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Natural “Pesticides” in Wheat: Is There a Role in Gluten Sensitivity and Celiac Disease?

By: Peter Olins, PhD on December 19, 2012

The critical role of wheat gluten in celiac disease is well-known, but it is not clear if other components of wheat might cause undesirable health effects. Non-celiac “gluten sensitivity” is becoming more accepted by clinicians, but in reality it hasn’t been proven if this is really an effect of gluten, or if it is caused by other substances in wheat.

The latest paper by Y. Junker and co-workers studied proteins in wheat called “amylase-trypsin inhibitors” (ATI), which as act as natural pesticides. They found that ATIs trigger “innate” immune responses that are similar to those caused by gluten. It is possible that consuming ATIs might be important in the early stages of celiac disease. Also, since ATIs provoke an immune response in the intestinal tissues of non-celiacs, it is conceivable that ATIs might also be the culprit in non-celiac gluten sensitivity.

Both “Innate” and “Adaptive” Immunity are Important in Celiac Disease

Innate Immunity

This is a simple defense system designed to protect us against invasive organisms, such as bacteria. It is an ancient system, and is even present in primitive organisms like insects. In the human intestine, the innate immune system recognizes simple molecular patterns on bacteria, and mounts an immediate protective response. It’s really our first line of defense in determining “friend or foe”. This molecular recognition process involves proteins, called toll-like receptors (TLRs), which I will describe later.

Adaptive Immunity

This mechanism evolved much more recently, and involves both antibodies and T-cells that recognize very specific molecular surfaces. Adaptive immunity involves many steps, and initially takes days or weeks to develop. The big advantage is that the immune system “remembers” previous encounters, so that repeat infections can be warded off very rapidly.

Immune “Tolerance”

Underlying all of this is the concept of immune “tolerance”. The body needs to be able to identify its own proteins as being safe, but still be able to detect potential threats from the outside world. Normally, we have immune “tolerance” to proteins in our food, and to our own proteins. But somehow, for the very small fraction of people that develop celiac disease, this tolerance breaks down, resulting in a dramatic immune response to both dietary gluten and one of our own proteins (tissue transglutaminase 2). The reason for this breakdown of tolerance is unknown.

One current model for celiac disease is that it occurs in two steps. First, there is an innate immune response to gluten which primes the immune system. Next, specific peptides within gluten provoke an adaptive immune response, which ultimately leads to an inappropriate reaction to certain human proteins. Activation of TLR4 is thought to play a role in priming the adaptive immune system.

Non-Celiac Gluten Sensitivity

One theory for non-celiac gluten sensitivity (NCGS) is that this is an innate immune response to gluten—the only difference is that people with NCGS do not go on to develop full-blown celiac disease (Ref. 1).

As I discussed in a previous blog (Ref. 2), most of the preliminary research on non-celiac gluten sensitivity has involved putting people on a gluten-free diet, and monitoring symptoms. The problem is that these diets are not really specific for gluten, but are really just wheat-free. So, it is a still an open question whether this condition is really caused by gluten or something else in wheat. “Wheat sensitivity of unknown origin” might be a more accurate description.

Ref. 1: Bernardo, et al. Is gliadin really safe for non-coeliac individuals? Production of interleukin 15 in biopsy culture from non-coeliacs challenged with gliadin peptides. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1954879/

Ref. 2: A consensus on diagnosis of gluten sensitivity? Not so fast. 

Natural Pesticides Are Common in Plants

Natural Pesticides in Wheat—Gluten-Sensitivity—Celiac-Diease—Amylase/Trypsin InhibitorWe tend to think of pesticides as being chemicals that we apply to crops to protect them from insects. In reality, millions of years ago, plants developed a wide range of defense mechanisms to prevent being eaten by insects or animals, or to thwart attacks by fungi and other diseases (Ref. 3). This is especially true for the protection of seeds, since these are essential for the propagation of the species.


Many of these natural pesticides are also unpleasant to humans, and a key part of plant breeding over the past few thousand years has been to reduce the amount of these irritants, and make the foods safer and more palatable. One simple example is the high levels of irritants in such foods as chile peppers or onions, which causes a burning sensation when consumed. (Of course, some people choose to ignore the warning signals of these irritants!).

Enzyme Inhibitors

In addition to irritants, plants produce a wide range of different biological pesticides that prevent seeds from being digested by pests. For example, there is a large family of proteins called “amylase-trypsin inhibitors” (ATI’s). ATI proteins block the enzymes that are normally used to digest starch (amylase) and protein (trypsin) in food, and are toxic to many insects and other pests. Many seeds, including wheat, contain a wide range of enzyme inhibitors, such as ATI.

Ref. 3: Ames, BN, et al. Dietary pesticides (99.99% all natural) Proc Natl Acad Sci U S A. 1990 Oct;87(19):7777-81.

Latest Results: ATI’s Stimulate the Human Innate Immune System

In the latest paper by Y. Junker and co-workers (Ref. 4), they found that wheat ATI is not only an enzyme inhibitor, but was also recognized as being “foreign” in the intestine, activating the innate immune system. Two ATI proteins were purified from wheat, and tested for their biological activity on three types of human immune cells: macrophages, monocytes and dendritic cells. In all three cases, ATIs provoked an immune response.

To investigate the mechanism, ATIs were fed to mice, and the researchers detected an inflammatory response in cells from both blood and intestine. The response in the intestine was shown to be dependent on the “Toll-Like Receptor 4″ (TLR4), which normally provides a defense against bacteria.

To test the relevance of this to humans, the two wheat ATIs were tested for their activity on human intestinal biopsy samples. The authors found a similar stimulation of the innate immune system as they had seen in mice. Remarkably, this effect was seen with the biopsy samples from both celiac patients and normal subjects. The effect of ATIs on the tissue from normal people was unexpected. While there is no direct evidence at the moment, the authors speculate that this newly described response to ATI might actually be the trigger for non-celiac gluten sensitivity.

Ref. 4: Junker, Y, et al. Wheat amylase trypsin inhibitors drive intestinal inflammation via activation of toll-like receptor 4. http://www.ncbi.nlm.nih.gov/pubmed/23209313 J. Exp. Med 2012, Dec. 3.

Amylase/Trypsin Inhibitors Also Present in Many Other Seeds

As I mentioned earlier, many plants produce a variety of natural pesticides. In addition to the three grains important in celiac disease (wheat, barley and rye), ATIs are also found in a wide range of other seeds used for food—including soy, quinoa, buckwheat, peas, and corn. In a recent abstract by Zevallos and co-workers (Ref. 5), ATIs were purified from a wide range of food seeds. When the ATIs were added to a cell culture assay, there was an increase in the production of interleukin 8, which is an indicator of innate immune system activation. (A similar response has been seen in a different study, triggered by gluten). The highest interleukin 8 stimulation was found for wheat, barley and rye ATIs, but a significant effect was also found for the other seeds tested. This is a very preliminary result, but it is conceivable that this might account for the intolerance that a few people have to a variety of seeds other than the familiar wheat/barley/rye trio.

Ref. 5: Zevallos, V. et al. 2012. Purifikation, Charakterisierung und biologische Wirkungen von alpha-Amylase-/Trypsininhibitoren aus Pflanzen, Effektoren von intestinalen TLR4 bei Zöliakie  https://www.thieme-connect.com/ejournals/abstract/10.1055/s-0032-1323857


This research is still in its early stages, and much more work will need to be done to determine if there is a cause-and-effect relationship between ATIs and human intestinal health. However, the potential implications of this work are intriguing:

  • While most people can eat wheat without any problems, the non-celiac “gluten sensitivity” found in some people might actually be caused by ATIs rather than gluten.
  • The research supports earlier research suggesting that ALL people might have some level of reaction to wheat, not just celiacs.
  • It is possible that stimulation of the innate immune system by ATI could be an early trigger for celiac disease. If this true, then it opens up the possibility of a completely different set of approaches for preventing or treating celiac disease, in parallel with the current focus on gluten.
  • Long-term, it might be possible to breed varieties of wheat that have reduced ATI content, and therefore might be safer.
  • The significance of the presence of ATIs in a variety of seeds other than wheat is not known. However, it is conceivable that it might account for the mysterious sensitivity to these seeds that has occasionally been described.

These ideas remain speculative at the moment. However, in my opinion, this work could represent a major breakthrough in the research on gluten sensitivity and celiac disease!



9 comments to Natural Pesticides in Wheat: Is There a Role in Gluten Sensitivity and Celiac Disease?

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  • Rob Warren

    My son suffers from ibs which seems to be partially alleviated by a gluten-free diet, which we could not understand as he didn’t have coeliac disease. After reviewing the Junker et al research, ATIs have become the number one suspects for his type of ibs.

    I was wondering if an amylase/trypsin supplement would assist or simply mask the problem? Clearly it would be best to remove the inflammatory ATIs from the diet. Is there a list of foods with high ATI levels?

    • Peter

      Thanks, Rob.
      I should stress that the role of ATIs in causing intestinal damage or symptoms is still preliminary, and has not been proven in a clinical trial. However, the results obtained in the laboratory are very suggestive.

      As far as I know, the response to ATIs is not a direct consequence of their normal enzyme inhibiting activities, so I doubt whether a supplement would help. ATIs appear to be recognized as “foreign”, rather like to way that the body can detect molecules on the surface of bacteria.

      Regarding your son, while this is not intended to be a medical site, I should mention that there are a significant number of people who have “non-celiac gluten sensitivity” (although the exact numbers are not known). Many people have assumed that this response is a specific reaction to gluten, but as I point out in my article, a gluten-free diet involves changes in a large number of proteins, not just gluten. It is quite plausible that we are really dealing with a collection of different food sensitivities, and gluten may only be the culprit in a portion the cases.

  • Amber

    I have Celiac and was wondering if wheat amylase inhibitor is a gluten free product or should i avoid it.

    • Peter Olins

      Amylase trypsin inhibitor is a protein naturally present in all wheat. I’m not sure that I understand your question.

  • Ann

    In addition to the naturally occuring pesticides in wheat, I read that some farmers have been using roundup and other pesticides prior to harvest to increase seed yield. I can no longer eat wheat or dairy as the proteins seem to bother my system. I tested negative for celiacs, but I have a rash on my back when I eat wheat. I developed an autoimmune disease, but I am now in remission after modifying my diet. Humans have been cosuming grains for thousands of years. Are the newer crops bred to contain more of these ATIs? Do the synthetic pesticides combine with the natural pesticides to cause an unfavorable reaction? Do GMOs play a role? I’ve also noticed that I don’t have as many issues when I eat bread while vacationing in Europe. It seems strange that there is such a massive increase in gluten related issues over the last several years.

    • Peter Olins

      Thanks, Ann. You raise so many different topics, it’s hard to know where to start, but I’l take them one by one.

      —Glyphosate. I can see no evidence or rationale for why the herbicide glyphosate is relevant to celiac disease. Please feel free to pursue this in the comment section of my blog on this topic: http://ultimateglutenfree.com/2014/02/does-glyphosate-cause-celiac-disease-actually-no/ It is quite unusual for glyphosate to be applied to wheat in the U.S. (despite some alarming claims that you may see on the Internet), and I’m not aware of any evidence of significant dietary exposure in this country.

      —Since you develop a rash when eating wheat, the most obvious interpretation is that you could have an allergy. Allergies to plant-based foods are relatively common, and I hope that you are working with a physician if this continues to be a problem. It’s intriguing that you seem to have less of a problem in Europe, but if I were in your shoes, I would tend to avoid foods that provoke an allergic reaction.

      —You raise an interesting question about whether the levels of ATI’s have increased in newer crops. I don’t know if this is true, but I’ll look into it.

      —Regarding synthetic versus natural pesticides, that was the underlying theme of the blog. We tend to lump a wide range of substances under the general heading of “pesticides”, but this is just for convenience, and is not meaningful. A pesticide is simply a substance that reduces the activity of an organism that is attacking a food that we care about. ATI’s are enzyme inhibitors that probably interfere animal pests, such as mice or insects. Other pesticides reduce the activity of bacteria or fungi, while others (herbicides) kill plants. I can’t think of any reason why such a wide range of molecules would interact, and I don’t think that there is anything special about “synthetic” versus “natural” pesticides.

      —As far as the increase in gluten-related disorders, there is only limited information about celiac disease, but it appears to have peaked around the turn of the 21st century. There is very little information about non-celiac disease, despite the huge anti-gluten dietary fad that has consumed the U.S.

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