The identification of effective tumor-suppressing neoantigens using a tumor-reactive TIL TCR-pMHC ternary complex.

Neoantigens are ideal targets for cancer immunotherapy because they are expressed de novo in tumor tissue but not in healthy tissue and are therefore recognized as foreign by the immune system. Advances in next-generation sequencing and bioinformatics technologies have enabled the quick identification and prediction of tumor-specific neoantigens; however, only a small fraction of predicted neoantigens are immunogenic. To improve the predictability of immunogenic neoantigens, we developed the in silico neoantigen prediction workflows VACINUSpMHC and VACINUSTCR: VACINUSpMHC incorporates physical binding between peptides and MHCs (pMHCs), and VACINUSTCR integrates T cell reactivity to the pMHC complex through deep learning-based pairing with T cell receptors (TCRs) of putative tumor-reactive CD8 tumor-infiltrating lymphocytes (TILs). We then validated our neoantigen prediction workflows both in vitro and in vivo in patients with hepatocellular carcinoma (HCC) and in a B16F10 mouse melanoma model. The predictive abilities of VACINUSpMHC and VACINUSTCR were confirmed in a validation cohort of 8 patients with HCC. Of a total of 118 neoantigen candidates predicted by VACINUSpMHC, 48 peptides were ultimately selected using VACINUSTCR. In vitro validation revealed that among the 48 predicted neoantigen candidates, 13 peptides were immunogenic. Assessment of the antitumor efficacy of the candidate neoepitopes using a VACINUSTCR in vivo mouse model suggested that vaccination with the predicted neoepitopes induced neoantigen-specific T cell responses and enabled the trafficking of neoantigen-specific CD8 + T cell clones into the tumor tissue, leading to tumor suppression. This study showed that the prediction of immunogenic neoantigens can be improved by integrating a tumor-reactive TIL TCR-pMHC ternary complex.

Effects of transition programmes to adulthood for adolescents and young adults with CHD: a systematic review with meta-analysis.

BACKGROUND: The increased survival rate among individuals with CHD has sparked interest in their transition to adult healthcare. Although there is a general agreement on the importance of transition interventions, the empirical evidence supporting them is insufficient. Therefore, this study aimed to conduct a systematic review and meta-analysis of transition interventions for adult healthcare in adolescents and young adults. METHODS AND RESULTS: A literature search was conducted for studies comparing the quantitative effects of transition interventions with control groups, published up to March 15, 2023, in major databases (CENTRAL, Embase, PubMed, Web of Science, CINAHL, KISS, and KMbase), major clinical trial registries, academic journal sites related to the topic, and grey literature databases. Ten studies involving a total of 1,297 participants were identified. Transition interventions proved effective in enhancing disease-related knowledge (Hedge's g = 0.89, 95% CI = 0.29-1.48) and self-management (Hedge's g = 0.67, 95% CI = 0.38-0.95), as well as reducing loss to follow-up (OR = 0.41, 95% CI = 0.22-0.77). The certainty of evidence for the estimated values of each major outcome was low or very low. CONCLUSIONS: This study supports the implementation of transition interventions by demonstrating that they can improve patients' disease knowledge and self-management, while also promoting treatment continuity. However, since the available data on transition interventions for adolescents and young adults with CHD remain limited, the widespread adoption of structured transition interventions in the future may alter the conclusions of this study. REGISTRATION: URL: https://www.crd.york.ac.uk/PROSPERO. Unique identifier: CRD42023399026.

Defining a TCF1-expressing progenitor allogeneic CD8+ T cell subset in acute graft-versus-host disease.

Graft-versus-host disease (GvHD) is a severe complication of hematopoietic stem cell transplantation driven by activated allogeneic T cells. Here, we identify a distinct subset of T cell factor-1 (TCF1)+ CD8+ T cells in mouse allogeneic and xenogeneic transplant models of acute GvHD. These TCF1+ cells exhibit distinct characteristics compared to TCF1- cells, including lower expression of inhibitory receptors and higher expression of costimulatory molecules. Notably, the TCF1+ subset displays exclusive proliferative potential and could differentiate into TCF1- effector cells upon antigenic stimulation. Pathway analyses support the role of TCF1+ and TCF1- subsets as resource cells and effector cells, respectively. Furthermore, the TCF1+ CD8+ T cell subset is primarily present in the spleen and exhibits a resident phenotype. These findings provide insight into the differentiation of allogeneic and xenogeneic CD8+ T cells and have implications for the development of immunotherapeutic strategies targeting acute GvHD.

Development and evaluation of case video-based debriefing on a simulation of high-risk neonatal care for nursing students in South Korea: a mixed-methods study.

BACKGROUND: The debriefing process after health care simulations should provide a psychologically safe learning environment for nursing students. Case video-based debriefing on a simulation of high-risk neonatal care can help nursing students feel psychologically safe and make learning more effective. In this study, we developed case video-based debriefing materials for a simulation of high-risk neonatal care for nursing students in South Korea and evaluated their effects. METHODS: This mixed-methods study, consisting of a survey and an in-depth interview, was conducted between August and December 2022. The participants were 27 nursing students for the development of the case video-based debriefing and 51 nursing students for the evaluation of its effects (25 in the experimental group and 26 in the control group) at a university in South Korea. A case video-based debriefing on a simulation of high-risk neonatal care was developed, and the experimental group took part in case video-based debriefing. The participants' self-efficacy, critical thinking, state anxiety, and satisfaction with practice were examined. The experimental group's learning experiences were explored. Quantitative data were analyzed using the chi-square test, the unpaired t-test, and repeated-measures analysis of variance. Qualitative content analysis was conducted. RESULTS: In the experimental group, critical thinking and satisfaction with practice increased to a greater extent than in the control group. However, the changes in self-efficacy and state anxiety were not significantly different between the experimental and control groups. Four categories were extracted from nursing students who participated in the case video-based debriefing: "learning facilitated by the simulation," "expanded learning," "safe learning environment," and "efficient utilization of case videos." CONCLUSIONS: Case video-based debriefing on a simulation of high-risk neonatal care effectively enhanced nursing students' critical thinking and satisfaction with practice, and it will be utilized to improve nursing students' competency in high-risk neonatal care.

Development and evaluation of a pediatric nursing competency-building program for nursing students in South Korea: a quasi-experimental study.

PURPOSE: The present study aimed to develop and examine the effectiveness of a pediatric nursing competency-building program for nursing students. METHODS: This was a quasi-experimental study with a nonequivalent control group pretest-posttest design conducted between October and December 2021. The participants included 40 nursing students (20 each in the experimental and control groups) at a university in a South Korean city. The pediatric nursing competency-building program integrated problem-based learning and simulation into clinical field practice. The experimental group participated in the program, while the control group did not. Data were analyzed using the x2 test, the independent t-test, and repeated-measures analysis of variance. RESULTS: Pediatric nursing competency and clinical performance showed a greater increase in the experimental group than in the control group. However, the change in problem-solving ability in the experimental group was not significantly different from that in the control group. CONCLUSION: The pediatric nursing competency-building program effectively improved students' pediatric nursing competency and clinical performance.

Development of a Project-Based Learning Program on High-Risk Newborn Care for Nursing Students and Its Effects: A Quasi-Experimental Study.

Project-based learning (PjBL) allows nursing students to participate in real problem-solving, construct knowledge, and improve their nursing skills in the process of accomplishing meaningful projects. This study was conducted to develop a PjBL program on high-risk newborn care for nursing students and evaluate its effects. A quasi-experimental study using a nonequivalent control group pretest-posttest design was employed between June and December 2021. The participants were 45 nursing students (24 in the experimental group and 21 in the control group). A PjBL program involving the creation of an educational video clip about high-risk newborn care for nursing students was developed, and the experimental group took part in PjBL. The participants' nursing competency for high-risk newborns, self-leadership, and practicum-related stress were assessed. In the experimental group, nursing competency for high-risk newborns increased and practicum-related stress decreased to a greater extent than in the control group. However, the change in self-leadership was not significantly different between the experimental and control groups. PjBL effectively improved students' nursing competency for high-risk newborns and decreased their practicum-related stress. PjBL will be utilized to enhance nursing students' expertise in high-risk newborn care.

Establishment of a mechanism-based in vitro coculture assay for evaluating the efficacy of immune checkpoint inhibitors.

Cancer immunotherapy, which blocks immune checkpoint molecules, is an effective therapeutic strategy for human cancer patients through restoration of tumor-infiltrating (TI) cell function. However, evaluating the efficacy of immune checkpoint inhibitors (ICIs) is difficult because no standard in vitro assay for ICI efficacy evaluation exists. Additionally, blocking a particular immune checkpoint receptor (ICR) is insufficient to restore T cell functionality, because other ICRs still transduce inhibitory signals. Therefore, limiting inhibitory signals transduced via other ICRs is needed to more accurately assess the efficacy of ICIs targeting a particular immune checkpoint. Here, we introduce a newly developed in vitro coculture assay using human peripheral blood mononuclear cells (hPBMCs) and engineered human cancer cell lines. We enriched CD8+ T cells from hPBMCs of healthy donors through low-dose T cell receptor stimulation and cytokine (human IL-2 and IL-7) addition. These enriched CD8+ T cells were functional and expressed multiple ICRs, especially TIM-3 and TIGIT. We also established immune checkpoint ligand (ICL) knockout (KO) cancer cell lines with the CRISPR-Cas9 system. Then, we optimized the in vitro coculture assay conditions to evaluate ICI efficacy. For example, we selected the most effective anti-TIM-3 antibody through coculture of TIM-3+CD8+ T cells with PD-L1-/-PVR-/- cancer cells. In summary, we developed a mechanism-based in vitro coculture assay with hPBMCs and ICL KO cancer cell lines, which could be a useful tool to identify promising ICIs by providing reliable ICI efficacy information.

Development of a protocol for guidance in the pediatric nursing practicum in South Korea: a methodology study.

PURPOSE: Practical training in pediatric nursing gives students the opportunity to apply nursing knowledge to children in a realistic clinical context. Clinical practice faculty (CPF) and clinical nursing instructor (CNI) have played important roles in the pediatric nursing practicum. This study was conducted to develop a protocol to guide clinical practicum in pediatric nursing. METHODS: A service design methodology was employed between August 2020 and May 2021 at four universities and four hospitals in South Korea. The participants were five CPFs, five CNIs, five nursing college graduates, and 60 nursing students. The service design process had four phases: discovery, definition, development, and delivery. Data were collected through self-report questionnaires, in-depth interviews, and observations. The data were analyzed using content analysis and descriptive statistics. RESULTS: The participants reported needs for providing concrete guidance and explanation, nursing practice experience, and a link between school and the clinical field. A protocol was developed to fulfill the participants' needs. The protocol comprised detailed information, teaching methodology, and partnership to guide students in the pediatric nursing practicum. CONCLUSION: The protocol developed in this study can be used to provide guidance for students' clinical practice in the field of pediatric nursing.

Educational needs for practicing neonatal intensive care among Korean nursing students.

PURPOSE: This study was conducted to investigate the educational needs for practicing neonatal intensive care among Korean nursing students. METHODS: An explorative, sequential, mixed-methods design was used. Qualitative content analysis was conducted of in-depth interviews of six nursing students, five clinical practice faculty members, and five nurses in the neonatal intensive care unit. The results of a survey of 174 nursing students were analyzed quantitatively. RESULTS: Nursing students, clinical practice faculty members, and nurses wanted opportunities for direct nursing practice and education in school during neonatal intensive care practice. In terms of specific educational content, nursing students expressed the highest observation-related educational needs for communication with medical team members, and they expressed the highest practice-related educational needs for operating medical equipment used for neonatal intensive care. The nursing students' needs with regard to the method of practice education were highest for orientation from the head nurses. CONCLUSION: Communication and operating medical equipment were found to be areas with high educational needs for practicing neonatal intensive care among Korean nursing students. Further research is needed to develop an educational framework and setting for practicing neonatal intensive care that would meet their needs.

Combination of PD-L1 and PVR determines sensitivity to PD-1 blockade.

Expression of immune checkpoint ligands (ICLs) is necessary to trigger the inhibitory signal via immune checkpoint receptors (ICRs) in exhausted T cells under tumor immune microenvironment. Nevertheless,to our knowledge, ICL expression profile in cancer patients has not been investigated. Using previously reported RNA-seq data sets, we found that expression of ICLs was patient specific but their coexpression can be patterned in non-small-cell lung cancers (NSCLCs). Since the expression of PD-L1 and poliovirus receptor (PVR) among various ICLs was independently regulated, we could stratify the patients who were treated with anti-PD-1 later into 4 groups according to the expression level of PD-L1 and PVR. Of interest, high PVR and low PVR expressions in PD-L1-expressing patients enriched nonresponders and responders to PD-1 blockade, respectively, helping in further selection of responders. Using a genetically engineered cancer model, we also found that PVR-deficient and PD-L1-sufficient tumor-bearing mice were highly sensitive to anti-PD-1 therapy, whereas PVR-sufficient and PD-L1-deficient tumor-bearing mice were resistant to anti-PD-1 therapy. Taken together, our study provides a concept that combinatorial expression patterns of PVR and PD-L1 are key determinants for PD-1 blockade and furthermore suggest a better therapeutic usage of immune checkpoint blockades (ICBs).

Needs for Post-hospital Education among Parents of Infants and Toddlers with Congenital Heart Disease.

PURPOSE: This study was conducted to investigate the educational needs of parents of infants and toddlers with congenital heart disease (CHD) after hospital discharge. METHODS: Qualitative content analysis was conducted of in-depth interviews of eight parents, and the results of an online survey of 171 parents were analyzed quantitatively. RESULTS: Only 16.4% of parents reported that they had received education after hospital discharge on how to provide care for a child with CHD at home. The main reason why parents did not receive education on this topic was that they did not have sufficient opportunities or information (75.5%). In addition, 97.1% of parents stated that they needed educational programs that would be available at home after discharge. In terms of specific educational content, parents expressed the highest needs for education on the symptoms of CHD and ways to cope with them, the prognosis of CHD, and the growth and development of infants and toddlers with CHD. CONCLUSION: The study showed that parents' educational needs were high in many ways. However, the information and educational opportunities offered after discharge were insufficient compared to those needs. Further research is needed to develop post-hospital educational programs that meet their needs.

Combination of TLR1/2 and TLR3 ligands enhances CD4(+) T cell longevity and antibody responses by modulating type I IFN production.

Despite the possibility of combining Toll-like receptor (TLR) ligands as adjuvants to improve vaccine efficacy, it remains unclear which combinations of TLR ligands are effective or what their underlying mechanisms may be. Here, we investigated the mechanism of action of L-pampo, a proprietary adjuvant composed of TLR1/2 and TLR3 ligands. L-pampo dramatically increased humoral immune responses against the tested target antigens, which was correlated with an increase in follicular helper T cells and the maintenance of antigen-specific CD4(+) T cells. During the initial priming phase, in contrast to the induction of type I interferon (IFN) and pro-inflammatory cytokines stimulated by polyI:C, L-pampo showed a greatly diminished induction of type I IFN, but not of other cytokines, and remarkably attenuated IRF3 signaling, which appeared to be critical to L-pampo-mediated adjuvanticity. Collectively, our results demonstrate that the adjuvant L-pampo contributes to the promotion of antigen-specific antibodies and CD4(+) T cell responses via a fine regulation of the TLR1/2 and TLR3 signaling pathways, which may be helpful in the design of improved vaccines.

Cisplatin induces tolerogenic dendritic cells in response to TLR agonists via the abundant production of IL-10, thereby promoting Th2- and Tr1-biased T-cell immunity.

Although many advantageous roles of cisplatin (cis-diamminedichloroplatinum (II), CDDP) have been reported in cancer therapy, the immunomodulatory roles of cisplatin in the phenotypic and functional alterations of dendritic cells (DCs) are poorly understood. Here, we investigated the effect of cisplatin on the functionality of DCs and the changes in signaling pathways activated upon toll-like receptor (TLR) stimulation. Cisplatin-treated DCs down-regulated the expression of cell surface molecules (CD80, CD86, MHC class I and II) and up-regulated endocytic capacity in a dose-dependent manner. Upon stimulation with various TLR agonists, cisplatin-treated DCs showed markedly increased IL-10 production through activation of the p38 MAPK and NF-κB signaling pathways without altering the levels of TNF-α and IL-12p70, indicating the cisplatin-mediated induction of tolerogenic DCs. This effect was dependent on the production of IL-10 from DCs, as neither DCs isolated from IL-10-/- mice nor IL-10-neutralized DCs generated tolerogenic DCs. Interestingly, DCs that were co-treated with cisplatin and lipopolysaccharide (LPS) exhibited a decreased immunostimulatory capacity for inducing the proliferation of Th1- and Th17-type T cells; instead, these DCs contributed to Th2-type T cell immunity. Furthermore, in vitro and in vivo investigations revealed a unique T cell population, IL-10-producing CD3+CD4+LAG-3+CD49b+CD25-Foxp3- Tr1 cells, that was significantly increased without altering the Foxp3+ regulatory T cell population. Taken together, our results suggest that cisplatin induces immune-suppressive tolerogenic DCs in TLR agonist-induced inflammatory conditions via abundant IL-10 production, thereby skewing Th cell differentiation towards Th2 and Tr1 cells. This relationship may provide cancer cells with an opportunity to evade the immune system.

A small molecule inhibitor for ATPase activity of Hsp70 and Hsc70 enhances the immune response to protein antigens.

The ATPase activities of Hsp70 and Hsc70 are known to be responsible for regulation of various biological processes. However, little is known about the roles of Hsp70 and Hsc70 in modulation of immune responses to antigens. In the present study, we investigated the effect of apoptozole (Az), a small molecule inhibitor of Hsp70 and Hsc70, on immune responses to protein antigens. The results show that mice administered with both protein antigen and Az produce more antibodies than those treated with antigen alone, showing that Az enhances immune responses to administered antigens. Treatment of mice with Az elicits production of antibodies with a high IgG2c/IgG1 ratio and stimulates the release of Th1 and Th2-type cytokines, suggesting that Az activates the Th1 and Th2 immune responses. The observations made in the present study suggest that inhibition of Hsp70 and Hsc70 activities could be a novel strategy designing small molecule-based adjuvants in protein vaccines.

Peptidylarginine deiminase inhibition impairs Toll-like receptor agonist-induced functional maturation of dendritic cells, resulting in the loss of T cell-proliferative capacity: a partial mechanism with therapeutic potential in inflammatory settings.

Cl-amidine, which is a small-molecule inhibitor of PAD, has therapeutic potential for inflammation-mediated diseases. However, little is known regarding the manner by which PAD inhibition by Cl-amidine regulates inflammatory conditions. Here, we investigated the effects of PAD inhibition by Cl-amidine on the functioning of DCs, which are pivotal immune cells that mediate inflammatory diseases. When DC maturation was induced by TLR agonists, reduced cytokine levels (IL-6, IL-1ß, and IL-12p70) were observed in Cl-amidine-treated DCs. Cl-amidine-treated, LPS-activated DCs exhibited alterations in their mature and functional statuses with up-regulated antigen uptake, down-regulated CD80, and MHC molecules. In addition, Cl-amidine-treated DCs dysregulated peptide-MHC class formations. Interestingly, the decreased cytokines were independent of MAPK/NF-κB signaling pathways and transcription levels, indicating that PAD inhibition by Cl-amidine may be involved in post-transcriptional steps of cytokine production. Transmission electron microscopy revealed morphotypical changes with reduced dendrites in the Cl-amidine-treated DCs, along with altered cellular compartments, including fragmented ERs and the formation of foamy vesicles. Furthermore, in vitro and in vivo Cl-amidine treatments impaired the proliferation of naïve CD4(+) and CD8(+) T cells. Overall, our findings suggest that Cl-amidine has therapeutic potential for treating inflammation-mediated diseases.