Deciphering regulation of FOXP3 expression in human conventional T cells.

FOXP3 is a lineage-defining transcription factor that controls differentiation and maintenance of suppressive function of regulatory T cells (Tregs). Foxp3 is exclusively expressed in Tregs in mice. However, in humans, FOXP3 is not only constitutively expressed in Tregs; it is also transiently expressed in stimulated CD4+CD25- conventional T cells (Tconvs).

Modular pooled discovery of synthetic knockin sequences to program durable cell therapies.

Chronic stimulation can cause T cell dysfunction and limit the efficacy of cellular immunotherapies. Improved methods are required to compare large numbers of synthetic knockin (KI) sequences to reprogram cell functions. Here, we developed modular pooled KI screening (ModPoKI), an adaptable platform for modular construction of DNA KI libraries using barcoded multicistronic adaptors.

HLA-C*07:985:01:02Q, a novel allele with an acceptor splice site mutation.

HLA-C*07:985:01:02Q differs from HLA-C*07:985:01 by one nucleotide substitution at the Intron 1 splicing acceptor site.

Genetic load in incomplete lupus erythematosus.

Objective: Patients with incomplete lupus erythematosus (ILE) have lupus features but insufficient criteria for SLE classification. Some patients with ILE transition to SLE, but most avoid major organ involvement. This study tested whether the milder disease course in ILE is influenced by reduced SLE risk allele genetic load.

HLA-DQA1*03:03:01:16Q, a novel allele with an acceptor splice site mutation.

DQA1*03:03:01:16Q differs from DQA1*03:03:01:01 by one nucleotide at the Intron 3 splicing acceptor site.

The effect of low-dose IL-2 and Treg adoptive cell therapy in patients with type 1 diabetes.

BACKGROUNDA previous phase I study showed that the infusion of autologous Tregs expanded ex vivo into patients with recent-onset type 1 diabetes (T1D) had an excellent safety profile. However, the majority of the infused Tregs were undetectable in the peripheral blood 3 months postinfusion (Treg-T1D trial). Therefore, we conducted a phase I study (TILT trial) combining polyclonal Tregs and low-dose IL-2, shown to enhance Treg survival and expansion, and assessed the impact over time on Treg populations and other immune cells.

XYZeq: Spatially resolved single-cell RNA sequencing reveals expression heterogeneity in the tumor microenvironment.

Single-cell RNA sequencing (scRNA-seq) of tissues has revealed remarkable heterogeneity of cell types and states but does not provide information on the spatial organization of cells. To better understand how individual cells function within an anatomical space, we developed XYZeq, a workflow that encodes spatial metadata into scRNA-seq libraries. We used XYZeq to profile mouse tumor models to capture spatially barcoded transcriptomes from tens of thousands of cells.

Regulatory T cell control of systemic immunity and immunotherapy response in liver metastasis.

Patients with cancer with liver metastasis demonstrate significantly worse outcomes than those without liver metastasis when treated with anti-PD-1 immunotherapy. The mechanism of liver metastases-induced reduction in systemic antitumor immunity is unclear. Using a dual-tumor immunocompetent mouse model, we found that the immune response to tumor antigen presence within the liver led to the systemic suppression of antitumor immunity.

Test performance evaluation of SARS-CoV-2 serological assays.

Background: Serological tests are crucial tools for assessments of SARS-CoV-2 exposure, infection and potential immunity. Their appropriate use and interpretation require accurate assay performance data.

Method: We conducted an evaluation of 10 lateral flow assays (LFAs) and two ELISAs to detect anti-SARS-CoV-2 antibodies.

Unexpected Short-Tandem-Repeat Patterns in Posttransplant Chimerism Testing: Investigation of 3 Cases with Help from Forensic Science.

Chimerism testing by short tandem repeats (STRs) is used to monitor engraftment after allogeneic hematopoietic stem cell transplantation (HSCT). Generally, STR alleles are stable and transferred from parent to child or from donor to recipient. However, 3 cases did not follow this norm.

Pooled Knockin Targeting for Genome Engineering of Cellular Immunotherapies.

Adoptive transfer of genetically modified immune cells holds great promise for cancer immunotherapy. CRISPR knockin targeting can improve cell therapies, but more high-throughput methods are needed to test which knockin gene constructs most potently enhance primary cell functions in vivo. We developed a widely adaptable technology to barcode and track targeted integrations of large non-viral DNA templates and applied it to perform pooled knockin screens in primary human T cells.

Chromatin accessibility promotes hematopoietic and leukemia stem cell activity.

Chromatin organization is a highly orchestrated process that influences gene expression, in part by modulating access of regulatory factors to DNA and nucleosomes. Here, we report that the chromatin accessibility regulator HMGN1, a target of recurrent DNA copy gains in leukemia, controls myeloid differentiation. HMGN1 amplification is associated with increased accessibility, expression, and histone H3K27 acetylation of loci important for hematopoietic stem cells (HSCs) and leukemia, such as HoxA cluster genes.

Restoration of MYC-repressed targets mediates the negative effects of GM-CSF on RUNX1-ETO leukemogenicity.

Granulocyte macrophage-colony-stimulating factor (GM-CSF) signaling regulates hematopoiesis and immune responses. CSF2RA, the gene encoding the α-subunit for GM-CSF, is significantly downregulated in t(8;21) (RUNX1-ETO or RE) leukemia patients, suggesting that it may serve as a tumor suppressor. We previously reported that GM-CSF signaling is inhibitory to RE leukemogenesis.

Hip function and gait in patients treated for bladder exstrophy.

Patients with bladder exstrophy demonstrate pubic diastasis with external rotation of the ilia and posterolateral orientation of the acetabula. Iliac osteotomy is performed to approximate the pubis and thus facilitate the genitourinary repair. The role of iliac osteotomy in the correction of gait abnormalities is controversial.

Fracture of polyethylene tibial component in a total knee replacement. A case report.

Aseptic loosening is the most common mode of failure for total knee replacements. Component breakage is rare, usually attributable to errors of design or manufacture. A case is presented in which rotational malplacement of the femoral component resulted in accelerated wear and eventual fracture of the polyethylene tibial component.

Fracture dislocation of the femoral head treated by open reduction and internal fixation.

Two patients who sustained posterior and anterior fracture dislocations of the femoral head, respectively, are reported. Each case was treated by open reduction and internal fixation of the fracture fragments. With this operative approach excellent results have been achieved on medium-term followup.

Dislocated patella associated with below-knee amputation in adolescent patients.

One-third of the adolescent patients followed up in our prosthetic clinic have developed patellar dislocation. Four cases are described, and radiographs are evaluated for femoral-tibial alignment, patellar height, and tibial length. Patella alta was present in each patient.

A simple method of hip arthrodesis.

A technique of intraarticular hip arthrodesis using a cancellous screw to provide compression across the joint is introduced. We present four patients in whom this method was successful, resulting in solid fusion and pain relief at follow-up.

Recurrent posterior dislocation of the shoulder: treatment using a bone block.

Acute posterior dislocations of the shoulder are uncommon. Recommended surgical treatments for recurrent posterior dislocation include soft-tissue advancement, posterior glenoid osteotomy, rotational osteotomy of the humerus, and posterior bone block. The posterior bone-block procedure successfully prevented recurrent dislocation in our series of five patients.

Confirmation of the posterolateral technique to decompress and fuse thoracolumbar spine burst fractures.

A posterolateral technique to decompress retropulsed bone from the spinal canal has been employed in nine patients, confirmed with intraoperative canal inspection and pre- and postoperative computed axial tomographic (CAT) evaluation. This technique has allowed reduction of retropulsed bony elements from the spinal canal, assessment of canal decompression and reestablishment of vertebral body alignment. The approach permits posterior fusion at the same operation.