Search bioRxiv⌕ Search

Biology subjects

Popescu, I.

Publications and source records attributed to Popescu, I..

2 recordsLinked to original sources

A two-compartment system for subfreezing temperatures preservation of large volumes of organic matter in an isochoric system

This is a technology paper on the design of and test results from an 11-liter isochoric (constant volume) chamber, for the preservation of large, organs in a supercooled state. Details of the design are given, as well as a proof that the chamber is isochoric. Five repeats show that in this large chamber, ice nucleation of steam distilled water occurs at - 2 {degrees}C within less than 12 hours, in all the repeats. An examination of the experimental results suggests that the ice nucleation starts on the inner walls of the isochoric chamber. A new two compartment isochoric chamber was designed to reduces the probability of ice nucleation on the walls of the chamber. In the two-compartment system, the biological matter and the preservation fluid are introduced in a sealed low-density polyethylene bag, and placed in the center of the isochoric chamber, in such a way that the bag does not touch the walls. The space between the inner walls of the isochoric chamber and the outer walls of the bag are filled with a fluid with a composition that does not freeze at the storage temperature. Three repeat experiments with steam distilled water and with in vitro pig liver show that with this technique, the system remained supercooled, without any ice nucleation for the duration of the experiments. Experiments were voluntary terminated at 48 hours of supercooling. This new technology may hold promise for long term preservation of large biological organs in a supercooled state, without the use of any chemical additives.

bioengineering↗

CD4+ T cell lymphopenia and dysfunction in severe COVID-19 disease is autocrine TNF-α/TNFRI-dependent

Lymphopenia is common in severe COVID-19 disease, yet the mechanisms are poorly understood. In 148 patients with severe COVID-19, we found lymphopenia was associated with worse survival. CD4+ lymphopenia predominated, with lower CD4+/CD8+ ratios in severe COVID-19 compared to recovered, mild disease (p<0.0001). In severe disease, immunodominant CD4+ T cell responses to Spike-1(S1) produced increased in vitro TNF-, but impaired proliferation and increased susceptibility to activation-induced cell death (AICD). CD4+TNF-+ T cell responses inversely correlated with absolute CD4+ counts from severe COVID-19 patients (n=76; R=-0.744, P<0.0001). TNF- blockade including infliximab or anti-TNFRI antibodies strikingly rescued S1-specific CD4+ proliferation and abrogated S1-AICD in severe COVID-19 patients (P<0.001). Single-cell RNAseq demonstrated downregulation of Type-1 cytokines and NF{kappa}B signaling in S1-stimulated CD4+ cells with infliximab treatment. Lung CD4+ T cells in severe COVID-19 were reduced and produced higher TNF- versus PBMC. Together, our findings show COVID-19-associated CD4+ lymphopenia and dysfunction is autocrine TNF-/TNFRI-dependent and therapies targeting TNF- may be beneficial in severe COVID-19. One Sentence SummaryAutocrine TNF-/TNFRI regulates CD4+ T cell lymphopenia and dysfunction in severe COVID-19 disease.

immunology↗