Science
Inflammation, innate immunity &
measurement cell activation
Functional cell response test
The Alcat Test measures changes in vital white blood cells while confronted with foods or other items. Using this biological approach it is examined whether and how strongly immune cell, especially granulocytes, respond.
– Granulocytes are a subgroup of white blood cells and well known for proinflammatory and highly toxic reactions.
– They represent 60-85% of the total population of the circulating leukocytes in human blood.
– They use receptors to recognise antigen-non-specific danger signals and determine whether a pathogen or a substance (incl. molecules inside the body) may be dangerous or non-dangerous. Such external and internal threats are neutralised by inflammatory reactions.
The acute effects of inflammation as a result of the cellular defence processes affect the entire body and its immune balance – everyone knows this; It is the principle of action if you catch the flu, for example. Scientists are convinced that the chronification of inflammatory processes (silent inflammation) is one of the causes of many diseases.
Study ’21, click here…
Non-IgE food allergy: Mitochondrial & nuclear DNA released from food allergen-activated neutrophils.
→ Explanation
Study ’21, click here…
Longitudinal trial shows significant WEIGHT LOSS and body composition improvement with the Alcat Test
→ Abstract
More studies – study section↓
See below – Yale School of Medicine (2 publications), Univ. of N. Illinois, Texas, Pavia, Camerino etc.↓
Innate immunity activation
The innate immune cells use various “killing mechanisms”:
- Phagocytosis (“eating”/ingesting)
- Degranulation (Vesicles release toxic mediators to neutralise microorganisms or endogenous “dangers”)
- NETosis (Cell burst; forming extracellular traps to bind pathogens or molecules. ETs consist of networks of the cell’s inner fibres, including DNA).
Video link: A neutrophil chasing bacteria (example phagocytosis)
Structure of the immune system (simplified)
"The Coarse Filter" - 1. Immune barrier
Protection of epithelial surfaces and sensing of the environment: Gastrointestinal / Skin /Respiratory
The first barrier of the immune system acts like a “coarse filter” by capturing, destroying, or deactivating potential pathogens by mechanical and biological means.
These include mucus, which can trap and help expel invaders; lytic enzymes in saliva; including, lysozyme and phospholipase A2; body
fluids such as tears and urine that flush microorganisms and other substances out of the system; hydrochloric acid in the stomach; antimicrobial peptides produced by the skin
and respiratory tract such as β-defensin; and, IgA antibodies in the intestines.
"FIREWALL" - 2. Immune barrier
“Firewall” – Non-specific Immune System innate, cellular defense without memory; Inhibition of antigen immune recognition, protection of epithelial surfaces and sensing of the environment
The second barrier, the innate immune defence, is the focus of the Alcat Test. It is evolutionarily the oldest part of our targeted immune defence and represents the adaptation to a life with pathogens. Furthermore, the innate immune system not only distinguishes between “own” and “foreign”, but also between “harmless” and “dangerous”. The non-specific immune system forms a kind of forefront, an initial “firewall”, which is supposed to eliminate invading pathogens or endogenous dangerous particles within minutes or to initiate an inflammation for continoued defence. Especially the neutrophil granulocytes (NG), with about 70-80% the largest leucocyte population, are active as acute cells and play an important role in the non-specific defence against harmful particles. They release inflammation mediators, free radicals and oxidative enzymes to a high degree and help to initiate an inflammatory reaction, which should make it easier for the body to fight the problem quickly and effectively. Neutrophils constantly patrol the body and can recognise what is harmless or dangerous based on certain molecular structural patterns.
Firewall and Alcat Test
Neutrophile Granulozyten (NG / grösste Untergruppe der Immunzellen) neutralisieren Pathogene (Erreger) oder körperinterne “Gefahrenmoleküle” auf vielerlei Weise, vornehmlich mit zelltoxischen Sauerstoff- oder Stickstoffradikalen, durch Produktion von Säuren, wie z.B. hypochloriger Säure oder der Aktivierung von speziellen Enzymen, wie z.B. der Myeloperoxidasen.
When granulocytes are activated by a structure acting as an antigen, special pattern recognition receptors in the cell membrane or cytoplasm of immune cells trigger an inflammation cascade and the immune system is put on alert. Cytoplasmic granules, i.e. small membrane-enveloped vesicles containing various substances, especially inflammation mediators, are formed inside the cell.
Both the resulting swelling of the cells and the subsequent release of the reactive substances and inflammation mediators (degranulation) up to the complete lysis of the cell lead to a measurable change in the form and number of cells. Simultaneously with these processes, the immune cells communicate with each other and with the rest of the immune system. (see cell graphic below)
Harmful particles and other debris must be constantly removed by phagocytosis. The largest group of phagocytizing cells are neutrophils. However, they are not involved in the processes of type I food allergy, but are used for effective initial control in acute infections and injuries. Chronic activation of the cells can lead to disease symptoms due to persistent inflammatory reactions and oxidative stress.
"Targeted Defense" - 3. Immune barrier
Specific Immune System acquired, adaptive defense with “memory”; Differentiation and regulation of atopic inflammation
The third barrier, the adaptive immune system, consists of “special units” that recognise pathogens with strain-specific and variable antigens (mostly proteins) and form corresponding antibodies. It has memory, as opposed to the non-specific immune system, and forms the basis of our adaptability to new or altered pathogens.
The innate, non-specific and the adaptive, specific immune system work closely together and are mutually dependent. Only through a well-coordinated interaction is the complex immune reaction of the body possible.
Inflammation /disease process
Malfunction in the “firewall” – chronic inflammatory diseases
The gut-associated immune system hosts almost 80% of the immunologically active cells. They distinguish between “friend or foe” and protect against infections by viruses, bacteria, etc. In particular, a damaged intestinal barrier can lead to malfunctions of the cellular immune defence and to chronic inflammation – especially if an irritating substance is consumed regularly. Foods, additives, medicines, moulds and even herbs may alert the immune cells if interpreted as “dangerous”.
Scientists at Yale University demonstrate that the Alcat test identifies food items that result in release of inflammatory markers and activation of innate immune cells (2).
Round 45 studies and observational applications of the Alcat Test show significantly positive health outcomes for a wide range of inflammatory conditions through dietary changes based on the Alcat Test results.
“The Alcat Test ist precise and effective”
Prof. Dr. W. Z. Mehal, Yale School of Medicine; Prof. of Medicine (Digestive Diseases); Director of Yale Weight Loss Program; Director of Fatty Liver Disease Program
Alcat Test / Measurement of diet-induced immune cell activation
For more in-depth literature, you are welcome to request the Scientific Dossier on the Alcat Test.
It contains literature and studies related to the Alcat Test, including double-blind validation studies on clinical and analytical sensitivity and specificity, observational studies and experimental studies on mechanisms of action or mechanistic studies. Furthermore, the scientific context and outdated classification attempts in allergological systems are discussed, and much more.
How are cell reactions measured?
Measurement method (impedance-flowcytometry)
Electrical impedance flow cytometry (blood cell analysis) is used as the traditional standard method for counting cells – also known as the Coulter principle. The principle is used in almost every haematological analyser. It involves passing whole blood between two electrodes through an opening that is so narrow that only one cell can pass through at a time. With the help of hydrodynamic focusing, the cells are sent through the opening cell by cell.
Why is the impedance method used as measuring technique for the Alcat Test?
Cell changes are detected using specialized automated impedance-flow cytometry, a form of “non-invasive realtime cell monitoring.” The measuring principle of the ROBOCat II device is based on the fact that during the passage of a cell through an electric field, the resistance (as a function of the cells) will change in proportion to the cell’s volume (impedance). The system’s processor will capture the change and express it as a pulse where its amplitude is directly proportional to the cell’s volume. The number and size distribution of the cells is displayed graphically. For relative analysis, the degree of deviation of the measurement to the control curve (baseline/negative control) is determined mathematically using computer algorithms. It is then categorized into four response levels: severe, moderate, mild, or negative.
According to the current state of science, it is used to analyse early cellular responses and is considered superior to other available methods for analysing cellular reactions. The analysis of cell reactions with conventional flow cytometers can miss early adverse cell reactions.
We refer to the work of Prof. M. Cooper, a pioneer in labelfree technology for “continuous non-invasive real-time cell monitoring.” More information on measurement technologies for cells: https://en.wikipedia.org/wiki/Hematology_analyzer
How accurate is the test?
The Alcat Test shows precise reliability in clinical and analytical validations. Independent double-blind studies, the highest validation standard of a laboratory procedure, have been performed at institutes and universities. In the last 2 years, these included peer-reviewed publications in scientific journals by researchers at Yale University, among others. According to these studies by independent researchers, the Alcat Test demonstrates a high benefit for a variety of complaints .
For this innovation, as well as for its customer service, the company was awarded the independent Frost & Sullivan Award 2016 as “Best Food Intolerance Testing Provider in North America”.
How does the test work?
Cellular activation naturally induces certain characteristic changes of cell size, volume, and number (see cell graph). These cell changes are detected fully-automatically using special impedance flow cytometry – called “non-invasive-live-cell-monitoring” – an indication that the cells are alive and are confronted with the stimulus in this moment.
The sensitive analytical device records the number and size distribution of the cells before (base curve) and after stimulation (reaction curve) with a substance and determines the degree of deviation of the measured values with algorithmic precision. Reactions are classified into four reaction levels: strong, moderate, mild, negative.
Effects of leukocyte activation
Defence reactions are associated with characteristic changes in cell size or number. So-called impedance flow cytometers are very precise and sensitive. The cell graph below illustrates the typical morphological changes of an activated immune cell:
(A) Trigger recognition via receptors
The immune system may be divided functionally into an unspecific (innate) and a specific (acquired) defence, which consists of both cellular and humoral (soluble) components. Both defence systems are closely linked and work in synergy.
The non-specific defence, already functioning to a large extent at birth and therefore also known as innate immune system, is directed against frequently occurring surface structures of exogenous factors (e.g. pathogens), which are called PAMPs = pathogen associated molecular patterns. In connection with chronic inflammatory diseases, however, the DAMPs = danger/damage associated molecular patterns are the subject of intensive current research. These include internal molecules such as for example cell debris, DNA adducts, or heat shock proteins etc. but also food particles or molecules, UV, stress, and many others.
Both PAMPs and DAMPs activate granulocytes, macrophages or dendritic cells via pattern recognition receptors (PRR) – specific surface receptors – such as TLR, NOD-like receptors and others.
(B) Flattening of the cell
The activation of immune cells (here neutrophilic granulocytes) leads to certain characteristic changes in size, shape and volume.
As a result, the immune cells flatten and the cell nucleus membrane begins to disintegrate. At this stage, cell-to-cell communication is active. If cells of the non-specific immune system recognise PAMPs or DAMPs, they are phagocyted and destroyed. At the same time, messenger agents are released which can attract further immune cells, trigger an inflammatory reaction or activate the specific defence by recruiting lymphocytes.
(C) Volume increase of the cell
Loss of segmentation of the cell nucleus; The granules (with pro-inflammatory mediators)
dissolve and the nuclear plasma mixes with the cytoplasm.
(D) Cellburst
Degranulation or NET-formation (neutrophil extracellular traps); Cellburst (after 2 hours) results in degranulation and the release of reactive oxygen species (ROS), pro-inflammatory cytokines and NET (neutrophil extracellular traps) formation.
Granulocytes can expel so-called extracellular traps (ET), extracellular fibres of granule proteins and chromatin, so that pathogens or internal danger molecules can be bound and eliminated.
The formation of ETs by neutrophil/eosinophil granulocytes or mast cells is an important mechanism of innate immune defence. During ET, the immune cell bursts and releases its toxic contents. A special “trap” is created, consisting of decondensed chromatin fibres containing a mixture of antimicrobial mediators from the granules including released pro-inflammatory DNA and reactive oxygen molecules. NETs can rapidly disarm and immobilize pathogens until they can be removed by macrophages.
For the first time, a team of researchers at Yale University has identified elevated cell-free DNA (cfDNA) supernatants associated with food reactions, which may be indicative of NETs. Further research related to NETs associated with food sensitivity is under current investigation.
Studies and literature
Alcat Test primary literature (DBOC, analytical sensitivity/specificity, others)
+++ UPDATE: Publications 2021+++
– B. König et al; Studies of mitochondrial and nuclear DNA released from food allergen-activated neutrophils. Implications for non-IgE food allergy; Allergy Asthma Proc.; 2021 May 1;42(3):e59-e70.doi: 10.2500/aap.2021.42.210021; https://pubmed.ncbi.nlm.nih.gov/33980341/
– F. B. Willis et al; Food Allergen Elimination for Obesity Reduction; a Longitudinal, Case-Control Trial; DOI:10.31488/bjg.1000122; https://britishjournalofgastroenterology.com/food-allergen-elimination-for-obesity
Double-blind studies on the Alcat Test
(1) Ali et al; Efficacy of individualised diets in patients with irritable bowel syndrome: a randomised controlled trial; Yale Scholl of Medicine; BMJ Open Gastroenterol. 2017 Sep 20;4(1):e000164. doi: 10.1136/bmjgast-201 (https://bmjopengastro.bmj.com/content/4/1/e000164 )
(2) Garcia-Martinez, I., Weiss, T.R., Yousaf, M.N. et al. A leukocyte activation test identifies food items which induce release of DNA by innate immune peripheral blood leucocytes. Yale School of Medicine; Nutr Metab (Lond) 15, 26 (2018). https://doi.org/10.1186/s12986-018-0260-4
(3) Lukaszuk I.M, Shokrani M, Ghosh Roy P, Hoppensteadt J, and Josephine Umoren; Effects of Antigen Leukocyte Cellular Activation Test-Based Diet on Inflammation, Body Composition, and Medical Symptoms; Northern Illinois University; Alternative and Complementary Therapies VOL. 24, NO. 5; 11 Oct 2018 https://doi.org/10.1089/act.2018.29183.jml
(4) Buck Willis F, Ram Shanmugam, Sarah A Curran. Food Allergen Eliminations for Obesity Reduction: A Comparison Study with Therapeutic Exercise. University of Texas; Food Sci Nutr Res. 2018; 1(1): 1-6.; https://scivisionpub.com/pdfs/food-allergen-eliminations-for-obesity-reduction-a-comparison-study-with-therapeutic-exercise-580.pdf
(5) Michele Di Stefano, Eugenia Vittoria Pesatori, Giulia Francesca Manfredi, Mara De Amici, Giacomo Grandi, Alessandro Gabriele, Davide Iozzi, Giuseppe Di Fede; Non-Celiac Gluten Sensitivity in patients with severe abdominal pain and bloating: The accuracy of ALCAT 5; University of Pavia; Clin Nutr ESPEN; 2018 Dec;28:127-131. DOI: 10.1016/j.clnesp.2018.08.017 https://www.ncbi.nlm.nih.gov/pubmed/30390869/
(6) Pierluigi Pompei, Iolanda Grappasonni, Stefania Scuri, Fabio Petrelli, Enea Traini, Sacha Sorrentino, Giuseppe Di Fede: A Clinical Evidence of a Correlation Between Insulin Resistance and the ALCAT Food Intolerance Test; University of Camerino; Altern Ther Health Med; 2019 Mar;25(2):22-38. https://www.ncbi.nlm.nih.gov/pubmed/30990791
(7) Pietschmann, N. “Food Intolerance: Immune Activation Through Diet-associated Stimuli in Chronic Disease,” Altern Ther Health Med, vol. 21, no. 4, pp. 42-52, 2015 Jul-Aug 2015
(8) “High Correlation of the Alcat Test Results with Double Blind Challenge (DBC) in Food Sensitivity“; Fell, Brostoff & Pasula, Präsentation der Studiendurchführung und Ergebnisse beim 45. Annual Congress of the American College of Allergy and Immunology, Los Angeles vom 12. – 16. November 1988 und anschließend Veröffentlichung in den Annals of Allergy.
(9) “Alcat a new test for food induced problems in medicine?“ Fell et al., Präsentation der Studiendurchführung und Ergebnisse beim Jahrestreffen der American Academy of Otolaryngic Allergy, Washington DC, 1. Oktober 1988
(10) ”Alcat® – a new cellular test for food sensitivity“; Fell, Brostoff & Soulsby, Präsentation der Studiendurchführung und Ergebnisse beim Jahrestreffen der American In-Vitro Allergy & Immunology Society, August 1990, Toronto, Canada
(11) ”Cellular responses to food in irritable bowel syndrome – an investigation of the Alcat Test“; Fell, Soulsby & Brostoff, Publikation der zusammengefassten Studien-Ergebnisse im Journal of Nutritional Medicine, Vol. 2, Nr. 2, 1991
(12) “Diagnostic Value of Alcat Test in intolerance to food additives compared with double blind placebo controlled (DBPC) oral challenges“ L. Hoj, J Allerg Clin Immun 1 (3); 1996
Reproducibility studies:
(13) “Reproducibility of the Alcat Test”; Studie von Dr. Paul Potter an der Universität Kapstadt, Johannesburg, Südafrika 1994.
(14) “Reproducibility of the Antigen Leukocyte Cellular Antibody Test (Alcat) – Statistical Analysis, Summary Statistics & Scientific Report“, University of the Range Free State in Bloemfontein, Südafrika, Dr. WML Neetling and Dr. AM Kachelhoffer von Januar – April, 1998.
(15) “Parexel Medstat Final Statistical Report – Study of the Alcat Test in 10 subjects”, Dr. Per Fuglerud, Parexel Norwegen, Nov. 1999
(16) Study Comparing Alcat Test Results With Flow Cytometry and Microscop, Dr. Gitte Jensen, NIS Labs (Natural Immune System) Oregon, USA, 2009
Alcat studies – other
(17) ”Evaluation of Alcat Test Results in the Non-IgE Mediated Pathology of the Skin” DeAmici et al., Studiendurchführung und -bericht der Universität von Pavia, Italien. Presented at the 30th Congress of the European Academy of Allergy and Clinical Immunology, 11 – 15 June 2011 – Istanbul, Turkey. (Poster Presentation, Abstract #553)
(18) ”Alcat Test Results in the Treatment of Gastrointestinal Symptoms” Berardi L. et al., Studiendurchführung und -bericht der Universität von Pavia, Italien. Presented at the 30th Congress of the European Academy of Allergy and Clinical Immunology, 11 – 15 June 2011 – Istanbul, Turkey. (Poster Presentation, Abstract #552)
(19) ”Rational management of food intolerance in an elite soccer club” Angelini et al., Journal of the International Society of Sports Nutrition 2011, 8(Suppl 1):36
(20) ”Alcat Test Identifies Food Intolerance in Patients with Gastrointestinal Symptoms” Berardi et al., Report of the XXVIII Congress of the European Academy of Allergy & Clinical Immunology, European Journal of Allergy and Clinical Immunology, Supplement 90, Volume 64, 2009, pg. 490.
(21) “Food Intolerance in Patients with Cutaneous Diseases: Diagnostic Value of the Alcat Test” Berardi et al., Report of the XXVIII Congress of the European Academy of Allergy and Clinical Immunology, European Journal of Allergy and Clinical Immunology, Supplement 90, Volume 64, 2009, pg. 490.
(22) “The Effect of the Alcat Test Diet Therapy for Food Sensitivity in Patients with Obesity” Akmal et al., Middle East Journal of Family Medicine. April 2009 – Vol. 7, Issue 3.
(23) ” IMS Health Economics and Outcomes Research – Influence of Food Intolerance in Migraines: Final Report of Statistical Results” Immunological Center of Cataluna, Version 3, December 28, 2006.
(24) “A Comparison of the Alcat Test for Food Reactions Amongst 2 Population Sub-Groups” Studienpräsentation von Dr. DH Sandberg und Dr. MJ Pasula, 45th Annual Congress of the American College of Allergy and Immunology, Los Angeles, CA: November 12 – 16, 1998, publiziert in den Annals of Allergy.
(25) “The Short Term Efficacy of the Alcat Test of Food Sensitivities to Facilitate Changes in Body Composition and Self-Reported Disease Symptoms: A Randomized Controlled Study” Kaats et al. in Am J of Bariatric Med, Spring 1996: 18 – 23.
(26) “El test Alcat de sensibilidad a los alimentos y su interés en Medicina Estética Cabo-Soler JR. Alcuni Particolari Della Dieta In Medicina Estetica (Comments On Diets In Esthetic Medicine)”. Abstract of 14th Med Day of Esthetical Medicine & Dermatological Survey. Venice, Italy, Sep. 22 – 23, 1995. Published in the proceedings.
(27) “Outcome Study in 353 Consecutive Patients Following The Alcat Diet”, Studie von Dr. Lene Hoj in Kopenhagen, Allergy Clinic Charlottenlund, Dänemark 1998. Non-Published.
(28) “Prevalence of food allergy and intolerance in children based on MAST CLA and Alcat Tests” Buczylko et al., Rocz Akad Med Bialymst. 1995; 40(3):452 – 456.
(29) “Alcat Test Results in the Treatment of Respiratory and Gastrointestinal Symptoms, Arthritis, Skin and Central Nervous System” Mylek et al., Rocz Akad Med Bialymst. 1995; 40(3): 625 – 629.
(30) “Food Intolerance in Patients with Angioedema and Chronic Urticaria. An investigation by RAST and Alcat Test Studie von Dr. Lene Hoj, präsentiert beim XVI European Congress of Allergy and Clinical Immunology”, Madrid, Spanien: June 25 – 30, 1995 und publiziert im European Journal of Allergy and Clinical Immunology – Supplement, No. 26, Vol. 50, 1995.
(31) “Multiple Pathogenic Mechanisms in Food Sensitivity Reaction In-Vitro”. Pasula MJ, Puccio SG, 4th International Symposium on Immunological and Clinical Problems of Food Allergy, Milan, Italy. November 5 – 9, 1989. Abstract Symposium Book, pg. 37.
(32) “Influence of Food antigens on Volumes of Circulating White Blood Cells and Platelets Aggregation Studienpräsentation beim 4. Symposium on Immunological and Clinical Problems of Food Allergy”, Mailand, Italien, 5. – 9. November 1989
(33) “The Alcat Test – A Guide and Barometer in the Therapy of Environmental and Food Sensitivities”. Dr. BA Solomon, Environmental Medicine, Vol. 9, Number 2, 1992:2 – 6
(34) “Pilot Study into the Effect of Naturally Occurring Pharmacoactive Agents on the Alcat Test”. Fell, PJ. American Otolaryngic Allergy Association Annual Meeting, September 27, 1991, Kansas City, MO. Published in the proceedings.
(35) “Inhibitory Effect of Sodium Cromoglycate on Granulocyte Response to Food Antigens In-Vitro”. Fell PJ, Sandberg DH, Pasula MJ. 47th Annual meeting of the American College of Allergy & Immunology, November 10 – 14, 1990, San Francisco, CA. Publ. in proceedings.
(36) “Gastrointestinal Complaints Related to Diet”, DH Sandberg, International Pediatrics, Vol. 5 No. 1, 1990:23 – 9.
(37) “South African Outcome Study randomisierte Studie an 274 Patienten”, Dr. Jan Geldenhuys, Johannesburg, Südafrika, 1997
(38) “Allergie alimentari. Tecniche diagnostiche a confronto [Food allergy: comparison of diagnostic techniques]”, Mancini S, Fierimonte V, Iacovoni R, Spaini A, Viarani P, Pichi A., Minerva Pediatr. 1995 May;47(5):159-63 [Italian]
(39). “Technical Study Comparing The Alcat Methodology With Activation Of Granulocytes Following Challenge With Zymosan”. Studie von Dr. Cristina Mele der Universität von Rom.
(40) “Autism – a multidisciplinary approach to treatment”, Kotsanis et al. 1994. Diese Studie wurde unter der Leitung von Dr. Constantine A. Kotsanis durchgeführt. Die Ergebnisse wurden auf dem Jahrestreffen der American Academy of Otolaryngic Allergy 1994 präsentiert und stehen auf der Webseite des Kotsianis Instituts zur Verfügung. https://www.kotsanisinstitute.com
(41) “Controversial antigen leucocyte cellular antibody test (Alcat): a non specific inhibitory effect of alpha glycoproteins”, Kedryna & Guminska, Med Sci Monit 1999; 5(2):BR193 – 197.
(42) “Ogni intervento comincia a tavola”, Mele Cristina, Medici Oggi, Maggio 2002: 210 – 213
(43) “Evaluation of the cytotoxic food test and the Alcat (antigen leukocyte cellular antibody test)”. Pol Merkuriusz Lek. 1997 Feb;2(8):154 – 9.
(44) “The Alcat Test: in vitro procedure for determining food sensitivities”, Pasula MJ., Folia Med Cracov. 1993; 34(1 –4):153 –7.
(45) “Pharmacoactive Compounds in Foods – The effect on the Alcat Test in Healthy volunteers and patients suffering from Migraine” Fell PJ, Brostoff J, Pasula M. AAOA News 9:2:29.
Alcat Test primary/secondary literature on Innate Immunity Activation & Diseases
(1) Sollid, L.M. and Jabri, B. “Triggers and drivers of autoimmunity: lessons from coeliac disease,” Nat Rev Immunol, vol. 13, no. 4, pp.294-302, 04 2013.
[2] Garcia-Martinez, I., Weiss, T.R., Ali, A. and Mehal, W.Z. “The ALCAT Test Predicts the Release of DNA and Myeloperoxidase by Innate Immune Peripheral Blood Leukocytes via a PKC Dependent Pathway,” presented at the he International Congress on Integrative Medicine and Health (ICIMH), Las Vegas, Nevada, USA, 2016.
[3] Ali, A. et al. “Efficacy of individualised diets in patients with irritable bowel syndrome: a randomised controlled trial.” BMJ Open Gastroenterol, vol. 4, no 1, p. e000164, Sep 2017.
[4] Kau, A.L., Ahern, P.P., Griffin, N.W., Goodman, A.L. and Gordon, J.I. “Human nutrition, the gut microbiome and the immune system,” Nature, vol. 474, no. 7351, pp. 327-36, Jun 2011.
[5] Berardi, L., De Amici, M., Vignini, A., Mantegna, G. and Mosca, M. “Alcat Test identifies food intolerance in patients with gastrointestinal symptoms,” presented at the The XXVIII European Academy of Allergy and Clinical Immunology Congress Warsaw, Poland, 2009.
[6] Pietschmann, N. “Food Intolerance: Immune Activation Through Diet-associated Stimuli in Chronic Disease,” Altern Ther Health Med, vol. 21, no. 4, pp. 42-52, 2015 Jul-Aug 2015.
[7] Berardi, L., De Amici, M., Vignini, A., Torre, C. and Mosca, M. “Food intolerance in patients with cutaneous diseases: diagnostic value of the alcat test,” Allergy: European Journal of Allergy and Clinical Immunology, vol. 64, p. 490, 2009.
[8] Buczyłko, K. et al. “Prevalence of food allergy and intolerance in children based on MAST CLA and ALCAT tests,” Rocz Akad Med Bialymst, vol. 40, no. 3, pp. 452-6, 1995.
[9] Mancini, S., Fierimonte, V., Iacovoni, R., Spaini, A., Viarani, P., and Pichi, A. “[Food allergy: comparison of diagnostic techniques],” inita), Minerva Pediatr, vol. 47, no. 5, pp. 159-63, May 1995.
[10] Sharma, S. et al. “Association of variants in innate immune genes with asthma and eczema,” Pediatr Allergy Immunol, vol. 23, no. 4, pp. 315-23, Jun 2012.
[11] Sweeney, C.M., Tobin, A.M. and Kirby, B. “Innate immunity in the pathogenesis of psoriasis,” Arch Dermatol Res, vol. 303, no. 10, pp. 691-705, Dec 2011.
[12] Mylek, D. “ALCAT Test results in the treatment of respiratory and gastrointestinal symptoms, arthritis, skin and central nervous system,” Rocz Akad Med Bialymst, vol. 40, no. 3, pp. 625-9, 1995.
[13] Rashtak, S., Snyder, M.R., Pittock, S.J., Wu, T.T., Gandhi, M.J. and Murray, J.A. “Serology of celiac disease in gluten-sensitive ataxia or neuropathy: role of deamidated gliadin antibody,” J Neuroimmunol, vol. 230, no. 1-2, pp. 130-4, Jan 2011.
[14] Vitte, J., Michel, B.F., Bongrand, P. and Gastaut, J.L. “Oxidative stress level in circulating neutrophils is linked to neurodegenerative diseases,” J Clin Immunol, vol. 24, no. 6, pp. 683-92, Nov 2004.
[15] Jyonouchi, H. “Food allergy and autism spectrum disorders: is there a link?,” Curr Allergy Asthma Rep, vol. 9, no. 3, pp. 194-201, May 2009.
[16] Jyonouchi, H., Geng, L., Streck, D.L. and Toruner, G.A. “Children with autism spectrum disorders (ASD) who exhibit chronic gastrointestinal (GI) symptoms and marked fluctuation of behavioral symptoms exhibit distinct innate immune abnormalities and transcriptional profiles of peripheral blood (PB) monocytes,” J Neuroimmunol, vol. 238, no. 1-2, pp. 73-80, Sep 2011.
[17] Samaroo, D. et al. “Novel immune response to gluten in individuals with schizophrenia,” Schizophr Res, vol. 118, no. 1-3, pp. 248-55, May 2010.
[18] Dickerson, F., Stallings, C., Origoni, A., Vaughan, C., Khushalani, S. and Yolken, R. “Markers of gluten sensitivity in acute mania: a longitudinal study,” Psychiatry Res, vol. 196, no. 1, pp. 68-71, Mar 2012.
[19] Pan, A. et al. “Depression and risk of stroke morbidity and mortality: a meta-analysis and systematic review.” JAMA, vol. 306, no 11, pp.1241-9, Sep 2011.
(20) P., Samuel, L.J., Miller, E.R. and Szanton, S.L. “Depression and oxidative stress: results from a meta-analysis of observational studies,” Psychosom Med, vol. 76, no. 1, pp. 12-9, Jan 2014.
[21] Rybka, J. et al. “Interplay between the pro-oxidant and antioxidant systems and proinflammatory cytokine levels, in relation to iron metabolism and the erythron in epression,” Free Radic Biol Med, vol. 63, pp. 187-94, Oct 2013.
[22] Liu, T. et al. “A Meta-Analysis of Oxidative Stress Markers in Depression,” PLoS One, vol. 10, no. 10, p. e0138904, 2015.
[23] Bajpai, A., Verma, A.K., Srivastava, M. and Srivastava, R. “Oxidative stress and major depression,” J Clin Diagn Res, vol. 8, no. 12, pp. CC04-7, Dec 2014.
[24] Galecki, P. “Oxidative Stress inDepression,” in Systems Biology of Free Radicals and Antioxidants, I. Laher, Ed. Germany: Springer-Verlag Berlin Heidelberg, 2014, pp. 2369-2395.
[25] Holgate, S.T. “Innate and adaptive immune responses in asthma,” Nat Med, vol. 18, no. 5, pp. 673-83, May 2012.
[26] Finn, P.W. and Bigby, T.D. “Innate immunity and asthma,” Proc Am Thorac Soc, vol. 6, no. 3, pp. 260-5, May 2009.
[27] Ghani, A., Mehal, W.Z. and Ali, A. “Food reactivity on the Alcat leukocyte activation test is associated with up-regulation of CD11b on T cells,” vol. 20, N. H. Yale School of Medicine, CT, USA, Ed., ed: The Journal of Alternative and Complementary Medicine, 2014, pp. A35-A36.
[28] Akmal, M., Khan, S.A. and Khan, A.Q. “The Effect of The ALCAT Test Diet Therapy for Food Sensitivity in Patient’s with Obesity,” MIDDLE EAST JOURNAL OF FAMILY MEDICINE, vol. 7, no. 3, 2009.
[29] Odegaard, J.I. and A. Chawla, A. Connecting type 1 and type 2 diabetes through innate immunity,” Cold Spring Harb Perspect Med, vol. 2, no. 3, p. a007724, Mar 2012.
[30] Tremellen, K. and Tunc, O. “Macrophage activity in semen is significantly correlated with sperm quality in infertile men,” Int J Androl, vol. 33, no. 6, pp. 823-31, Dec 2010.
[31] Bastard, J.P. et al. “Recent advances in the relationship between obesity, inflammation, and insulin resistance,” Eur Cytokine Netw, vol. 17, no. 1, pp. 4-12, Mar 2006.
[32] Miesel, R., Hartung, R., and Kroeger, H. “Priming of NADPH oxidase by tumor necrosis factor alpha in patients with inflammatory and autoimmune rheumatic diseases,” nflammation, vol. 20, no. 4, pp. 427-38, Aug 1996.
[33] Fitzpatrick, A.L. et al. “Leukocyte telomere length and cardiovascular disease in the cardiovascular health study,” Am J Epidemiol, vol. 165, no. 1, pp. 14-21, Jan 2007.
[34] Tlaskalová-Hogenová, H. et al. Involvement of innate immunity in the development of inflammatory and autoimmune diseases,” Ann NY Acad Sci, vol. 1051, pp. 787-98, Jun 2005.
[35] Li, L. et al. “[Changes of neutrophil myeloperoxidase in coronary circulation among patients with acute coronary syndrome],” (in chi), Zhonghua Xin Xue Guan Bing Za Zhi, vol. 33, no. 12, pp. 1106-8, Dec 2005.
[36] Lin, W.W. and Karin, M.”A cytokine-mediated link between innate immunity, inflammation, and cancer,” J Clin Invest, vol. 117, no. 5, pp. 1175-83, May 2007.
[37] Poon, B.Y., Ward, C.A., Cooper, C.B., Giles, W.R., Burns, A.R. and Kubes, P. “alpha(4)-integrin mediates neutrophil-induced free radical injury to cardiac myocytes,” J Cell Biol, vol. 152, no. 5, pp. 857-66, Mar 2001.
[38] Nascimento, G.G., Leite, F.R., Correa, M.B., Horta, B.L., Peres, M.A. and Demarco, F.F. “Relationship between periodontal disease and obesity: the role of life-course events,” Braz Dent J, vol. 25, no. 2, pp. 87-9, 2014.
[39] Bostanci, N. et al. “Expression and regulation of the NALP3 inflammasome complex in periodontal diseases,” Clin Exp Immunol, vol. 157, no. 3, pp. 415-22, Sep 2009.
[40] Geering, B., Stoeckle, C., Conus, S. and Simon, H.U. “Living and dying for inflammation: neutrophils, eosinophils, basophils,” Trends Immunol, vol. 34, no. 8, pp. 398-409, Aug 2013.
(41)Jensen, G. “Study Comparing Alcat Test Results With Flow Cytometry and Microscop.” Oregon USA: NIS Labs (Natural Immune System) 2009.
[42] Mehal, W.Z. “CELLS ON FIRE,” Sci Am, vol. 312, no. 6, pp. 44-9, Jun 2015.
[43] Hou, W. et al. “Strange attractors: DAMPs and autophagy link tumor cell death and immunity,” Cell Death Dis, vol. 4, p. e966, Dec 2013.
[44] Ueki, S., Melo, R.C., Ghiran, I., Spencer, L.A., Dvorak, A.M. and Weller, P.F. “Eosinophil extracellular DNA trap cell death mediates lytic release of free secretion-competent eosinophil granules in humans,” Blood, vol. 121, no. 11, pp. 2074-83, Mar 2013.
[45] Garcia-Martinez, I., et al., A leukocyte activation test identifies food items which induce release of DNA by innate immune peripheral blood leucocytes. Nutr Metab (Lond), 2018. 15: p. 26.
(46) Willis, F.B. et al. Food Allergen Eliminations for Obesity Reduction: A Comparison Study with Therapeutic Exercise, et. al, Editor. 2018: Food Sci Nutr Res.
[47] Di Stefano, M., et al., Non-Celiac Gluten Sensitivity in patients with severe abdominal pain and bloating: The accuracy of ALCAT 5. ClinNutr ESPEN, 2018. 28: p. 127-131.
[48] Lukaszuk, J.M., Effects of Antigen Leukocyte Cellular Activation Test-Based Diet on Inflammation, Body Composition, and Medical Symptoms, e. al, Editor. 2018: Alternative and Complementary Therapies.
[49] Pompei, P., et al., A Clinical Evidence of a Correlation Between Insulin Resistance and the ALCAT Food Intolerance Test. Altern Ther Health Med, 2019. 25(2): p. 22-38.
(50) Sapone, A. et al. “Divergence of gut permeability and mucosal immune gene expression in two gluten-associated conditions: celiac disease and gluten sensitivity,” BMC Med, vol. 9, p. 23, Mar 2011.
[51] Schuppan, D., Junker, Y. and Barisani, D. “Celiac disease: from pathogenesis to novel therapies,” Gastroenterology, vol. 137, no. 6, pp. 1912-33, Dec 2009.
[52] Fasano, A., Sapone, A., Zevallos, V. and Schuppan, D. “Nonceliac gluten sensitivity,” Gastroenterology, vol. 148, no. 6, pp. 1195-204, May 2015
[53] Boyce, J.A. et al. “Guidelines for the diagnosis and management of food allergy in the United States: report of the NIAID-sponsored expert panel,” J Allergy Clin Immunol, vol. 126, no. 6 Suppl, pp. S1- 58, Dec 2010.
[54] Ellis, A. and Linaker, B. “Non-coeliac gluten sensitivity?” The Lancet, vol. 311, no. 8078 pp. 1358-1359, 1978.
[55] Di Stefano, M., et al., Non-Celiac Gluten Sensitivity in patients with severe abdominal pain and bloating: The accuracy of ALCAT 5. Clin Nutr ESPEN, 2018. 28: p. 127-131.
Mode of action & basic research on Innate Immunity & Inflammation
(1) Garcia-Martinez, I., Weiss, T.R., Yousaf, M.N. et al. A leukocyte activation test identifies food items which induce release of DNA by innate immune peripheral blood leucocytes. Yale School of Medicine; Nutr Metab (Lond) 15, 26 (2018).
(2) Rosales, C. and Uribe-Querol, E. Phagocytosis: A Fundamental Process in Immunity. Biomed Res Int, 2017. 2017: p. 9042851.
[3] Geering, B., et al. Living and dying for inflammation: neutrophils, eosinophils, basophils. Trends Immunol, 2013. 34(8): p. 398-409.
[4] Rosenberg, H.F., J.C. Masterson, and G.T. Furuta. Eosinophils, probiotics, and the microbiome. J Leukoc Biol, 2016. 100(5): p. 881-888.
[5] Nicholls, S.J. and S.L. Hazen. Myeloperoxidase and cardiovascular disease. Arterioscler Thromb Vasc Biol, 2005. 25(6): p. 1102-11.
[6] Brinkmann, V., et al.. Neutrophil extracellular traps kill bacteria. Science, 2004. 303(5663): p. 1532-5.
[7] Wartha, F., et al. Neutrophil extracellular traps: casting the NET over pathogenesis. Curr Opin Microbiol, 2007. 10(1): p. 52-6.
[8] Lögters, T., et al. The clinical value of neutrophil extracellular traps. Med Microbiol Immunol, 2009. 198(4): p. 211-9.
[9] Medina, E. Neutrophil extracellular traps: a strategic tactic to defeat pathogens with potential consequences for the host. J Innate Immun, 2009. 1(3): p. 176-80.
[10] Branzk, N. and V. Papayannopoulos. Molecular mechanisms regulating NETosis in infection and disease. Semin Immunopathol, 2013. 35(4): p. 513-30.
[11] Hahn, S., et al. Modulation of neutrophil NETosis: interplay between infectious agents and underlying host physiology. Semin Immunopathol, 2013. 35(4): p. 439-53.
[12] Saffarzadeh, M. and K.T. Preissner. Fighting against the dark side of neutrophil extracellular traps in disease: manoeuvres for host protection. Curr Opin Hematol, 2013. 20(1): p. 3-9.
[13] Yipp, B.G. and P. Kubes. NETosis: how vital is it? Blood, 2013. 122(16): p. 2784-94.
[14] Zawrotniak, M. and M. Rapala-Kozik. Neutrophil extracellular traps (NETs) – formation and implications. Acta Biochim Pol, 2013. 60(3): p. 277-84.
[15] Manda, A., et al. Neutrophil extracellular traps in physiology and pathology. Cent Eur J Immunol, 2014. 39(1): p. 116-21.
[16] Pinegin, B., N. Vorobjeva, and V. Pinegin. Neutrophil extracellular traps and their role in the development of chronic inflammation and autoimmunity. Autoimmun Rev, 2015. 14(7): p. 633-40.
[17] Gupta, S. and M.J. Kaplan. The role of neutrophils and NETosis in autoimmune and renal diseases. Nat Rev Nephrol, 2016. 12(7): p.402-13
[18] Castanheira, F.V.S. and P. Kubes. Neutrophils and NETs in modulating acute and chronic inflammation. Blood, 2019. 133(20): p. 2178-2185.
[19] Lacy, P. Mechanisms of degranulation in neutrophils. Allergy Asthma Clin Immunol, 2006. 2(3): p. 98-108.
[20] Jorch, S.K. and P. Kubes. An emerging role for neutrophil extracellular traps in noninfectious disease. Nat Med, 2017. 23(3): p. 279-287.
[21] Delgado-Rizo, V., et al. Neutrophil Extracellular Traps and Its Implications in Inflammation: An Overview. Front Immunol, 2017. 8: p. 81.
[22] Masuda, S., et al. NETosis markers: Quest for specific, objective, and quantitative markers. Clin Chim Acta, 2016. 459: p. 89-93.
[23] Yang, H., et al. New Insights into Neutrophil Extracellular Traps: Mechanisms of Formation and Role in Inflammation. Front Immunol, 2016. 7: p. 302.
[24] McEwen, B.J. Eosinophils: a review. Vet Res Commun, 1992. 16(1): p. 11-44.
[25] Muniz, V.S., R. Baptista-Dos-Reis and J.S. Neves. Functional extracellular eosinophil granules: a bomb caught in a trap. Int Arch Allergy Immunol, 2013. 162(4): p. 276-82.
[26] Ueki, S., et al. Eosinophil extracellular DNA trap cell death mediates lytic release of free secretion-competent eosinophil granules in humans. Blood, 2013. 121(11): p. 2074-83.
[27] Falcone, F.H., D.I. Pritchard, and B.F. Gibbs. Do basophils play a role in immunity against parasites? Trends Parasitol, 2001. 17(3): p. 126-9.
[28] Bianchi, M.E. DAMPs, PAMPs and alarmins: all we need to know about danger. J Leukoc Biol, 2007. 81(1): p. 1-5.
[29] Fink, S.L. and B.T. Cookson. Apoptosis, pyroptosis, and necrosis: mechanistic description of dead and dying eukaryotic cells. Infect Immun, 2005. 73(4): p. 1907-16.
[30] van Schaarenburg, R.A., et al. C1q Deficiency and Neuropsychiatric Systemic Lupus Erythematosus. Front Immunol, 2016. 7: p. 647.
[31] Amarante-Mendes, G.P., et al. Pattern Recognition Receptors and the Host Cell Death Molecular Machinery. Front Immunol, 2018. 9: p. 2379.
[32] Krug, S.M., J.D. Schulzke, and M. Fromm. Tight junction, selective permeability, and related diseases. Semin Cell Dev Biol, 2014. 36: p.166-76
Webinars, publications, lectures, research, news, symptom center, and more
Cell Science Systems offers a wide range of educational materials for both healthcare professionals and patients. Click on the button to open in a new window the education homepage of Cell Science Systems, Corp. (https://cellsciencesystems.com/education/news/).
+++ Disclaimer +++
The Alcat Test is to be understood as a nutritional concept and health consultation of complementary nutritional therapy and is NOT a disease diagnosis or healing method. Nutrition alone is NOT a substitute for careful diagnosis and medical care in case of illness.
The Alcat Test should therefore be carried out in consultation with a healthcare professional in the case of a pre-existing illness. This practitioner can integrate the Alcat Test into the treatment plan in an integrative and compatible manner. However, a healthy diet according to the Alcat principle can easily be used for prevention.
We emphasize that the Alcat Test does NOT measure parameters of the specific immune system, i.e. no antibody levels are measured and it is not an LTT /lymphocyte transformation test, but cell-mediated food-related hypersensitivities are examined, which are mainly mediated by the innate immune system.
Thus, the Alcat Test does NOT diagnose food allergies (type 1/IgE – allergies have to be avoided!) and NO enzymatically caused intolerances to e.g. lactose, fructose or histamine.
It is possible that the innate immune system may be activated at the same time despite existing classical allergy or intolerance and therefore does not exclude cell-mediated pro-inflammatory reactivity as measured by the Alcat Test.
The Alcat Test is NOT an IgG test – we explicitly distance ourselves from antibody tests of IgG class(es).