Primary Ciliary Dyskinesia in a Portuguese Bronchiectasis Outpatient Clinic.

Primary ciliary dyskinesia (PCD) is a rare hereditary condition characterized by decreased mucociliary clearance of the airways and a compromised reproductive system, resulting in male and female infertility. Several mutations with varied clinical and pathological features have been documented, making diagnosis a challenging process. The purpose of this study is to describe the clinical and pathological features of Portuguese patients with PCD and to examine their genetic variants. A retrospective observational analysis was conducted with patients who were being monitored at a bronchiectasis outpatient clinic in 2022 and had a confirmed or high-likelihood diagnosis of PCD. In total, 17 patients were included in the study, with 12 (66.7%) having PCD confirmed and 5 (29.4%) having a high-likelihood diagnosis. Furthermore, 12 patients were subjected to transmission electron microscopy (TEM), with 7 (58.3%) exhibiting one hallmark defect. Genetic test data was obtained for all 17 patients, with 7 of them (41.2%) displaying a pathogenic/likely pathogenic mutation in homozygosity. To summarize, PCD is an uncommon but significant hereditary illness with consequences regarding morbidity and mortality. Despite the lack of a specific treatment, it is critical to confirm the diagnosis with genetic testing in order to effectively manage the disease and its accompanying disorders.

Δ133p53α enhances metabolic and cellular fitness of TCR-engineered T cells and promotes superior antitumor immunity.

BACKGROUND: Tumor microenvironment-associated T cell senescence is a key limiting factor for durable effective cancer immunotherapy. A few studies have demonstrated the critical role of the tumor suppressor TP53-derived p53 isoforms in cellular senescence process of non-immune cells. However, their role in lymphocytes, in particular tumor-antigen (TA) specific T cells remain largely unexplored. METHODS: Human T cells from peripheral blood were retrovirally engineered to coexpress a TA-specific T cell receptor and the Δ133p53α-isoform, and characterized for their cellular phenotype, metabolic profile and effector functions. RESULTS: Phenotypic analysis of Δ133p53α-modified T cells revealed a marked reduction of the T-cell inhibitory molecules (ie, CD160 and TIGIT), a lower frequency of senescent-like CD57+ and CD160+ CD8+ T cell populations, and an increased number of less differentiated CD28+ T cells. Consistently, we demonstrated changes in the cellular metabolic program toward a quiescent T cell state. On a functional level, Δ133p53α-expressing T cells acquired a long-term proliferative capacity, showed superior cytokine secretion and enhanced tumor-specific killing in vitro and in mouse tumor model. Finally, we demonstrated the capacity of Δ133p53α to restore the antitumor response of senescent T cells isolated from multiple myeloma patients. CONCLUSION: This study uncovered a broad effect of Δ133p53α isoform in regulating T lymphocyte function. Enhancing fitness and effector functions of senescent T cells by modulation of p53 isoforms could be exploited for future translational research to improve cancer immunotherapy and immunosenescence-related diseases.

Epstein-Barr Virus-Associated With Lymphoepithelial Carcinoma: A Rare Tumor of the Larynx.

Lymphoepithelial carcinoma of the larynx is a rare tumor, as this histological entity is mostly diagnosed in nasopharynx. However, it may be present in other non-nasopharyngeal sites and it is extremely rare in the larynx. The authors present a case of a 59-year-old man who presented to the Otorhinolaryngology-Head and Neck Surgery Department complaining of a long-standing dysphonia, odynophagia, and dysphagia. The clinical examination revealed a laryngeal tumor involving the right epiglottis, right aryepiglottic fold, and ipsilateral false vocal fold. It presented with ispilateral neck lymph node extension. Multiple biopsies of the laryngeal lesion were performed under local anesthesia and the histological examination showed a poorly differentiated squamous cell carcinoma. After discussing the case in a multidisciplinary tumor board, a total laryngectomy with a bilateral neck dissection was performed and the histological specimen showed a lymphoepithelial carcinoma. Although immunostaining with LMP-1 antibody was negative, in situ hybridization for Epstein-Barr virus was positive. He underwent adjuvant chemoradiation. He is now at 9-months follow-up period, with no evidence of disease. Lymphoepithelial carcinoma of the larynx is an extremely rare disease, with an aggressive pattern. Epstein-Barr virus-associated lymphoepithelial carcinoma has been exceptionally reported. A correct diagnosis and close collaboration with pathologist is crucial to achieve the best treatment strategy. We present this case to discuss the clinical and histology findings and the different therapeutic aspects of this uncommon histological subtype carcinoma.

Active Life: a project for a safe hospital-community transition after arthroplasty.

OBJECTIVE: To define the criteria for the continuity of care to elderly people submitted to arthroplasty. METHOD: This is a qualitative study, inserted in the constructivist paradigm, whose methodological option fell on research-action. The participants were the health professionals of an orthopedic service and of the community care teams in the area of the hospital. RESULTS: The different techniques allowed us to identify the difficulties in the safe transition from the hospital to the community. At this level, two categories of criteria for continuity of care emerged: criteria associated with the risk of ineffective management of the therapeutic regimen, and criteria associated with the knowledge and level of competence of the informal caregiver. FINAL CONSIDERATIONS: An elderly person undergoing arthroplasty (hip or knee) has functional alterations that affect their capacity for self-care and may lead to dependence, our findings allowed the design of an algorithm to facilitate clinical decision making and promote a safe hospital-community transition.

Active Life: a project for a safe hospital-community transition after arthroplasty / Vida Ativa: projeto para uma transição hospital-comunidade segura após artroplastia / Vida Activa: proyecto para una transición hospital-comunidad segura después de una artroplastia

ABSTRACT Objective: To define the criteria for the continuity of care to elderly people submitted to arthroplasty. Method: This is a qualitative study, inserted in the constructivist paradigm, whose methodological option fell on research-action. The participants were the health professionals of an orthopedic service and of the community care teams in the area of the hospital. Results: The different techniques allowed us to identify the difficulties in the safe transition from the hospital to the community. At this level, two categories of criteria for continuity of care emerged: criteria associated with the risk of ineffective management of the therapeutic regimen, and criteria associated with the knowledge and level of competence of the informal caregiver. Final Considerations: An elderly person undergoing arthroplasty (hip or knee) has functional alterations that affect their capacity for self-care and may lead to dependence, our findings allowed the design of an algorithm to facilitate clinical decision making and promote a safe hospital-community transition.

RESUMO Objetivo: Definir os critérios para a continuidade da assistência à pacientes idosos submetidos à artroplastia. Método: Estudo qualitativo, inserido no paradigma construtivista, utilizando a pesquisa-ação como opção metodológica. Os participantes do estudo foram os profissionais de saúde de um serviço ortopédico e das equipes de saúde comunitária na área de cobertura do hospital. Resultados: As diferentes técnicas utilizadas nos permitiram identificar as dificuldades para uma transição segura do hospital para a comunidade. Duas categorias de critérios para a continuidade da assistência surgiram: critérios associados ao risco de gestão ineficiente do regime terapêutico, e critérios associados ao conhecimento e nível de competência do cuidador informal. Considerações finais: Um idoso submetido a artroplastia (quadril ou joelho) sofre alterações funcionais que afetam a sua capacidade de autocuidado e podem levar à dependência. Nossos achados permitiram o desenvolvimento de um algoritmo para facilitar a tomada de decisão clínica e promover uma transição segura do hospital para a comunidade.

RESUMEN Objetivo: Definir los criterios para la continuidad de la atención a pacientes ancianos sometidos a la artroplastia. Método: Estudio cualitativo, fundamentado en el paradigma constructivista, utilizando la investigación-acción como opción metodológica. Los participantes del estudio fueron los profesionales de salud de un servicio ortopédico y de los equipos de salud comunitaria en el área de cobertura del hospital. Resultados: Las diferentes técnicas utilizadas permitieron identificar las dificultades para una transición segura del hospital hasta la comunidad. Dos categorías de criterios para la continuidad de la atención surgieron: criterios asociados al riesgo de gestión ineficiente del régimen terapéutico, y criterios asociados al conocimiento y nivel de competencia del cuidador informal. Consideraciones finales: Un anciano sometido a artroplastia (cadera o rodilla) sufre alteraciones funcionales que afectan su capacidad de autocuidado y pueden conducir a la dependencia. Nuestros hallazgos permitieron el desarrollo de un algoritmo para facilitar la toma de decisión clínica y promover una transición segura del hospital hasta la comunidad.

A Potent Tumor-Reactive p53-Specific Single-Chain TCR without On- or Off-Target Autoimmunity In Vivo.

Genetic engineering of T cells with a T cell receptor (TCR) targeting tumor antigen is a promising strategy for cancer immunotherapy. Inefficient expression of the introduced TCR due to TCR mispairing may limit the efficacy and adversely affect the safety of TCR gene therapy. Here, we evaluated the safety and therapeutic efficiency of an optimized single-chain TCR (scTCR) specific for an HLA-A2.1-restricted (non-mutated) p53(264-272) peptide in adoptive T cell transfer (ACT) models using our unique transgenic mice expressing human p53 and HLA-A2.1 that closely mimic the human setting. Specifically, we showed that adoptive transfer of optimized scTCR-redirected T cells does not induce on-target and off-target autoimmunity. Furthermore, ACT resulted in full tumor protection and led to a long-lived effective, antigen-specific memory T cell response in syngeneic and xenograft models. Taken together, the study demonstrated that our scTCR specific for the broadly expressed tumor-associated antigen p53(264-272) can eradicate p53+A2.1+ tumor cells without inducing off-target or self-directed toxicities in mouse models of ACT. These data strongly support the improved safety and therapeutic efficacy of high-affinity p53scTCR for TCR-based immunotherapy of p53-associated malignancies.

Needle in the external auditory canal: an unusual complication of inferior alveolar nerve block.

Inferior alveolar nerve block is used to anesthetize the ipsilateral mandible. The most commonly used technique is one in which the anesthetic is injected directly into the pterygomandibular space, by an intraoral approach. The fracture of the needle, although uncommon, can lead to potentially serious complications. The needle is usually found in the pterygomandibular space, although it can migrate and damage adjacent structures, with variable consequences. The authors report an unusual case of a fractured needle, migrating to the external auditory canal, as a result of an inferior alveolar nerve block.

Nitric oxide promotes resistance to tumor suppression by CTLs.

Many human tumors express inducible NO synthetase (NOS2), but the roles of NO in tumor development are not fully elucidated. An important step during tumor development is the acquisition of apoptosis resistance. We investigated the dose-dependent effects of endogenously produced NO on apoptosis using ecdysone-inducible NOS2 cell lines. Our results show that short-term NOS2 expression enhances CD95-mediated apoptosis and T cell cytotoxicity dose dependently. Furthermore, we could show that during chronic exposure to NO, besides the primary cytotoxic NO effect, there is selection of cell clones resistant to NO that show cross-resistance to CD95-induced apoptosis and the killing by CTLs. We propose that NO production could initially act as an autocrine suicide or paracrine killing mechanism in cells undergoing malignant transformation. However, once failed, the outcome is fatal. NO promotes tumor formation by enhancing the selection of cells that can evade immune attack by acquiring apoptosis resistance.

Targeting positive regulatory domain I-binding factor 1 and X box-binding protein 1 transcription factors by multiple myeloma-reactive CTL.

Growing evidence indicates that multiple myeloma (MM) and other malignancies are susceptible to CTL-based immune interventions. We studied whether transcription factors inherently involved in the terminal differentiation of mature B lymphocytes into malignant and nonmalignant plasma cells provide MM-associated CTL epitopes. HLA-A*0201 (A2.1) transgenic mice were used to identify A2.1-presented peptide Ag derived from the plasma cell-associated transcriptional regulators, positive regulatory domain I-binding factor 1 (PRDI-BF1) and X box-binding protein 1 (XBP-1). A2.1-restricted CTL specific for PRDI-BF1 and XBP-1 epitopes efficiently killed a variety of MM targets. PRDI-BF1- and XBP-1-reactive CTL were able to recognize primary MM cells from A2.1(+) patients. Consistent with the expression pattern of both transcription factors beyond malignant and nonmalignant plasma cells, PRDI-BF1- and XBP-1-specific CTL activity was not entirely limited to MM targets, but was also associated with lysis of certain other malignancies and, in defined instances, with low-to-intermediate level recognition of a few types of normal cells. Our results also indicate that the A2.1-restricted, PRDI-BF1- and XBP-1-specific human CD8(+) T cell repertoire is affected by partial self tolerance and may thus require the transfer of high-affinity TCR to break tolerance. We conclude that transcription factors governing terminal cellular differentiation may provide MM- and tumor-associated CTL epitopes.

Cooperation of human tumor-reactive CD4+ and CD8+ T cells after redirection of their specificity by a high-affinity p53A2.1-specific TCR.

Efficient immune attack of malignant disease requires the concerted action of both CD8+ CTL and CD4+ Th cells. We used human leukocyte antigen (HLA)-A*0201 (A2.1) transgenic mice, in which the mouse CD8 molecule cannot efficiently interact with the alpha3 domain of A2.1, to generate a high-affinity, CD8-independent T cell receptor (TCR) specific for a commonly expressed, tumor-associated cytotoxic T lymphocyte (CTL) epitope derived from the human p53 tumor suppressor protein. Retroviral expression of this CD8-independent, p53-specific TCR into human T cells imparted the CD8+ T lymphocytes with broad tumor-specific CTL activity and turned CD4+ T cells into potent tumor-reactive, p53A2.1-specific Th cells. Both T cell subsets were cooperative and interacted synergistically with dendritic cell intermediates and tumor targets. The intentional redirection of both CD4+ Th cells and CD8+ CTL by the same high-affinity, CD8-independent, tumor-specific TCR could provide the basis for novel broad-spectrum cancer immunotherapeutics.

Control of organ transplant-associated graft-versus-host disease by activated host lymphocyte infusions.

BACKGROUND: Prolonged persistence of donor-derived T cells after organ transplantation has been proposed to improve long-term allograft survival. However, surviving transplant-derived T cells are also able to mediate devastating graft-versus-host disease (GvHD). Currently, GvHD after organ transplantation is usually refractory to conventional therapy and the disease outcome fatal. METHODS: Graft-reactive host T cells were generated ex vivo from a patient suffering from a severe and refractory liver-transplant-associated GvHD. To control GvHD, activated alloreactive host T cells were repetitively retransferred into the patient (activated host lymphocyte infusion [aHLI]). RESULTS: Adoptive transfer of ex vivo activated alloreactive host T cells (aHLI) led to the control and complete resolution of severe GvHD without inducing allograft rejection. CONCLUSIONS: aHLI opens a novel therapeutic window to control solid-organ transplant-associated GvHD while preserving allograft integrity.

Partial tyrosinase-specific self tolerance by HLA-A*0201-restricted cytotoxic T lymphocytes in mice and man.

The human tyrosinase (hTyr) (369-377) cytotoxic T lymphocyte (CTL) epitope is presented by malignant melanoma and various nontransformed cells in association with human leukocyte antigen (HLA)-A*0201 (A2.1) and used for vaccination-based immunotherapy of melanoma patients. Its mouse homologue, mTyr (369-377), is naturally processed and bound by A2.1 with equivalent efficacy and thus enabled us to explore the effect of self tolerance on Tyr-specific T cells in different lines of A2.1 transgenic (Tg) mice and man. We found that self Tyr-reactive CTL in Tg mice and, importantly, in man were affected by partial tolerance resulting in only residual T lymphocytes of higher avidity for self Tyr along with low-avidity T cells to be present in the periphery. Immunizing mice with the xenogeneic nonself Tyr peptide facilitated the generation of self Tyr-reactive CTL. As compared to Tyr-reactive CTL induced by high amounts of the self Tyr epitope, however, the nonself antigen (Ag) had no effect on improving the avidity of self Tyr-specific mouse and human T cells. Depleting mice of CD25(+) T cells with and without CTL-associated Ag 4 (CTLA-4) blockade demonstrated that tolerance of Tyr-specific CTL was not regulated by CD4(+)CD25(+) T regulatory cells (Treg) or CTLA-4. Our studies have important implications for the design of anti-Tyr-based immunotherapeutics.

The effect of the interferon-gamma-inducible processing machinery on the generation of a naturally tumor-associated human cytotoxic T lymphocyte epitope within a wild-type and mutant p53 sequence context.

The human wild-type (wt) p53.264-272 peptide is a universal tumor antigen and recognized by HLA-A*0201 (A2.1)-restricted CTL. Generation of this epitope by constitutive 20S proteasomes is prevented by a p53 R to H hotspot mutation at the C-terminal flanking residue 273. We report on the impact of the interferon-gamma (IFN-gamma)-inducible proteasomal activator PA28 (11S regulator) and the immunoproteasome on the in vitro and cellular processing of wt and mutant (mut) p53 substrates. We found that production of the antigenic 264-272 peptide from wt p53 by constitutive as well as immunoproteasomes is accelerated and amplified by the PA28 activator. PA28 and (immuno)proteasomes were not capable to reconvert the resistance of epitope release from mut p53. Maximum and accelerated antigen production in vitro and on the cellular level required the IFN-gamma-inducible interaction of immunoproteasomes and PA28. We conclude that efficient processing of p53.264272 from wt p53 is governed by the proteasome/PA28 complex. These studies have important implications for p53-specific cancer immunotherapy and demonstrate that the effects of the immunoproteasome and PA28 are influenced by the individual epitope and its flanking sequence context.