depth and scientific quality of the work.  I 
certainly believe too that this has worldwide implications and therefore needs to be carefully
scrutinized by the scientific community.
      I am reporting on the detailed scientific information disclosed in the 
world patent (#WO9205760) issued to Elaine DeFreitas and Brendan Hilliard, inventors
assigned to Wistar Institute.  This patent was applied for in August 1991 
after the PNAS article was published.
     The title of the patent is "Method and Compositions for Diagnosing and 
Treating Chronic Fatigue Immunodysfunction Syndrome.  The abstract reads as follows: 
"The present invention provides compositions and methods for diagnosis, 
treatment and prophylaxis of Chronic Fatigue Immunodysfunction Syndrome (CFIDS) based
on the detection of the presence of a novel CFIDS-associated virus, CAV, in 
the body fluids or tissues of a patient."
      In the first page of the patent disclosure, the following is stated:  
"The invention described herein was made in the course of work under grants or awards from 
The United States National Institutes of Health, the Department of Health and 
Human Services."
      The inventors cover the working case definition of CFIDS and various 
outbreaks associated with the illness.  The inventors then provide information 
associated with the field of retrovirology, disclosing various families and specific viruses 
associated with each of them.
      The summary of the invention is as follows:  "The present invention 
provides a novel, substantially isolated Chronic Fatigue Immunodeficiency
Syndrome-associated virus, hereafter referred to by the name CAV.  
Polynucleotide sequences of CAV and polypeptides of CAV are useful as diagnostic reagents in 
the diagnosis of CFIDS patients.  Polynucleotide sequences of CAV and polypeptide
sequences of CAV are useful in therapeutic or vaccinal compositions for the
treatment or prevention of CFIDS.  Also disclosed by this invention are 
methods and assays for diagnosing and/or treating CFIDS patients.  Antibodies to CAV
antigenic regions and in vitro cells containing CAV polynucleotide sequences 
or polypeptides are also described."
      The inventors go on to report two major CAV DNA nucleotide sequences as
well as electron photomicrographs of T-cells and B-cells infected with the 
CAV.  In the initial descriptive reference to retroviruses in this patent, the 
inventors state:
"CAV may be morphologically characterized as a retrovirus, particularly a 
non-C retrovirus which is capable of infecting humans.  Electron microscopy of viral
particles formed in infected human cell cultures suggests that CAV is a 
non-C-type retrovirus because of its diameter, morphology, formation and location of
intracellular virions.  More specifically, CAV-infected cells could be 
characterized by electron-dense circular virions, some with electron-luscent cores and 
others with electron-dense cores, associated with the rough endoplasmic reticulum and 
inside large abnormally distended mitochondria in the cells.  All particles are the 
same shape and size, 46-50 nm.  No extracellular virus is observed.  No forms 
budding from the cytoplasmic membranes are observed.  Thus, CAV-infected cells could
also be characterized by the presence of intracytoplasmic particles....  The 
apparent location of its virions in the mitochondria distinguishes CAV from HIV." 
[Mr.Cocchetto's emphasis here.]
      The inventors then provide additional characteristics of the retrovirus 
such as its ability to infect both T and B-cells and that the primer binding site is for 
the transfer RNA, or tRNA, of lysine indicating that CAV is a non-C type retrovirus.
 The inventors examined low molecular weight sas proteins and found the 
presence of p11-12, p13-14, and p27-28.  Classes of primate and nonprimate animal
retroviruses have such characteristically sized sas proteins.
      The inventors disclose that the virus has the ability to induce the 
presence of viral pap proteins in the nucleus and cytoplasm of cells which it infects.  
This characteristic of viral pap protein localization also indicates a non-C type 
retrovirus. Summaries of correlations of CFIDS retrovirus to known retroviruses are 
included with extensive descriptions and explanations.  Full disclosure of the methods 
appear to be very specific and extensive.  The entire patent is approximately 40 
pages.  If the NIH ignored the depth of this work, since they chose to fund Sidney 
Grossberg, who only had a theory, then the NIH dropped the ball on this one and the 
agency should be held accountable!       The inventors even state "The ability to 
screen blood samples infected by CAV enables producers and distributors of blood
products, e.g. the American Red Cross, to identify and discard donated blood
samples which are intended for use in transfusions or in the isolation of 
plasma, therapeutically useful blood proteins and blood cells.  If unscreened, the 
use of such blood and blood-derived products could contribute to the spread of CFIDS."  
The implications here are staggering!
      The inventors mention various cell lines including T-cell 
lymphoblastoid and B-cell lymphoblastoid lines as well as a macrophage monocyte cell line that 
have all been identified to support the growth of CAV.  They then disclose the primer
sequences for CAV and then state that "body fluids of CFIDS patients have 
shown reactivity with antigens of HTLV-I by Western blot....  Moreover, the 
majority of CFIDS patients have serum antibodies to a P27 protein on the HTLV-I Western
blot.  P27 is presumably a product of the tax gene."  "In still another 
aspect, the invention provides a diagnostic method for detecting CAV in a patient sample 
by a conventional reverse transcriptase assay as described in Example 10 below.  
This assay may be performed on body fluids of a suspected CFIDS patient, using a
polyriboadenylate template primer and the divalent cation Mn++.  No other 
known human retrovirus employs this primer or cation in this assay."
     Of course, all inventors identify their test kit - one that is necessary 
for hospitals, doctors, etc. to officially diagnose the patient as having this illness.  
"The methods, probes, primers, and antibodies described herein may be efficiently utilized 
in the assembly of a diagnostic test kit, which may be used by health care providers 
for the diagnosis and/or treatment of CFIDS."
     The inventors also discuss the details of a CFIDS vaccine and the vaccine
composition!  Furthermore, they disclose that "For performance of these
experiments, patient body fluid samples were obtained from clinical practices 
in North Carolina and New York.  The investigators were all blinded by coded
samples in each experiment."
      Under the heading "Morphometric Analysis of CFIDS Retrovirus" the 
inventor disclose: "Electron-dense circular virions, some with electron-luscent cores 
and others with electron-dense cores, were seen associated with the rough 
endoplasmic reticulum and inside large abnormally distended mitochondria inside the 
cells.  All particles were the same shape and size, 46-50 nm. No extracellular virus was
observed.  No forms budding from the cytoplasmic membranes were observed. 
These observations suggest that CAV is a non-C type animal retrovirus for 
three reasons:  First, human C-type viruses like HTLV-I and HTLV-II do not appear to
form intracellular virions.  The only human C-type forming intracellular 
particles is HIV and these are found intracisternally in conjunction with budding forms. 
Circular C-type virions are usually formed as the virus buds from the cell's
cytoplasmic membrane.  Second, neither HTLV-I, II, nor HIV virions have ever
been found inside mitochondria.  Third, the diameter and morphology of these
virions suggest that they may be Primate D-type retroviruses or spuma 
      Extensive test results are disclosed at this point and the inventors 
reveal:  "The results of the same PCR analyses of blood samples from adult CFIDS patients 
was compared with persons with whom they live or closely associate, e.g. roommates
and friends (called Exposure Controls).  Nonexposure controls are healthy 
persons selected at random who have not come into contact with CFIDS patients nor
experienced symptoms associated with CFIDS."  The inventors report their data
from CFIDS patients including pediatric CFIDS patients!  To quote the patent, 
"the positive results seen in the Exposure Controls support the possibility that 
this CAV is capable of casual transmission to non-infected persons, as is the case
with  many non-human retroviruses."  [Author's emphasis here.]  Now, if the
NIH ignored this last comment, then something is dramatically wrong with the
agency that is supposed to protect and safeguard the welfare of the citizens 
of the United States!  Again, the implications here are just staggering!  This is 
especially alarming in light of the testing, revealed by the inventors, which continues 
as follows:
      Since the inventors ran four different tests on each patient, exposure 
control, and non-exposure control, then I will report on the high values from each test 
group. For the first group, the patients tested with a positivity of 82%, exposure 
controls at 43%, and non-exposure controls at 0%.  With the first group, there were 11 
patients, 14 exposure controls, and 4 non-exposure controls.
      With the pediatric group, the patients tested with a positivity of 74%, 
exposure controls at 43%, and non-exposure controls at 0%.  With the pediatric group, 
the sample size was 19 patients, 7 exposure controls, and 4 non-exposure controls.
      The inventors then disclose more PCR work, citing "partial viral DNA 
sequence was obtained by the procedure described below from CFIDS patient NY1-12 using
the HTLV-II crap specific primers g2-1 and g-2-2 of Table III...  Figs. 1A 
and 1B illustrate the partial putative CAV viral DNA sequences obtained.  Upon 
analysis on GenBank and EMBL, the putative CAV sequences of Figs. 1A and 1B have not
been found to be significantly similar to the sequences of any known 
retrovirus. Thus, these sequences suggest that CAV may not be identified as any other
known human or animal virus."  [Author's emphasis.]
      At this point, the inventors disclose several other tests completed on 
patient, exposure, and non-exposure controls.  These were primarily specific protein 
and retroviral tests and probes.  Additional testing reveals the following 
results with corresponding comments by the inventors: 
      tRNA primer techniques using sense and antisense methods revealed that 
10 out of 10 CFIDS patient DNA samples showed the same sized products using the
primer for the monkey D-type retrovirus (MPMV).  The inventors suggest that 
these results, from this test, imply that CAV "is either a type of lentivirus, 
primate D-type retrovirus, or Foamy (Spuma) virus, all of which us a tRNA lysine primer."
      Characterization of cracr proteins of CAV reveals that "animal 
retroviruses that have been shown to express cracr proteins of these molecular weights are:  
primate D-type retroviruses; primate C-type; lentiviruses (EIAV but not HIV); mouse 
B-type (MMTV); avian C-type retroviruses, and perhaps Foamy (Spuma) viruses.
     Location of crap proteins in the nucleus reveals that "more than 50% of 
patient samples tested (and none of controls) revealed cells staining for crap 
proteins.  Most importantly, the staining is found in both the cytoplasm and nucleus of the 
positive cells. The only known retroviruses to display nuclear staining for viral 
proteins are the Foamy virus group."
      The last test was for reverse transcriptase (RT) with the inventors 
"CAV appears to prefer a template-primer of polyzA-oligo-(dT) with Mn++. 
Among the retroviruses that show the same RT characteristics as that of CAV
(polyzA-oligo(dT) template-primer and Mn++ preferences) are the Spuma (foamy)
virus and the monkey D-type retroviruses."
      Any way you cut this, the only conclusion that can be reached is that 
this work is very thorough and extensive.  It has been funded by the NIH!  And I 
believe that, much like the work revealed by Grossberg's patent (also funded by the NIH), 
the NIH certainly has more than a singular idea about what is happening to us as
patients, all the while denying the existence of retroviral involvement and 
not providing details to outside scientists for additional examination and perhaps
subsequent replication!  Any retrovirus that can invade the mitochondria 
directly indicates trouble!  Why?  Because the mitochondria are the energy powerhouses 
in the body and a direct infection of them spells major trouble --- alteration of
mitochondrial function and dysfunctional energy production!  This could very 
well account for the patient's lack of stamina and that 'F-word', fatigue!  
     As far as I'm concerned here, there needs to be a criminal investigation 
of the NIH regarding why they refused to fund upon submission of all this data as 
well as the involvement of the NIH in Grossberg's work. They are supposed to fund 
based on productivity and Grossberg had none in comparison.  Maybe then, some heads
will roll and we'll begin to get some real answers!  After all, each and 
every patient certainly deserves this and so much more!

[Ed. Note: Dr. DeFrietas presented much of this work at the Albany Medical Convention in 1991. She also submitted a paper of the work to the PNAS three times but was turned down. Why? Were the same people at the NIH who refused to fund her threatening the publication in some way? The refusal to fund her along with the CFIDS Assoc. pulling her funding lost us more than a decade of work!]

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