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Gifford, D.

Publications and source records attributed to Gifford, D..

2 recordsLinked to original sources

Extreme temperature exposure has negative demographic consequences for Sulfolobus acidocaldarius

Microorganisms inhabiting geothermal springs and volcanic systems experience fluctuating temperatures that can periodically exceed their upper thermal limits, but the demographic consequences of such exposure remain poorly understood. Here, we investigated demographic responses of the thermophilic archaeon Sulfolobus acidocaldarius to an extreme temperature (94.1{degrees}C) under two regimes: sustained exposure varying in duration, and episodic exposure interspersed with recovery at a permissive temperature (75{degrees}C). Under sustained exposure, populations showed no detectable loss of viability after 15 min but declined thereafter, decreasing by approximately five orders of magnitude after 120 min. Under episodic exposure, populations remained viable across nine exposure-recovery cycles but declined in density with successive cycles. Similar responses were observed for three strains, including a DNA mismatch repair knockout ({Delta}nucS), indicating that mismatch repair deficiency did not affect viability or recovery. Together, these results demonstrate that S. acidocaldarius can withstand brief and repeated exposure to near-boiling temperatures, with mortality determined primarily by cumulative exposure duration rather than a fixed thermal threshold.

microbiology↗

Deep mapping of the TCR-antigen interface using pMHC-pseudotyped viruses and yeast display

T cell receptor (TCR) specificity is central to the efficacy of T cell therapies, yet scalable methods to map how TCR sequences shape antigen recognition remain limited. To address this, we introduce VelociRAPTR, a library-on-library approach that combines yeast-displayed TCR libraries with pMHC-displaying virus-like particles (pMHC-VLPs) to rapidly screen millions of TCR-antigen interactions. We show that pMHC-VLPs efficiently bind TCRs on yeast and generate equivalent data to recombinantly produced pMHC protein. We then apply VelociRAPTR to screen 47 million variants of the A6 and 868 TCRs against 92 pMHCs simultaneously, mutating both the CDR3 loops and cognate peptides. The resulting CDR3-pMHC maps reveal biased recognition patterns, where mutations to CDR3 loops can selectively constrain or broaden specificity to peptide analogs. These insights provide a foundation for engineering TCRs with defined pMHC binding profiles and improving models that predict TCR-antigen interactions, including the prediction of off-target recognition. By coupling the scale of yeast display with the modularity of VLPs, VelociRAPTR offers a generalizable strategy for generating deep, high-throughput protein- protein interaction data.

immunology↗