PD1+TIGIT+2B4+KLRG1+ Cells Might Underlie T Cell Dysfunction in Patients Treated with BCMA-Directed Chimeric Antigen Receptor T Cell Therapy.

Chimeric antigen receptor T cell (CAR-T) therapy has shown rapid, frequent, and deep responses in patients with relapsed/refractory multiple myeloma (RRMM). However, relapse frequently occurs following CAR-T therapy, and the cause of this resistance is not well defined. Among the potential mechanisms of resistance, T cell intrinsic factors may be an important source of failure. Here we used spectral flow cytometry to identify the changes in T cell phenotypes in bone marrow aspirates at different stages of multiple myeloma progression, including cases that relapsed after anti-BCMA CAR-T therapy. We identified completely different T cell phenotypes in RRMM and post CAR-T relapse cases compared to healthy donors and earlier stages of multiple myeloma, novel double-negative CD3+ T cells in RRMM and CAR-T relapsed cases, and differences in CD8 T cell phenotype at the baseline between peripheral blood and bone marrow from healthy donors. We found that the majority of T cells in RRMM patients and significant T cell subsets in post-CAR-T relapsed patients expressed multiple coinhibitory markers, including PD1, TIGIT, 2B4, and KLRG1.

Acute evaluation of the acute vestibular syndrome: differentiating posterior circulation stroke from acute peripheral vestibulopathies.

This review article aims to provide an evidence-based approach to evaluating the patient who presents with acute prolonged, spontaneous vertigo in the context of the acute vestibular syndrome (AVS). Differentiation of posterior circulation stroke (PCS) presenting as an AVS has been regarded as an important diagnostic challenge for physicians involved in acute care. Current evidence suggests that a targeted approach to history taking and physical examination with emphasis on the oculomotor examination, more specifically the HINTS (Head Impulse/Nystagmus/Test-of-skew) examination battery, yields a higher sensitivity for the diagnosis of PCS than even standard magnetic resonance imaging with diffusion-weighted imaging. However, most studies have only validated the utility of the HINTS examination when performed by experts, who interpret the most powerful component of HINTS, namely the head impulse test (HIT), considerably different to the novice. Several investigations useful in the differentiation of the AVS are becoming more accessible and portable, such as videooculography with Frenzel goggles and video head impulse testing (vHIT), which allows for the quantitative assessment of the HIT. In clinical practice, vHIT has already become accepted as standard of care in the evaluation of AVS.

Dosimetric experience with 2 commercially available multilumen balloon-based brachytherapy to deliver accelerated partial-breast irradiation.

The purpose of this work was to report dosimetric experience with 2 kinds of multilumen balloon (MLB), 5-lumen Contura MLB (C-MLB) and 4-lumen MammoSite MLB (MS-MLB), to deliver accelerated partial-breast irradiation, and compare the ability to achieve target coverage and control skin and rib doses between 2 groups of patients treated with C-MLB and MS-MLB brachytherapy. C-MLB has 5 lumens, the 4 equal-spaced peripheral lumens are 5 mm away from the central lumen. MS-MLB has 4 lumens, the 3 equal-spaced peripheral lumens are 3 mm away from the central lumen. In total, 43 patients were treated, 23 with C-MLB, and 20 with MS-MLB. For C-MLB group, 8 patients were treated with a skin spacing < 7 mm and 12 patients with rib spacing < 7 mm. For MS-MLB group, 2 patients were treated with a skin spacing < 7 mm and 5 patients with rib spacing < 7 mm. The dosimetric goals were (1) ≥ 95% of the prescription dose (PD) covering ≥ 95% of the target volume (V(95%) ≥ 95%), (2) maximum skin dose ≤ 125% of the PD, (3) maximum rib dose ≤ 145% of the PD (if possible), and (4) the V(150%) ≤ 50 cm(3) and V(200%) ≤ 10 cm(3). All dosimetric criteria were met concurrently in 82.6% of C-MLB patients, in 80.0% of MS-MLB patients, and in 81.4% of all 43 patients. For each dosimetric parameter, t-test of these 2 groups showed p > 0.05. Although the geometric design of C-MLB is different from that of MS-MLB, both applicators have the ability to shape the dose distribution and to provide good target coverage, while limiting the dose to skin and rib. No significant difference was observed between the 2 patient groups in terms of target dose coverage and dose to organs at risk.

RpL22e, but not RpL22e-like-PA, is SUMOylated and localizes to the nucleoplasm of Drosophila meiotic spermatocytes.

Duplicated ribosomal protein (Rp) gene families often encode highly similar or identical proteins with redundant or unique roles. Eukaryotic-specific paralogues RpL22e and RpL22e-like-PA are structurally divergent within the N terminus and differentially expressed, suggesting tissue-specific functions. We previously identified RpL22e-like-PA as a testis Rp. Strikingly, RpL22e is detected in immunoblots at its expected molecular mass (m) of 33 kD and at increasing m of ~43-55 kD, suggesting RpL22e post-translational modification (PTM). Numerous PTMs, including N-terminal SUMOylation, are predicted computationally. Based on S2 cell co-immunoprecipitations, bacterial-based SUMOylation assays and in vivo germline-specific RNAi depletion of SUMO, we conclude that RpL22e is a SUMO substrate. Testis-specific PTMs are evident, including a phosphorylated version of SUMOylated RpL22e identified by in vitro phosphatase experiments. In ribosomal profiles from S2 cells, only unconjugated RpL22e co-sediments with active ribosomes, supporting an extra-translational role for SUMOylated RpL22e. Ectopic expression of an RpL22e N-terminal deletion (lacking SUMO motifs) shows that truncated RpL22e co-sediments with polysomes, implying that RpL22e SUMOylation is dispensable for ribosome biogenesis and function. In mitotic germ cells, both paralogues localize within the cytoplasm and nucleolus. However, within meiotic cells, phase contrast microscopy and co-immunohistochemical analysis with nucleolar markers nucleostemin1 and fibrillarin reveals diffuse nucleoplasmic, but not nucleolar RpL22e localization that transitions to a punctate pattern as meiotic cells mature, suggesting an RpL22e role outside of translation. Germline-specific knockdown of SUMO shows that RpL22e nucleoplasmic distribution is sensitive to SUMO levels, as immunostaining becomes more dispersed. Overall, these data suggest distinct male germline roles for RpL22e and RpL22e-like-PA.

Expression of ribosomal protein L22e family members in Drosophila melanogaster: rpL22-like is differentially expressed and alternatively spliced.

Several ribosomal protein families contain paralogues whose roles may be equivalent or specialized to include extra-ribosomal functions. RpL22e family members rpL22 and rpL22-like are differentially expressed in Drosophila melanogaster: rpL22-like mRNA is gonad specific whereas rpL22 is expressed ubiquitously, suggesting distinctive paralogue functions. To determine if RpL22-like has a divergent role in gonads, rpL22-like expression was analysed by qRT-PCR and western blots, respectively, showing enrichment of rpL22-like mRNA and a 34 kDa (predicted) protein in testis, but not in ovary. Immunohistochemistry of the reproductive tract corroborated testis-specific expression. RpL22-like detection in 80S/polysome fractions from males establishes a role for this tissue-specific paralogue as a ribosomal component. Unpredictably, expression profiles revealed a low abundant, alternative mRNA variant (designated 'rpL22-like short') that would encode a novel protein lacking the C-terminal ribosomal protein signature but retaining part of the N-terminal domain. This variant results from splicing of a retained intron (defined by non-canonical splice sites) within rpL22-like mRNA. Polysome association and detection of a low abundant 13.5 kDa (predicted) protein in testis extracts suggests variant mRNA translation. Collectively, our data show that alternative splicing of rpL22-like generates structurally distinct protein products: ribosomal component RpL22-like and a novel protein with a role distinct from RpL22-like.