Assessing the Mechanism of Immunotherapy for Allergy and Allergic Asthma: Effect of Viral Respiratory Infections on Pathogenesis and Clinical Course of Asthma and Allergy
Principal Investigator: Ronald Rabin, MD
Office / Division / Lab: OVRR / DBPAP / LI
One in five Americans have allergies or asthma. Our laboratory regulates biologics intended for diagnosis and treatment of allergies and asthma (most commonly allergenic extracts for "allergy shots"). Only a few allergenic extracts are standardized for potency to insure consistent dosing among manufactured lots of allergenic extracts. Our lot release lab provides reagents to manufacturers to insure uniform potency testing, and validates manufacturers' potency measurements with our own. Currently, allergy extracts contained unmodified allergens, which modify hay fever or asthma, but are not effective for treatment of food allergy, and risk dangerous allergic reactions. FDA is evaluating novel products for treatment of food and environmental allergy, many of which different mechanisms of action. Our research program insures that scientists and reviewers have the appropriate knowledge base to evaluate these novel therapies, and strives to expand the number of allergenic extracts that are standardized for potency.
Our laboratory has two overall projects: The first project explores environmental factors that trigger allergic diseases and asthma. One factor is respiratory syncytial virus (RSV). RSV may infect lower airways of infants, which increases disease severity and may trigger asthma. We seek to understand why, in addition to their location, infection of lower airways with RSV triggers asthma. We use in vitro culture systems to study biological responses to RSV by cells from the upper and lower airways. In particular, we focus on the balance between expression and responses to interferons (IFN), which protect cells from infection, and pro-inflammatory cytokines, which may be detrimental to infection. We propose that characterizing cellular responses to RSV infection will reveal novel insights towards the pathogenesis of allergic asthma.
Our second project is to characterize house dust mite (HDM) allergens. Indoor allergens, including HDM are a second environmental factor that triggers allergic diseases and asthma. HDM allergy is complex because as many as 32 proteins contribute to the allergic response; how many are clinically important is not known. Allergenic extracts used for diagnosis or treatment of HDM allergy may vary widely in their content of allergenic proteins depending on differences in methods of culturing HDM, source materials, or extraction methods. The current method of standardization of HDM extracts for potency does not measure the concentration of individual allergens and cannot detect qualitative or differences in individual allergen content among HDM allergen extracts. We propose that measurement of all clinically relevant allergenic proteins may reveal clinically important differences among HDM preparations that may inform patient care and provide direct benefit to public health. Current methods aren't feasible, so we are developing the use of mass spectrometry to biochemically characterize and standardize HDM extracts. We believe that standardization of HDM extracts by MS will provide a benchmark to standardize other complex allergenic extracts, thus expanding the number of extracts that are standardized for potency.
Asthma and atopy are quintessential diseases for which genetics loads the gun and environment pulls the trigger. GWAS studies reveal associations of asthma with polymorphisms in genes associated with type 2 adaptive responses and the epithelial cell barrier. Two viral pathogens that cause the common cold are also environmental factors associated with asthma: respiratory syncytial virus (RSV) and rhinovirus (RV). Both RSV and RV can infect lower airways, and asthma concurrent with either virus is associated with poor expression of types I or III interferons (IFN).
Our laboratory uses in vitro tissue culture models of respiratory epithelium to compare the cellular factors that determine mild versus severe RSV infection, and which components of the IFN response locally contain RSV infection. Human type I IFNs include IFN and twelve subtypes of IFN-alpha, all of which signal through the ubiquitously expressed IFNAR1/2 receptor complex. There are four type III IFNs, all of which signal through a receptor dimer comprised of IFNLR, selectively expressed by cells of epithelial lineage, and IL10RB. Although they signal through distinct receptors, types I and III IFN share a common signaling pathway that induces autocrine and paracrine expression of more than 600 genes, collectively referred to as interferon response genes (ISG). Many ISGs code for proteins that directly inhibit viral replication and protect against spread of viral infection to neighboring cells.
In addition to IFNs, viral pathogens simultaneously induce expression of NF-kB dependent pro-inflammatory cytokines. For example, IL-6 drives STAT3 mediated IL-17 expression, which in turn drives canonical NF-B mediated expression of IL-6, CXCL8 and CCL2--a positive feedback loop that contributes to immune-mediated pathology of viral lower respiratory infections. Because the same pattern recognition receptors (RIG-I or MDA5) activate IFN and proinflammatory cytokine expression, they may be considered a point of bifurcation. Properly balanced, viral infection is locally contained. Improperly balanced, prolonged expression of IFNs or more likely, high expression of pro-inflammatory cytokines mediates autoinflammatory tissue destruction and promotes allergic sensitization or asthma.
Of interest, some allergenic proteins mimic or enhance the pro-inflammatory response. For example, Der p 2, a house dust mite (HDM) allergen, enhances TLR signaling and NF-kB activation in response to LPS. Because they are ubiquitous in American homes, infants are exposed to HDM early. In genetically susceptible individuals, HDM are among the first environmental allergens to elicit allergic disease, and may initiate the allergic march towards multiple allergies and allergic asthma.
We focus on factors that determine successful local immunity to respiratory viruses, and that shift the response from one that is primarily anti-viral towards a one that is pro-inflammatory. Our Specific Aims are:
- Define cellular factors that direct the balance between expression of IFN versus pro-inflammatory cytokines in response to viral infection.
- Define non-redundant functions of types I versus type III IFNs, and of individual IFN subtypes
- Mass Spectrometry for characterization and rapid, quantitative, and definitive standardization of potency of house dust mite allergenic extracts
Clin Exp Allergy 2020 Jun;50(6):741-51
Measurement of German cockroach (GCr) allergens and their isoforms in allergen extracts with mass spectrometry.
Mindaye ST, David NA, Esfahani SAZ, Schal C, Matsui EC, Ronald RL, Slater JE
Allergy 2019 Dec;74 Suppl 108:3-25
Perspectives in allergen immunotherapy: 2019 and beyond.
Pfaar O, Agache I, de Blay F, Bonini S, Chaker AM, Durham SR, Gawlik R, Hellings PW, Jutel M, Kleine-Tebbe J, Klimek L, Kopp MV, Nandy A, Rabin RL, van Ree R, Renz H, Roberts G, Salapatek AM, Schmidt-Weber CB, Shamji MH, Sturm GJ, Virchow JC, Wahn U, Willers C, Zieglmayer P, Akdis CA
J Allergy Clin Immunol 2019 Oct;144(4):1140
Regulation of allergen immunotherapy products in Europe and the United States.
Rabin RL, Bridgewater J, Slater JE
J Virol 2019 Oct 29;93(22):e01216-19
Lack of activation marker induction and chemokine receptor switch in human neonatal myeloid dendritic cells in response to human respiratory syncytial virus.
Le Nouen C, Hillyer P, Levenson E, Martens C, Rabin RL, Collins PL, Buchholz UJ
PLoS One 2019 Sep 6;14(9):e0221159
Gender differences in innate responses and gene expression profiles in memory CD4 T cells are apparent very early during acute simian immunodeficiency virus infection.
George J, Johnson RC, Mattapallil MJ, Renn L, Rabin R, Merrell DS, Mattapallil JJ
J Interferon Cytokine Res 2019 May;39(5):283-92
S27 of IFNalpha1 contributes to its low affinity for IFNAR2 and weak antiviral activity.
Sharma N, O'Neal AJ, Gonzalez C, Wittling M, Gjinaj E, Parsons LM, Panda D, Khalenkov A, Scott D, Misra S, Rabin RL
Front Immunol 2019 May 15;10:1019
IRF1 maintains optimal constitutive expression of antiviral genes and regulates the early antiviral response.
Panda D, Gjinaj E, Bachu M, Squire E, Novatt H, Ozato K, Rabin RL
J Allergy Clin Immunol 2019 Mar;143(3):1176-82.e5
Developmental regulation of type 1 and type 3 IFN production and risk for infant infections and asthma development.
Holt PG, Mok D, Panda D, Renn L, Fabozzi G, deKlerk NH, Kusel MM, Serralha M, Hollams EM, Holt BJ, Sly PD, Rabin RL
J Virol 2018 Aug 16;92(17):e00518-18
Strand-specific dual RNA-seq of bronchial epithelial cells infected with influenza A/H3N2 viruses reveals splicing of gene segment 6 and novel host-virus interactions.
Fabozzi G, Oler AJ, Liu P, Chen Y, Mindaye S, Dolan MA, Kenney H, Gucek M, Zhu J, Rabin RL, Subbarao K
J Virol 2018 Jul 17;92(15):e02202-17
Differential responses by human respiratory epithelial cell lines to respiratory syncytial virus reflect distinct patterns of infection control.
Hillyer P, Shepard R, Uehling M, Krenz M, Sheikh F, Thayer KR, Huang L, Yan L, Panda D, Luongo C, Buchholz UJ, Collins PL, Donnelly RP, Rabin RL
Allergy 2018 Jun;73(6):1196-205
Macrophages-common culprit in obesity and asthma.
Sharma N, Akkoyunlu M, Rabin RL
Allergy 2018 Apr;73(4):816-26
Allergen manufacturing and quality aspects for allergen immunotherapy in Europe and the United States: an analysis from the EAACI AIT Guidelines Project.
Bonertz A, Roberts G, Slater JE, Bridgewater J, Rabin RL, Hoefnagel M, Timon M, Pini C, Pfaar O, Sheikh A, Ryan D, Akdis C, Goldstein J, Poulsen LK, van Ree R, Rhyner C, Barber D, Palomares O, Pawankar R, Hamerlijnk D, Klimek L, Agache I, Angier E, Casale T, Fernandez-Rivas M, Halken S, Jutel M, Lau S, Pajno G, Sturm G, Varga EM, van Wijk RG, Bonini S, Muraro A, Vieths S
Allergy 2018 Jan;73(1):64-76
Challenges in the implementation of EAACI guidelines on allergen immunotherapy: a global perspective on the regulation of allergen products.
Bonertz A, Roberts G, Hoefnagel M, Timon M, Slater J, Rabin R, Bridgewater J, Pini C, Pfaar O, Akdis C, Goldstein J, Poulsen LK, van Ree R, Rhyner C, Barber D, Palomares O, Sheikh A, Pawankar R, Hamerlijnk D, Klimek L, Agache I, Angier E, Casale T, Fernandez-Rivas M, Halken S, Jutel M, Lau S, Pajno G, Sturm G, Maria Varga E, van Wijk RG, Bonini S, Muraro A, Vieths S
Clin Exp Allergy 2017 Dec;47(12):1661-70
Accurate quantification of 5 German cockroach (GCr) allergens in complex extracts using multiple reaction monitoring mass spectrometry (MRM MS).
Mindaye ST, Spiric J, David NA, Rabin RL, Slater JE
Ann Allergy Asthma Immunol 2017 May;118(5):531-6
Allergenic extracts to diagnose and treat sensitivity to insect venoms and inhaled allergens.
Khurana T, Bridgewater JL, Rabin RL
Clin Exp Allergy 2017 May;47(5):604-17
Mass spectrometry to complement standardization of house dust mite and other complex allergenic extracts.
Spiric J, Reuter A, Rabin RL
Front Immunol 2017 Apr 21;8:459
Type I and Type III interferons display different dependency on mitogen-activated protein kinases to mount an antiviral state in the human gut.
Pervolaraki K, Stanifer ML, Munchau S, Renn LA, Albrecht D, Kurzhals S, Senis E, Grimm D, Schroder-Braunstein J, Rabin RL, Boulant S
Virology 2017 Apr;504:63-72
Respiratory syncytial virus infection induces a subset of types I and III interferons in human dendritic cells.
Hillyer P, Mane VP, Chen A, Dos Santos MB, Schramm LM, Shepard RE, Luongo C, Le Nouen C, Huang L, Yan L, Buchholz UJ, Jubin RG, Collins PL, Rabin RL
Allergy 2017 Jul;72(7):1035-42
Allergen Exposure Chambers (AEC): harmonizing current concepts and projecting the needs for the future--an EAACI position paper.
Pfaar O, Calderon MA, Andrews CP, Angjeli E, Bergmann KC, Bonlokke JH, de Blay F, Devillier P, Ellis AK, Gerth van Wijk R, Hohlfeld J, Horak F, Jutel M, Jacobs RL, Jacobsen L, Kaul S, Larche M, Larenas-Linnemann D, Mosges R, Nolte H, Patel P, Peoples L, Rabin RL, Rather C, Salapatek AM, Sigsgaard T, Thaarup S, Yang J, Zieglmayer P, Zuberbier T, Demoly P
Cell Immunol 2016 Dec;310:156-64
Early treatment with reverse transcriptase inhibitors significantly suppresses peak plasma IFNalpha in vivo during acute simian immunodeficiency virus infection.
George J, Renn L, Verthelyi D, Roederer M, Rabin RL, Mattapallil JJ
J Interferon Cytokine Res 2016 Oct;36(10):589-98
Distinct patterns of expression of transcription factors in response to interferon beta and interferon lambda1.
Novatt H, Theisen TC, Massie T, Massie T, Simonyan V, Voskanian-Kordi A, Renn LA, Rabin RL
Allergy 2016 Oct;71(10):1414-24
Standardization of allergen products: 3. validation of candidate european pharmacopoeia standard methods for quantification of major birch allergen bet v 1.
Kaul S, Zimmer J, Dehus O, Costanzo A, Daas A, Buchheit KH, Asturias JA, Barber D, Carnes J, Chapman M, Dayan-Kenigsberg J, Doring S, Fuhrer F, Hanschmann KM, Holzhauser T, Ledesma A, Moingeon P, Nony E, Pini C, Plunkett G, Reese G, Sandberg E, Sander I, Strecker D, Valerio C, van Ree R, Vieths S
J Virol 2015 Aug 1;89(15):7567-83
Different temporal effects of Ebola virus VP35 and VP24 proteins on the global gene expression in human dendritic cells.
Ilinykh PA, Lubaki NM, Widen SG, Renn LA, Theisen TC, Rabin RL, Wood TG, Bukreyev A
J Vis Exp 2015 Mar 24;(97):e52650
High-throughput quantitative real-time RT-PCR assay for determining expression profiles of types I and III interferon subtypes.
Renn LA, Theisen TC, Navarro MB, Mane VP, Schramm LM, Kirschman KD, Fabozzi G, Hillyer P, Puig M, Verthelyi D, Rabin RL