A luminescence-based method to assess antigen presentation and antigen-specific T cell responses for in vitro screening of immunomodulatory checkpoints and therapeutics.

Investigations into the strength of antigen-specific responses in vitro is becoming increasingly relevant for decision making in early-phase research of novel immunotherapeutic approaches, including adoptive cell but also immune checkpoint inhibitor (ICI)-based therapies. In the latter, antigen-specific rapid and high throughput tools to investigate MHC/antigen-specific T cell receptor (TCR) activation haven't been implemented yet. Here, we present a simple and rapid luminescence-based approach using the human papillomavirus 16 (HPV16) E711-20 peptide as model antigen and E7-TCR transgenic Jurkat.NFAT-luciferase reporter cells. Upon E7 peptide pulsing of HLA-A2+ cell lines and macrophages, an effector to target ratio dependent increase in luminescence compared to non-pulsed cells was observed after co-incubation with E7-TCR expressing Jurkat, but not with parental cells. Analogous experiments with cells expressing full-length HPV16 identified that E7-specific activation of Jurkat cells enabled detection of endogenous antigen processing and MHC-I presentation. As proof of concept, overexpression of established checkpoints/inhibitory molecules (e.g., PD-L1 or HLA-G) significantly reduced the E7-specific TCR-induced luminescence, an effect that could be restored after treatment with corresponding targeting antagonistic antibodies. Altogether, the luminescence-based method described here represents an alternative approach for the rapid evaluation of MHC-dependent antigen-specific T cell responses in vitro. It can be used as a rapid tool to evaluate the impact of the immunosuppressive tumor microenvironment or novel ICI in triggering effective T cell responses, as well as speeding up the development of novel therapeutics within the immune-oncology field.

The novel use of cryopreserved human allograft in extensive Hurley stage III hidradenitis suppurativa.

INTRODUCTION: HS is a debilitating dermatologic condition in which apocrine sweat glands become occluded, leading to severe inflammation. Treatment usually ranges from conservative management to surgical intervention with the goal of treating existing lesions while reducing the rate of recurrence, progression, and scarring. Depending on the surface area involved, autologous skin grafting may be difficult when donor sites are limited due to the extent of disease, previous surgery, or scarring. This case report examines the efficacy of cryopreserved human allograft as a surgical treatment of extensive HS. CASE REPORT: A 37-year-old man presented with severe, refractory Hurley stage III HS in which cryopreserved human allograft was used to aid in wound contracture and granulation tissue formation. In addition, its use improved contour deformities and served as a bridge to autologous skin grafting, minimizing donor site size and morbidity. CONCLUSIONS: While autologous skin grafting is necessary for final wound closure, the use of cryopreserved human allograft provides biologic wound management that aids as a bridge to autologous skin grafting. As such, the authors advocate its use as a tissue scaffold in the management of severe, extensive HS and other dermatologic conditions requiring skin excision.

Xerostomia - A Comprehensive Review with a Focus on Mid-Life Health.

Xerostomia is defined as the subjective complaint of a dry mouth. Xerostomia is common in menopausal women owing to the hormonal changes which take place during midlife. Studies show a higher incidence of oral cancer in the postmenopausal period which substantiates the theory of estrogen deficiency in carcinogenesis. Radiotherapy in the treatment of these cancers can lead to oral dryness. Other etiological factors of xerostomia include systemic diseases commonly occurring in middle-aged individuals and xerogenic drugs. Saliva plays a pivotal role in the maintenance of oropharyngeal health and xerostomia can severely impair the quality of life. The aim of this review was to provide vital information pertaining to the etiology, signs, diagnosis, and treatment of xerostomia with an emphasis on midlife health. The articles for this review were obtained from PubMed Central, Google Scholar, EBSCO, Science Direct, Medknow, Scopus, EMBASE, Web of Science, and authorized textbooks published between 1988 and 2021.

DSP107 combines inhibition of CD47/SIRPα axis with activation of 4-1BB to trigger anticancer immunity.

BACKGROUND: Treatment of Diffuse Large B Cell Lymphoma (DLBCL) patients with rituximab and the CHOP treatment regimen is associated with frequent intrinsic and acquired resistance. However, treatment with a CD47 monoclonal antibody in combination with rituximab yielded high objective response rates in patients with relapsed/refractory DLBCL in a phase I trial. Here, we report on a new bispecific and fully human fusion protein comprising the extracellular domains of SIRPα and 4-1BBL, termed DSP107, for the treatment of DLBCL. DSP107 blocks the CD47:SIRPα 'don't eat me' signaling axis on phagocytes and promotes innate anticancer immunity. At the same time, CD47-specific binding of DSP107 enables activation of the costimulatory receptor 4-1BB on activated T cells, thereby, augmenting anticancer T cell immunity. METHODS: Using macrophages, polymorphonuclear neutrophils (PMNs), and T cells of healthy donors and DLBCL patients, DSP107-mediated reactivation of immune cells against B cell lymphoma cell lines and primary patient-derived blasts was studied with phagocytosis assays, T cell activation and cytotoxicity assays. DSP107 anticancer activity was further evaluated in a DLBCL xenograft mouse model and safety was evaluated in cynomolgus monkey. RESULTS: Treatment with DSP107 alone or in combination with rituximab significantly increased macrophage- and PMN-mediated phagocytosis and trogocytosis, respectively, of DLBCL cell lines and primary patient-derived blasts. Further, prolonged treatment of in vitro macrophage/cancer cell co-cultures with DSP107 and rituximab decreased cancer cell number by up to 85%. DSP107 treatment activated 4-1BB-mediated costimulatory signaling by HT1080.4-1BB reporter cells, which was strictly dependent on the SIRPα-mediated binding of DSP107 to CD47. In mixed cultures with CD47-expressing cancer cells, DSP107 augmented T cell cytotoxicity in vitro in an effector-to-target ratio-dependent manner. In mice with established SUDHL6 xenografts, the treatment with human PBMCs and DSP107 strongly reduced tumor size compared to treatment with PBMCs alone and increased the number of tumor-infiltrated T cells. Finally, DSP107 had an excellent safety profile in cynomolgus monkeys. CONCLUSIONS: DSP107 effectively (re)activated innate and adaptive anticancer immune responses and may be of therapeutic use alone and in combination with rituximab for the treatment of DLBCL patients.

Duplex Shiny app quantification of the sepsis biomarkers C-reactive protein and interleukin-6 in a fast quantum dot labeled lateral flow assay.

Fast point-of-care (POC) diagnostics represent an unmet medical need and include applications such as lateral flow assays (LFAs) for the diagnosis of sepsis and consequences of cytokine storms and for the treatment of COVID-19 and other systemic, inflammatory events not caused by infection. Because of the complex pathophysiology of sepsis, multiple biomarkers must be analyzed to compensate for the low sensitivity and specificity of single biomarker targets. Conventional LFAs, such as gold nanoparticle dyed assays, are limited to approximately five targets-the maximum number of test lines on an assay. To increase the information obtainable from each test line, we combined green and red emitting quantum dots (QDs) as labels for C-reactive protein (CRP) and interleukin-6 (IL-6) antibodies in an optical duplex immunoassay. CdSe-QDs with sharp and tunable emission bands were used to simultaneously quantify CRP and IL-6 in a single test line, by using a single UV-light source and two suitable emission filters for readout through a widely available BioImager device. For image and data processing, a customized software tool, the MultiFlow-Shiny app was used to accelerate and simplify the readout process. The app software provides advanced tools for image processing, including assisted extraction of line intensities, advanced background correction and an easy workflow for creation and handling of experimental data in quantitative LFAs. The results generated with our MultiFlow-Shiny app were superior to those generated with the popular software ImageJ and resulted in lower detection limits. Our assay is applicable for detecting clinically relevant ranges of both target proteins and therefore may serve as a powerful tool for POC diagnosis of inflammation and infectious events.

Successful peripheral nerve field stimulation for thoracic radiculitis following Brown-Sequard syndrome.

OBJECTIVE: The objective of this study is to present a novel approach to the treatment of thoracic radiculitis following Brown-Sequard syndrome with peripheral nerve field stimulation (PNFS). Furthermore, we endeavor to discuss the role of PNFS in the management of refractory neuropathic pain conditions including post-traumatic and post-surgical neuropathy particularly with regards to the post-surgical spine. MATERIALS AND METHODS: Presented is a 57-year-old man with history of thoracic microdiscectomy resulting in Brown-Sequard syndrome presented with chronic post-operative thoracic radicular pain radiating to the abdomen, refractory to conservative management. The patient underwent three intercostal nerve blocks from T7 to T9 with transient symptomatic relief. The patient's options were limited to chemomodulation, neuromodulation, or selective intercostal nerve surgical neurectomy. He subsequently underwent a PNFS trial and reported >75% pain reduction. Permanent percutaneous PNFS electrodes were implanted subcutaneously over the right T7 and T9 intercostal nerves and replicated the trial results. RESULTS: Neuromodulation produced pain relief with >90% improvement in pain compared with baseline both during the trial and following permanent implantation of the PNFS system. CONCLUSION: Chronic radicular pain may be difficult to manage in the post-surgical patient and often requires the use of multiple therapeutic modalities. In this case, we successfully utilized PNFS as it demonstrated greater technical feasibility when compared with dorsal column stimulation and repeat surgery; therefore, it may be considered for the management of post-surgical neuropathy. Further controlled studies are needed to evaluate the efficacy of PNFS as a treatment option.

Outcomes of reconstructive surgery in pediatric oncology patients: review of 10-year experience.

BACKGROUND: The purpose of our study was to review the role of reconstructive surgery in the management of pediatric oncology patients and to assess patients' outcomes, including functional status. METHODS: We evaluated 177 children with cancer who underwent reconstructive surgery at our institution between 1999 and 2008. RESULTS: The mean age was 12.1 years, and the mean follow-up duration was 27.3 months. The most common tumor pathology was sarcoma (49.7%), and the most common reconstruction site was the head and neck (41.8%). Nearly half of all patients underwent preoperative (44.1%) and/or postoperative (45.8%) chemotherapy. Immediate reconstruction was performed in 84.7% of patients. Free tissue transfer (33.9%) was the most common form of reconstruction, and the fibula flap (58.4%) was the most common free flap used. Additional surgery (for any reason) was required in 41.8% of patients. In general, functional outcomes were excellent: 78.4% of head and neck reconstruction patients tolerated a regular diet postoperatively, 72.0% of upper extremity reconstruction patients experienced no postoperative functional deficits, and 70.6% of lower extremity reconstruction patients achieved ambulatory status (without assistance). Of the 177 patients, 74.6% had no evidence of disease at last known follow-up. CONCLUSIONS: Reconstructive surgery in children with cancer is complex and often requires multiple procedures, although treatment usually results in excellent functional outcomes. A multidisciplinary approach is essential in the treatment of these patients, who are still in their growth phase, to optimize their functional capacity, quality of life, and overall survival.