Clarifying Misconceptions about LRA Testing

We read with interest a recent web post comparing and contrasting various methods of detecting food and chemical sensitivities. We applaud the effort to provide an in-depth review of what can be a confusing subject. With regard to the technical aspects of the LRA by ELISA/ACT® testing, we would like to clarify a few details here so patients, practitioners and readers are well informed about this procedure and its clinical significance.

In the description of the LRA testing method the writer states, “the assumption is that this increase in volume is caused by the activation of the major histocompatibility complex (MHC) of the white blood cell, which triggers an immune response. However, there is no research provided to support this assumption.”

It is important to understand that the LRA does NOT use change in lymphocyte volume as its measurement index. The methods that use devices to detect such changes have high variance on split sample comparisons and have limited clinical value. We would also point out that it is NOT the MHC complex, per se, that is activated. The mechanism for Lymphocyte Response Assay (LRA) has been known for some time, although we are the first to make it clinically reproducible. When a lymphocyte is presented with an antigen to which it was previously sensitized a series of kinase induced phosphorylation occurs. There are several results including that the glycocalyx becomes phosphorylated and as a result the highly charged glycoproteins push away from each other. This creates the halo of light seen to reflect cell activation. Phosphorylated signals induce nuclear cell division and proliferation. (1)

There are 3 types of delayed reactions (2). B Lymphocytes are directly responsible for Type 2 and 3 reactions; T cells are responsible for type 4 non-antibody reactions. Type 2 delayed reactions that are antibody based are also associated with lymphocytes since B type of lymphocytes produce the Immunoglobulins involved. Therefore, only a functional test like LRA measures types 2, 3, and 4 concurrently and directly.

LRA cell cultures are reproducible with a variance of less than 3% for over 30 years on consecutive blind split samples – consecutive data shown in Table 1 presented at the American Society for Investigative Pathology (ASIP) conference in 2016 (3).

Table 1. Consecutive blind split Lymphocyte Response Assay (LRA) samples showing high split sample reproducibility.

# items

# items matched #  items unmatched (%) items matched (%) items unmatched Time (years)
4138 4050 88 97.60±3.00% 2.25±2.75%


Test precision is measured best by consecutive blind split samples analyzed over at least 2 years as shown in this table. By contrast other cell culture methods that assess lymphocytic white blood cell function, report 15-30% variance. LRA by this method is a documented advance in precision, sensitivity, specificity and predictive significance.

A different post lists as a weakness of LRA testing that it does not look at granulocytes and therefore does not measure Type I reactions. In Type I reactions histamine sensitizes neutrophils (type of granulocyte) which are responsible for the Type I or IgE (immediate) allergic reaction. (4) The post is correct that the LRA does not measure these reactions because they are totally different in mechanism compared to delayed reactions. There are many types of white blood cells involved in immune reactions beyond lymphocyte and granulocyte subsets. Because lymphocytes are the key cells involved in all 3 types of delayed reactions, the LRA testing is devoted to lymphocytes.

In evaluating the studies of LRA testing, the writer presented the use of supplements and a support group in the fibromyalgia study as “horribly confounding.” This study was designed to be comprehensive and it achieved meaningful results as noted in the journal (5). This study showed more improvement over six months than any previous comprehensive intervention. A single variable study would have been entirely inappropriate in such a community based multi-variable experiment.

The community-based outcome studies in diabetes and fibromyalgia show documented substantially better outcomes using the results of the LRA testing compared to best standard of care (5,6). There is more clinical and basic research in support of LRA than any other functional immunology tests. More studies are always needed. In the meantime, LRA tests lead in both technology and clinical application. We encourage you to dive deeper into any of the technical aspects of LRA testing, I invite you to contact us to schedule a conversation.

Sincere Regards,

Russell Jaffe, MD, Ph.D., CCN

  1. Peter J. Delves, Seamus J. Martin, Dennis R. Burton, Ivan M. Roitt. 13thEd Roitt’s Essential Immunology
  2. Brostoff J, Challacombe SJ, Food Allergy and Intolerance. Saunders Ltd.; 2ndedition(2002)
  3. Lynch A E, Jaffe R Lymphocyte Response Assay: Report on Precision of a Novel Cell Culture Test2016, The FASEB Journal:4.
  4. Lynch A E, Jaffe R, “Lymphocyte Response Assay: Report on Precision of Novel Cell Culture Test, Experimental Biology,” ASIP, San Diego, CA, 2016.
  5. Alcañiz L,Vega AChacón PEl Bekay RVentura IAroca RBlanca MBergstralh DTMonteseirin J.Histamine production by human neutrophils. FASEB J. 2013 Jul;27(7):2902-2910.
  6. Deuster PA and Jaffe R. A Novel Treatment for fibromyalgia improves Clinical Outcomes in a Community Based Study.J Musculo Pain 1998; 6:133-149.
  7. Jaffe R, Mani J, DeVane J, Mani H. Tolerance loss in diabetics: Association with foreign antigen exposure. Diabetic Medicine: J British Diabetic Association2006Aug; 23(8): 924-925.