Stereotypic T cell receptor clonotypes in the thymus and peripheral blood of Myasthenia gravis patients.

Myasthenia Gravis (MG) patients with anti-acetylcholine receptor (AChR) antibodies frequently show hyperplastic thymi with ectopic germinal centers, where autoreactive B cells proliferate with the aid of T cells. In this study, thymus and peripheral blood (PB) samples were collected from ten AChR antibody-positive MG patients. T cell receptor (TCR) repertoires were analyzed using next-generation sequencing (NGS), and compared with that of an age and sex matched control group generated from a public database. Certain V genes and VJ gene recombination pairs were significantly upregulated in the TCR chains of αß-T cells in the PB of MG patients compared to the control group. Furthermore, the TCR chains found in the thymi of MG patients had a weighted distribution to longer CDR3 lengths when compared to the PB of MG patients, and the TCR beta chains (TRB) in the MG group's PB showed increased clonality encoded by one upregulated V gene. When TRB sequences were sub-divided into groups based on their CDR3 lengths, certain groups showed decreased clonality in the MG group's PB compared to the control group's PB. Finally, we demonstrated that stereotypic MG patient-specific TCR clonotypes co-exist in both the PB and thymi at a much higher frequency than that of the clonotypes confined to the PB. These results strongly suggest the existence of a biased T cell-mediated immune response in MG patients, as observed in other autoimmune diseases.

Selective Control of Novel TiO2 Nanorods: Excellent Building Blocks for the Electron Transport Layer of Mesoscopic Perovskite Solar Cells.

Novel TiO2 nanorods (NRs) with various lengths of 70-200 nm and uniform widths of 46-48 nm are selectively synthesized by a solvothermal reaction under a basic environment. The length of TiO2 NRs is reproducibly tuned by varying the concentration of tetramethylammonium hydroxide (TMAH), while the NRs in the pure anatase phase are grown in the [001] direction, caused by the preferential binding affinity of TMAH to the TiO2 (101) facet. TiO2 NRs of various lengths are then applied to form the electron transporting layer (ETL) of mesoscopic perovskite solar cells (PSCs). We found that PSC devices with NRs exhibit superior photovoltaic (PV) performance to those with conventional 46 nm-sized TiO2 nanoparticles (NP46). Particularly, the PSC with TiO2 NRs of 110 nm length (NR110) exhibits the optimum PV conversion efficiency (PCE): the average PCE is 22.64% with a VOC of 1.137 V, a JSC of 24.60 mA·cm-2, and a FF of 80.96%, while the champion PCE is 23.18%. In addition, the PSC with NR110 (PSC-NR110) reveals significantly improved long-term stability in air with a relative humidity of 40-50%. In 1000 h, its PCE is reduced by only 9% whereas that of PSC with NP46 decreases by 25%. The PSC properties analyzed by impedance spectroscopy and J-V curve measurements under dark conditions and at various light intensities provide evidence that PSC-NR110 has fewer defects and shows significantly reduced charge recombination. We discuss the advantages of NR structures in preparing the ETL of PSC devices and also explain why the charge recombination is suppressed.

Enhanced Phase Stability of Compressive Strain-Induced Perovskite Crystals.

Control of strain in perovskite crystals has been considered as an effective strategy to ensure the phase stability of perovskite films where a compressive strain is particularly preferred over a tensile strain due to a lowered Gibbs free energy by the unit cell contraction effect. Here we adapt the strategy of strain control into perovskite solar cells in which the compressive strain is applied by utilizing a thermal expansion difference between the perovskite film and an adjacent layer. Poly(4-butylphenyldiphenylamine), with a higher thermal expansion coefficient compared to that of perovskite, is employed as a substrate for perovskite crystal growth at 100 °C, followed by cooling to room temperature. The applied compressive strain at the interface, as a result of a greater contraction of the polymer compared to the perovskite film, is confirmed by grazing incidence X-ray diffraction showing a red peak shift with increasing secondary angle. The compressive strain-induced perovskite film shows relatively constant absorbance spectra as a function of time. In the meantime, the absorbance spectra of a film without strain control exhibit a gradual decay with developing an Urbach tail. Importantly, the effect of strain engineering is remarkably prominent in the long-term photovoltaic performance. The photocurrent drops by 41% over 911 h without controlling strain, which is significantly improved by employing compressive strain, showing only a 6% drop in photocurrent from a shelf-stability test without encapsulation. It is also noted that an S-shaped kink appears in the current-voltage curves since 579-h-long storage for the device without strain control, leading to unreliable and overestimated fill factor and conversion efficiency. On the other hand, a 16% increase in fill factor with a stable performance is derived over 911 h from the compressive strain-induced device.

Induction of Anti-Aquaporin 5 Autoantibody Production by Immunization with a Peptide Derived from the Aquaporin of Prevotella melaninogenica Leads to Reduced Salivary Flow in Mice.

Sjögren's syndrome (SS) is an autoimmune disease characterized by dryness of the mouth and eyes. The glandular dysfunction in SS involves not only T cell-mediated destruction of the glands but also autoantibodies against the type 3 muscarinic acetylcholine receptor or aquaporin 5 (AQP5) that interfere with the secretion process. Studies on the breakage of tolerance and induction of autoantibodies to these autoantigens could benefit SS patients. To break tolerance, we utilized a PmE-L peptide derived from the AQP5-homologous aquaporin of Prevotella melaninogenica (PmAqp) that contained both a B cell "E" epitope and a T cell epitope. Repeated subcutaneous immunization of C57BL/6 mice with the PmE-L peptide efficiently induced the production of Abs against the "E" epitope of mouse/human AQP5 (AQP5E), and we aimed to characterize the antigen specificity, the sequences of AQP5E-specific B cell receptors, and salivary gland phenotypes of these mice. Sera containing anti-AQP5E IgG not only stained mouse Aqp5 expressed in the submandibular glands but also detected PmApq and PmE-L by immunoblotting, suggesting molecular mimicry. Characterization of the AQP5E-specific autoantibodies selected from the screening of phage display Ab libraries and mapping of the B cell receptor repertoires revealed that the AQP5E-specific B cells acquired the ability to bind to the Ag through cumulative somatic hypermutation. Importantly, animals with anti-AQP5E Abs had decreased salivary flow rates without immune cell infiltration into the salivary glands. This model will be useful for investigating the role of anti-AQP5 autoantibodies in glandular dysfunction in SS and testing new therapeutics targeting autoantibody production.

Amplification of a minimally biased antibody repertoire for in vitro display using a universal primer-based amplification method.

Immune hosts are valuable sources for antibody discovery. To construct in vitro display antibody libraries from immune repertoires, singleplex or multiplex PCR amplification were employed using primers targeting multiple immunoglobulin genes. However, during this process, the B cell receptor repertoire is distorted due to interactions between multiple target genes and primers. To minimize this alternation, we devised a new method for harvesting immunoglobulin genes and tested its performance in rabbit variable heavy chain (VH) and variable kappa light chain (VK) genes. Double-stranded cDNA was synthesized using primers containing V/J gene-specific regions and universal sequence parts for in vitro display. VH and VK gene libraries were obtained through subsequent PCR amplification using primers with universal sequences. Next-generation sequencing analysis confirmed that universal PCR libraries had more diverse VH and VK clonotypes, and a less biased clonal distribution, than conventional singleplex or multiplex gene-specific PCR libraries.