The Crohnie

NOD1 expression in the eye and functional contribution to IL-1beta-dependent ocular inflammation in mice.
Rosenzweig HL, Galster KT, Planck SR, Rosenbaum JT.
Casey Eye Institute, Oregon Health & Science University, Portland,
Oregon 97239, USA

http://www.ncbi.nlm.nih.gov/pubmed/19074813

PURPOSE: NOD1 plays an important role in host defense and recognizes the minimal component of bacterial cell walls, meso-diaminopimelic acid (iE-DAP). Polymorphisms in NOD1 are associated with autoinflammatory diseases characterized by uveitis such as Crohn’s disease and sarcoidosis. NOD1 is homologous to NOD2, which is responsible for an autosomal dominant form of uveitis. Nonetheless, the role of NOD1 in intraocular inflammation has not been explored. The induction of uveitis by iE-DAP in mice and the potential contribution of interleukin (IL)-1beta were investigated. METHODS: BALB/c mice or mice deficient in caspase-1 or IL-1R1 and their congenic controls were injected intravitreally with iE-DAP or saline. The time course, dose response, and contribution of IL-1beta to ocular inflammation were quantified by intravital video microscopy, histology, and immunohistochemistry. NOD1 and IL-1beta were measured in eye tissue by immunoblotting and ELISA. RESULTS: NOD1 protein is expressed in the eye and promotes ocular inflammation in a dose- and time-dependent fashion. The authors previously defined the role of IL-1beta in NOD2 uveitis and tested whether NOD1 and NOD2 used similar mechanisms. Treatment with iE-DAP significantly increased IL-1beta, which was caspase-1 dependent. However, in contrast to NOD2, caspase-1 and IL-1R1 were essential mediators of iE-DAP-induced uveitis, suggesting that NOD1 and NOD2 induce ocular inflammation by distinct mechanisms involving IL-1beta. CONCLUSIONS: These findings demonstrate that NOD1 is expressed within the eye and that its activation results in uveitis in an IL-1beta-dependent mechanism. Characterizing the differences between NOD1 and NOD2 responses may provide insight into the pathogenesis of uveitis.

Liverpool, UK – 10 December 2007: Scientists at the University of Liverpool have found how a bacterium, known to cause illness in cattle, may cause Crohn’s disease in humans.

http://www.liv.ac.uk/newsroom/press_releases/2007/12/crohns-disease.htm

Crohn’s is a condition that affects one in 800 people in the UK and causes chronic intestinal inflammation, leading to pain, bleeding and diarrhoea.

The team found that a bacterium called Mycobacterium paratuberculosis releases a molecule that prevents a type of white blood cell from killing E.coli bacteria found in the body. E.coli is known to be present within Crohn’s disease tissue in increased numbers.

It is thought that the Mycobacteria make their way into the body’s system via cows’ milk and other dairy products. In cattle it can cause an illness called Johne’s disease – a wasting, diarrhoeal condition. Until now, however, it has been unclear how this bacterium could trigger intestinal inflammation in humans.

Professor Jon Rhodes, from the University’s School of Clinical Sciences, explains: “Mycobacterium paratuberculosis has been found within Crohn’s disease tissue but there has been much controversy concerning its role in the disease. We have now shown that these Mycobacteria release a complex molecule containing a sugar, called mannose. This molecule prevents a type of white blood cells, called macrophages, from killing internalised E.Coli.”

Scientists have previously shown that people with Crohn’s disease have increased numbers of a ’sticky’ type of E.coli and weakened ability to fight off intestinal bacteria. The suppressive effect of the Mycobacterial molecule on this type of white blood cell suggests it is a likely mechanism for weakening the body’s defence against the bacteria.

Professor Rhodes added: “We also found that this bacterium is a likely trigger for a circulating antibody protein (ASCA) that is found in about two thirds of patients with Crohn’s disease, suggesting that these people may have been infected by the Mycobacterium.”

The team is beginning clinical trials to assess whether an antibiotic combination can be used to target the bacteria contained in white blood cells as a possible treatment for Crohn’s disease.

The research was funded by Core and the Medical Research Council and is published in Gastroenterology.

Live Mycobacterium avium subspecies paratuberculosis (MAP) has been cultured from retail milk purchased from stores in California, Minnesota and Wisconsin.

This means that American consumers are being exposed to live bacteria that are known to cause Inflammatory Bowel Disease (Johne’s Disease) in a wide range of animals, including dairy and beef cattle, and is suspected of being a cause of human Crohn’s Disease.

The most important points are

1. From May 2002 through April 2003, milk was purchased from stores in California, Minnesota and Wisconsin – three of the USA’s top 5 milk-producing states. (The other two are New York and Pennsylvania.)

2. Milk was tested for presence of viable MAP, using methodologies created in the 1990’s by British researchers, to study the presence of MAP in retail milk in the UK. It has been known since 1998 that United Kingdom dairy products are contaminated with live MAP.

3. Of 702 US samples tested, 2.8 percent contained viable MAP – that is, MAP bacteria that was alive, capable of multiplying and establishing infection, and capable of causing Inflammatory Bowel Disease in susceptible species.

4. Rate of positives was similar among states, but there was a seasonal effect. More positive samples were found during July, August and September.

This study confirms what we in the Paratuberculosis Awareness and Research Association have long believed: that American consumers are eating and drinking food that contains a live and dangerous bacterium, through the medium of MAP-contaminated dairy products.

On average 2.8% of milk cartons were found to be contaminated. Assuming that the average milk consumer drinks from a single carton of milk per day, this means that the average milk consumer is exposed to live paratuberculosis on average ten times a year. Consuming from 2 different milk cartons per day, 20 times a year, etc. This applies particularly to children, who are encouraged to consume milk, for the calcium and protein content. An average American child living in Minnesota, California or Wisconsin, if they consume from one milk carton per day, will have been exposed to live paratuberculosis up to 100 times by their tenth birthday.

The published results apply only to milk. Although research has shown that the food treatment methodologies used to manufacture other dairy products, such as cheese, chocolate, whey, etc, are incapable of destroying MAP, no US research has sought to determine the percentage of these retail dairy products also contaminated with live MAP. The majority of Wisconsin milk is used for cheese manufacture. Recent scientific results have shown that the methods to manufacture cheddar cheese do not kill paratuberculosis.

To this date, the food safety regulators in the United States, the Food & Drug Admninistration (FDA), have taken no action on the presence of live paratuberculosis in milk, dairy and beef products. The time has now come for the FDA to revise its policy of inaction, and to act immediately to protect American consumers from this dangerous bacterium.

If you believe that the US Government should put the interests of the American public before the interests of American Dairy and Beef Industries, and act to eradicate MAP from human food, please visit the PARA web site for steps you can take to help.

1. Original datasheet from American retail milk study
http://www.johnes.org/newsfiles/109216471862392.html

2. Articles about live paratuberculosis contamination in human food
http://www.crohns.org/map_food

3. Articles about the relationship between Mycobacterium avium subspecies paratuberculosis and Crohn’s Disease.
http://www.crohns.org/articles
http://www.crohns.org/research
http://www.crohns.org/treatment

4. Government agencies with responsibility for regulating Food Safety.
http://www.crohns.org/governments

5. PARA’s work to get the US Congress to help address this problem.
http://www.crohns.org/congress

6. What you can do to help.
http://www.crohns.org/help

I’d like to draw your attention to a major success in PARA’s drive to find a cure for Crohn’s Disease.

Since the National Institute of Allergy and Infectious Diseases (NIAID) hosted a workshop on possible infectious causes of Crohn’s Disease in 1998, attended by all of the PARA Board of Directors, including myself, PARA directors Cheryl Miller and Karen Meyer have worked tirelessly with both NIAID staff and with Crohn’s Disease researchers to obtain funding for those researchers who wish to investigate infectious causes of Crohn’s Disease.

The first spectacular success in this ongoing campaign to find a cure for Crohn’s Disease is the provision of US$1.8 million in funding for researchers who are investigating a infectious cause for Crohn’s Disease, with a strong emphasis on research into Mycobacterium avium subspecies paratuberculosis (MAP).

Further information from the PARA web site

http://www.crohns.org/

Among the researchers who have received research funding in this round are two members of PARA’s Scientific Advisory Council, Dr. Saleh Naser and Dr. Norman Pace.

http://www.crohns.org/council/naser.htm
http://www.crohns.org/council/pace.htm

The NIH funding involves not only the National Institute of Allergy and Infectious Diseases (NIAID), but also involves the National Institute of Diabetes, Digestive and Kidney Diseases (NIDDK), the National Institute of Health that has more traditionally been associated with Crohn’s Disease research.

Specifically, the NIDDK has funded another member of the PARA Scientific Advisory Council, Dr. Fouad El-Zaatari, to continue his long and detailed research program on the role of paratuberculosis in Crohn’s Disease. Dr. El-Zaatari has received funding to continue, among other research, his pioneering work on “In-Situ Hybridization”.

The latter is a technique whereby paratuberculosis bacteria which are present in gut tissue can be labelled and made detectable by attaching, for example, x-ray opaque metal atoms to them. This technique has potential for use as a clinical diagnostic technique, particularly because it can detect the cell-wall-deficient forms of MAP which are believed to be involved in the pathogenesis of Crohn’s Disease.

More details on Dr. El-Zaatari and his work from the following URLs

http://www.crohns.org/para/el-zaatari.htm
http://www.paratuberculosis.org/members/el-zaatari.htm

More information on In-Situ Hybridization from

http://www.paratuberculosis.org/proc6/abst5_2.htm
http://www.paratuberculosis.org/proc7/abst6_p7.htm

You can read more from the following links
NIAID: Crohn’s Disease: Is There a Microbial Etiology? Recommendations for a Research Agenda
NIAID: Recommendations for a Research Agenda
PARA’s efforts benefit Crohn’s sufferers – NIH allocates $1.8 million to study infectious cause of Crohn’s

Momentous developments are afoot!!

I attended the UK Government meeting on MAP in milk last Wednesday, and wrote the following report.

====================================================
London, 5th Dec 2001.
UK Government adopts comprehensive strategy for eliminating MAP from milk.

The UK government today adopted a comprehensive strategy to prevent human exposure to the bacterium Mycobacterium avium subspecies paratuberculosis (MAP). MAP is believed by a growing number of scientists to be a cause of Crohn’s Disease, a lifelong, debiliating and incurable bowel disease suffered mainly by the young.

The Advisory Committee on the Microbiological Safety of Food (ACMSF), which advises the UK Government Food Standards Agency, today approved a comprehensive program of measures aimed at eliminating MAP from retail milk, as purchased by consumers. Previous research commissioned by the ACMSF showed that live MAP could be cultured from approximately 2% of retail milk on sale in the United Kingdom.

The strategy adopted by the ACMSF shows that the UK Government is taking the issue of MAP and Crohn’s Disease extremely seriously. As the ACMSF says in its strategy document: “…. the Agency has put to one side the question of whether or not there is a link between MAP and Crohn’s Disease. The Agency believes that precautionary action to reduce human exposure to MAP should start now and should not be dependent on waiting for the link to be proven.”

Among the raft of measures approved by the ACMSF are:

– Increasing pasteurisation times from 15 seconds to 25 seconds. Although some dairies had voluntarily adopted this extended pasteurisation time in 1998, the more stringent conditions will now become standard government recommendation.

– Stricter quality monitoring of pasteurisation plants. Due to the potential for MAP to survive pasteurisation because of defective or improperly operated pasteurisation machinery, dairies and farms will be closely monitored to ensure that they are complying with regulations.

– Improvement of on-farm milking practices. Because a likely route for MAP to infect milk is faecal contamination, on-farm milking practices are to be closely studied to find the most effective method to prevent this contamination.

– Elimination of MAP infection from herds. The ACMSF is initiating a multi-pronged effort to eliminate MAP from herds of food animals, including improvement of existing diagnostics, a national survey to determine the prevalence of MAP infection in UK dairy herds, and development of a improved vaccination methods to protect animals from the infection.

– Alternative pasteurisation technologies. The ACMSF is coordinating several research projects which are assessing the effectiveness of several novel pasteurisation methods against MAP. The methods being studied include high-pressure homogenisation, double pasteurisation, microfiltration and bactofugation.

The timetable by which these measures will be implemented will be finalised in another ACMSF meeting, to be held in London in January
2002.

PARA greatly welcomes these developments, and commends the UK Government on its willingness to act in the best interests of its citizens and the best interests of the public health. However, there are some further measures which PARA would like to see the UK Government undertake.

o Labelling of extended pasteurisation. Since it is not possible for the UK Government to mandate 25 second pasteurisation for all UK milk, for reasons of European regulation, there will still be some 15 second pasteurised milk for sale in the UK. In order that Crohn’s Disease patients and their families be able to differentiate between 25 second and 15 second pasteurised milk, it is vital that the pasteurisation time be labelled on retail milk containers.

o Elimination of MAP from beef. Milk is not the only route for transmission of MAP to the human population. MAP can also be transmitted through beef from infected cows, and there is evidence to believe that the standard temperatures used for cooking of beef will not effectively kill the organism. Although the comprehensive strategy to deal with MAP in milk is a welcome start, it does not deal with the whole MAP problem.

Paratuberculosis Awareness & Research Association is non-profit organisation of Crohn’s Disease patients, their families and friends who are dedicated to the following goals

1. To promote awareness of the disease-causing potential of the bacterium Mycobacterium paratuberculosis in the national community of sufferers of Crohn’s Disease and Inflammatory Bowel Disease; in medical, veterinary and food research communities; in governmental agencies and in the public in general.

2. To promote clinical trials of therapy effective against MAP as treatment for Crohn’s Disease.

3. To promote mandated national testing programs to ensure that the milk/dairy, beef and other products on our grocery shelves are free of contamination with Mycobacterium avium subspecies paratuberculosis.

For further information, please visit the PARA web site at

http://www.crohns.org/

Steve Merkel is a member of the PARA board of directors. Steve decided to approach his Congressman, Dennis Kucinich, about MAP and Crohn’s Disease. Rep. Kucinich decided that this issue was far too important to ignore, and thus began PARA’s first initiatives on Capitol Hill.

If it wasn’t for Rep. Kucinich, it is quite possible that the NIAID would not have become involved, and thus the MAP/CD hypothesis would still have significant perception problems.

Rep. Kucinich is far more likely to succeed in advancing the cause of MAP/CD research if he has support from other Representatives in the house, and from Senators.

The only way that is going to happen is if other Congressmen and Senators get involved.

The only way that other Congressmen and Senators are going to get involved is if YOU ask them to! Yes, YOU!

If Steve Merkel had waited for someone else to the job for him, then no-one would have contacted Rep. Kucinich, and he would not now be fighting our corner in DC.

The same goes for each and every one of your Congressmen and Senators. If you don’t contact them, who will? How could get they possibly get involved if no-one asks them to?

Contact your elected representatives today. Phone them, email them, fax them, write them, whatever it takes. You can find contact details for your elected representatives, along with materials which you can send to them, at this URL

http://www.crohns.org/congress/

As promised a few months back, I’ve managed to digitize the the BBC TV programs about Mycobacterium avium subspecies paratuberculosis (MAP), Crohn’s Disease and MAP contamination of milk and water. Apologies for the delay, there’s been quite a few technical hitches along the way. You can access them from

http://www.crohns.org/media/

Also on that page, you will find a U.K. government interim report on the MAP contamination of retail milk in the UK. The results are definitive and beyond all doubt: MAP bacteria are alive and growing in our retail milk.

MAP is in the milk supply, and it’s in the water supply. Antibiotics effective against MAP make Crohn’s Disease better. When will the US and other Governments wake up to the suffering of Crohn’s Disease, and how it could be prevented?

If you want to do something about this terrible situation, please join PARA and add your voice to the growing chorus calling for change.

As some of you may be aware, there is a wealth of evidence which suggests that some cases, and possibly a majority of cases, of Crohn’s Disease are caused by infection with Mycobacterium avium subspecies paratuberculosis.

To date, research in this field has been severely hampered by chronic lack of funding.

In order to address this funding shortfall, Paratuberculosis Awareness & Research Association (PARA) has assembled a proposal for the U.S. Government. The premise is simple: We want funding for research to develop better diagnostics and treatments for Crohn’s Disease!

We intend to raise awareness of this promising but neglected research avenue. In order for our efforts to succeed, we need as much support as possible.

We are asking you (yes, you!) to please join us in this most important endeavour. It is only by raising awareness of this issue on Capitol Hill that we can hope to achieve our goals.

Please (please, please, please) visit the following page, where you will find

o The contents of PARA’s submission to the U.S. Government.
o Instructions on how to contact the members of the U.S. Government committees responsible for funding allocation
o Instructions on how to contact your elected representatives, if you are a U.S. citizen.

http://www.crohns.org/congress/support.htm

Many of you reading this message have suffered for many years because of Crohn’s Disease. We ask you to please consider spending 30 minutes to join with us in helping to bring an end to Crohn’s Disease. It’s in your own best interest, and the interest of all of our relatives, friends and family.

Please feel free to post copies of this message on other Crohn’s Disease boards/forums/chats.

A group of immunologists at UCLA who were investigating antibody responses in IBD (both Crohn’s and UC) focussed on the antibody known as “pANCA”, which is proposed to be of diagnostic use in IBD.

The investigators conducted a search for microbial antigens that pANCA binds to. The surprising result they found was that, in a percentage of Crohn’s patients, pANCA reacts strongly with a protein which is shared among several species of mycobacteria.

The importance of this is that the team doing the investigation were “mainstream” immunologists who started out looking at the immune responses of Crohn’s Disease and found mycobacteria, rather than starting out with mycobacteria and finding immune responses.

As the investigators themselves conclude: “The association of HupB- binding serum IgA with IBD provides new evidence for the association of a mycobacterial species with Crohn’s disease. ”

Abstract available from
http://www.ncbi.nlm.nih.gov/pubmed/10569769

Full abstract repeated at the bottom this post

================================================================

Journal: Infect Immun 1999 Dec;67(12):6510-7
Title: Identification of a novel mycobacterial histone H1 homologue (HupB) as an antigenic target of pANCA monoclonal antibody and serum immunoglobulin A from patients with Crohn’s disease.
Authors: Cohavy O, Harth G, Horwitz M, Eggena M, Landers C, Sutton C, Targan SR, Braun J
Address: Department of Pathology and Laboratory Medicine, Los Angeles,
California 90095, USA.

Abstract: pANCA is a marker antibody associated with inflammatory bowel disease (IBD), including most patients with ulcerative colitis and a subset with Crohn’s disease. This study addressed the hypothesis that pANCA reacts with an antigen(s) of microbial agents potentially relevant to IBD pathogenesis. Using a pANCA monoclonal antibody, we have previously identified the C-terminal basic random-coil domain of histone H1 as a pANCA autoantigen. BLAST analysis of the peptide databases revealed H1 epitope homologues in open reading frames of the Mycobacterium tuberculosis genome. Western analysis of extracts from six mycobacterial species directly demonstrated reactivity to a single, conserved approximately 32-kDa protein. Direct protein sequencing, followed by gene cloning, revealed a novel 214-amino-acid protein, an iron-regulated protein recently termed HupB. Sequence analysis demonstrated its homology with the mammalian histone H1 gene family, and recombinant protein expression confirmed its reactivity with the 5- 3 pANCA monoclonal antibody. Binding activity of patient serum immunoglobulin G (IgG) to HupB did not correlate with reactivity to histone H1 or pANCA, indicating the complex character of the pANCA antigen. However, anti-HupB IgA was strongly associated with Crohn’s disease (P < 0.001). These findings indicate that the 5-3 pANCA monoclonal antibody detects a structural domain recurrent among mycobacteria and cross-reactive with a DNA-binding domain of histone H1. The association of HupB-binding serum IgA with IBD provides new evidence for the association of a mycobacterial species with Crohn’s disease.

PMID: 10569769, UI: 20038316

Source: The London Times, 25th January 2000.
http://www.the-times.co.uk/news/pages/tim/2000/01/25/timnwsnws02030.html?999

Crohn’s linked to bacteria in milk

BY IAN MURRAY, MEDICAL CORRESPONDENT

CROHN’S disease is almost certainly caused by bacteria found in milk – even if pasteurised – and drinking water supplies, according to research by a world expert on the chronic illness. John Hermon-Taylor of St George’s Medical School in Tooting, South London, says that up to 55 per cent of dairy herds in Western Europe and America are infected with the bacteria, which can survive the pasteurisation process. Water supplies become infected as the droppings from herds seep into the soil, down into natural aquifers.

The organism is called MAP (Mycobacterium avium subspecies paratuberculosis), which is difficult to detect and destroy. The normal pasteurisation process involves heating milk to 72C for 15 seconds, but to be sure of killing MAP it would need to be heated to that level for twice as long.

Crohn’s disease is not fatal but causes chronic diarrhoea, persistent abdominal pain, weight loss, tiredness and mental problems. Because it is not notifiable, the number of people affected can only be estimated but it is thought that up to 80,000 suffer from it in Britain, with between 4,000 and 8,000 new cases every year. The cost to the nation of sufferers’ medical care is estimated to be GBP 240 million a year.

Professor Hermon-Taylor, who has researched the illness for 20 years, said: “If there were no MAP I believe there would be almost no Crohn’s disease. It is certainly responsible for between 60 per cent and 90 per cent of all cases and I would think that it is more likely to be 90 per cent.”

His study, funded by Action Research, the medical charity, shows that MAP can live undetected in cattle for years. Infected cows secrete the bacteria into their milk and on to their pastures. Tests have proved that MAP causes chronic infection of the intestines of many animals, including four types of primates. American studies have isolated MAP from the breast milk of women with Crohn’s disease but not in women who do not have the illness.

“The problems caused by MAP in the milk supply constitute a public health disaster of tragic proportions, for which a range of remedial measures are urgently needed and for which the Government must take responsibility,” Professor Hermon-Taylor said.

He wants to see the Government reverse its decision to allow the sale of unpasteurised milk and to increase the stringency of the pasteurising process. Dairy herds ought to be tested for the infection and the illness ought to notifiable, he said.

Professor Hermon-Taylor said that anybody worried about catching the disease from milk could be certain of killing any MAP by heating it to 80C and then allowing it to cool before drinking it.