The Patented Mediator Release Test (MRT®): A Comprehensive Blood Test for Inflammation Caused by Food and Food-Chemical Sensitivities

Mark J. Pasula, PhD

Introduction

There are a wide range of chronic inflammatory conditions where food and food-chemical sensitivities play either a primary or secondary role in generating inflammation and symptoms (Table 1). Fully addressing food sensitivities can have a major impact on the speed and completeness of clinical outcomes. It can also improve the effectiveness of other therapies, as a chronic source of inflammation has been eliminated.

Definition of Food Sensitivity

Food sensitivities can be defined as any inflammation-generating reaction against a specific food or food component that does not involve Type 1 IgE mediated hypersensitivity or food-related autoimmunity. The inflammatory process associated with food sensitivities is significantly more complex than IgE mediated food allergy. Multiple triggering mechanisms and pathways, multiple classes of reacting white cells, a vast number of proinflammatory mediators, and a wide array of symptoms and conditions make sensitivities a highly complex category of adverse food reaction. (Diagram 1)

Innate Immunity Governs Reactions in Gluten Sensitivity

In 2011, Fasano, et al, published a study documenting a new type of adverse reaction to gluten. 1 Termed “Gluten Sensitivity” (GS), it established that GS was distinctly different than celiac disease in several important ways:

  1. Expression of TLR-2 shows that gluten sensitivity is governed by innate immunity
  2. Gut permeability in GS was significantly less than in both controls and celiac patients
  3. Low to moderate levels of inflammation were detected but no tissue damage was visible
  4. There was less infiltration of intraepithelial lymphocytes

The historical importance of this study cannot be overstated. Until this study, the innate immune system had been fully ignored by allergy researchers as having any involvement in diet-induced inflammation. Estimates of the prevalence of GS are 6-8 times greater than celiac disease, affecting approximately 15-20 million Americans.

It is worth noting that the most commonly ordered blood test to help identify culprit food items -food-specific IgG – is a response of the adaptive immune system as is Type 4 delayed type hypersensitivity governed by sensitized T-cells (lymphocyte transformation).

It is also worth noting that the clinical presentations of the GS patients in this study (brain fog, joint and muscle pain, headaches, diarrhea, etc.) were consistent with the presentations of food sensitivity sufferers reported for decades. Further research has confirmed the findings of innate involvement in gluten sensitivity. 2-4

Inflammation and Symptoms Depend on Mediator Release

All clinical and subclinical effects brought about by food-induced inflammation are the direct result of proinflammatory and proalgesic mediator release from various white blood cells. Without the release of histamine, cytokines, prostaglandins, leukotrienes, etc., there is no tissue damage, no pain receptor activation, no smooth muscle contraction, nor any other negative effect associated with diet-induced inflammation. This is true for any form of food-induced inflammation, i.e. food allergy, food-related autoimmunity, or food sensitivity, and whether or not potential mechanisms are elevated.

Identifying Trigger Foods and Food-Chemicals with Antibodies

Food sensitivity management starts with identification of trigger foods and food-chemicals. Therapy typically involves some form of elimination of offending substances, i.e. rotation or avoidance diets. The more precisely practitioners are able to identify and remove inflammation-generating food items, the greater the clinical value of the method.

Most blood tests designed to identify sensitive foods and food-chemicals are typically limited to either a single mechanism, or a specific part of the inflammatory process, which may or may not be involved in actual inflammation and consequently may or may not be clinically relevant. For example, food-specific antibodies other than IgE have not shown a strong correlation with inflammation or symptoms.

IgE, by its function as a trigger of mast cell degranulation in allergy, has an acceptable, though not excellent correlation with both the degree of the inflammatory response and the severity of clinical effects. Other antibody tests (IgG, IgM, IgA) have not demonstrated an acceptable correlation with inflammation or clinical symptomology in adverse food reactions.5-8

The Function of Non-IgE Antibodies

The function of non-IgE food-specific antibodies appears to be related more to the clearing of food antigens and macromolecules via immune complexes, rather than a direct inflammation-producing role. If any, the inflammatory effects of food-specific IgG, IgM or IgA are more likely the result of factors related to the in vivo environment, such as the production of too many smaller immune complexes or complexes that deposit on tissue, eliciting an aggressive reaction by immune cells. But insight into the sizes of immune complexes or whether or not they deposit on tissue cannot be gleaned from quantifying how much food-specific IgG, IgA, or IgM is produced. In addition, in IgE mediated inhalant allergy, elevated levels of allergen-specific IgG has shown anti-inflammatory properties. 9, 10

Other limitations of non-IgE antibody tests are that they don’t offer testing for food-chemical reactions, an area that is often clinically significant. Thus, the information provided by most commercially available food sensitivity blood tests is of limited clinical value.

White Cells Are an Immunologic Endpoint

White cells play a critical role in food-induced inflammation. Neutrophils, monocytes, eosinophils, mast cells and various lymphocytes release mediators in pathogenic reactions. Neutrophils and monocytes/macrophages are first responders in innate pathways (Figure 1). 18-25 Other cells, such as tissue mast cells, eosinophils, and lymphocytes are involved in reactions related to both adaptive and innate pathways. Whether reactions are governed by innate or adaptive pathways, mediator release from white cells are the immunologic “endpoint” of all food-induced inflammatory reactions.

Figure 1 represents a simplified in-vivo reaction of cells involved in the innate inflammatory reaction. When “sensitive” food antigens cross the tight junctions, neutrophils and macrophages are typically the first cells to react. They engage in the destruction of offending pathogens/antigens, ultimately releasing various cytokines and other proinflammatory mediators.

Elevation of various inflammatory markers (HS-CRP, IL-15, PGE3, etc.) as well as white cell involvement has been documented in many food sensitivity related conditions. Neutrophils and eosinophils were confirmed active in ulcerative colitis and diarrhea predominant IBS via measurement of myeloperoxidase and eosinophil cationic protein. 11 Eosinophils were shown to infiltrate the colon in some cases of chronic constipation.27, 28 Killer and Natural Killer cells were shown to be active in food-induced migraine. 14, 15

Mediator Release Causes Volumetric Changes in White Cells

During an inflammatory reaction, phagocytic cells identify offending antigen as foreign, attacking and engulfing, and then eliminating the antigen. During that process, whatever cells are involved release cytoplasm and chemical mediators, causing a volumetric change in reacting cells. This is true of all cells whenever they release mediators. Volumetric changes are typically subtle, in the range of 5-30 fl. [Figure 2]

Figure 2 shows a peritoneal mast cell degranulating. Release of mediators invariably affects volumetric changes in reacting cells.

The Patented Mediator Release Test (MRT®)

The Mediator Release Test (MRT®) developed by Oxford Biomedical Technologies, is a new functional blood test designed to identify sensitive foods and food-chemicals. It utilizes 2 advanced methods of measurement – flow cytometry and ribbon impedance. MRT is the only blood test in the world that tests cellular endpoint reactions to foods, chemicals and other foreign substances, quantifying the degree of the inflammatory response and simultaneously determining which types of cells are reacting. MRT provides the highest therapeutic value of any commercially available food sensitivity blood test.

The Advantage of Ribbon Impedance over Normal Impedance

The ribbon method is a patented form of impedance-based measurement, developed by Oxford Biomedical Technologies. The ribbon method eliminates the baseline and threshold associated with every other impedance-based sizing technology (see Figure 3).16, 17

Figure 3 shows the difference in precision between typical impedance based counting and sizing technologies which rely on a baseline and threshold and the patented ribbon method, which is able to detect the true volume of each cell.

Impedance based methods that employ a baseline and threshold have 3 main shortcomings:

  1. They improperly size cells when dissimilar sized particles traverse the aperture near each other
  2. Impedance technologies that rely on a baseline begin their sizing from the level of the threshold, not from the when the actual start of the particle passing through the aperture occurs
  3. They are only capable of measuring size changes (2-D) not volumetric changes (3-D)

The ribbon method is extremely precise, measuring the entire flow of liquid and cells millions of times per second as they pass through the aperture. Because the ribbon identifies the true starting point and ending point of each cell (see Figure 3), it is able to provide a volumetric determination of all tested cells. Incorporation of the ribbon method gives MRT superior precision and accuracy over other impedance-based cell sizing methods of food sensitivity testing.

Flow Cytometry Provides Vital Clinical Data

Impedance based sizing methods are incapable of differentiating between different types of similar sized cells. (Figure 4) The use of advanced flow cytometry in MRTIII is another area that helps provide vital clinical information to practitioners. Cytometry allows the measurement of all types of circulating white cells simultaneously (neutrophils, monocytes, eosinophils, lymphocytes). This is important because different types of white cells can act independently of each other.

Figure 4 shows where the different cell types are sized using impedance based sizing methods. Due to very similar sizes of various peripheral granulocytes, impedance based technologies are incapable of distinguishing between different classes of reacting cells.

For example, monocytes, which comprise one to seven percent of white cells, can strongly react to an antigen independent of any other group of white cells. So can any group of cells, i.e. lymphocytes, neutrophils, eosinophils, etc.

If neutrophils, which comprise between 55-75% of total white cells have a cumulative three percent reaction (which would be non-reactive) and monocytes have a cumulative 50% reaction (which would be strongly reactive), the clinically significant monocyte reaction would not be recorded as clinically significant because the cumulative reaction of the monocytes is small relative to the total cumulative reaction (or non-reaction) of the neutrophils.

However, because MRT is able to distinguish between reacting groups of cells, it is the only food sensitivity testing technology that is able to deliver the most complete picture of cellular reactivity. (Figure 5) Thus, MRT provides practitioners and their patients the highest level of clinical utility.

Figure 5 shows that the patented MRT, with the use of flow cytometry, is the only technology able to distinguish between different types of white cell reactions. This provides unparalleled therapeutic information for practitioners.

Advantages of MRT over Other Food Sensitivity Tests

MRT offers several important advantages over other sensitivity-based blood tests:

  1. MRT is a functional measure of sensitivity-based inflammatory responses, not just a measurement of a potential trigger which may or may not have clinical relevance.
  2. Because MRT is an endpoint test, it is able to account for the widest range of triggering mechanisms involved in sensitivity reactions, accounting for both innate and adaptive pathways.
  3. MRT can test reactions to foods, food-chemicals and other substances.
  4. Because it is a 3 dimensional volumetric measurement, MRT is able to reliably quantify the degree of the inflammatory response. This is important because MRT provides insight into dose-dependent reactions and subclinical reactions. These are clinically meaningful but virtually impossible to identify without MRT.
  5. Because MRT combines patented impedance based technology with advanced flow cytometry, MRT is the only food sensitivity blood test that is able to identify different types of cellular reactions independently and simultaneously (monocytes, neutrophils, eosinophils, lymphocytes). This has great importance as different white cells can react independently and exclusively of each other.

MRT has overcome most of the immunologic and/or clinical challenges related to identification of trigger substances. Thus, MRT provides practitioners and their patients a new level of insight into food-induced inflammatory processes that will improve therapeutic outcomes and make the designing of anti-inflammatory eating plans easier and more clinically effective.
For more information about MRT and food sensitivities, please visit www.betterbloodtest.com or call 1-888-669-5327.

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