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Background: Interferon (IFN)-β is an effective therapy for relapsing-remitting multiple sclerosis, yet its mechanism of action remains ill-defined.Objectives: Our objective was to characterize the role of IFN-β in immune regulation in experimental autoimmune encephalomyelitis (EAE).Methods: IFN-β +/+ and IFN-β—/— mice were immunized with myelin oligodendrocyte glycoprotein peptide in the presence or absence of IFN-β, to induce EAE. Disease pathogenesis was monitored in the context of incidence, time of onset, clinical score, and immune cell activation in the brains, spleens and lymph nodes of affected mice.Results: Compared with IFN-β+/+ mice, IFN-β—/ — mice exhibited an earlier onset and a more rapid progression of EAE, increased numbers of CD11b+ leukocytes infiltrating affected brains and an increased percentage of Th17 cells in the central nervous system and draining lymph nodes. IFN-β treatment delayed disease onset and reduced disease severity. Ex vivo experiments revealed that the lack of IFN-β results in enhanced generation of autoreactive T cells, a likely consequence of the absence of IFN-β-regulated events in both the CD4+ T cells and antigen-presenting dendritic cells. Gene expression analysis of IFN-β-treated bone marrow macrophages (CD11b +) identified modulation of genes affecting T cell proliferation and Th17 differentiation.Conclusions: We conclude that IFN-β acts to suppress the generation of autoimmune-inducing Th17 cells during the development of disease as well as modulating pro-inflammatory mediators. [ABSTRACT FROM AUTHOR]

Home health care (HHC) is growing rapidly and yet health and safety conditions of HHC clinicians are poorly understood. Study of this workforce presents unique challenges because it is decentralised, often part-time and mobile. As part of a larger project on sharps injuries and blood exposures in HHC, this paper addresses the challenges of recruiting a large cohort of HHC nurses and aides and describes novel cross-sectional survey methodology. Recruitment was conducted with cooperation from eight HHC agencies and two labour unions. Intensive personal contacts and a financial incentive ($25) were employed. Some groups of HHC clinicians could be contacted only by mail, while others were contacted during a promotional 'mini-fair' at their agency. A total of 1772, 18-page health and safety surveys were distributed and 1225 usable surveys were collected. This 69% overall response rate is better than that in many recent health surveys. Survey returns were highest (67DS91%) where promotional events were held. The mailing-only strategy generated lower response rates (53DS55%), despite the same financial incentive. Despite the challenges of reaching out to the decentralised HHC workforce, adequate response to a detailed health survey is possible, using appropriate techniques and with the close cooperation of employers and labour unions. [ABSTRACT FROM AUTHOR]

Interferon (IFN)-α and IFN-β are critical mediators of host defense against microbial challenges, directly interfering with viral infection and influencing both the innate and adaptive immune responses. IFNs exert their effects in target cells through the activation of a cell-surface receptor, leading to a cascade of signaling events that determine transcriptional and translation regulation. Understanding the circuitry associated with IFN-mediated signal transduction that leads to a specific biological outcome has been a major focus of our laboratory. Through the efforts of graduate students, postdoctoral fellows, a skilled research technologist, and important collaborations with investigators elsewhere, we have provided some insights into the complexity of the IFN system—and the elegance and simplicity of how protein-protein interactions define biological function. [ABSTRACT FROM AUTHOR]

Background: Recombinant human interleukin-2 (rhIL-2, aldesleukin) is Food and Drug Administration approved for the treatment of metastatic melanoma and renal cell carcinoma and has achieved durable response in a subset of patients. However, its utility as an immunotherapeutic drug is limited by undesirable activation of immune suppressive regulatory T cells (Tregs) and a short half-life requiring frequent high dose administration, leading to unacceptable toxicities. We have engineered MDNA11, a long-acting IL-2 superkine, to overcome these limitations by (1) modifying receptor selectivity in favor of anti-cancer immune cells to increase therapeutic efficacy and (2) fusion to human albumin to extend the pharmacokinetic (PK) profile, circumventing the need for frequent dosing., Methods: MDNA11 was evaluated using in vitro and in vivo studies including: binding analyses to measure receptor affinity, IL-2 pathway signaling, PK studies in mice, and efficacy studies in syngeneic tumor models as single agent and in combination with immune checkpoint inhibitors. Finally, the safety and pharmacodynamic profile of MDNA11 was assessed in non-human primate (NHP)., Results: Binding studies with MDNA11 demonstrated increased affinity for IL-2Rβ (CD122) and no binding to IL-2Rα (CD25). As a result, MDNA11 exhibits reduced/limited Treg stimulation while triggering an enhanced activation of natural killer and naïve CD8 T cells compared with rhIL-2. When administered to animals with pre-established tumors, MDNA11 controlled tumor growth in a monotherapy setting and in combination with anti-PD1 or anti-CTLA4 to induce durable tumor clearance with a once weekly dosing regimen. In a NHP model, MDNA11 was well tolerated while triggering durable and potent immune responses including expansion of lymphocytes without significant effect on Tregs and eosinophils, the latter been linked to an increased risk of vascular leak syndrome., Conclusion: MDNA11 is a next generation long-acting IL-2 immunotherapeutic with a highly favorable pharmacodynamic profile that translates to a strong therapeutic efficacy in preclinical tumor models and a strong and durable immune response in NHP., Competing Interests: Competing interests: RM, CG, MAM, FM and MDT are employees of Medicenna and declare competing financial interest. LBP, PL and PS are consultants and were compensated by Medicenna for their work on the studies in this paper., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: Home care (HC) services are crucial to the health and social wellbeing of older adults, people with disabilities, and the chronically ill. Although the HC sector is growing rapidly in the USA, there is high job turnover among the HC aide workforce. HC provides an important alternative to facility-based care, yet it has often been overlooked within the larger health care system: most recently, in COVID-19 pandemic planning. The objective of the study was to characterize qualitatively the impact of the COVID-19 pandemic on three key HC stakeholders: clients, aides, and agency managers., Methods: The study included 37 phone interviews conducted during April - November 2020: HC clients (n = 9), aides (n = 16), and agency managers (n = 12). All interviews were audio recorded and transcribed verbatim. Qualitative analysis of the transcripts followed the grounded theory approach. The interview transcriptions were coded line-by-line into hierarchical themes with NVivo 12 software which allowed weighting of themes based on the number of interviews where they were coded., Results: Fear of infection and transmission among HC clients and aides were strong themes. Infection prevention and control became the top priority guiding day-to-day business operations at agencies; sourcing adequate personal protective equipment for staff was the most urgent task. HC aides expressed concerns for their clients who showed signs of depression, due to increased isolation during the pandemic. The disappearance of comforting touch - resulting from physical distancing practices - altered the expression of compassion in the HC aide-client care relationship., Conclusions: The findings suggest that the pandemic has further increased psychosocial job demands of HC aides. Increased isolation of clients may be contributing to a wider public health problem of elder loneliness and depression. To support the HC stakeholders during the on-going COVID-19 pandemic, for future pandemic planning or other health emergencies, it is important to improve HC aide job retention. This action could also ease the serious care services shortage among the growing population of older adults., (© 2021. The Author(s).)

Single-molecule and single-cell analysis techniques have opened new opportunities for characterizing and analyzing heterogeneity within biological samples. These detection methods are often referred to as digital assays because the biological sample is partitioned into many small compartments and each compartment contains a discrete number of targets (e.g. cells). Using digital assays, researchers can precisely detect and quantify individual targets, and this capability has made digital techniques the basis for many modern bioanalytical tools (including digital PCR, single cell RNA sequencing, and digital ELISA). However, digital assays are dominated by optical analysis systems that typically utilize microscopy to analyze partitioned samples. The utility of digital assays may be dramatically enhanced by implementing cost-efficient and portable electrical detection capabilities. Herein, we describe a digital electrical impedance sensing platform that enables direct multiplexed measurement of single cell bacterial cells. We outline our solutions to the challenge of multiplexing impedance sensing across many culture compartments and demonstrate the potential for rapidly differentiating antimicrobial resistant versus susceptible strains of bacteria.

The Adisintegrin and metalloprotease domain-containing (ADAM) family of proteins is involved in cell adhesion, migration, proteolysis, and signaling. Many ADAMs are required for reproduction; however, the role of Adam6 has remained largely unknown. In the course of humanizing the mouse immunoglobulin heavy chain (IgH) locus, we generated Adam6-deficient mice that demonstrate severe subfertility. We decided to elucidate the role of ADAM6 in fertility and explore the underlying mechanisms. Despite normal sperm development and motility, Adam6-deficient mice display diminished male fertility, have abnormal sperm adhesion, and most importantly cannot transition from uterus to oviduct. To test whether ADAM6 is required for sperm's binding to extracellular matrix (ECM) components, we used a panel of ECM components and showed that unlike normal sperm, Adam6-deficient sperm cannot bind fibronectin, laminin, and tenascin. Reintroduction of Adam6 into these deficient mice repaired sperm interaction with ECM, restored male fertility, and corrected the sperm transport deficit. Together, our data suggest that ADAM6, either alone or in complex with other proteins, aids sperm transport through the female reproductive tract by providing a temporary site of attachment of sperm to ECM components prior to ascent into the oviduct., (© The Author(s) 2018. Published by Oxford University Press on behalf of Society for the Study of Reproduction.)

Test disk electrodes were fabricated from carbon nanotubes (CNT) using the Carbon Nanotube Templated Microfabrication (CNT-M) technique. The CNT-M process uses patterned growth of carbon nanotube forests from surfaces to form complex patterns, enabling electrode sizing and shaping. The additional carbon infiltration process stabilizes these structures for further processing and handling. At a macroscopic scale, the electrochemical, electrical and magnetic properties, and magnetic resonance imaging (MRI) characteristics of the disk electrodes were investigated; their microstructure was also assessed. CNT disk electrodes showed electrical resistivity around 1 Ω·cm, charge storage capacity between 3.4 and 38.4 mC/cm 2 , low electrochemical impedance and magnetic susceptibility of -5.9 to -8.1 ppm, closely matched to that of tissue (∼-9 ppm). Phantom MR imaging experiments showed almost no distortion caused by these electrodes compared with Cu and Pt-Ir reference electrodes, indicating the potential for significant improvement in accurate tip visualization., (Copyright © 2018 Elsevier Inc. All rights reserved.)

We present methods to fabricate high-capacity redox electrodes using thick membrane or fiber casting of conjugated polymer solutions. Unlike common solution casting or printing methods used in current organic electronics, the presented techniques enable production of PEDOT:PSS electrodes with high charge capacity and the capability to operate under applied voltages greater than 100 V without electrochemical overoxidation. The electrodes are shown integrated into several electrokinetic components commonly used in automated bioprocess or bioassay workflows, including electrophoretic DNA separation and extraction, cellular electroporation/lysis, and electroosmotic pumping. Unlike current metal electrodes used in these applications, the high-capacity polymer electrodes are shown to function without electrolysis of solvent (i.e., without production of excess H + , OH - , and H 2 O 2 by-products). In addition, each component fabricated using the electrodes is shown to have superior capabilities compared with those fabricated with common metal electrodes. These innovations in electrokinetics include a low-voltage/high-pressure electroosmotic pump, and a "flow battery" (in which electrochemical discharge is used to generate electroosmotic flow in the absence of an applied potential). The novel electrodes (and electrokinetic demonstrations) enable new applications of organic electronics within the biology, health care, and pharmaceutical fields.

Home care aides risk musculoskeletal injury because they lift and move clients; the body weight of most adults exceeds the NIOSH recommended limit for lifting. Methods to reduce manual patient lifting in institutional settings are often technically or economically infeasible in home care. Our goal was to identify suitable, safe, low-technology transfer devices for home care use. Sixteen experienced home care aides performed client transfers from wheelchair to bed (upward) and bed to wheelchair (downward) in a simulated home care environment (laboratory), using four different slide boards and by hand without a device. Aides' hand forces were measured during client transfers; aides also evaluated usability of each board. Hand forces exerted while using slide boards were mostly lower than in manual transfer, and forces were lower in downward versus upward transfers. Aides judged a board with a sliding mechanism easier to use than boards without a sliding mechanism. Practitioner Summary: This paper provides quantitative biomechanical measurements showing that slide boards reduced the hand forces needed by home care aides to transfer clients from bed to wheel chair and vice versa, compared to manual lifting. Using a semi-quantitative usability survey, aides identified boards with a sliding mechanism easiest to use.

Microelectrode arrays of carbon nanotube (CNT)/carbon composite posts with high aspect ratio and millimeter-length were fabricated using carbon-nanotube-templated microfabrication with a sacrificial "hedge". The high aspect ratio, mechanical robustness, and electrical conductivity of these electrodes make them a potential candidate for next-generation neural interfacing. Electrochemical measurements were also demonstrated using an individual CNT post microelectrode with a diameter of 25 μm and a length of 1 mm to perform cyclic voltammetry on both methyl viologen and dopamine in a phosphate-buffered saline solution. In addition to detection of the characteristic peaks, the CNT post microelectrodes show a fast electrochemical response, which may be enabling for in vivo and/or in vitro measurements. The CNT post electrode fabrication process was also integrated with other microfabrication techniques, resulting in individually addressable electrodes.

Background: Home care aides perform personal care and homemaking services in client homes, including cleaning and disinfection (C&D). Although C&D are performed to remove soil and dust, they are increasingly performed for infection prevention. Many C&D products contain respiratory irritants. The objective of this study was to evaluate 2 commercial products for C&D effectiveness on common household surfaces in seniors' homes., Methods: Two C&D visits were conducted in 46 seniors' homes. One visit applied a bleach-containing cleaning product and the other applied an environmentally preferable product. Before and after C&D, the study team performed organic soil bioluminometer measurements on surfaces and collected cotton swab and wipe samples for total bacteria count, Staphylococcus aureus, and Clostridium difficile identification., Results: Both products removed microorganisms from tested surfaces. S aureus was found in 7 households, 1 strain of which was methicillin-resistant. Both products removed S aureus from all surfaces. Bleach-containing products removed somewhat more soil than environmentally preferable products, although results were statistically significant for only 1 surface., Conclusions: The study showed similar, not identical, C&D performance for 2 cleaning products with potentially different consequences for respiratory health. Additional research is needed to develop robust recommendations for safe, effective C&D in home care., (Copyright © 2018 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.)

In the United States, home health care (HHC) is a rapidly growing industry and home infusion therapy is a rapidly growing market. HHC can present substantial occupational safety and health (OSH) risks. This article summarizes major OSH risks relevant to home infusion therapy by illustrating them through real-life scenarios collected systematically using qualitative research methods by the National Institute for Occupational Safety and Health-funded research projects at the University of Massachusetts Lowell. The need for home infusion therapy will continue to grow in the future, and safety interventions to prevent or minimize OSH risks are essential.

Background: Nurses in the cardiac intensive care unit often attend professional education opportunities. However, it is difficult to share this information among nursing staff. Varying schedules, different shifts, and patient acuity limit the amount of time available for peer-to-peer sharing of educational information. A review of the literature revealed scant research on blogging for peer-to-peer education in general and particularly in nursing., Objectives: To explore nurses' perception of the effectiveness of using a blog as a forum to provide peer-to-peer sharing of relevant professional education., Methods: Using a simple, free blogging website, the unit's nursing practice council developed a private blog for educational information sharing among the nursing staff. An online survey was administered to the unit's staff 15 months after the blog was implemented., Results: Most respondents indicated that they thought the blog is an effective way to share professional education (86%), keeps them abreast of evidence-based practice (81%), and has led to practice change (59%). Nearly 80% of respondents agreed that they are more likely to attend professional conferences, and 62% would consider contributing blog posts., Conclusion: The survey results suggest that blogging may be an effective method of peer-to-peer sharing of education, although more rigorous research is required in this area., (©2017 American Association of Critical-Care Nurses.)

Background: Home healthcare is one of the fastest growing sectors in the United States. Percutaneous injuries from sharp medical devices (sharps) are a source of bloodborne pathogen infections among home healthcare workers and community members. Sharps use and disposal practices in the home are highly variable and there is no comprehensive analysis of the system of sharps procurement, use and disposal in home healthcare. This gap is a barrier to effective public health interventions. The objectives of this study were to i) identify the full range of pathways by which sharps enter and exit the home, stakeholders involved, and barriers for using sharps with injury prevention features; and ii) assess the leverage points for preventive interventions., Methods: This study employed qualitative research methods to develop two systems maps of the use of sharps and prevention of sharps injuries in home healthcare. Twenty-six in-depth interview sessions were conducted including home healthcare agency clinicians, public health practitioners, sharps device manufacturers, injury prevention advocates, pharmacists and others. Interview transcripts were audio-recorded and analyzed thematically using NVIVO qualitative research analysis software. Analysis of supporting archival material also was conducted. All findings guided development of the two maps., Results: Sharps enter the home via multiple complex pathways involving home healthcare providers and home users. The providers reported using sharps with injury prevention features. However, home users' sharps seldom had injury prevention features and sharps were commonly re-used for convenience and cost-savings. Improperly discarded sharps present hazards to caregivers, waste handlers, and community members. The most effective intervention potential exists at the beginning of the sharps systems maps where interventions can eliminate or minimize sharps injuries, in particular with needleless treatment methods and sharps with injury prevention features. Manufacturers and insurance providers can improve safety with more affordable and accessible sharps with injury prevention features for home users. Sharps disposal campaigns, free-of-charge disposal containers, and convenient disposal options remain essential., Conclusions: Sharps injuries are preventable through public health actions that promote needleless treatment methods, sharps with injury prevention features, and safe disposal practices. Communication about hazards regarding sharps is needed for all home healthcare stakeholders.

Background: Home care (HC) aide is the fastest growing occupation, yet job hazards are under-studied. This study documents the context of HC aide work, characterizes occupational safety and health (OSH) hazards, and identifies preventive interventions using qualitative methods., Methods: We conducted 12 focus groups among aides and 26 in-depth interviews comprising 15 HC agency, union, and insurance company representatives as well as 11 HC recipients in Massachusetts. All focus groups and interviews were audio-recorded, transcribed, and coded with NVIVO software., Results: Major OSH concerns were musculoskeletal disorders from client care tasks and verbal abuse. Performing tasks beyond specified job duties may be an OSH risk factor. HC aides' safety and clients' safety are closely linked. Client handling devices, client evaluation, care plan development, and training are key interventions for both aides' and clients' safety., Conclusions: Promoting OSH in HC is essential for maintaining a viable workforce., (© 2013 Wiley Periodicals, Inc.)

Monocyte chemoattractant protein-1 (MCP-1)/CCL2 is a chemokine regulating the recruitment of monocytes into sites of inflammation and cancer. MCP-1 can be produced by a variety of cell types, such as macrophages, neutrophils, fibroblasts, endothelial cells, and epithelial cells. Notably, macrophages produce high levels of MCP-1 in response to proinflammatory stimuli in vitro, leading to the hypothesis that macrophages are the major source of MCP-1 during inflammatory responses in vivo. In stark contrast to the hypothesis, however, there was no significant reduction in MCP-1 protein or the number of infiltrating macrophages in the peritoneal inflammatory exudates of myeloid cell-specific MCP-1-deficient mice in response to i.p injection of thioglycollate or zymosan A. Furthermore, injection of LPS into skin air pouch also had no effect on local MCP-1 production in myeloid-specific MCP-1-deficient mice. Finally, myeloid-specific MCP-1-deficiency did not reduce MCP-1 mRNA expression or macrophage infiltration in LPS-induced lung injury. These results indicate that non-myeloid cells, in response to a variety of stimulants, play a previously unappreciated role in innate immune responses as the primary source of MCP-1.

MCP-1/CCL2 plays a critical role in monocyte recruitment into sites of immune responses and cancer. However, the role of other MCPs remains unclear. In this study, we generated a novel MCP-1-deficient (designated as MCP-1(Delta/Delta)) mouse model by deleting a 2.3-kb DNA fragment from the mouse genome using the Cre/loxP system. MCP-1 was not produced by LPS-activated MCP-1(Delta/Delta) macrophages; however, the production of MCP-3, coded by the immediate downstream gene, was significantly increased. In contrast, macrophages from another mouse line with a neo-gene cassette in intron 2 produced a significantly lower level of MCP-1 and MCP-3. Decreased MCP-3 production was also detected in previously generated MCP-1-deficient mice in which a neo-gene cassette was inserted in exon 2 (designated as MCP-1 knockout (KO)). Altered MCP-1 and/or MCP-3 production was also observed in vivo in each mouse model in response to i.p. injection of thioglycolate or zymosan. The up- and down-regulation of MCP-3 production in MCP-1(Delta/Delta) and MCP-1 KO mice, respectively, provided us with a unique opportunity to evaluate the role for MCP-3. Despite the increased MCP-3 production in MCP-1(Delta/Delta) mice, thioglycolate- or zymosan-induced monocyte/macrophage accumulation was still reduced by approximately 50% compared with wild-type mice, similar to the reduction detected in MCP-1 KO mice. Thus, up-regulated MCP-3 production did not compensate for the loss of MCP-1, and MCP-3 appears to be a less effective mediator of monocyte recruitment than MCP-1. Our results also indicate the presence of other mediators regulating the recruitment of monocytes in these models.

Home health care is one of the fastest growing industries in the United States. Approximately 20,000 provider agencies deliver home health care services to 7.6 million individuals with acute illness, long-term health conditions, permanent disability, or terminal illness. The home health care setting poses many challenges that likely increase the risk of sharps injuries. Home health nurses face unique challenges in preventing and reporting sharps injuries in the home. This article examines the nature of and risk factors for sharps injuries in the home health care setting, the scope of the problem, the legislative and regulatory framework relevant to sharps injuries, and the role of occupational health nurses in promoting a culture of safety to prevent sharps injuries and bloodborne pathogen exposures.

Progressive demyelination in multiple sclerosis (MS) reflects the negative balance between myelin damage and repair due to physical and molecular barriers, such as astrocytic glial scars, between oligodendrocytes and target neurons. In this paper, we show that combination therapy with paclitaxel (Taxol) plus the universal methyl-donor, vitamin B12CN (B12CN), dramatically limits progressive demyelination, and enhances remyelination in several independent, immune and nonimmune, in vivo and in vitro model systems. Combination therapy significantly reduced clinical signs of EAE in SJL mice, as well as the spontaneously demyelinating ND4 transgenic mouse. Astrocytosis was normalised in parallel to ultrastructural and biochemical evidence of remyelination. The combination therapy suppressed T cell expansion, reduced IFN-gamma, while enhancing IFN-beta and STAT-1 expression, STAT-1 phosphorylation and methylation of STAT-1 and MBP in the brain. Paclitaxel/B12CN has nearly identical effects to the previously described combination of IFN-beta/ B12CN, whose clinical usefulness is transient because of IFN-neutralising antibodies, not observed (or expected) with the present drug combination. This report provides a mechanistic foundation for the development of a new therapeutic strategy in humans with MS.

Tumour necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a member of the TNF superfamily, which is capable of inducing apoptosis in many cell types, including tumour and virus-infected cells, but rarely in normal cells. Expression of TRAIL mRNA and TRAIL receptors has previously been detected in neutrophils; however, the expression of TRAIL protein and the regulation of TRAIL and TRAIL receptor expression in these cells remain unknown. Here we report, for the first time, that neutrophils constitutively express TRAIL protein on their cell surface and that the TRAIL protein is shed during culture. TNF-alpha is a down-regulator of TRAIL expression, whereas IFN-gamma up-regulates the expression of TRAIL. Neutrophils did not express a detectable level of TRAIL-R1 or -R4, but constitutively expressed a low, but substantial, level of TRAIL-R2 and a high level of TRAIL-R3. Although the level of TRAIL-R2 was not significantly altered during culture under different experimental conditions, approximately 30% of TNF-alpha-treated cells rapidly lost their high-level TRAIL-R3 expression, whereas the majority of IFN-gamma-treated cells retained a high level of TRAIL-R3 expression. Anti-TRAIL neutralizing antibody significantly inhibited neutrophil apoptosis during cultures in medium alone, or in the presence of TNF-alpha or IFN-gamma. Thus, our study identified human neutrophils as a cellular source of TRAIL and suggests that neutrophil-derived TRAIL may play a role in immune surveillance. Our results also suggest a role for the TRAIL/TRAIL receptor system in neutrophil apoptosis.

Discoidin domain receptor 1 (DDR1) is a receptor tyrosine kinase activated by collagen. DDR1 is constitutively expressed in a variety of normal and transformed epithelial cells and plays a role in cell migration and differentiation through as yet unidentified signaling pathways. We previously reported inducible expression of DDR1 in human leukocytes and suggested a role for the DDR1a isoform in leukocyte migration through extracellular matrix. Here, we evaluated the contribution of DDR1 in the differentiation of the human monocytic THP-1 cells overexpressing these isoforms and of primary macrophages. Interestingly, collagen activation of DDR1b, but not DDR1a, further promoted phorbol ester-induced differentiation of THP-1 cells as determined by reduced cell proliferation and up-regulated expression of HLA-DR, CD11c, CD14, and CD40. Collagen activation of DDR1b also induced the recruitment and phosphorylation of Shc and subsequent phosphorylation of p38 mitogen-activated protein (MAP) kinase and its substrate ATF2. A p38 MAP kinase inhibitor, SB203580, completely inhibited DDR1b-mediated HLA-DR expression. Activation of DDR1 endogenously expressed on macrophages also up-regulated their HLA-DR expression in a p38 MAP kinase-dependent manner. Thus, DDR1b in response to collagen transduces signals that promote maturation/differentiation of HLA-DR-positive antigen-presenting cells and contributes to the development of adaptive immunity in a tissue microenvironment.

Although integrins are crucial for migration of leukocytes through endothelium, integrin-independent mechanisms appear to take over and mediate the migration of leukocytes through extracellular matrix (ECM) in a three-dimensional tissue microenvironment. Discoidin domain receptor (DDR) 1 is a receptor tyrosine kinase activated by collagen, the most abundant ECM protein. In the present study, we detected that peripheral blood mononuclear cells (PBMC) and polymorphonuclear neutrophils were induced to express DDR1 after incubation in RPMI 1640. The expression level of DDR1 in PBMC was increased further by stimulation with tumor necrosis factor-alpha, interleukin-1beta, granulocyte-macrophage colony-stimulating factor, lipopolysaccharide, or phytohemagglutinin, but not with interferon-gamma. In vivo, DDR1 mRNA was detectable in mononuclear leukocytes infiltrating human renal tumor tissue. Among three DDR1 isoforms, DDR1alpha was the major transcript in leukocytes. Functionally, overexpression of either DDR1alpha or DDR1beta in THP-1 cells resulted in increased adherence to collagen-coated plates in a beta1-integrin independent manner. However, only DDR1alpha-, but not DDR1beta-, overexpressing cells exhibited marked pseudopod extension and migrated successfully through three-dimensional collagen lattices. Consequently, we propose that the interaction of DDR1alpha with collagen of the ECM results in a requisite intracellular signaling that enables leukocytes to migrate in a tissue microenvironment and participate in host defense.