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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 1,423 records · Page 79
All sourcesNASA NTRSDOE OSTI
DOE OSTI2019

Development of a Squeeze Semisolid High-Pressure Die Casting Process for Magnesium Structural Parts

High-pressure die casting is the most common method used to produce magnesium castings, due to the excellent balance of cost and properties for high production volumes with limitations in terms of final mechanical properties. A newly developed process based on employing low injection speeds, in a range slightly over the standard semisolid speeds with a modified die, with thick gates and high die temperatures has been developed. Despite working with speeds that are not in the lamellar flow, the obtained parts present very low porosity, allowing the use thermal treatments to increase the ductility without producing blisters. The demonstration has been performed in a AM60B magnesium body joint produced by the squeeze casting process. Finally, the microstructure and the mechanical properties of as-cast and T4 heat-treated samples were studied. The results indicate the improvement of the mechanical properties in T4 heat-treated parts; specifically, a 40% improved deformation-to-failure, 40-60% improved penetration force and 90% of energy absorption were possible to obtain employing the newly developed SC + T4 process.

36 MATERIALS SCIENCE↗
DOE OSTI2019

Development of Texture in Ultrafine-Grained Low-Carbon Steel Processed through Equal-Channel Angular Pressing

Developments of microstructures and texture are investigated for a low-carbon steel deformed by equal-channel angular pressing (ECAP) at room temperature for equivalent strain (ε{sub vm}) of 16.8. ECAP at ε{sub vm} = 0.6 reduces grain size by two orders of magnitude. With increasing strain, grain size reduces further and high-angle grain boundary (HAGB) fraction improves by grain subdivision and its change in orientation. ECAP of low-carbon steel up to ε{sub vm} = 16.8 can produce ultrafine grains of 0.2 µm size with HAGB fraction of 82%. The as-received steel has low texture index. Microstructural changes strongly influence texture development. It increases with equivalent strain as grain subdivision continues. Texture index fluctuates at intermediate strain range due to randomization of initial texture. Texture index reaches saturation at the high equivalent strain. On ECAP at low to intermediate strain ε{sub vm} = 0.6-6, components of both α (F{sub θ}, J{sub θ}, and J-bar {sub θ}) and γ (D{sub 1θ}, D{sub 2θ}, and E{sub θ}) fibers are developed though all of them deviate from their ideal positions. At higher strain (ε{sub vm} = 9-16.8), the γ fiber components (D{sub 1θ}, D{sub 2θ}, E{sub θ}) are strengthened, and the E{sub θ} component reaches the highest intensity of ≈ 17 at ε{sub vm} = 12. There is no texture symmetry at low to high strain level. The presence of pearlite in low-carbon steel enhances strongly the deviation of texture components from their ideal positions in Euler’s space as well as it increases the intensities of the components more than that of IF steel for a similar equivalent strain.

36 MATERIALS SCIENCE↗
DOE OSTI2024

Multiplexed Quantitative Proteomics in Prostate Cancer Biomarker Development

Prostate cancer (PCa) is the most common non-skin cancer among men in the United States. However, the widely used protein biomarker in PCa, prostate-specific antigen (PSA), while useful for initial detection, its use alone cannot detect aggressive PCa and can lead to overtreatment. This chapter provides an overview of PCa protein biomarker development. It reviews the state-of-the-art liquid chromatography-mass spectrometry-based proteomics technologies for PCa biomarker development, such as enhancing the detection sensitivity of low-abundance proteins through antibody-based or antibody-independent protein/peptide enrichment, enriching post-translational modifications such as glycosylation as well as information-rich extracellular vesicles, and increasing accuracy and throughput using advanced data acquisition methodologies. This chapter also summarizes recent PCa biomarker validation studies that applied those techniques in diverse specimen types, including cell lines, tissues, proximal fluids, urine, and blood, developing novel protein biomarkers for various clinical applications, including early detection and diagnosis, prognosis, and therapeutic intervention of PCa.

Prostate cancer, SRM, PRM, DIA, protein biomarker↗
DOE OSTI2018

Novel epoxy-silica nanoparticles to develop non-enzymatic colorimetric probe for analytical immuno/bioassays

We have developed a novel method to develop epoxy silica nanoparticles (EfSiNP) in a single pot. High surface coverage of epoxy functional groups between 150 and 57000 molecules per particles (~10 13 –10 16 molecules/mL of 200 nm EfSiNPs) was achieved for different preparation conditions. We then created a red colored probe by conjugating Fuchsin dye to the epoxy functionalities of EfSINPs. Anti-mouse IgG was co-immobilized with Fuchsin and their ratios were optimized for achieving optimum ratios by testing those in functional assays. Dye to antibody ratios were in good negative correlation with a coefficient of -1.00 measured at a confidence level of over 99%. We employed the developed non-enzymatic colorimetric immunonanoprobe for detecting mouse IgG in a direct immunoassay format. We achieved a sensitivity of 427 pg/mL with the assay.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2022

Development of grain-scale slip activity and lattice rotation fields in Inconel 718

Using a combination of in-situ high-resolution digital image correlation (HR-DIC), Heaviside-DIC method (H-DIC), and crystal plasticity finite element (CPFE), we investigate the evolution of intragranular lattice rotations and slip activity during monotonic and cyclic loading in a high performance, polycrystalline face centered cubic material. The CPFE employs a quasi-3D model microstructure, which is a highly resolved mirror representation of the experimental in-situ test sample. In agreement, the measurements and calculations reveal that most grains, regardless of their size and lattice orientation, develop intragranular lattice rotation gradients that span the grain. For a small cluster of grains on the deformed material, we perform HR-DIC analysis of slip lines to demonstrate agreement in the active slip systems and changes in this local slip activity across the individual grains. The combined analysis reveals that deforming grains are divided into sub-granular regions of uniform lattice rotation and these regions are most often associated with only one or two active slip systems. The gradient lines that divide them correspond to changes in the predominant slip system. The model is used to examine the evolution of intragranular lattice rotation in a single fully reversed tension-compression cycle. The calculations indicate that intragranular gradients intensify during the reverse loading path as nearest neighboring regions appear to shed lattice rotation, increasing the lattice rotation in some regions, while shutting down rotation in neighboring regions. So these findings provide insight into the irreversible changes that develop within deforming grains at the scale of the grain, particularly the heterogeneous development of intragranular lattice rotation in early stages of deformation, which could serve as precursors to localization.

36 MATERIALS SCIENCE↗
DOE OSTI2021

Towards developing multiscale-multiphysics models and their surrogates for digital twins of metal additive manufacturing

Artificial intelligence (AI) embedded within digital models of manufacturing processes can be used to improve process productivity and product quality significantly. The application of such advanced capabilities particularly to highly digitalized processes such as metal additive manufacturing (AM) is likely to make those processes commercially more attractive. AI capabilities will reside within Digital Twins (DTs) which are living virtual replicas of the physical processes. DTs will be empowered to operate autonomously in a diagnostic control capacity to supervise processes and can be interrogated by the practitioner to inform the optimal processing route for any given product. The utility of the information gained from the DTs would depend on the quality of the digital models and, more importantly, their faster-solving surrogates which dwell within DTs for consultation during rapid decision-making. In this article, we point out the exceptional value of DTs in AM and focus on the need to create high-fidelity multiscale-multiphysics models for AM processes to feed the AI capabilities. We identify technical hurdles for their development, including those arising from the multiscale and multiphysics characteristics of the models, the difficulties in linking models of the subprocesses across scales and physics, and the scarcity of experimental data. We discuss the need for creating surrogate models using machine learning approaches for real-time problem-solving. We further identify non-technical barriers, such as the need for standardization and difficulties in collaborating across different types of institutions. We offer potential solutions for all these challenges, after reflecting on and researching discussions held at an international symposium on the subject in 2019. Here, we argue that a collaborative approach can not only help accelerate their development compared with disparate efforts, but also enhance the quality of the models by allowing modular development and linkages that account for interactions between the various sub-processes in AM. A high-level roadmap is suggested for starting such a collaboration.

36 MATERIALS SCIENCE↗
DOE OSTI2019

Development of the multigroup cross section library for the CASL neutronics simulator MPACT: Verification

The MPACT neutronics module of the Consortium for Advanced Simulation of Light Water Reactors (CASL) core simulator is a 3D whole core transport code being developed for the CASL toolset, known as the Virtual Environment for Reactor Analysis (VERA). Key characteristics of the MPACT code include a subgroup method for resonance self-shielding and a whole-core transport solver with a 2D/1D synthesis method. Depletion capability is supported by coupling SCALE-ORIGEN with MPACT. A new ENDF/B-VII.1 MPACT 51-group library has been developed by using the AMPX/SCALE and CASL-XSTools code packages based on the procedure introduced in the companion paper, “Development of the Multigroup Cross Section Library for the CASL Neutronics Simulator MPACT: Method and Procedure.” The cross section library generation procedure, cross section library, and MPACT transport capability have been verified by performing various benchmark calculations and comparing the benchmark results to the continuous-energy (CE) Monte Carlo results. Furthermore, the code-to-code comparison for the benchmarks shows that the CASL neutronics simulator MPACT and its 51-group library are working reasonably.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗
DOE OSTI2019

Development of high efficiency window air conditioner using propane under limited charge

The residential air conditioner market has a significant portion of window air conditioners (WAC). Due to the compact size and small refrigerant charge, WACs are most tolerant of flammable refrigerants. Hydrocarbon (HC) refrigerants are natural substances and have significantly better environmental impacts and lower costs than conventional refrigerants like R-410A and R-22. The paper aimed to develop a WAC prototype using propane (R-290) and verify the performance metrics in the laboratory. The key of developing the WAC using a flammable refrigerant is to control the refrigerant charge under the limit of building safety regulation with the performance meeting the Energy Star standard. To minimize the charge, compact heat exchangers were utilized. To achieve the target efficiency, we collaborated with a leading rotary compressor manufacturer to develop a compressor prototype specifically optimized for R-290, able to achieve superior energy efficiency. In addition, we conducted model-based investigations to identify further charge reduction potential. Finally, with using microchannel heat exchangers to replace the fin-and-tube condenser and evaporator, the system charge can be decreased to 150 g.

42 ENGINEERING↗
DOE OSTI2021

Development of a cosmic ray oriented trigger for the fluorescence telescope on EUSO-SPB2

The Extreme Universe Space Observatory on a Super Pressure Balloon 2 (EUSO-SPB2), in preparation, aims to make the first observations of Ultra-High Energy Cosmic Rays (UHECRs) from near space using optical techniques. EUSO-SPB2 will prototype instrumentation for future satellite-based missions, including the Probe of Extreme Multi-Messenger Astrophysics (POEMMA) and K-EUSO. The payload will consist of two telescopes. The first is a Cherenkov telescope (CT) being developed to quantify the background for future below-the-limb very high energy (E 10 PeV) astrophysical neutrino observations, and the second is a fluorescence telescope (FT) being developed for detection of UHECRs. The FT will consist of a Schmidt telescope, and a 6192 pixel ultraviolet camera with an integration time of 1.05 s. The first step in the data acquisition process for the FT is a hardware level trigger in order to decide which data to record. In order to maximize the number of UHECR induced extensive air showers (EASs) which can be detected, a novel trigger algorithm has been developed based on the intricacies and limitations of the detector. Finally, the expected performance of the trigger has been characterized by simulations and, pending hardware verification, shows that EUSO-SPB2 is well positioned to attempt the first near-space observation of UHECRs via optical techniques.

79 ASTRONOMY AND ASTROPHYSICS↗
DOE OSTI2015

Analysis of cardiomyocyte movement in the developing murine heart

The precise assemblage of several types of cardiac precursors controls heart organogenesis. The cardiac precursors show dynamic movement during early development and then form the complicated heart structure. However, cardiomyocyte movements inside the newly organized mammalian heart remain unclear. We previously established the method of ex vivo time-lapse imaging of the murine heart to study cardiomyocyte behavior by using the Fucci (fluorescent ubiquitination-based cell cycle indicator) system, which can effectively label individual G1, S/G2/M, and G1/S-transition phase nuclei in living cardiomyocytes as red, green, and yellow, respectively. Global analysis of gene expression in Fucci green positive ventricular cardiomyocytes confirmed that cell cycle regulatory genes expressed in G1/S, S, G2/M, and M phase transitions were upregulated. Interestingly, pathway analysis revealed that many genes related to the cell cycle were significantly upregulated in the Fucci green positive ventricular cardiomyocytes, while only a small number of genes related to cell motility were upregulated. Time-lapse imaging showed that murine proliferating cardiomyocytes did not exhibit dynamic movement inside the heart, but stayed on site after entering the cell cycle. - Highlights: • We directly visualized cardiomyocyte movement inside the developing murine heart. • Cell cycle related genes were upregulated in the proliferating cardiomyocytes. • Time-lapse imaging revealed that proliferating murine cardiomyocytes stayed in place. • Murine ventricular cardiomyocytes proliferate on site during development.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2015

Non-absorbable mesoporous silica for the development of protein sequestration therapies

While our understanding of the molecular events leading to disease onset and progression have increased exponentially, our capacity to therapeutically intervene in these events with new chemical diversity has clearly fallen short of that pace. In the quest to readdress this situation, the drug discovery sector is slowly but increasingly exploring sources of alternative chemical matter, such as the ones provided by material science and nanotechnology. While new functional nano-sized materials hold great promise for the future, our lack of understanding of the long term safety implications associated with systemic exposure as well as the unclear regulatory path ahead hamper their present impact in drug development. Paradoxically, the exploitation of novel, functionally active micron-sized, synthetic, non-absorbable chemical matter, for the treatment or prevention of a number of epidemiologically significant conditions remains clearly underexplored. A combination of pre-existing evidence and future potential indicates that micron-sized mesoporous silica materials could be an untapped source of new drug candidates. These are free from both the dreaded high attrition associated with small molecule drug discovery and the uncertainties of nano-size technologies. This, together with the coming of age of synthetic methodologies to control particle size and shape; pore size and geometry; surface chemistry, bioconjugation and formulation, open up exciting possibilities to exploit this novel chemistry-biology therapeutic interface. - Highlights: • The development of functionally active micron-sized particles in medicine is underexplored. • Mesoporous materials offer the advantage of nanostructured particles in the micron size. • Non-absorbable drugs based on such particles for enzyme inhibition are being developed. • Several conditions can be targeted such as obesity, sepsis or celiac disease.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2016

Rho GTPase protein Cdc42 is critical for postnatal cartilage development

Cdc42, a small Rho GTPase family member, has been shown to regulate multiple cellular functions in vitro, including actin cytoskeletal reorganization, cell migration, proliferation, and gene expression. However, its tissue-specific roles in vivo remain largely unknown, especially in postnatal cartilage development, as cartilage-specific Cdc42 inactivated mice die within a few days after birth. In this study, we investigated the physiological functions of Cdc42 during cartilage development after birth using tamoxifen-induced cartilage-specific inactivated Cdc42 conditional knockout (Cdc42 {sup fl/fl}; Col2-CreERT) mice, which were generated by crossing Cdc42 flox mice (Cdc42 {sup fl/fl}) with tamoxifen-induced type II collagen (Col2) Cre transgenic mice using a Cre/loxP system. The gross morphology of the Cdc42 cKO mice was shorter limbs and body, as well as reduced body weight as compared with the controls. In addition, severe defects were found in growth plate chondrocytes of the long bones, characterized by a shorter proliferating zone (PZ), wider hypertrophic zone (HZ), and loss of columnar organization of proliferating chondrocytes, resulting in delayed endochondral bone formation associated with abnormal bone growth. Our findings demonstrate the importance of Cdc42 for cartilage development during both embryonic and postnatal stages. - Highlights: • Tamoxifen-induced cartilage specific inactivated Cdc42 mutant mice were generated. • Cdc42 mutant mice were shorter limbs and body. • Severe defects were found in growth plate chondrocytes.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2016

NSrp70 is significant for embryonic growth and development, being a crucial factor for gastrulation and mesoderm induction

NSrp70 (nuclear speckle-related protein 70), a recently discovered protein and it belongs to the serine/arginine (SR) rich related protein family. NSrp70 is recognized as an important splicing factor comprising RNA recognition motif (RRM) and arginine/serine (RS)-like regions at the N- and C-terminus respectively, along with two coiled coil domains at each terminus. However, other functions of NSrp70 remain unelucidated. In this study, we investigated the role of NSrp70 in Xenopus embryogenesis and found that its maternal expression plays a critical role in embryonic development. Knockdown of NSrp70 resulted in dramatic reduction in the length of developing tadpoles and mild to severe malformation in Xenopus embryos. In addition, knockdown of NSrp70 resulted in an extremely short axis by blocking gastrulation and convergent extension. Further, animal cap assays along with activin A treatment revealed that NSrp70 is an essential factor for dorsal mesoderm induction as knockdown of NSrp70 caused a dramatic down-regulation of dorsal mesoderm specific genes and its loss significantly shortened the elongation region of animal caps. In conclusion, NSrp70 is crucial for early embryonic development, influencing gastrulation and mesoderm induction. - Highlights: • NSrp70 is required for proper length and dorso-anterior structures. • NSrp70 is essential for gastrulation movement during embryogenesis. • NSrp70 is crucial component for dorsal mesoderm induction.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Histone methyltransferase Setdb1 is indispensable for Meckel's cartilage development

The histone methyltransferase Setdb1 represses gene expression by catalyzing lysine 9 of histone H3 trimethylation. Given that the conventional knockout of Setdb1 is embryo-lethal at the implantation stage, its role in craniofacial development is poorly understood. Here, we investigated the role of Setdb1, using conditional knockout mice—in which Setdb1 was deleted in the Meckel's cartilage (Setdb1 CKO)—and the mouse chondrogenic cell line ATDC5—in which Setdb1 was inhibited by siRNA. Deletion of Setdb1 in Meckel's cartilage, the supportive tissue in the embryonic mandible, led to its enlargement, instead of the degeneration that normally occurs. Chondrocytes from the Meckel's cartilage of Setdb1 CKO mice showed increased size. Furthermore, at embryonic days 16.5 and 18.5, part of the perichondrium was disrupted and mineralization was observed in the Meckel's cartilage. Proliferation analysis showed that inhibition of Setdb1 caused increased proliferation in chondrocytes in the Meckel's cartilage as well as in ATDC5 cells. Quantitative RT-PCR showed decreased expression of chondrogenic genes, such as Sox9, Mmp13, Collagen II, and Aggrecan, as a result of Setdb1 inhibition in ATDC5 cells. Along with these phenomenons, SMAD-dependent BMP signaling was significantly increased by the loss of Setdb1 in both the Meckel's cartilage of Setdb1 CKO mice and ATDC5 cells. Therefore, the abnormal development of Meckel's cartilage in Setdb1 CKO mice is partly due to the enhanced SMAD-dependent BMP signaling. Overall, to our knowledge, the present study is the first to show that epigenetic regulation by Setdb1 is indispensable for the embryonic development of Meckel's cartilage. - Highlights: • Deletion of Setdb1 led to enlarged Meckel's cartilage. • Chondrocytes from the Meckel's cartilage of Setdb1 mutant showed increased in size. • Part of the perichondrium was disrupted and mineralization was observed in the Meckel's cartilage of Setdb1 mutant. • Deletion of Setdb1 increases proliferation in vivo and in vitro. • Deletion of Setdb1 enhances SMAD-dependent BMP signaling in vivo and in vitro.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Development of an aptamer-ampicillin conjugate for treating biofilms

Biofilm formation involves the development of extracellular matrix and initially depends on adherence and tropism by flagellar movement. With the widespread development of antibiotic resistance and tolerance of biofilms, there is a growing need for novel anti-infective strategies. No currently approved medications specifically target biofilms. Aptamers are single-stranded nucleic acid molecules that may bind to their targets with high affinity and affect the target functions. We developed a bifunctional conjugate by linking an aptamer targeting bacterial flagella with ampicillin. We investigated its influence on biofilm prevention and dissolution by ultraviolet–visible spectrophotometry, inverted microscopy, and atomic force microscopy. This conjugate had distinctive antibacterial activity. Notably, the conjugate was more active than either component, and thus had a synergistic effect against biofilms.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Loss of PiT-2 results in abnormal bone development and decreased bone mineral density and length in mice

Highlights: • PiT-2 deficiency decreased mineral density in craniofacial and long bones in mice. • Bone length was reduced in long bones in PiT-2 deficient mice. • Trabecular and cortical bone mineral density were decreased in PiT-2 deficient mice. • Static bone morphometric parameters were altered in PiT-2 deficient mice. • PiT-2 deficiency altered craniofacial bone morphology. Normal bone mineralization requires phosphate oversaturation in bone matrix vesicles, as well as normal regulation of phosphate metabolism via the interplay among bone, intestine, and kidney. In turn, derangement of phosphate metabolism greatly affects bone function and structure. The type III sodium-dependent phosphate transporters, PiT-1 and PiT-2, are believed to be important in tissue phosphate metabolism and physiological bone formation, but their requirement and molecular roles in bone remain poorly investigated. In order to decipher the role of PiT-2 in bone, we examined normal bone development, growth, and mineralization in global PiT-2 homozygous knockout mice. PiT-2 deficiency resulted in reduced vertebral column, femur, and tibia length as well as mandibular dimensions. Micro-computed tomography analysis revealed that bone mineral density in the mandible, femur, and tibia were decreased, indicating that maintenance of bone function and structure is impaired in both craniofacial and long bones of PiT-2 deficient mice. Both cortical and trabecular thickness and mineral density were reduced in PiT-2 homozygous knockout mice compared with wild-type mice. These results suggest that PiT-2 is involved in normal bone development and growth and plays roles in cortical and trabecular bone metabolism feasibly by regulating local phosphate transport and mineralization processes in the bone. Further studies that evaluate bone cell-specific loss of PiT-2 are now warranted and may yield insight into complex mechanisms of bone development and growth, leading to identification of new therapeutic options for patients with bone diseases.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Development of a novel cellulase biosensor that detects crystalline cellulose hydrolysis using a transcriptional regulator

Highlights: • A novel biosensor for detecting cellulolytic activity is developed using cellobiose-dependent transcriptional regulator. • CBGESS shows quantitative response to cellobiose with hyper sensitivity and specificity. • Cellulase activity including hydrolysis of crystalline cellulose is measured by CBGESS via intracellular fluorescence. Successful utilization of cellulose as renewable biomass depends on the development of economically feasible technologies, which can aid in enzymatic hydrolysis. In this study, we developed a whole-cell biosensor for detecting cellulolytic activity that relies on the recognition of cellobiose using the transcriptional factor CelR from Thermobifida fusca and transcriptional activation of its downstream gfp reporter gene. The fluorescence intensity of whole-cell biosensor, which was named as cellobiose-detectible genetic enzyme screening system (CBGESS), was directly proportional to the concentration of cellobiose. The strong fluorescence intensity of CBGESS demonstrated the ability to detect cellulolytic activity with two cellulosic substrates, carboxymethyl cellulose and p-nitrophenyl β-D-cellobioside in cellulase-expressing Escherichia coli. In addition, CBGESS easily sensed crystalline cellulolytic activity when commercial Celluclast 1.5L was dropped on an Avicel plate. Therefore, CBGESS is a powerful tool for detecting cellulolytic activity with high sensitivity in the presence of soluble or insoluble cellulosic substrates. CBGESS may be further applied to excavate novel cellulases or microbes from both genetic libraries and various environments.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Distorted antibody repertoire developed in the absence of pre-B cell receptor formation

Highlights: • Leaky B cells in SLC-deficient mice circumvent the pre-BCR checkpoint. • Antibody repertoires differing in VDJ-usage exist in leaky B cells in λ5{sup −}/{sup −} mice. • Leaky B cells do not originate from B1 or particular B2 cells. • The CDR-H3 middle loop in λ5{sup −}/{sup −} bone marrow B cells is enriched in arginines. The pre-B cell receptor (pre-BCR), consisting of the μ heavy chain (μHC) and the surrogate light chain (SLC, Vpre-B and λ5), plays important roles during B cell development. The formation of the pre-BCR, which enables the nascent immunoglobulin HC to associate with the SLC, is considered a prerequisite for B cell development. However, a significant number of peripheral mature (leaky) B cells exist in SLC-deficient mice. These leaky B cells develop in the absence of pre-BCR and do not undergo the pre-BCR checkpoint. The antibody repertoires of leaky B cells thus reflect the absence of pre-BCR function. To investigate how the absence of the pre-BCR is circumvented by these leaky-B cells and examine the effect of the pre-BCR checkpoint on the antibody system, we analyzed the antibody repertoires of λ5-deficient (λ5{sup −/−}) mice using next-generation sequencing. In λ5{sup −/−} mice, spleen B cells displayed different patterns of VDJ-usage, relative to those in wild-type (WT) mice. Moreover, leaky B cells were neither derived from unusual B2 cells, characterized by particular LC gene rearrangements in the absence of pre-BCR signaling, nor from B1 cells, originating from different B cell progenitors. Analysis of the CDR-H3 amino acid sequences of μ-chain repertoires revealed that certain bone marrow B cells with particular CDR-H3 profiles undergo clonal expansion in λ5{sup −/−} mice. Part of these CDR-H3s contain arginine(s) in the middle of the CDR-H3 loop in λ5{sup −/−} mice, whereas few arginine(s) exist in this middle loop in WT CDR-H3s in the absence of clonal expansion. This CDR-H3 feature in λ5{sup −/−} mice presumably reflects the role of the pre-BCR in autoantibody regulation, since arginine(s) are often found in the antigen-binding site of autoantibodies. Here, we present a unique viewpoint on the role of pre-BCR, by assessing the whole antibody repertoire formed in SLC-deficient mice.

60 APPLIED LIFE SCIENCES↗