Against the grain
If you have a chronic illness - or are just feeling run down - the answer could lie in your bread bin. Jerome Burne reveals why more of us should steer clear of wheat
Tuesday September 17, 2002
The Guardian
If you suffer from a condition such as osteoporosis, Crohn's disease, rheumatoid arthritis or depression, you're unlikely to blame your breakfast cereal. After all, intolerance of wheat, or coeliac disease (CD), is a an allergic reaction to a protein called gluten, thought to affect only about one in 1,000 people. But now two American clinicians, James Braly and Ron Hoggan, have published a book, Dangerous Grains, claiming that what was thought to be a relatively rare condition may be more widespread than was previously thought. Braly and Hoggan suggest that gluten intolerance does not just affect a few people with CD, but as much as 2-3% of the population. They claim that gluten sensitivity (GS) is at the root of a proportion of cases of cancer, auto-immune disorders, neurological and psychiatric conditions and liver disease. The implication is that the heavily wheat-based western diet - bread, cereals, pastries, pasta - is actually making millions of people ill.
Your doctor, if asked about CD, would tell you that it involves damage to the gut wall, which makes for problems absorbing certain nutrients, such as iron, calcium and vitamin D. As a result, you are more likely to develop conditions such as osteoporosis and anemia, as well as a range of gastrointestinal problems. Children who have it are often described as "failing to thrive". The proof that you have CD comes when gut damage shows up in a biopsy. The treatment, which has a high rate of success,is to remove gluten - found in rye and barley as well as wheat - from your diet.
But if Braly and Hoggan are right, the problem is far more widespread than the medical profession believes. Coeliac disease, they suggest, should be renamed "gluten sensitivity" and, in an appendix to the book, they claim that no fewer than 192 disorders, ranging from Addison's disease and asthma to sperm abnormalities, vasculitis, rheumatoid arthritis and hyperthyroidism, are "heavily overrepresented among those who are GS".
Dangerous Grains contains more than a dozen case histories of people who have recovered from a wide variety of chronic conditions - back pain, chronic fatigue, the auto-immune disorder lupus - simply by following a gluten-free diet. Both authors claim great personal benefits from such a change. "After eliminating gluten grains," writes Hoggan, "I realised how uncomfortable and chronically ill I had been for most of my life."
If you are someone who has visited a clinical nutritionist or a naturopath, this will come as no great surprise. One of their most common suggestions is temporarily to remove wheat from the diet to see if it makes a difference. In fact, so widespread has talk of a wheat allergy become that last November the Flour Advisory Board felt impelled to issue a statement warning of the dangers of this idea. Professor Tom Sanders, head of nutrition and dietetics at King's College, London, was quoted as saying: "Unless you suffer from coeliac disease, a very rare condition, cutting wheat out of your diet is extremely unwise."
Sanders certainly represents the mainstream medical view, but there is good evidence - such as the work of Dr Harold Hin, a GP from Banbury in Oxfordshire - to suggest that it may be in need of revision. Over the course of a year, Hin carried out a blood test on the first 1,000 patients who came to his surgery complaining of symptoms that might indicate CD, such as anaemia or being "tired all the time". Thirty proved positive and a diagnosis of CD was confirmed by a biopsy.
This indicated that CD was showing up at a rate of three per 100 - 30 times more than expected. Significantly, all but five had no gastrointestinal symptoms. "Underdiagnosis and misdiagnosis of coeliac disease," Hin concluded in an article for the British Medical Journal in 1999, "are common in general practice and often result in protracted and unnecessary morbidity."
More recently, a large research programme carried out by the University of Maryland Center for Celiac Research in Baltimore has confirmed Hin's findings. Scientists there tested 8,199 adults and children. Half the sample had various symptoms associated with CD and, of those, one in 40 of the children tested positive for CD and one in 30 of the adults.
But it wasn't just those who seemed ill who were having problems with wheat. Far more worrying was what the Maryland researchers found when they tested the other half of the sample, who were healthy volunteers, selected at random. Among kids under 16, one in 167 had CD, while the rate among the adults was even higher - one in 111.
If those proportions are true for the American population in general, this means that 1.8m adults and 300,000 children have undiagnosed CD - people who, sooner or later, are going to develop vague symptoms of feeling generally unwell, for which they will be offered various drugs that are unlikely to make much difference. Ultimately, they are at higher risk of a range of chronic diseases.
There seems, therefore, to be good evidence that CD is underdiagnosed. But Braly's and Hoggan's proposition is more radical than that. They believe that the immune reaction to gluten that damages the gut in CD can also cause problems almost anywhere else in the body. The evidence for this is a test involving a protein found in gluten called gliadin. When the body has an immune reaction, it makes antibodies. The test for anti-gliadin antibodies is known as AGA and people who test positive to AGA often have no sign of gut damage.
In fact, according to Dr Alessio Fasano, who carried out the University of Maryland research, "Worldwide, CD 'out of the intestine' is 15 times more frequent than CD 'in the intestine'." Braly estimates that between 10% and 15% of the US and Canadian populations have anti-gliadin antibodies, putting them at risk of conditions as varied as psoriasis, multiple sclerosis, jaundice, IBS and eczema.
The idea of gluten causing damage to parts of the body other than the gut is supported by another UK practitioner, Dr M Hadjivassiliou, a neurologist at the Royal Hallamshire Hospital in Sheffield. He ran an AGA test on patients who had "neurological dysfunction" with no obvious cause and found that more than half tested positive. What is more, only a third of the positive group had any evidence of CD gut damage. In other words, while the gluten antibodies can damage the bowels, they can also cause problems elsewhere. In this case, it was the cerebellum, or the peripheral nervous system.
So if a reaction to gluten can cause problems in the brain, might it also be linked to immune disorders? Braly and Hoggan certainly think so, and claim considerable clinical success in treating patients for conditions such as Addison's disease, lupus, rheumatoid arthritis and ulcerative colitis with a gluten-free diet. In fact, almost all the body's systems can be affected (see below). So if you suffer from a chronic condition that doesn't seem to respond to treatment, cutting out wheat for a while seems worth a try.
Are you gluten sensitive?
If you suffer from any of the following, the possibility that you are GS may be worth investigating.
· Upper respiratory tract problems such as sinusitis, "allergies", "glue
ear"
· Symptoms related to malabsorption of nutrients such as anaemia and fatigue (lack of iron or folic acid), osteoporosis, insomnia (lack of calcium)
· Bowel complaints: diarrhoea, constipation, bloating and distention, spastic colon, Crohn's disease, diverticulitis
· Autoimmune problems: rheumatoid arthritis, bursitis, Crohn's disease
· Diseases of the nervous system: motor neurone disease, certain forms of epilepsy
· Mental problems: depression, behavioural difficulties, ADD
Contacts
University of Maryland Center for Celiac Research: 001-800-492 5538.
Website: www.celiaccenter.org
For help on starting a gluten-free diet, visit www.inside-story.com
UK website for sufferers of coeliac disease: www.coeliac.co.uk
A good US website: www.celiac.com
York Nutritional Laboratory (01904 410410) conducts tests for various food sensitivities including gluten.
· A longer version of this article appears in the September edition of the newsletter Medicine Today, website: www.medicine-today.co.uk Dr Braly will be lecturing in London on October 2. Call 0208 877 9993 for details. Dangerous Grains is published by Avery, price £11.99.
Special reports
Medicine and health
Tory Mead's child's story
We decided to have William tested and this is what turned up:
Seven times the allowable reference range for mercury (the reference
range is 1-3, William had 22) Myelin basic protein antibodies (William’s immune system was attacking his own brain tissue)
Immune dysfunction
Yeast (Candida) overgrowth
Elevated Measles titer
Dramatically low Magnesium and Zinc
Elevated Copper
Extremely low IGg antibodies (plasma)
Malnutrition
All of these findings are "classic" markers for the autistic child. In children with ADHD/ADD or Pervasive Development Disorder-Not Otherwise Specified (PDD-NOS) the clinical findings may be less dramatic. They are, nonetheless, likely to be present in one degree or another. Recent findings
have demonstrated deficiencies in essential metabolic processes in 99% of children on the spectrum (all neurobehavioral disorders). Despite these findings, very few clinicians have even bothered to ask why the autistic children seem to have these dramatic and similar lab presentations.
Ockem’s Razor
William of Ockem was an English medieval scientist who first observed that, in weighing competing causes for a scientific phenomenon, the hypothesis with the least amount of variables was usually the correct one. He described cutting off extraneous theories and ultimately arriving at the core solution. Put another way, the scientific theory, which has multiple factors, is less likely to be truly causative than a theory, which has, at its root, one simple explanation. This process has been called "Ockem’s Razor." Thus, in analyzing any scientific phenomenon, it may be of value to look for the solution that has the least amount of variables.
In the case of autism spectrum, recent developments suggest a simple theory: a group of children are predisposed, either genetically or through environmental insult, with a lack of defense mechanisms to deal with heavy metals and other environmental insults, in particular mercury. All but one of the childhood vaccines until recently contained a preservative known as Thimerosal, which is 49% mercury, by weight. Thimerosal has been listed for years as a pesticide with the EPA. Mercury is the second most toxic substance on the planet after Uranium. The children were exposed via the vaccine schedule to 237.5 mcg . This amount greatly exceeds the EPA allowable amount. Recent studies by the Pfeiffer Clinic, led by Dr. Bill Walsh, indicate that 99% of affected children show significant deficiencies of Metallathionein (MT). MT is a protein, along with glutathione, responsible for detoxification, especially mercury. It is also the principle transporter of zinc throughout the body. MT is present in the gut and in the blood brain barrier. A number of studies have been conducted on genetically altered mice, whose MT factor has been removed. These studies are significant in many different respects. Perhaps, most significant is the inability of these mice to process heavy metals and the complete deterioration of their immune response.
The good news is that there appears to be the possibility of restoring the MT system. This therapy, along with enzymes, and glycoproteins, has shown dramatic effect in reversing the effects of the exposure and to improve immune functioning. In short, it appears that some children have a predisposition to reduced levels of MT. Whether this is genetic or the result of some intrauterine insult is still unclear. What is clear is that, despite representations to the contrary, both industry and the government have known for years that thimerosal is extraordinarily toxic. As the controversy continues more light will be shed on the role of industry and government in covering up this information and in seeking accountability. As the late Supreme Court Justice Potter Stewart said, "sunlight is the best
disinfectant".
William’s Treatment
The Gut
Piece by piece, we began to treat each of the issues, peeling away the skin of the onion. First, we had to get the gut "in order." The conventional wisdom is that any serious effort in treating the mercury or immune issues should begin with the gut. Unfortunately, many of the gut issues are difficult to resolve without removing the mercury and getting the immune system intact. A leaky swollen gut cannot effectively remove heavy metals. Similarly, a compromised immune system cannot effectively combat serious gut bugs. Given the persistence of the "bugs" (like Clostridia Difficile and Candida Albicans) it is unlikely you will get complete resolution of the issues right away. We placed William on anti-fungals (Diflucan, Ketokonazole, Nizoral, and Nystatin) We also placed William on Yeast Control, grapefruit seed extract, and garlic oil. No one treatment is completely effective. The most important aspect of treating gut bugs is rotating the treatment.
We also use probiotics. These are beneficial microbes, which combat overgrowth of "bad" bugs. Although it is somewhat controversial, using Sacchromyces Boulardi, a form of yeast, which excretes toxins to other yeasts, has been reported as effective. The trick is not to use it exclusively for a long period of time. Most important, yeast and gut bugs will respond very favorably if you starve them. Indeed, it appears that the most effective long-term solution to killing gut bugs is to address the diet. For William, this meant that, in addition to limiting Casein and Gluten, William is on a high protein, low carbohydrate diet. We avoid potatoes, white rice, all sugar, fruit juices and dried fruit. We have chicken, beef, beans (green, pinto, black, garbanzo, white and all legumes). We also have started to use alternative flours such as Amaranth, Quinoa and Tapioca. These excellent flours are bean, not grain flours and are extremely high in amino acids and protein. Finally, we have substituted Stevia for sweetener. Stevia is an herb from South America, which has been used for centuries. It is many tens of times more sweet than sugar (a little goes a long way!) and is not metabolized as sugar.
A final note on diet. One good signal your child may be suffering from yeast is a craving for sweets or carbohydrates. If they throw fits because they are not getting fries, or if they have tantrums because they are not getting a fruit punch drink, rest assured, it is because they are, in the words of the street "Jonesing" for some sugar. The yeast is telling them it needs to be fed, and it is making them feel uncomfortable. Anyone who has tried quitting cigarettes has had a pretty good taste of this experience.
Finally, we have supplemented with a concentrated fish protein which has been used very effectively to heal gut lesions and decubitus ulcers. As the yeast is removed from the gut, the fish protein promotes healing and growth of new intestinal tissue.
A STORY OF HOPE, RECOVERY AND AUTISM
By George And Tory Mead
(End Of Series)
Because of recurrent diarrhea and gut inflammation, many kids have demonstrated classic malnutrition symptoms. The essence of good health is good nutrition. The essence of good nutrition is providing the body with the elements ( Zinc, Magnesium, Selenium, Molybdenum, and others) fatty acids ( omega 3 fatty acids and others) and Vitamins ( A, B6, B12, C, E, and others) that it needs to maintain optimum function. A deficiency in any one of these substances has a chain reaction effect on the entire system. For instance, the absence of zinc, affects the uptake of vitamin A as well as the functioning of the immune system. The absence of Vitamin A has an entirely different set of symptoms associated with zinc deficiency. Without the presence of zinc in the liver, retinol is not produced and vitamin A is not metabolized.
Affected children do not have good intestinal absorption function. Therefore, a substantial number of the nutrients "normal" people take for granted are not absorbed. Laboratory tests for children with autism, ADHD / ADD and other spectrum disorders almost invariably show extremely low zinc, extremely high copper, and very low magnesium, and other trace elements needed for metabolism. On top of this, the children are on an extremely restricted diet, at least to begin with. It is therefore essential to rigorously address the deficiencies with supplements. It is also essential the supplements be casein and gluten free.
There are numerous studies, which show that supplementation with B2, B6 and B12 as well as trace elements will have significant benefits for the child with autism and ADD/ADHD. Some of the supplements are packaged specially for the autistic child. Typically children will have a supplement list which will include:
- Vitamins B2, B6 and B12
- Vitamin C
- Zinc
- Magnesium
- Omega 3 fatty acids
- Molybdenum
- Selenium
Finally, Dr, Rosemary Waring, a research scientist and physician in Great Britain has found that the children suffer from problems of sulfation. In other words, the body’s natural detoxification pathways, heavily moderated by sulfur are deficient. Studies have shown that supplementation of sulfate minerals can have a dramatic effect. For instance, bathing with magnesium sulfate (Epsom Salts) can have an extremely beneficial effect. Similarly, children who get transdermal zinc sulfate also seem to do better. Sulfation is also greatly aided by molybdenum and selenium. Lastly, transdermal glutathione is an essential part of the sulfation chain.
Glycoproteins
We were well into our recovery program with William, who had been making steady, but sometimes, slow progress. The idea of "progress" with a supposedly lifelong, incurable disease is certainly an interesting one. William began seeing a naturopath who had quietly been treating children for several years with glycoproteins. Without getting into complex biochemistry, glycoproteins are molecules on the outside of the cell wall. They were discovered in the 1950’s, and for many years were called "ornamentals" because the scientific community did not understand what they did. They must therefore have simply been "ornamental,"or useless. Several decades later, it was discovered these molecules, of which there are eight essential types, combine in 1.2 million different combinations, and are the principal way cells tell each other what to do. Subsequently, scientists learned how to "synthesize" glycolproteins, which occur naturally only in vine-ripened fruits and vegetables.
Without an adequately functioning glycoprotein system, you can pour supplements down your child. If the cell doesn’t know it needs zinc for instance, it will not take it up to manufacture glutathione. Glycoproteins help this function. Glycoproteins are safe, because they are what the body uses to metabolize. They are also essential to maximize metabolism.
We placed William on glycoproteins and within two weeks he was putting on weight ( he is now squarely in the middle of his height and weight percentiles). He lost the pallor and chalkiness and his expressive speech and generative behavior increased dramatically. William has now started imaginative play and actively interacts with his sister. He appears, for all purposes, to be doing a lot of the "chelation and sulfation" work himself.
In short, we believe the use of glycoproteins has been an integral part of William’s progress. Those children we know who have started them have also shown dramatic progress.
THE ROAD AHEAD
We plan to continue giving William the best chance at a full recovery. The road has been filled with people telling us why what we are doing will not work. To them we say that we would rather have done something and found that it was not helpful, than not to have done anything. Everything we have done seems to have helped William. Because most of the therapies are nutritionally-based, there is very little you can do which can be harmful.
In the immortal words of the American poet, John Greenleaf Whittier: "Of all sad words of mouth or pen, the saddest are these: it might have been."
AUTISM - THE GUT CONNECTION
================================================================
In 1971, Goodwin, et al, studied malabsorption and cerebral dysfunction in autistic children and reported that 40% (6/15) of the children in the study had bulky, odorous or loose stools, or diarrhea.
Since that time numerous studies have confirmed the gastrointestinal disorder-autism link. (which is linked to the mercury article above, simply becuase DPPIV enzymes are turned off by mercury in the gatrointestinal tract)
In 1999, pediatrician Karoly Horvath, et al, of the University of Maryland School of Medicine, performed gastrointestinal evaluation on 36 severely autistic children and found they often showed signs of chronic inflammation in the esophagus, stomach, and duodenum, and, because of enzyme deficiencies, had trouble digesting and absorbing carbohydrates - possibly the cause of the chronic loose stools and gas.
Dr. Horvath stated that "Although gastrointestinal symptoms frequently accompany the manifestations of autism, little attention has been paid to this aspect."
In a more recent study by Wakefield, et al, published in the American Journal of Gastroenterology in September 2000, colonoscopies were performed on 60 autistic children who also had gastrointestinal symptoms such as stomach pain, constipation, bloating, and diarrhea. The study found much greater evidence of intestinal lesions in autistic children than in healthy or non-autistic children with similar digestive problems. Over 90% of autistic children showed clinical evidence of chronic enterocolitis (an inflammation of the mucous membrane of the intestine), such as lymphoid nodular hyperplasia - greater than six times that found in non-autistic children with inflammatory bowel disease.
Although researchers are not certain what causes this condition, in a recent issue of Medical Hypotheses, Mark A. Brudnak, Ph.D., N.D., constructed a theory that could explain how the condition could develop and progress.
Dr. Brudnak pointed out that childhood vaccinations have been implicated in the onset of autism, and subsequent prognosis has implicated diet. A strong gut-brain connection is also apparent.
Dr. Brudnak speculates that in early childhood, sensitivity to a vaccine, or a reaction to a mycobacterial infection, could disrupt pivotal molecular mechanisms that regulate genetic expression - how specific genes in the body switch "on" or "off". This may trigger malfunctioning of the immune and gastrointestinal systems, particularly in gut-associated lymphoid tissue. As a result, proteins are no longer properly broken down in the digestive tract and cells in gut tissue die off prematurely as the gut lining becomes "leaky" and unable to repair itself. Casein, gluten, and other compounds in the diet may then permeate into the bloodstream. Their activated by-products, called exorphins, could act directly on the brain to trigger opioid-like effects associated with autistic symptoms.
Dr. Brudnak's theory could explain why enzyme therapy (which improves the gut's ability to break down proteins) and probiotics (supplementation with beneficial gut microbes that help repair the intestinal lining) have both produced positive clinical results in autistic children, as these therapies restore healthy gut barrier function.
Whatever the cause, the link is clear, and the researchers recommend that autistic children undergo gastrointestinal evaluation.
What does the Acellular DTaP Vaccine have to do with Gluten?
Many Autistic children cannot digest the protein from gluten. It causes gastrointestinal, and
neurological symptoms.
There is an interesting connection between the gliadin peptide, (gluten) and pertactin. Pertactin is the protein used in the acellular DTaP vaccine. Following is an explanation of the connection, proceeding with various CDC information, and medical abstracts.
http://www.eurosur.org/celiacos/Shan.pdf
The primary sequence of the 33-mer gliadin peptide also had homologs among a few nongluten proteins. Among the strongest homologs were internal sequences from pertactin (a highly immunogenic protein from Bordetella pertussis) and a mammalian protein tyrosine phosphatase of unknown function. In both cases, available information suggested that this homology could have biological relevance. For example, the region of pertactin that is homologous to the 33-mer gliadin peptide is known to be part of the immuno-dominant segment of the protein (34). In the case of the phosphatase homolog, the protein is known to undergo vesicular trafficking into
the cytoplasmic Golgi (35). By analogy with the current understanding of how gliadin peptides
are acquired by HLA-DQ2 via a tTGase-mediated pathway (3), we hypothesized that these Pro-Gln-rich segments of both pertactin and the phosphatase are likely to be high-affinity tTGase substrates.
To test this hypothesis, we synthesized the corresponding peptides and measured the selectivity
of tTGase for these. As predicted, both peptides were found to be good substrates of
tTGase (36). Studies by Pastan and co-workers showed that tTGase plays a key role in
receptor-mediated endocytosis of several biologically important proteins (37). Therefore,
we therefore propose that the biological activities of both pertactin and the phosphatase
depend on tTGase-mediated trafficking. Analogous to gliadin in Celiac Sprue, pertactin
elicits a vigorous antibody response because it reacts with tTGase on the extracellular surface of antigen presenting cells to produce a long-lived intermediate that is internalized via endocytosis and presented to the immune system via the class II MHC-mediated pathway. If this analogy holds, then, drawing upon the Celiac Sprue analogy, one could predict that tTGase-mediated endocytosis might be a highly effective mechanism for oral vaccination with the use of immunogenic peptide epitopes, as long as they are intrinsically resistant to the action of
pepsin, trypsin, chymotrypsin, and elastase.
By flanking such epitopes with proteolytically stable high-affinity tTGase substrates [e.g., the sequence PQPQLPYPQPQLP from gliadin (21)], they could be protected from exposure to potent pancreatic and intestinal exopeptidases and would therefore have sufficiently long half-lives to permit efficient stimulation of the gut-associated lymphoid system. Secondary structural studies using circular dichroism spectroscopy on the 33-mer gliadin peptide and its homologs from pertactin and the tyrosine phosphatase reveal that these peptides have strong type II
polyproline helical character (fig. S2).
In addition to reinforcing the proteolytic resistance of these peptides, the type II polyproline helical conformation is typical of peptides bound to class II MHC proteins and is likely to enhance their binding to these proteins (38, 39). The abundance and location of proline
residues is a crucial factor contributing to the resistance of the 33-mer gliadin peptide to
gastrointestinal breakdown. Therefore, we hypothesized that a prolyl endopeptidase could catalyze breakdown of this peptide, thereby diminishing its toxic effects. Preliminary in vitro studies with short gliadin peptides and the prolyl endopeptidase (PEP) from Flavobacterium meningosepticum supported this hypothesis (21).
The ability of this PEP to cleave the 33-mer gliadin peptide was evaluated in vitro (Fig. 4A) and in vivo using the rat intestinal perfusion model (Fig. 4B). In the latter assay, the synergistic effect of PEP and BBM peptidases was evident. Moreover, the T cell stimulatory potential of PEP treated peptide was shown to decrease rapidly (Fig. 4C). Given the preference of PEP for
Pro-Xaa-Pro tripeptides (40) and the abundance of this motif in immunogenic peptides from gluten (41), these results highlight the potential of detoxifying gluten in Celiac Sprue patients by peptidase therapy.
References and Notes
1. D. Schuppan, Gastroenterology 119, 234 (2000).
2. S. N. McAdam, L. M. Sollid, Gut 47, 743 (2000).
3. L. M. Sollid, Annu. Rev. Immunol. 18, 53 (2000).
4. M. Maki, P. Collin, Lancet 349, 1755 (1997).
5. G. Corrao et al., Lancet 358, 356 (2001).
6. The family of storage proteins in cereals also includes secalins from rye and hordeins from barley. For review see [H. Wieser, Bailliere’s Clin. Gastroenterol. 9, 191 (1995)].
7. W. K. Dicke, N. A. Weijers, J. H. van der Kramer, Acta Paediatr. 42, 34 (1953).
8. H. Arentz-Hansen et al., J. Exp. Med. 191, 603 (2000).
9. R. P. Anderson, P. Degano, A. J. Godkin, D. P. Jewell, A.V. Hill, Nature Med. 6, 337(2000).
10. W. Dieterich et al., Nature Med. 3, 797 (1997).
11. The extracellular tissue transglutaminase is the primary autoantigen in Celiac Sprue (10).
12. L. M. Sollid et al., J. Exp. Med. 169, 345 (1989).
13. The class II HLA DQ2 molecule is expressed in 90% of Celiac Sprue patients (12).
14. E. H. Arentz-Hansen, S. N. McAdam, . Molberg, C. Kristiansen, L.M. Sollid, Gut 46, 46 (2000).
15. Materials and methods are available as supporting online material on Science Online.
16. Single-letter abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.
17. H. Arentz-Hansen et al., Gastroenterology 123, 803 (2002).
18. L. Shan et al., unpublished data.
19. G. M. Gray. Handbook of Physiology, R.A. Frizzell, M. Field, Eds. (Oxford Univ. Press, New York, 1991), pp. 411-420.
20. D. J. Ahnen, A. K. Mircheff, N. A. Santiago, C. Yoshioka, G. M. Gray, J. Biol. Chem. 258, 5960 (1983).
21. F. Hausch, L. Shan, N. A. Santiago, G. M. Gray, C. Khosla, Am. J. Physiol., in press.
22. K. W. Smithson, G. M. Gray, J. Clin. Invest. 60, 665 (1977).
23. Molberg et al., Eur. J. Immunol. 31, 1317(2001).
24. J. L. Piper, G. M. Gray, C. Khosla, Biochemistry 41, 386 (2002).
25. For example, the speci.city of tTGase for the -gliadin - derived peptide PQPQLPYPQPQLPY is .vefold higher than that for its target peptide sequence in
.brinogen, its natural substrate (24).
26. kcat/KM440 min 1mM 1 for LQLQPFPQPQLPYPQPQLPYPQPQLPYPQPQPF
compared with kcat/KM 82 min 1mM 1 for PQPQLPY (24) and kcat/KM 350
min 1mM 1 for PQPQLPYPQPQLPY.
27. G. D’Argenio, I. Sorrentini, C. Ciacci, G. Mazzacca,
Enzyme 39, 227(1988).
28. T cell clones and T cell lines were established from small intestinal biopsies of adult HLA DQ2-positive Celiac Sprue patients as described (8). Stimulation of T cells by peptides was measured by a [3H]-thymidine incorporation assay. HLA DQ2 homozygous Epstein Barr virus-transformed B cell lines were used as antigen presenting cells.
29. J. J. Boniface et al., Immunity 9, 459 (1998).
30. J. R. Cochran, T. O. Cameron, L. J. Stern. Immunity 12, 241 (2000).
31. K. Falk et al., J. Exp. Med. 191, 717 (2000).
32. M. Stienekemeier et al., Proc. Nat. Acad. Sci. U.S.A. 98, 13872 (2001).
http://www.nature.com/cgi-taf/Gateway.taf?g=5&file=/drugdisc
/res_high/articles/nrd1118.
html&file&_UserReference=
Gluten intolerance:
Inhibitors for human TGM2 enzyme
from Nature Reviews Drug Discovery
Melanie Brazil
Also known as coeliac sprue, gluten intolerance is a widely prevalent genetically determined condition that affects almost 1% of the population. At present, there are no therapeutic agents for this disease, and the only known treatment is a strict, lifelong gluten-free diet. In the March issue of Chemistry and Biology, new research describes the design of proteolytically stable peptide inhibitors of the enzyme involved in the production of autoantigens from gluten, tissue transglutaminase (TGM2).
Ingestion of gluten proteins, from the common food grains wheat, rye, and barley, by coeliac patients results in the flattening of the epithelial villous lining of the small intestine, which leads to malabsorption of nutrients, weight loss and a whole host of other symptoms, including intestinal malignancies. Several short proline- and glutamine-rich sequences, identified from wheat gluten, activate gluten-responsive T cells extracted from coeliac patients, but not those from control individuals. Interestingly, most of these peptides are also substrates of TGM2, which is also known to be the principal focus of the auto-antibody response in coeliac sprue.
Selective inhibition of TGM2 might be a useful therapeutic strategy for avoiding the immunotoxic response to dietary gluten, an idea supported by reports that mice deficient in TGM2 are viable and phenotypically normal. In the design of one of the inhibitors, Hausch et al. replaced the glutamine in the immunodominant gluten peptide with a 6-diazo-5-oxo-norleucine residue to obtain a highly active and tight-binding inhibitor of TGM2 with low cellular toxicity. The inhibitor inactivated TGM2 by binding within the enzyme's active site, and so could interfere with the disease pathogenesis. Experiments demonstrated that the inhibitor interfered with enzyme activity in cultured cells, and effectively inhibited TGM2-mediated differentiation of an established cell model of intestinal enterocyte maturation.
This potent and selective TGM2 inhibitor will be a valuable tool for further research into coeliac disease, although whether it will be effective in humans remains to be seen. Interestingly, aberrant TGM2 activity is believed to play a role in neurological disorders such as Alzheimer's, Parkinson's and Huntington's diseases, indicating potentially wider use for any ultimately approved TGM2 inhibitors.
References
ORIGINAL RESEARCH PAPER
Hausch, F. et al. Design, synthesis and evaluation of gluten peptide analogs as selective inhibitors of human tissue transglutaminase. Chem. Biol. 10, 225-231 (2003)
FURTHER READING
Schuppan, D. A molecular warhead and its target. Tissue transglutaminase and celiac sprue. Chem. Biol. 10, 199-201 (2003)
Sollid, L. M. Coeliac disease: dissecting a complex inflammatory disorder. Nature Rev. Immunol. 2, 647-655 (2002)
http://www.cdc.gov/ncidod/eid/vol7no6/makinen.htm
Bordetella pertussis is the causative agent of pertussis (whooping cough), which is increasing in incidence in several countries despite high vaccination rates (1-5). One explanation for the increase might be the adaptation of B. pertussis bacteria to vaccine-induced immunity. Pertactin, a 69-kDa outer membrane protein, is an important virulence factor of B. pertussis.
http://www.cdc.gov/ncidod/eid/vol7no3_supp/mooi.htm
Changes in B. pertussis in a highly vaccinated population
In some countries with highly vaccinated populations such as Australia (5), Canada (6), and The Netherlands (7) (Figure 1), pertussis has reemerged. Such a phenomenon may have been caused by changes in the accuracy of notifications, decreases in vaccine coverage, or changes in vaccine quality. These possibilities have been excluded for The Netherlands (7), and we have proposed another possible cause: adaptation of B. pertussis to the vaccine. To investigate this hypothesis, B. pertussis strains collected in The Netherlands from 1949 to 1996 were characterized by DNA fingerprinting and sequencing of genes coding for surface proteins (8,9).
Initially, we studied changes in the B. pertussis population by IS1002-based DNA fingerprinting (9,10). Strains collected from 1949 to 1996 were stratified in periods of 5 to 8 years, and the frequency of fingerprint types in each period was determined (Figure 2). Widespread vaccination was introduced in The Netherlands in 1953, and we assumed that the B. pertussis population was not significantly affected by vaccination from 1949 to 1954 (defined as the prevaccination period in Figure 2). Notable differences were found between the populations from the prevaccination era and the subsequent period, both in the type and frequency of fingerprint types (e.g., the major fingerprint type found in strains collected from 1965-1972 [Ft-29] was absent during the prevaccination period). These qualitative observations were confirmed by the trend in genotypic diversity (Figure 2). Genotypic diversity decreased significantly after the introduction of vaccination and subsequently increased to prevaccination levels. In the 1980s, a second decrease in genotypic diversity occurred. Apart from sampling artifacts, a drop in genotypic diversity may be caused by a decrease in population size or clonal expansion. Indeed, we found that the reduction in genotypic diversity in the 1960s and 1980s was associated with the expansion of antigenically distinct strains.
In a second study of changes in the B. pertussis population, we investigated whether antigenic shifts had occurred in surface proteins (8). Very little polymorphism was observed in most proteins studied. However, two virulence factors, pertussis toxin and pertactin, were polymorphic. Interestingly, antibodies against these proteins correlate with protection against disease, which suggests they have an important role in inducing host immunity (12,13). Essentially, all DNA polymorphisms observed were nonconservative, indicating Darwinian selection. Three pertactin and three pertussis toxin variants were found in the Dutch B. pertussis population (Figure 3). Polymorphism in pertussis toxin was restricted to the S1 subunit (PtxS1), which carries the toxic activity. Variation in PtxS1 was observed in two regions. Significantly, one of the polymorphic residues has been implicated in binding to the T-cell receptor (14).
Polymorphism in pertactin was confined to a region comprised of tandem repeats located proximally to the RGD motif involved in adherence to host tissues (15). Regions with tandem repeats are known to undergo rapid variation due to slipped-strand mispairing during replication (16). Pertactin and PtxS1 variants, identical to those included in the Dutch whole-cell vaccine, were found in 100% of the strains from the 1950s, when the whole-cell vaccine was introduced in The Netherlands (Figure 4). However, nonvaccine types of pertactin and PtxS1 gradually replaced the vaccine types in later years and were found in 90% of strains collected from 1990 to 1996. These results suggest that vaccination has caused strains that are antigenically distinct from vaccine strains to be selected. The drop in genotypic diversity observed in the 1960s and the 1980s coincided with the emergence of nonvaccine-type pertussis toxin and pertactin variants, respectively, suggesting that the drop was caused by clonal expansion. Antigenic divergence between vaccine strains and clinical isolates was also observed in other countries with a long history of pertussis vaccination, such as Finland and the United States (17,18), and also in Italy, where vaccine coverage has varied considerably.
Discussion
Is polymorphism in pertactin and pertussis toxin driven by host immunity, or is it the result of random fixation due to genetic drift? The latter possibility is highly unlikely since essentially all DNA mutations we detected in the pertactin and pertussis toxin genes were nonconservative. In contrast, random genetic drift is characterized by a high degree of conservative mutations in protein coding regions (20). Further, the polymorphic regions interact directly with the immune system. The polymorphic region of pertactin induces a protective immune response (unpublished data). One of the polymorphic residues in PtxS1 has been implicated in binding to the T-cell receptor (14). Finally, the fact that the same temporal trends in allele frequencies are observed in geographically distinct regions such as Finland, the United States, and The Netherlands argues against random genetic drift. It is possible that the polymorphic loci we have identified are linked to other, as yet unknown, polymorphic loci that increase fitness of strains in vaccinated populations (hitchhiking).
Strains carrying nonvaccine-type pertactin or pertussis toxin variants were not found in the prevaccination era. Although the number of strains analyzed from this period was limited, these data suggest that the nonvaccine-type variants are not able to displace the vaccine-type strains in unvaccinated populations (i.e., they have a lower fitness level?, or reproductive rate, in unvaccinated communities). Alternatively, the nonvaccine-type strains may have evolved relatively recently. Consistent with the first hypothesis, we have observed that nonvaccine-type strains are less fit in naive mice than vaccine-type strains. In immune mice the difference in fitness between the two types of strains was much less pronounced (unpublished data). Thus vaccination has acted to shift the competitive balance between strains.
An important question to address is whether adaptation of the B. pertussis population has affected vaccine efficacy, i.e., contributed to the reemergence of B. pertussis. Animal experiments have indicated that variation in pertactin affects vaccine efficacy (unpublished data). Further, we found vaccine-type pertactin variants less frequently among vaccinated persons than among unvaccinated persons, which would be expected if the vaccine protects differentially against strains with distinct pertactin types (8). However, the extent to which polymorphism affects vaccine efficacy is probably dependent on the vaccine used. It is conceivable that the increase in fitness associated with nonvaccine types of pertactin and pertussis toxin in vaccinated populations is substantial enough to drive expansion of strains carrying these protein variants but that the effect is too small to result in a measurable drop in vaccine efficacy. Further studies are required to assess the effect of the observed adaptations on the efficacy of pertussis vaccines. In this period, when whole-cell vaccines are being replaced by acellular vaccines in many countries, continued strain surveillance is of paramount importance.
Frits R. Mooi is at the National Institute of Public Health and the Environment, The Netherlands, and the Eijkman-Winkler Laboratory of the University of Utrecht. His current interests are molecular epidemiology and evolution of Bordetella spp.
This research was supported by the PraeventieFonds, grant numbers 25-2545 and 28-2852, the Ministry of Health, Welfare and Culture, and the RIVM.
Address for correspondence: Frits R. Mooi, Research Laboratory for Infectious Diseases (LIO), National Institute of Public Health and the Environment, P.O. Box 1, 3720 BA, Bilthoven, The Netherlands. Fax: 31-30-274.4449, e-mail: fr.mooi@rivm.nl
http://www.pasteur.fr/recherche/RAR/RAR2001/Bordetel-en.html
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=
PubMed&list_uids=12695998&dopt=Abstract
J Infect Dis 2003 Apr 15;187(8):1200-5 Related Articles,Links
Bordetella pertussis protein pertactin induces type-specific antibodies: one possible explanation for the emergence of antigenic variants?
He Q, Makinen J, Berbers G, Mooi FR, Viljanen MK, Arvilommi H, Mertsola J.
Department of Human Microbial Ecology and Inflammation, National Public Health Institute, Turku, Finland. qiushui.he@ktl.fi
Divergence has been found between Bordetella pertussis vaccine strains and circulating strains. Polymorphism in pertactin (Prn) is essentially limited to region 1, which is made up of repeats. Today, the 3 most prevalent Prn variants are Prn1-3. Vaccine strains produce Prn1, whereas Prn2 is the predominant type found in circulating strains. We investigated how variation in region 1 affects the production of human serum antibodies. Individuals infected by Prn2 strains had significantly fewer antibodies to Prn1 did than those infected by Prn3 strains and those immunized with a booster dose of acellular vaccines containing Prn1. Moreover, in contrast to vaccine recipients and subjects infected by Prn3 strains, individuals infected by Prn2 strains had hardly any antibodies specific to the variable region of Prn1. These results indicate that conformational changes have occurred in the variable region of Prn, which may offer a possible explanation for the emergence of Prn2 strains in certain countries.
PMID: 12695998 [PubMed - indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&
db=PubMed&list_uids=11825003&dopt=Abstract
J Microbiol Immunol Infect 2001 Dec;34(4):243-51 Related Articles,Links
Characteristics and potency of an acellular pertussis vaccine composed of pertussis toxin, filamentous hemagglutinin, and pertactin.
Sheu GC, Wo YY, Yao SM, Chou FY, Hsu TC, Ju CL, Cheng Y, Chang SN, Lu CH.
Vaccine Development Center, Taipei, Taiwan, ROC.
In an attempt to develop a safer pertussis vaccine, we successfully purified 3 pertussis protective antigens-pertussis toxin, filamentous hemagglutinin, and a 69-kDa outer membrane protein (also named pertactin), from Bordetella pertussis strain ATCC 9340. The toxicity of pertussis toxin could be effectively reduced by the treatment with formaldehyde 0.07% while preserving of a high degree of immunogenicity. By mixing purified pertussis antigens with diphtheria and tetanus toxoids (DT), we have formulated a DT acellular pertussis (DTaP) vaccine. Toxicity studies on body-weight gain in mouse, histamine sensitization, lymphocyte promoting, and Chinese hamster ovary cell clustering tests suggested that this DTaP vaccine is safer than a whole cell vaccine produced in France (DTP[F]). The formulated vaccine elicited high levels of anti-pertussis toxin antibodies in both mice and monkeys. In mice, a 2-fold neutralization of anti-pertussis toxin antibodies was produced by DTaP compared with DTP(F) vaccine and an acellular vaccine manufactured in Japan (DTaP[J]). More importantly, in intracerebral challenge assay in mouse, this vaccine also provided a better protection than DTaP(J).
Publication Types:
Evaluation Studies
PMID: 11825003 [PubMed - indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=
PubMed&list_uids=11700340&dopt=Abstract
Microbiology 2001 Nov;147(Pt 11):2885-95 Related Articles,Links
Role of the polymorphic region 1 of the Bordetella pertussis protein pertactin in immunity.
King AJ, Berbers G, van Oirschot HF, Hoogerhout P, Knipping K, Mooi FR.
Research Laboratory for Infectious Diseases, National Institute of Public Health and the Environment, PO Box 1, 3720 BA Bilthoven, The Netherlands.
In several countries pertussis is re-emerging, despite a high vaccination coverage. It is suggested that antigenic divergence between Bordetella pertussis vaccine strains and circulating strains, in particular with respect to pertactin, has contributed to pertussis re-emergence. Polymorphism in pertactin is essentially limited to region 1, which is composed of repeats and is located adjacent to an Arg-Gly-Asp motif implicated in adherence. Evidence is provided for the immunological relevance of polymorphism in region 1. Region 1 was found to contain a B-cell epitope recognized in both humans and mice. Furthermore, variation in region 1 affected antibody binding and, in a mouse respiratory infection model, the efficacy of a whole-cell vaccine. Moreover, passive and active immunization indicated that region 1 confers protective immunity. An mAb directed against a linear conserved epitope conferred cross-immunity against isolates with distinct pertactin variants. The results indicate an important role of region 1 of pertactin in immunity.
PMID: 11700340 [PubMed - indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=
PubMed&list_uids=11672891&dopt=Abstract
Vaccine 2001 Nov 12;20(3-4):299-303 Related Articles,Links
Temporal nucleotide changes in pertactin and pertussis toxin genes in Bordetella pertussis strains isolated from clinical cases in Poland.
Gzyl A, Augustynowicz E, van Loo I, Slusarczyk J.
Department of Sera and Vaccines Evaluation, National Institute of Hygiene, Chocimska Street 24, 00-791 Warsaw, Poland. agzyl@pzh.gov.pl
Growing number of Bordetella pertussis infections in 1997-1998 in Poland overshadowed the successful national vaccination program. This situation prompted us to investigate if this shift reflects changes in the B. pertussis population. We investigated the possible divergence in genes encoding pertussis toxin subunit 1 (PtxS1) and pertactin (P.69) in B. pertussis population strains during the period of 1960-2000. The pertussis toxin and pertactin variants (ptxS1B and prn1) were found in strains used for production of the whole-cell pertussis vaccine (WCV) production in Poland. Results of the study indicate that the ptxS1A-allele replaced the vaccine variant in 69% in the 1960s, and in 100% in 1990s, and although the prn1-allele was found in all strains from the 1960s and 1970s, after 1995 was gradually replaced by prn2 and prn4 variants. Presumably, vaccination could affect the population structure of B. pertussis in Poland and resulted in antigenic shift in both genes analyzed. Our findings may have implications for the composition of polish WCV and the currently licensed acellular pertussis vaccines.
PMID: 11672891 [PubMed - indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=
PubMed&list_uids=11485646&dopt=Abstract
Emerg Infect Dis 2001;7(3 Suppl):526-8 Related Articles,Links
Adaptation of Bordetella pertussis to vaccination: a cause for its reemergence?
Mooi FR, van Loo IH, King AJ.
National Institute for Public Health and the Environment, Bilthoven, the Netherlands. fr.mooi@rivm.nl
In the Netherlands, as in many other western countries, pertussis vaccines have been used extensively for more than 40 years. Therefore, it is conceivable that vaccine-induced immunity has affected the evolution of Bordetella pertussis. Consistent with this notion, pertussis has reemerged in the Netherlands, despite high vaccination coverage. Further, a notable change in the population structure of B. pertussis was observed in the Netherlands subsequent to the introduction of vaccination in the 1950s. Finally, we observed antigenic divergence between clinical isolates and vaccine strains, in particular with respect to the surface-associated proteins pertactin and pertussis toxin. Adaptation may have allowed B. pertussis to remain endemic despite widespread vaccination and may have contributed to the reemergence of pertussis in the Netherlands.
PMID: 11485646 [PubMed - indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=
PubMed&list_uids=11023463&dopt=Abstract
J Infect Dis 2000 Nov;182(5):1402-8 Related Articles,Links
Polymorphism in Bordetella pertussis pertactin and pertussis toxin virulence factors in the United States, 1935-1999.
Cassiday P, Sanden G, Heuvelman K, Mooi F, Bisgard KM, Popovic T.
Division for Bacterial and Mycotic Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA. PCassiday@cdc.gov. PCassiday@cdc.gov
To elucidate the potential role of the etiologic agent in recent increases of pertussis incidence in the United States, we studied the polymorphism in pertactin and pertussis toxin, which are Bordetella pertussis proteins important for pathogenesis and immunity. We sequenced regions of their genes (prn and ptx) in 152 B. pertussis strains isolated from 1935 through 1999 and identified 2 prn sequences: prn1 (old), observed continuously since 1935, and prn2 (new), not recognized until 1981 but seen in 97% of tested isolates in 1999. There were 3 ptx S1 subunit sequences: ptxS1D (old) was identified in 3 strains (1935 and 1939); ptxS1B (old) represented 87% of the strains recovered during 1935-1974; and ptxS1A (new) was the most prevalent during 1975-1987 and 1989-1999 (64% and 78%, respectively). Potential association between vaccination and the observed shift from old to new types requires further study. Our results provide the basis for prospectively monitoring for changes among circulating B. pertussis that might have epidemiologic relevance.
PMID: 11023463 [PubMed - indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=
PubMed&list_uids=10600202&dopt=Abstract
Biologicals 1999 Jun;27(2):133-41 Related Articles,Links
Approaches to the control of acellular pertussis vaccines.
Corbel MJ, Xing DK, Bolgiano B, Hockley DJ.
Division of Bacteriology, National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, Hertfordshire, EN6 3QG, U.K. mcorbel@nibsc.ac.uk
The quality control of acellular pertussis vaccines presents particular problems related to the differences in composition and method of detoxification used in the various type of preparation. These vaccines are not amenable to potency assay by the active mouse protection test used for whole-cell pertussis vaccines and assurance of protective activity is problematic.In contrast, monitoring of these vaccines for safety is relatively straightforward and is centred on assays for the lipooligosaccharide endotoxin, active pertussis toxin and absence of reversion to toxicity of detoxified product. The absence of heat-labile toxin, tracheal cytotoxin and adenyl cyclase toxin is assumed provided that adequate validation of the process has been performed. Confirmation of the antigenic content of the detoxified bulk components is difficult to achieve by conventional binding assays based on monoclonal antibodies because of changes in accessibility of reactive sites post-toxoiding. However, single radial diffusion assay using polyclonal antisera permits estimation of pertussis toxoid (PT), filamentous haemagglutinin (FHA) and pertactin (P69). Dot blot immunoassay can be used for the fimbrial agglutinogens 2 and 3 (Fim 2 and 3) and potentially could also be used to check the composition of final filling lots for PT, FHA, P69 and Fim 2 and 3.Gel electrophoresis and immunoblotting can be applied to monitor purity of purified bulk components and the characteristics of these change after chemical detoxification. Electron microscopy provides a useful semi-quantitative supporting method for checking purity of bulk components. Physico-chemical examination, particularly CD and fluorescence spectroscopy, offer a means of monitoring the consistency of detoxified bulk components.No completely satisfactory method is available for monitoring potency. Immunogenicity assays may be useful for checking consistency but do not necessarily correlate with protection. At present, active protection against aerosol challenge offers the best prospect of a functional assay. Copyright 1999 The International Association for Biologicals.
PMID: 10600202 [PubMed - indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=
PubMed&list_uids=10592076&dopt=Abstract
Eur J Pediatr 1999 Dec;158(12):989-94 Related Articles,Links
Edelman K, Malmstrom K, He Q, Savolainen J, Terho EO, Mertsola J.
Department of Paediatrics, Turku University Hospital, FIN-20520, Turku, Finland.
Local reactions and pertussis toxin specific immunoglobulin E antibodies (PT-IgE) were investigated in healthy children following primary and booster immunization with a combined diphtheria tetanus acellular pertussis vaccine (DTPa) including pertussis toxin, filamentous haemagglutinin and pertactin. A primary series of DTPa was administered to 150 infants, and 104 of them received a booster dose of DTPa combined with inactivated polio vaccine at 2 years of age. PT-IgE was measured in serum samples from 72 children using a modified nitrocellulose RAST. Primary immunization was associated with low incidence of local reactions (1%-5%). After the booster dose 21% of children had a local reaction >/=20 mm. Local reactions after the booster dose tended to be more common in children who had experienced reaction at primary immunization. PT-IgE was detected in 18% and 86% of children following primary and booster vaccinations, respectively. Allergic and non-allergic children did not differ in PT-IgE responses. After primary immunization, elevated PT-IgE levels were found more often in children with a family history of allergy than in those without known allergy in the family. Children with local reactions had significantly higher pre- and post-booster PT-IgE levels and median post-booster pertactin IgG and diphtheria-IgG levels than children without local reactions. Conclusion Acellular pertussis immunization induces IgE antibodies to pertussis toxin, especially after booster vaccination. The higher median pre- and post-booster levels of pertussis toxin specific immunoglobulin E and post-booster levels of IgG to pertactin and diphtheria in children with local side-effects reflect a multifactorial immunological mechanism of such reactions.
PMID: 10592076 [PubMed - indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=
PubMed&list_uids=10463173&dopt=Abstract
Microbiology 1999 Aug;145 ( Pt 8):2069-75 Related Articles,Links
Antigenic variants in Bordetella pertussis strains isolated from vaccinated and unvaccinated children.
Mastrantonio P, Spigaglia P, van Oirschot H, van der Heide HG, Heuvelman K, Stefanelli P, Mooi FR.
Department of Bacteriology and Medical Mycology, Istituto Superiore di Sanita, Rome, Italy. pmastran@iss.it
Bordetella pertussis shows polymorphism in two proteins, pertactin (Prn) and the pertussis toxin (PT) S1 subunit, which are important for immunity. A previous study has shown antigenic shifts in these proteins in the Dutch B. pertussis population, and it was suggested that these shifts were driven by vaccination. The recent Italian clinical trial provided the opportunity to compare the frequencies of Prn and PT S1 subunit variants in strains isolated from unvaccinated children, and from children vaccinated with two acellular and one whole-cell pertussis vaccine. Four Prn variants (Prn1, Prn2, Prn3 and Prn5) were found in the 129 strains analysed. Prn1, Prn2 and Prn3 have been described previously, whereas Prn5 is a novel variant. Prn1, Prn2, Prn3 and Prn5 were found in, respectively, 6, 41, 51 and 2% of the strains. The B. pertussis strains used to produce the vaccines administered in the clinical trial were found to produce Prn1, or a type which differed from Prn1 in one amino acid. The frequency of the Prn1 variant was found to be lowest in the strains isolated from vaccinated groups, suggesting that Prn1 strains are more affected by vaccine-induced immunity than Prn2 and Prn3 strains. Only one PT S1 type (S1A) was observed in the examined strains, which was distinct from the types produced by the vaccine strains (S1B and S1D). The S1A type also predominates in the Dutch B. pertussis population. The genetic relationship among B. pertussis strains analysed by IS1002-based DNA fingerprinting revealed that three fingerprint types predominate, representing more than 70% of the strains. Prn2 strains showed a greater variety of fingerprint types compared to Prn3, suggesting that Prn3 has emerged more recently. The results are discussed in the light of vaccine-driven evolution.
Publication Types:
Clinical Trial
Controlled Clinical Trial
PMID: 10463173 [PubMed - indexed for MEDLINE]
Celiac Disease
What is celiac disease? Celiac disease is a disorder that causes problems in your intestines when you eat gluten, which is in wheat, rye, barley and oats. Gluten is like a poison to people with celiac disease, because it damages their intestines.
What does gluten do to people with celiac disease? Gluten damages the intestines. This damage keeps your body from taking in many of the nutrients in the food you eat. This includes vitamins, calcium, protein, carbohydrates, fats and other important nutrients. Your body can't work well without these nutrients.
How did I get celiac disease? Celiac disease runs in the family. You inherited the tendency to get this disease from your parents. If 1 member of your family has celiac disease, about 1 out of 10 other members of your family is likely to have it. You may have this tendency for a while without getting sick. Then something like severe stress, physical injury, infection, childbirth or surgery can "turn on" your celiac disease.
What happens to people with celiac disease? Celiac disease can cause different problems at different times:
An infant with celiac disease may have abdominal pain and diarrhea (even bloody diarrhea), and may fail to grow and gain weight.
A young child may have abdominal pain with nausea and lack of appetite, anemia (not enough iron in the blood), mouth sores and allergic dermatitis (skin rash).
A child could be irritable, fretful, emotionally withdrawn or excessively dependent.
In later stages, a child may become malnourished, with or without vomiting and diarrhea. This would cause the child to have a large tummy, thin thigh muscles and flat buttocks.
Teenagers may hit puberty late and be short. Celiac disease might cause some hair loss (a condition called alopecia areata).
What happens in adults with celiac disease? Adults who begin to be ill with celiac disease might have a general feeling of poor health, with fatigue, irritability and depression, even if they have few intestinal problems. One serious illness that often occurs is osteoporosis (loss of calcium from the bones). A symptom of osteoporosis may be night-time bone pain. About 5% of adults with celiac disease have anemia. Lactose intolerance (problem with foods like milk) is common in patients of all ages with celiac disease. It usually disappears when they follow a gluten-free diet.
Celiac disease sounds really serious! How can I control it? Celiac disease is serious. Fortunately you can control celiac disease just by not eating any gluten. By following the right diet, you can reverse the damage caused by celiac disease and you'll feel better. But if you "cheat" on your diet, the damage will come back, even if you don't feel sick right away.
You'll have to explain your problem and the gluten-free diet to your family members and ask for their support and help. It will take time for you and your family to learn how to avoid gluten in your diet. You can contact one of the celiac support groups listed at the end of this handout. These groups are excellent sources of information and advice. They'll help you find gluten-free foods and good recipes, and give you tips for successfully living with celiac disease.
How can I be sure I have celiac disease? National and local support groups
Celiac Sprue Association/United States of America, Inc.
P.O. Box 31700
Omaha, NE 68131-0700
Telephone: 402-558-0600
www.csaceliacs.org
American Celiac Society-Dietary Support Coalition
58 Musano Court
West Orange, NJ 07052
Telephone: 973-325-8837
Celiac Disease Foundation
13251 Ventura Blvd., #1
Studio City, CA 91604
Telephone: 818-990-2354
www.celiac.org
Gluten Intolerance Group of North America
15110 10th Ave. SW
Suite A
Seattle, WA 98166-1820
Telephone: 206-246-6652
New blood tests can help your doctor diagnose this disease. It's necessary to have these blood tests before you start a gluten free-diet. If you have dermatitis herpetiformis (an itchy, blistery skin problem), you have celiac disease. The diagnosis can be confirmed with a biopsy (taking a piece of tissue using a thin tube that is put into your intestines). The best confirmation, though, is if your symptoms go away when you follow a strict gluten-free diet.
What resources are there for people with celiac disease? In these cookbooks, the author, who has celiac disease herself, shares what she has learned about a gluten-free diet. Bette Hagman is the author. The publisher is Henry Holt and Co.
The Gluten-Free Gourmet: Living Well Without Wheat
More From the Gluten-Free Gourmet
The Gluten-Free Gourmet Cooks Fast and Healthy
Wheat-Free and Gluten-Free with No Fuss and Fat
This book is a general guide to living gluten-free:
Against the Grain: The Slightly Eccentric Guide to Living Well Without Gluten or Wheat, written by Jax Peters Lowell and published by Henry Holt and Co.
This handout provides a general overview on this topic and may not apply to everyone. To find out if this handout applies to you and to get more information on this subject, talk to your family doctor.
Visit familydoctor.org for information on this and many other health-related topics.
Copyright © 2001 by the American Academy of Family Physicians.
Permission is granted to print and photocopy this material for nonprofit educational uses. Written permission is required for all other uses, including electronic uses.
Saturday, February 3, 2007
'Autistic diet' getting a closer look
Wheat-, dairy-free plan proving successful for some
By CHERIE BLACK
P-I REPORTER
http://seattlepi.nwsource.com/local/302360_olympichouse03.html
When he was 3 years old, Matthew Sebastian was diagnosed with autism.
Four years later, he began having seizures, which are much more common in autistic children than in the broader population. Doctors told his parents that by the time their son reached puberty, his seizures would get worse and he would have to wear a helmet to protect his head.
High doses of two prescription anti-seizure medications controlled the attacks, but the effects of his autism still kept the small boy in constant motion. He slept poorly and displayed multiple violent daily outbursts, which eventually made him too dangerous to himself and his family to live at home.
Sebastian moved from Federal Way to a home in Seattle, which cared for autistic children in a residential setting. It was there, at the age of 10, that he received what his mother calls the treatment that saved her son's life.
Dubbed by some as the "autism diet," it is a gluten- and casein-free way of eating, often used by people diagnosed with the digestive disorder celiac disease. Gluten products such as wheat, rye and barley are eliminated, as are dairy products, which contain the protein casein.
For eight weeks, Sebastian was weaned off of his anti-seizure medication and placed on the diet. Now 20, he has been seizure-free and drug-free for the past 10 years. His violent behavior stopped.
"Matthew is the complete opposite of what he was before," said his mother Janet Sebastian. "That's why the diet works. His behaviors decreased dramatically over the years and now he's positive and happy."
Why the diet seems to work isn't completely understood. One theory involves the "leaky gut syndrome," in which the autistic child's body isn't able to process proteins found in wheat and dairy products, said Gary Stobbe, medical director of Seattle's Autism Spectrum Treatment and Research Center, a non-profit organization that diagnoses, treats and manages people with autism. The undigested chunks of protein get into the bloodstream and affect the brain. Another theory is the body's immune system is reacting to the proteins in the body.
"Nothing is determined for certain, and there is no set approach with the diet," he said. "In my practice, it is something we encourage in younger kids or if we see a kid not making progress with more conventional therapies."
Stobbe said for some children, especially the more severe autism cases and those with physical complaints, the diet works well. They are calmer, have better attention spans and have less severe behavioral disturbances.
Still no one knows whether this will work in the long term. So far, only anecdotal evidence from parents is available.
One study under way at the University of Rochester Medical Center in New York looks at the effects of the diet in autistic children between the ages of 2 1/2 and 4 1/2. Sponsored by the National Institute of Mental Health, it began in 2004 and should be completed in 2008.
Dr. Geraldine Dawson, director of the University of Washington's Autism Center, is waiting for data from more studies before she'll recommend the diet to her patients, but tells parents who have decided to try it to make sure a nutritionist is involved. She said about half the children seen at the center are on the diet, which has worked for some, and not others.
"While we wait to find out more, parents should watch their children," she said. "You end up restricting what they're eating and some children are suffering nutritionally."
But Sebastian and his family have no doubts. Sebastian now lives at Olympic House, opened in June in North Seattle for those with autism or celiac disease who are on the diet. The house is a part of Alpha Supported Living Services in Seattle, which has 16 homes for disabled adults.
Sebastian was joined by Jacob Al-Hakim, 24, who is also autistic and has celiac disease. Since being put on the diet, his mother, Cheryl Gere, said he is calm and making eye contact with people. Although he doesn't talk much, if at all, he interacts with people and is more aware, she said.
"Simply what they're eating could change their lives," she said.
A nutritionist helped create a rotating meal plan involving a main dish of pork, chicken, beef or fish accompanied by rice, fruit and vegetables. The main dishes are served for both lunch and dinner and changed each day. A rotating staff of at least two people is at the house 24 hours a day and has been trained on what and how to prepare the limited diet.
Gere buys groceries for the house once a week at Central Market in Shoreline and spends about $150 to $200, using money both men receive from the state and their part-time jobs that is allocated for food.
At the grocery store, she can't buy citrus, apples, potatoes, avocados, peppers or tomatoes. She stocks up on rice and rice cakes, yams, bananas and meat. They need special deodorant and shampoo. The tiniest cheat on the diet can cause behavioral problems.
"At one point on his medication, he never left the floor of his room; he was almost comatose," she said of her son. "He sings with us now. He's awake."
Sebastian and Al-Hakim's families say the diet brought back to them sons they thought were lost to the behavioral effects of autism. Sebastian, never without his dog, Holly, is a gold medalist in the Special Olympics and works part-time at a toy store. Al-Hakim recently ice skated for the first time.
"When he was born I wondered what he would become," Janet Sebastian said. "Look at him now."
Source: Autism Spectrum Treatment and Research Center
P-I reporter Cherie Black can be reached at 206-448-8180 or cherieblack@seattlepi.com.
