Check out @Steph_Gaglione's amazing preprint introducing a highly scalable (and inexpensive) approach to TCR synthesis that enables library-on-library screening of TCRs + antigens 👇!.

here's a peek of Dr. Perez showing this work - how diverse CAR libraries (inputs) can be leveraged to generate useful datasets that describe differential CAR phenotypes (outputs) at his thesis defense yesterday 👇. super lucky to be your lab mate & play a small part in this work!

An incredibly impressive body of work from the newly-minted PhD @calebrperez comparing clinical candidate CAR-T architectures head-to-head at single cell resolution, shedding light on mechanisms of differential CAR-T efficacy in vivo with insights into better construct design 🚗.

In sum, Dezfulian et al. have successfully re-centaured T cell antigen discovery around CD4+ T cells 😉.

The authors then applied their TScan-II methodology to uncover antigens in cancer, autoimmunity, & infectious disease across many different MHC class II alleles & TCRs. Their screens uncovered antigens recognized by clonally-expanded CD4+ T cells from Sjögren's syndrome patients

For their last trick🪄, the authors introduced the candidate human TCR antigen-binding variable region atop a murine TCR constant region resulting in a human-mouse hybrid TCR that reduces cross-reactivity with the endogenous TCR. They termed these chimeras "centaur T cells"🏃🐎

For the T cell side of the equation, they made use of the granule-releasing properties of primary CD8+ T cells, but introduced a CD4 co-receptor alongside a candidate T cell receptor (TCR) of interest. But what if the introduced TCR mispairs with the endogenous CD8+ T cell's TCR?

They also needed to introduce a reporter of T cell antigen recognition. The reporter is an infrared fluorescent protein (IFP) that only becomes fluorescent upon exposure to cytotoxic granzyme B (GzB), representing a functional & sortable readout of cognate CD4+ T cell recognition

But to do so, the authors needed to engineer both the antigen presenting cells (APCs), alongside the primary T cells used. For the APCs, they modified HEK-293 cells (eHEK-293-II) to lack MHC class I, but present MHC class II allele(s) of interest alongside appropriate machinery

TScan-II allows for genome-scale interrogation of MHC class II antigens using endogenous processing & presentation machinery of antigen libraries, alongside physiological T cell activation & degranulation (GzB) 👇.

Wrote a Spotlight piece together w/ lab mate @amandaochan & @MEBirnbaum covering an impressive new methodology from @elledge_stephen's group that enables MHC class II antigen discovery for CD4+ T cells called TScan-II.

new @runthelistpod episode is now out hosted on @MHMedical's AccessMedicine webpage!. best part about it?. you can search for key words & phrases to more easily listen to high-yield content 🤯.

RT @TheVinodLab: 1. ENTER #mRNAvaccines for #pancreaticcancer! Long-term #PDAC survivor story PART3. "Do #PDACs REALLY have vaccine neoant….

So excited to be at #SITC22 in Boston hearing from some of the giants in the field!

and there we have it - a "protein PROTAC" that does more than just get degraded in target cells:. it protects mice & ferrets from subsequent influenza A challenge demonstrating its ability to serve as a live-attenuated viral vaccine!. 💉💉💉

influenza A virus expressing M1-ALAPYIP protein + a protease (TEV) tag can be made in cells expressing TEVp, but degraded in target cells that *lack* TEVp, but express VHL. thus, they generated fully-assembled influenza A virus whose Achilles heel is its small degron handle

any alum from the @CraigMCrews lab would recognize this protein domain by ❤️:. ALAPYIP is derived from HIF-1a & the hydroxy-proline is a potent degron, even when conjugated to novel proteins (e.g. M1). VHL = E3 ubiquitin ligase that will degrade proteins containing this domain ->

the authors of this paper came up with a clever system to generate recombinant influenza A from mammalian cells, but have this live-attenuated virus be unable to complete more than 1 round of infection. the trick?. appending a VHL-targeting degron domain.

had to come out of Twitter hibernation to highlight this out-of-the box use case for #PROTACs:. live-attenuated virus vaccines. Flu vaccines have a whole host of difficulties from vaccine manufacturing to antigenic drift & this approach simplifies it all.

RT @BrownJHM: Thrombophilia & Anticoagulation (Handout from Run The List podcast .@runthelistpod) #BJHM #MedEd #FOAMed #Thrombophilia #Med….