TGF-ß specifies TFH versus TH17 cell fates in murine CD4+ T cells through c-Maf.

T follicular helper (TFH) cells are essential for effective antibody responses, but deciphering the intrinsic wiring of mouse TFH cells has long been hampered by the lack of a reliable protocol for their generation in vitro. We report that transforming growth factor-ß (TGF-ß) induces robust expression of TFH hallmark molecules CXCR5 and Bcl6 in activated mouse CD4+ T cells in vitro. TGF-ß-induced mouse CXCR5+ TFH cells are phenotypically, transcriptionally, and functionally similar to in vivo-generated TFH cells and provide critical help to B cells. The study further reveals that TGF-ß-induced CXCR5 expression is independent of Bcl6 but requires the transcription factor c-Maf. Classical TGF-ß-containing T helper 17 (TH17)-inducing conditions also yield separate CXCR5+ and IL-17A-producing cells, highlighting shared and distinct cell fate trajectories of TFH and TH17 cells. We demonstrate that excess IL-2 in high-density T cell cultures interferes with the TGF-ß-induced TFH cell program, that TFH and TH17 cells share a common developmental stage, and that c-Maf acts as a switch factor for TFH versus TH17 cell fates in TGF-ß-rich environments in vitro and in vivo.

Leveraging microRNAs for cellular therapy.

Owing to Karl Landsteiner's discovery of blood groups, blood transfusions became safe cellular therapies in the early 1900s. Since then, cellular therapy made great advances from transfusions with unmodified cells to today's commercially available chimeric antigen receptor (CAR) T cells requiring complex manufacturing. Modern cellular therapy products can be improved using basic knowledge of cell biology and molecular genetics. Emerging genome engineering tools are becoming ever more versatile and precise and thus catalyze rapid progress towards programmable therapeutic cells that compute input and respond with defined output. Despite a large body of literature describing important functions of non-coding RNAs including microRNAs (miRNAs), the vast majority of cell engineering efforts focuses on proteins. However, miRNAs form an important layer of posttranscriptional regulation of gene expression. Here, we highlight examples of how miRNAs can successfully be incorporated into engineered cellular therapies.

Forced expression of the non-coding RNA miR-17∼92 restores activation and function in CD28-deficient CD4+ T cells.

CD28 provides the prototypical costimulatory signal required for productive T-cell activation. Known molecular consequences of CD28 costimulation are mostly based on studies of protein signaling molecules. The microRNA cluster miR-17∼92 is induced by T cell receptor stimulation and further enhanced by combined CD28 costimulation. We demonstrate that transgenic miR-17∼92 cell-intrinsically largely overcomes defects caused by CD28 deficiency. Combining genetics, transcriptomics, bioinformatics, and biochemical miRNA:mRNA interaction maps we empirically validate miR-17∼92 target genes that include several negative regulators of T cell activation. CD28-deficient T cells exhibit derepressed miR-17∼92 target genes during activation. CRISPR/Cas9-mediated ablation of the miR-17∼92 targets Pten and Nrbp1 in naive CD28-/- CD4+ T cells differentially increases proliferation and expression of the activation markers CD25 and CD44, respectively. Thus, we propose that miR-17∼92 constitutes a central mediator for T cell activation, integrating signals by the TCR and CD28 costimulation by dampening multiple brakes that prevent T cell activation.

Cross-cultural adaptation and validation of the Ukrainian version of the ABILHAND-Kids questionnaire.

PURPOSE: To develop and cross-culturally validate the Ukrainian version of the ABILHAND-Kids questionnaire by testing its psychometric properties in a sample of Ukrainian children with cerebral palsy. METHODS: The ABILHAND-Kids questionnaire was translated into Ukrainian, cross-culturally adapted, and administered to 113 parents of children with cerebral palsy. The psychometric properties of the Ukrainian version and its cross-cultural validation were investigated through the Rasch rating scale model. RESULTS: One major misfit has been found for the item "Rolling up a sleeve of a sweater" that further was removed. The item "Putting on a backpack/schoolbag" was split into gender-specific items, separately for girls and for boys, as it was systematically easier for Ukrainian girls. All remaining items contributed to the definition of a unidimensional measure of manual ability. The internal consistency reliability of the scale was high (R = 0.95). No significant floor (4%) and ceiling effects (5%) were observed. Three major differential item functioning items were found across Belgium and Ukraine, highlighting the need to use the Ukrainian calibration of ABILHAND-Kids in Ukraine. CONCLUSION: The Ukrainian ABILHAND-Kids questionnaire has good psychometric properties for assessing manual ability in Ukrainian children with cerebral palsy, holding potential to be implemented in clinical practice nationwide.Implications for rehabilitationCerebral palsy impairs manual ability leading to decreased quality of life and participation.Professionals need valid and reliable tools to detect small changes of manual ability during rehabilitation.Metric properties and availability of the Ukrainian version of the ABILHAND-Kids questionnaire make it a useful tool in the assessment of children with cerebral palsy.

Influence of Spinal Manipulation on Muscle Spasticity and Manual Dexterity in Participants With Cerebral Palsy: Randomized Controlled Trial.

OBJECTIVES: The aim of this study was to investigate the short-term effects of spinal manipulation (SM) on wrist muscle spasticity and manual dexterity in participants with cerebral palsy (CP). METHODS: After baseline examination, 78 participants with spastic CP (7-18 years) without contractures or hyperkinetic syndrome were randomly allocated into 2 groups. The experimental group underwent SM to the cervical, thoracic, and lumbar spine, and the control group received sham SM. A second evaluation was performed 5 minutes postintervention. Wrist muscle spasticity was measured quantitatively with NeuroFlexor (Aggero MedTech AB, Solna, Sweden), a device assessing resistance to passive movements of different velocities. Between-group difference was calculated using the Mann-Whitney U test. Manual dexterity was evaluated by the Box and Block test. RESULTS: In the experimental group, muscle spasticity was reduced by 2.18 newton from median 5.53 with interquartile range 8.66 to median 3.35 newton with interquartile range 7.19; the difference was statistically significant (P = .002). In the control group, reduction in spasticity was negligible. The between-group difference in change of muscle spasticity was statistically significant (P = .034). Improvement of manual dexterity was not statistically significant (P = .28). CONCLUSIONS: These findings suggest that SM may, in the short term, help to reduce spasticity in participants with CP. Long-term effects of SM on muscle spasticity have yet to be studied.