Immune Engineering Laboratory

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					Immune Engineering Laboratory



      Jennifer A. Maynard
         Chemical Engineering
      University of Texas at Austin
Bio-molecular engineering
    Bio-centric perspective of chemical engineering




 Bioreactor         Cell as reactor           Products

                                      Interactions
                   Catalytic

     Structural                  Self-
                               assembly


Genes                               Mechanical
On/off            Proteins            work



   Complex
                               Therapeutics
   machines

                   Signaling
                                    Folds to active protein
                  Linear string
                  Amino acids


Questions                                Tools

-how do they perform function?           -directed evolution/ mutagenesis
-how can you manipulate function?        -crystallography
-how to make large quantities?           -biochemical/ biophysical
                                         -in vitro/in vivo assays
Immune system                                        Innate
                                                     immunity




                                                                            Antibodies
                          QuickTime™ and a
                TIFF (Uncompressed) decompressor
                   are needed to see this picture.




                                                         T cell receptors


                                                           TCR
Impact of simple physical insight…


                                            B   B


                                            B   B



       Kd 1-100 uM                      Kd ~4x better



      T cell surface with multiple identical TCRS



               Altman et al, Science 1996
I. Develop TCR technology

Ability to manipulate TCRs, library of different
TCRs

Reagents - track pMHCs in response to
Disease & vaccination

Basic - ability to study properties in vitro-
With low affinity techniques

Immunotherapeutics: target cancer/ virally
Infected cells, alter autoimmune disease

• synthetic, naïve libraries; grafted specificity
• Isolate low affinity (physiological)
• Marc Jenkins - immunologist
 Using scTCR system
Use this system to probe basic science issues and applications

Expressed other scTCRs: D10, DO11.10
      - block pMHC in vivo?
      - immunize with variable regions?
Nuclear Magnetic Resonance




                   15N   172GH HSQC
                   0.1-1 mM labeled protein,
                  expressed in minimal media
172GH: I-Au Crystal structure

                          • Peptide register
                            shift
                          • Vβ bias
                            (no Vα H-bonds
                            with MHC)
                          • Few specific TCR-
                            peptide contacts
                          • peptide contacts
                            restricted to 3 α/ β
                          • 3 β compressed,
                            consistent with
                            entropy
I. TCRs: Experimental Autoimmune
Encephalomyelitis

 • Murine model of multiple sclerosis
 • Inflammatory T cell response mediates
   destruction of nervous tissue
I. TCRs: Regulatory T Cells
• Certain T cells specifically down
  regulate immune response.

• CD4+CD25+ Tregs cells possess
  high affinity to self-peptides.

• Potential Autoimmunity Treatment

• Problems:
    – Low Cell Numbers
    – Wide-Ranging Specificities
                                      Roncarolo et al., 2001
• Proposal: Create specific
  regulatory cell line
II. Whooping Cough

• Highly contagious
respiratory infection
(Bordetella pertussis)

• Prolonged cough and
systemic symptoms

• Significant morbidity and
mortality in infants


                              US CDC
II. Anti-PTx antibodies
PTx: A-B5 toxin, adhesin, enzymatic activities
Inactivates Gi/o proteins to poison immune
  responses
II. Anti-PTx antibodies
Use antibody to optimize toxin
adjuvant
III. Bacterial production/ delivery system

             Bacteria      Cell
 Power of engineering

Understand underlying biology such that we can develop
design rules to predictably manipulate systems




          Basic science      Applied science