|
|
 |
![]() |
![]() |
![]() |
|
|
TCCC.UK - Breaking News - 1.
|
![]() |
|
Breaking News !!!!!!!!!!!!!
On the 25th May 2004 Prof. John Hermon Taylor gives a Presentation to the huge ASM - American Society of Microbiology Symposium in New Orleans, "It Is The Speech Of His Life " he says.
The Gastroenterologists have had their presentations , but will not be talking about MAP Bacteria and it's linkage to Crohns Disease. The reason being is they are being spoon fed by the large drug companies, who make vast fortunes from all their
Immunosuppressant drugs.
The Microbiologists however who have cottoned on to the link with MAP Bacteria and Crohns Disease have asked for Presentations on the Facts by Prof. John Hermon Taylor and others.
There is an American called Dr Jay Ellingson from the Marshfield Clinic - Wisconsin
USA, who has mirrored Prof John Hermon Taylor's work, taking random milk samples and people with Crohns. The National Academy of Sciences is involved , but the operation becomes "covert" at this stage, as it is apparently clear the U.S dept of Agriculture is uneasy.Is this really possible hundreds of thousands of people with Crohns, some with totally ruined lives, and yet responsible National Bodies are all watching their backs?????
Are we becoming Paranoid, or is this the TRUTH finally filtering through.Watch for the "Tidal Wave" as after the 25 May 2004, I am sure you will see it coming to the
UK.
Here in the UK, The University of East Anglia is co-ordinating a study, funded by DEFRA which is based on 270 Crohns Sufferers in 5 different areas of this Country : Liverpool, Leicester, Sheffield, Bristol and Norwich.
Questions asked of those :
Where you lived for more than 3 months including College and work.
Countries where you spent more than 7 days.
Holidays spent on Farms and when were you in contact with Farm animals.
What do you eat and drink.
Study will take 18 months
Explanation sheet with this study is dated July 2003.
Interesting Times ahead....................
Update : - There is a sort of "Gag order" on the study from the Marshfield Clinic - - Wisconsin- USA, but the TRUTH will out, filtering into the NAS Journal.
UPDATE 22/08/2004
Research now released and shows independently, support of our own Prof John Hermon Taylor @ St Georges Hospital, London.
|
|
|
 |
|
|
Prof. John Hermon-Taylor - Data.
Prof.John Hermon-Taylor - Data Supporting Map as a Causative Agent of Crohn’s Disease.
Prof John Hermon-Taylor reiterated some of the points raised by Dr. Collins regarding Johne's disease in animals particularly the broad range of histopathological types from pluribacillary to paucimicrobial like leprosy in humans. He also noted that in cattle infected with Map, fecal culture is falsely negative in about 20% of cases, and in infected sheep in 60-80% of cases. Laboratory culture is not a reliable method of detecting Map in animal, food, and environmental or human clinical samples. However, the Map genome contains 14-16 copies of the 1451bp DNA insertion element termed IS900 that is uniquely specific for this organism. In recent years IS900 has provided a convenient target for DNA-based detection systems.
DNA fingerprinting of 740 isolates of culturable Map from infected animals and humans around the world carried out by Dr Ivo Pavlik in the Veterinary Research Institute, Czech Republic, has so far identified 23 different genotypes. A multiplex PCR typing system for MAP is applicable to culturable as well as unculturable forms, supports the view that different strains of Map have emerged with individual host preferences for bovines, ovines, and for humans.
Prof John Hermon-Taylor then presented a detailed description of the occurrence of CD in the UK. Western Europe has a high incidence of the disease estimated conservatively to be 280,000 rising at the rate of 23,800 per year. He pointed out that certain "hot spots" occur and that they are coincident with areas that use water supplies that can be contaminated by agricultural run-off. Map could be periodically detected with the IS900 probe in these waters. He made the case for the possible spread of Map by contaminated waters or their aerosols and pointed out that standard water disinfection methods would not eliminate Map. From late 1990 to mid 1994 he carried out an extensive survey of MAP in retail whole pasteurized cows' milk widely obtained throughout central and southern England and South Wales. With peaks in January, February, March and September, October, November, an overall 7% of retail milk cartons tested positive. Some liquid cultures were seen to contain visible acid fast mycobacteria after 6 weeks of incubation and tested strongly positive with IS900 PCR. He concluded that in the U.K., the risk that MAP is being conveyed to the human population in retail milk supplies is very high. Recently, Dr. Irene Grant in the Department of Food Science, University of Belfast, Northern Ireland, has developed an immunomagnetic capture method for MAP applicable to milk sample processing. Using this, together with BACTEC liquid culture and IS900 PCR, she recently tested 31 cartons of pasteurized milk from 16 pasteurization plants and found 6 (19%) to be positive.
Prof.John Hermon-Taylor cited 6 studies each reporting residual culturability of Map at least once following exposure of spiked milk to experimental pasteurization at 65oC for 30 minutes or 72oC for 15 seconds, and two other studies which reported no culturable organisms remaining. He invoked the "viable but non-culturable" state to explain why individuals may be infected with food or environmental samples that fail to demonstrate the presence of "culturable" organisms. Such a state has been demonstrated in other bacterial species such as Vibrio cholerae. In general, the problem of identification of Map in samples can be attributed to the difficulty of culturing this organism. A recent study by Dr. Ivo Pavlik in the Czech Republic cultured 2 strains of Map from fresh tissues of 39 CD patients.
The detection rate of laboratory culture of Map in Crohn's disease has risen to about 30% when IS900 PCR has been applied to long term cultures. Recent work by Dr. Saleh Naser and colleagues from the University of Central Florida, using an improved liquid culture incubated for about 10 weeks followed by IS900 PCR, has so far detected Map in 6 of 7 Crohn's disease patients and none of 10 non-IBD controls.
Tissue preparation may facilitate the identification of Map by IS900 PCR applied directly to DNA extracts of Crohn's disease tissue. Using fresh frozen tissues from sheep with paucimicrobial Map and humans with Crohn's disease, it was shown that treatment with 6M guanidine HCl at 50oC or prolonged incubation in SDS with proteinase K, are not sufficient for Map. Reliable access to Map DNA in both sheep and human samples required dissolution of the sample in SDS proteinase K followed by mechanical disruption for 45 seconds using the Hybaid ribolyser system followed by nested PCR for IS900.
Prof.John Hermon-Taylor then summarized what is known about immunological detection of Map in humans. A number of protein antigens from Map have been identified by researchers and are recognized by a significantly greater proportion of CD patients than controls. Among these are two antigens of 35K and 36K molecular weight (see Dr. El-Zaatari’s transcript). In immunoblots, IgG recognition of either or both of these proteins was 92% by CD sera and 25% in controls (p<0.01); recognition of both proteins was seen in 75% of CD and none of the control sera. The Map specific insertion element IS900 that is the target of PCR diagnosis encodes a 43kDa protein on its positive strand. Studies using RT-PCR have identified p43 mRNA expression in association with inflammatory bowel disease in humans. A preliminary study suggested the presence of an epitope within the carboxy terminal portion of p43 that is recognized by CD sera. This has subsequently been confirmed in a blinded study using sera from 104 CD patients and 104 age/sex matched controls. IgG binding was elevated (p< 0.0001) in Crohn's disease compared with control normal subjects. Recent outcome analyses have reported substantial remission in active Crohn's disease following treatment with combinations of rifabutin and clarithromycin or azithromycin, drugs known to have enhanced activity against M. avium complex. Controlled trials are needed.
Prof John Hermon-Taylor summarized his view of the pathogenesis of CD as follows. Some strains of Map are pathogenic for humans and cause chronic granulomatous inflammation of the intestine. We are unlikely to make satisfactory progress in elucidating the causation of CD and the role of Map if we think CD is like tuberculosis. Although we do not yet know its pathogenic mechanisms, these are unlikely to involve the release of damaging microbial toxins, or of molecules which are directly granulomagenic. Map is present in low abundance within a critical population of target cells such as macrophages. It is much more likely to cause an intermittent immune disregulation within the gut wall and elsewhere, in humans with an inherited or acquired susceptibility. Together with an accompanying defect in mucosal integrity, this results in a chronic inflammatory and allergic response to bacteria and other constituents present in the fecal stream.
Clinical improvement can be achieved by suppressing or modulating the immune response itself or by reducing the intensity of the allergic component by diet. Some clinical improvement can also come from the use of general antimicrobial agents. However, without killing the causative agent, such therapeutic approaches are unlikely to achieve lasting resolution of the disease. New drugs and immunotherapeutics active against MAP are needed. By preventing infection, however, we can prevent CD in the first place. Strategies aimed at controlling Map in domestic food animals and water supplies may help in this regard.
Question : I'd like to explore the cell-mediated immune response. It's always bothered me that we haven't been able to detect a CMI reproducibly in CD. How could mycobacteria alter antigen presentation to deregulate the normal dendritic function. John, could you expound on how you think engulfing Map with M cell macrophages could deregulate the immune response, and maybe we could hear what might happen in leprosy with APC activity?
Answer : This is entirely putative but it's consistent with what we know and what we see. The model that I was sketching is not one like tuberculosis. It's one in which this organism, in a phenotype that we badly need to identify, becomes taken up by macrophages and I would guess it exists free in the cytoplasm. It's not in the bacillary form. It can just sit there and do nothing, like an inert mitochondrion. But if the relationship between the two is perturbed, and this can be a perturbation that is made more likely on the basis of an inherited or acquired susceptibility – a susceptibility which includes psychological stress factors known to perturb the immune system. Then, a disruption of the normal immunoregulatory functions of APC’s and macrophages can occur, which by analogy with other tissues leads to imperfections in the function of the overlying epithelial layer. An overreaction to normal flora or food results in an inflammatory process within the bowel wall. The best mimic of this is the IL10 knockout mice or the cadherin gene knockouts. I think the way that fits with the evidence that Dr. Bayless and others were discussing is that the immunologic reaction that characterizes the major portion of the disease process is not against MAP itself. It's against these other things. So if we apply non-specific inflammatory influences, we can temporally control the disease, the same with reducing the allergic component, the same with reducing the ambient microbiological flora, and that would fit. Of course, the signals that allow the organism and the cell to communicate are not known.
Comment : There is compelling data in animal models, that normal bacteria provide the antigenic drive, but if a mycobacteria could alter the host immune response to those bacteria, I could see how a very paucibacillary form of disease could cause inflammation. It's always been my personal hang-up with the whole mycobacterial theory, that how does an extremely paucibacillary, or paucimicrobial form of infection, end up causing the disease? We have to wrestle with that, I believe, in order to understand how a small infection might cause chronic inflammation.
Comment : Tubercular leprosy is a classical example. The paucibacillary form of the disease can't really be diagnosed on the presence of organisms. You look at the pathogenesis as manifested nerve damage, and so on. It happens to be granulomatous and it happens to have a very strong cellular immune response so it's easy to detect the presence of an invisible antigen immunologically, but you can't usually find organisms in those patients. Obviously it's possible, I think the difficulty here is that we can't find the organisms and we can't see a good T cell immune response, so we're sort of stuck without a good model. We have to create a new model that combines the lack of organisms, or very low number of organisms, or modified organisms, with a somewhat unusual cellular immune response. We have to think very creatively to come up with a new model.
Comment : There's no doubt that you can induce a cross-reacting self-perpetuating autoimmune response due to molecular mimicry from an inducing agent. The inducing agent goes away but the immune response continues. But you still should have an immune response against the original inducing antigen in that scenario.
2001.
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
