Basic Immunology

Developing a fundamental understanding of T cell activation and tolerance

  • Mechanobiology

    Mechanotransduction -- the conversion of mechanical forces to biochemical signals–has not been thoroughly studied in immune cells. It is our long-term goal to understand the biophysical basis of mechanotransduction in modulating T cell responses, which can strategically improve T cell functions in autoimmunity and cancer.

    • How do mechanical signals modulate activation of T cells?
    • What are the cytoskeletal pathways that facilitate mechanotransduction in T cells
    • What is the impact of extracellular matrix structure on T cell activation
  • Improving our understanding of the threshold of activation of T cells
  • Immune synapse
  • Advancing Microscopy
    • 3D Deconvolution -- real-time deconvolution using the NVIDIA GPU processor
  • Microfluidics
    • Capturing and delivering cells for high-resolution microscopy

Atomic Force Microscopy

Biology at the nanoscale

We have pioneered the use of AFM in studying immune cells.

  • Delivering molecular signals to single, live cells, with controlled force
    • Hu KK, Bruce MA, Butte MJ. Spatiotemporally and mechanically controlled triggering of mast cells using atomic force microscopy. Immunol Res. 58(2-3):211-7 (May 2014). PMID: 24777418.
  • Measuring beating cardiomyocytes
    • Ge X, Wang IE, Toma I, Sebastiano V, Liu J, Butte MJ, Reijo Pera R, Yang PC. Human amniotic mesenchymal stem cell-derived induced pluripotent stem cells may generate a universal source of cardiac cells. Stem Cells and Development. 2012 Oct 10;21(15):2798-808. doi: 10.1089/scd.2011.0435. Epub 2012 Jun 11. PMID 22530853.
    • Sun N, Yazawa M, Navarrete EG, Sanchez-Freire V, Hu S, Wang L, Han L, Lee A, Abilez OJ, Pavlovic A, Liu J, Hajjar RJ, Dolmetsch RE, Butte MJ, Ashley EA, Longaker MT, Robbins RC, Wu JC. Patient-Specific Induced Pluripotent Stem Cell as A Model for Familial Dilated Cardiomyopathy. Science Translational Medicine, 2012 Apr 18;4(130):130ra47. PMID 22517884.
    • Liu J, Sun N, Bruce MA, Wu JC, Butte MJ. Atomic Force Mechanobiology of Pluripotent Stem Cell-Derived Cardiomyocytes. PLoS ONE, 7(5):e37559. Epub 2012 May 18. PMID 22624048.
    • Wang IN, Wang X, Ge X, Anderson J, Ho M, Ashley E, Liu J, Butte MJ, Yazawa M, Dolmetsch RE, Quertermous T, Yang PC Apelin enhances directed cardiac differentiation of mouse and human embryonic stem cells. PLoS ONE, 2012; 7(6): e38328. Epub 2012 Jun 1. PMID 22675543.
  • Measuring the elastic modulus and viscoelasticity of biomimetic hydrogels
    • Serpooshan V, Zhao M, Metzler S, Weo K, Shah, PB, Wang, A, Mahmoudi M, Malkovskiy AV, Rajadas J, Butte MJ, Bernstein D, Ruiz-Lozano, P (2014). Use of bio-mimmetic 3D technology in therapeutics for heart disease. Bioengineered, 5(3), 0-1 (2014). PMID: 24637710.
    • Serpooshan V, Wei K, Zhao M, Metzler SA, Shah PB, Wang A, Mahmoudi M, Butte MJ, Bernstein D, Ruiz-Lozano P. Bioengineered Acellular Collagen Patch Attenuates Cardiac Remodeling and Improves Ventricular Function post Myocardial Infarction. Biomaterials. 34.36: 9048-9055 (2013). PMID: 23992980.
    • Wei K*, Serpooshan V*, Metzler SE, Zhao M, Shah PB, Kim PJ, Savtchenko A, Wang A, Diez-Cunado M, Cai W, Wei K, Spiering S, Zhu W, Butte MJ, Yang P, Bernstein D, Mercola M, Ruiz-Lozano P. Engineered Epicardium Activates Cardiac Regeneration. In press, Nature (2015).
  • Improving our understanding of exosomes and microvesicular particles
    • Kanada M, Bachmann MH, Hardy JW, Bronsart L, Wang A, Frimannson DO, Sylvester MD, Schmidt TL, Kaspar RL, Butte MJ, Matin AC, Contag CH. Differential fates of biomolecules delivered to target cells via extracellular vesicles. PNAS. 112.12 (2015): E1433-E1442.
  • Development of advanced AFM technologies for rapidly measuring cells
    • Vijayraghavan K, Wang A, Solgaard O, Butte MJ, Melosh NA. Measurement of Elastic Properties in Fluid Using High Bandwidth AFM Probes. Applied Physics Letters, 102, 103111 (2013).
    • Vijayraghavan K, Gellineau AA, Wang A, Butte MJ, Melosh NA, Solgaard O. Modeling, design, and fabrication of a high bandwidth AFM probe for imaging in air and water. J. Micromechanical Systems. 22(3):603-612. Jun 2013.
    • Wang A, Vijayraghavan K, Solgaard O, Butte MJ. Fast Stiffness Mapping of Cells Using High-Bandwidth Atomic Force Microscopy. Submitted.

Immune Diseases

Diaganosis of rare immune diseases

By elucidating the underlying cause of rare immune diseases, we learn more about the human immune system. Furthermore, we improve our ability to apply rational treatments.

  • PI3K
  • RNA modification diseases
  • Adult-onset immune deficiencies
  • Matthews AGW, Briggs CE, Yamanaka K, Small TN, Mooster JL, Bonilla FA, Oettinger MA, Butte MJ. Compound Heterozygous Mutation of Rag1 leading to Omenn Syndrome. PLoS ONE. 10(4): e0121489 (2015). PMID: 25849362
  • Mathieu A-L, … Butte MJ, … Belot A. PRKDC mutations associated with immunodeficiency, granuloma, and autoimmune regulator–dependent autoimmunity. J. Allergy and Clinical Immunology. 2015 Jun;135(6):1578-1588.e5. doi: 10.1016/j.jaci.2015.01.040. Epub 2015 Apr 2. PMID: 25842288

Food Allergy

Improving the diagnosis of food allergies

Diagnosis of food allergies currently does not provide information about the persistence or severity of food allergies. We are developing techniques to facilitate these measurements by advancing skin prick testing.

  • Sun W, Araci Z, Inayathullah M, Manickam S, Zhang X, Bruce MA, Marinkovich MP, Lane AT, Milla C, Rajadas J, Butte MJ. Polyvinylpyrrolidone microneedles enable delivery of intact proteins for diagnostic and therapeutic applications. Acta Biomaterialia, 9 (8), 7767-7774 (2013). PMID: 23648574.

We present a method of fabricating microneedles from polyvinylpyrrolidone (PVP) that enables delivery of intact proteins (or peptides) to the dermal layers of the skin. PVP is known to self-assemble into branched hollow fibers in aqueous and alcoholic solutions; we utilized this property to develop dissolvable patches of microneedles. Proteins were dissolved in concentrated PVP solution in both alcohol and water, poured into PDMS templates shaped as microneedles, and upon evaporation of solvent, formed into concentric, fibrous, layered structures. This approach of making PVP microneedles overcomes problems in dosage, uniform delivery, and stability of protein formulation as compared to protein-coated metallic microneedles or photo-polymerized PVP microneedles. Here we characterize the PVP microneedles and measured the delivery of proteins into skin. We show that our method of fabrication preserves the protein conformation. These microneedles can serve as a broadly useful platform for delivering protein antigens and therapeutic proteins to the skin, for example for allergen skin testing or immunotherapy.

Immune Diagnostics

Low cost counting of cells

We have an ongoing project using microfluidics tools for immunological diagnostics.

  • Newborn Screening: We are developing tools to identify newborns with serious, life-threatening immunodeficiencies (such as SCID).
    • SCID can be identified by low numbers of T cells in the peripheral blood
    • Maternal T cells in the fetal/newborn circulation can confound this count
  • Complete blood counting: Our tools will facilitate semi-quantitative counting of blood cells from a drop of blood.
    • Disaster settings
    • Military applications
    • Resource-poor (rural) settings
    • Home monitoring
We have two publications on this technology:
  • Daniel Garcia; Isaac Ghansah; John LeBlanc; Manish J. Butte. Counting cells with a low-cost integrated microfluidics-waveguide sensor. Biomicrofluidics. March 7, 2012.
  • LeBlanc J; Muller A; Prinz A; Butte MJ. Optical Planar Waveguide for Cell Counting. Applied Physics Letters. Jan 23, 2012.
First generation sensor 2010
Second generation sensor 2012
Third generation sensor 2013 curved sensor
Fourth generation sensor 2015 linear sensor