By Jessica Martin, PhD
Thomas Edison once reflected on his so-called “light bulb” moments: “During all those years of experimentation and research, I never once made a discovery. All my work was deductive, and the results I achieved were those of invention, pure and simple…I speak without exaggeration when I say that I have constructed 3,000 different theories in connection with the electric light, each one of them reasonable and apparently likely to be true. Yet only in two cases did my experiments prove the truth of my theory.”
So much for a romantic notion of science and discovery. True light bulb moments are few and far between. Actually doing science (or applying science as in Edison’s case) is a great act of perseverance that for every moment of inspiration, there are ninety-nine instances of feeling your way around in the darkness. Edison’s reflections are as true today as they were over a century ago. And by Edison’s definition, we have a lot of modern-day geniuses toiling in our research labs and clinics advancing our understanding of and working toward solutions to food allergy with a very small chance of paying off.
I was reminded of this recently in a casual exchange with Henry Ehrlich, who seems to value my experience in research science, which has shown me that negative findings or “failures” rarely make it beyond the walls of the lab in any formality. Peer-reviewed scientific journals generally don’t like to publish them – it isn’t sexy science that demands to be read. And let’s face it—scientists are people, too. Hypotheses going unsupported can really be a blow to the ego especially when it seemed like such a great idea before the months to even years of “bench work”. And don’t get me started on scientific funding. With many good ideas going unfunded these days, negative results do not bode well for a scientist’s future support. It is easier to trash negative lines of research before there is enough detail and rigor required for a peer-reviewed publication and move on to more promising ones.
But can’t we salvage something from the negative? Shouldn’t other scientists working on the same problem learn from these experiments? How often do scientists working in the same field, toward the same goals, and toiling just as hard in their labs, end up testing the same hypotheses only to get the same “negative” result without each other’s knowledge? Isn’t this a waste of valuable resources, both time and money? Couldn’t we reach our ultimate goals faster if we formally shared the negative results of solidly executed experiments that yielded less than desirable findings? [By the way, science writer Matt Shipman wrote an in-depth piece earlier this year on SciLogs that interviews the editorial director of PLOS One, chronicling a multitude of reasons why negative findings rarely get published http://www.scilogs.com/communication_breakdown/negative-results-plos-one/].
While I am a food-allergy Mom, I trained as a scientist first, and so it was with great interest that I read a peer-reviewed article that showed negative results in a series of food-allergy-related experiments. Publication of negative results is definitely not the norm for most scientists (and the journals that will ultimately review and publish them). However, this well-respected journal in the field of immunology undoubtedly made scientists circa 1999 question that a “simple” cure – a fusion of gene therapy and immunotherapy – was on the horizon for food allergy. Most importantly, they likely spared some of their colleagues from repeating similar experiments.
In the late 1990s, Xiu-Min Li and colleagues at Mount Sinai set out in an attempt to “fool the immune system” by combining gene therapy and immunotherapy. The researchers hypothesized that they could prevent sensitization to peanut protein in mice by first injecting them with a DNA vector containing peanut DNA (gene for the highly allergenic Ara h2 protein). For their experimental control – mice injected with a DNA vector lacking Ara h2 – they hypothesized would show no protection against peanut sensitization. Three weeks following the DNA injection, the researchers would attempt to sensitize both sets of mice to peanut protein. The idea was that the DNA from an allergenic portion of the peanut (Ara h2), would get incorporated into the mouse’s cells, and these cells in turn would make the Ara h2 protein. They thought that if the mouse’s own cells were expressing and releasing the Ara h2 protein, the mouse’s immune system would deem Ara h2 harmless, and fail to become sensitized to Ara h2. After all, why would the immune system attack a substance made by the mouse’s very own cells? Yet, the scientists never ended up testing their original hypothesis. Much to their surprise, when they were going to attempt sensitizing the mice to peanut protein, they discovered that the mice receiving the Ara h2-containing DNA vector were already highly allergic (control mice were not allergic). So much for a “simple” solution to food allergy. The immune system was definitely not “fooled.”
According to Dr. Li, at the time they submitted their findings to the Journal of Immunology, the editor felt their “negative” findings were incredibly important to publish to the larger scientific community. So important and well-executed in fact that the paper sailed past the first round of peer-review (a rarity in science) and into the published realm quite quickly (the paper was received at the journal on 9/17/1998 and was accepted for publication on 11/11/1998). Dr. Li explained that during the mid to late 1990’s a lot of scientific exploration was aimed front and center at gene therapy’s potential cure for a multitude of maladies, including allergy. In fact, before this paper was published, there were two peer-reviewed publications showing successful DNA vector-allergen immunizations preventing allergy in other rodent models[2,3]. Dr. Li and colleagues’ research proved this was not going to be such an easy endeavor. In fact, while gene therapy holds great promise, the potential perils of this technology have halted many clinical trials in their tracks. (For more information on the challenges of gene therapy, see the following link http://learn.genetics.utah.edu/content/tech/genetherapy/gtchallenges/)
As to how many scientists altered the trajectory of their research as a result of this one “negative” publication is difficult to say. As a future food allergy parent (I was a senior in high school at the time, and children, let alone food allergies, were nowhere on the radar!), I am thankful Dr. Li and colleagues performed the rigorous studies required for a peer-reviewed publication. Likewise, I am just as thankful to the journal editor who realized the importance of this work to the larger scientific community. After all, “the definition of insanity is doing the same thing over and over again and expecting a different result” – a quote often attributed to Einstein. Then again, without this publication, other researchers may not have known the “insanity” of their endeavors.
As for Dr. Li and colleagues, these “negative” findings certainly altered the path of their research trajectory. They attempted modified versions of immunotherapy involving genetic manipulation of the DNA for allergenic peanut proteins in bacterial cells instead of mouse cells[4,5]. These bacterial cells, once heat-killed, were then administered to peanut-sensitized mice. This method got around potential issues with direct DNA injection into mice, while simultaneously serving to stimulate and tip the immune system toward a non-allergic phenotype. While results of these experiments were encouraging, Dr. Li was simultaneously experimenting with Chinese herbs to treat both food allergy and allergic asthma. Her already extensive publication record on this topic continues to grow[8,9].The success of Chinese herbs to treat both food allergy and asthma in allergic mice has led directly to current clinical trials in humans. Henry’s upcoming book* documents the amazing story behind the years of laboratory toil and the promise of a solution, not engineered in a lab, but rather growing in remote parts of China, just waiting to be harvested. Rather than trying to “fool” the immune system, Dr. Li’s current approach attempts to “educate” the immune system.
With that lab success, I come back to Thomas Edison because Dr. Li’s story is the universal experience of laboratory science and exploration. Her words are remarkably similar to Edison’s over a century ago –“Before I had the idea to test Chinese herbs, we had tested at least 10 different types of immunotherapy.”
I am so happy that she not only shares her successes, but also the “negative” results. Immunotherapy still certainly holds promise, but “fooling” the immune system into a lasting food allergy cure will require a lot of engineering and likely many more failures before true successes, although one can always hope differently. So, please scientists and journals alike, find a way to share those “negative” results with colleagues. Those affected by food allergy await reversing them once and for all.
*Now in print
2. Hsu CH, Chua KY, Tao MH, Lai YL, Wu HD, et al. (1996) Immunoprophylaxis of allergen-induced immunoglobulin E synthesis and airway hyperresponsiveness in vivo by genetic immunization. Nat Med 2: 540-544.
3. Raz E, Tighe H, Sato Y, Corr M, Dudler JA, et al. (1996) Preferential induction of a Th1 immune response and inhibition of specific IgE antibody formation by plasmid DNA immunization. Proc Natl Acad Sci U S A 93: 5141-5145.
4. Li XM, Srivastava K, Grishin A, Huang CK, Schofield B, et al. (2003) Persistent protective effect of heat-killed Escherichia coli producing “engineered,” recombinant peanut proteins in a murine model of peanut allergy. J Allergy Clin Immunol 112: 159-167.
5. Li XM, Srivastava K, Huleatt JW, Bottomly K, Burks AW, et al. (2003) Engineered recombinant peanut protein and heat-killed Listeria monocytogenes coadministration protects against peanut-induced anaphylaxis in a murine model. J Immunol 170: 3289-3295.
7. Li XM, Huang CK, Zhang TF, Teper AA, Srivastava K, et al. (2000) The chinese herbal medicine formula MSSM-002 suppresses allergic airway hyperreactivity and modulates TH1/TH2 responses in a murine model of allergic asthma. J Allergy Clin Immunol 106: 660-668.
Jessica Martin earned a Ph.D. in Neuroscience from Oregon Health and Science University in 2011. She lives in the Portland, Oregon area with her husband Jason and their sons where she teaches undergraduate biology and anatomy and physiology at Portland Community College. Although not currently engaged in cutting-edge laboratory science, she continues to actively research current findings in allergy and immunology, where she writes about some of those findings on her blog, The Food Allergy Sleuth. She aspires to eventually return to the trenches of doing laboratory science in allergy and immunology, but for now, her life is happily filled to the brim with being a Mom, an educator, a writer, and as most food allergy sufferers and parents already know, part-time cook.