“…2 The magnitude of the response and range of secreted cytokines depends on crystal’s chemistry. To assess the ability of natural human kidney stones to induce secretion of cytokines and chemokines in human macrophages, we employed the Proteome Profiler Antibody Arrays that allow detecting multiple proteins simultaneously.…”
Section: Resultsmentioning
confidence: 99%
“…Inorganic crystalline material such as CaOx can induce strong immune responses in human macrophages – responses that are directly controlled by crystal chemistry. 2 Macrophages including renal tissue macrophages scavenge dead cells through phagocytosis and are critical to maintaining healthy kidneys. 3–5 In addition, experimental studies in hyperoxaluric rodents show that renal macrophages frequently surround CaOx crystals that have migrated into the interstitium.…”
Purpose
In murine and human hyperoxaluric conditions, macrophages can be seen surrounding renal calcium oxalate (CaOx) crystal deposits. We hypothesize that macrophages play a role in degrading and destroying these deposits and investigated inflammatory response and phagocytic mechanisms when macrophages are exposed to human kidney stones and inorganic crystals.
Materials and Methods
Human monocytes were differentiated into resting, fully-differentiated macrophages by treating with recombinant human M-CSF or GM-CSF for 6 days. After confirming phenotype by flow cytometry, macrophages were exposed for 20 hours to fragments of sterile human CaOx stones or CaOx crystals. Crystal uptake was determined, and supernatant cytokine and chemokine profiles were analyzed using antibody arrays. qRT-PCR was used to validate mRNA profile expression.
Results
Under direct-vision fluorescent microscopy, activated human macrophages were noted to surround both stone fragments and synthesized crystals and destroy them in a step-by-step process that involved clathrin-mediated endocytosis and phagocytosis. An inflammatory cascade was released by macrophages, including chemokines CCL2, CCL3, interleukin-1 receptor antagonist (IL-1ra), complement component C5/C5a and IL-8. The response patterns to stone and crystal material was dependent on macrophage phenotype and activation status.
Conclusions
In our in vitro study, macrophages differentiated with M-CSF displayed a greater ability to phagocytize crystal deposits than those treated with GM-CSF. Following clathrin-mediated endocytosis, macrophages released a number of cytokines crucial for inflammatory immune response, suggesting that tissue macrophages play an important role in preventing kidney stone disease by removing and digesting interstitial renal crystal deposits.
“…2 The magnitude of the response and range of secreted cytokines depends on crystal’s chemistry. To assess the ability of natural human kidney stones to induce secretion of cytokines and chemokines in human macrophages, we employed the Proteome Profiler Antibody Arrays that allow detecting multiple proteins simultaneously.…”
Section: Resultsmentioning
confidence: 99%
“…Inorganic crystalline material such as CaOx can induce strong immune responses in human macrophages – responses that are directly controlled by crystal chemistry. 2 Macrophages including renal tissue macrophages scavenge dead cells through phagocytosis and are critical to maintaining healthy kidneys. 3–5 In addition, experimental studies in hyperoxaluric rodents show that renal macrophages frequently surround CaOx crystals that have migrated into the interstitium.…”
Purpose
In murine and human hyperoxaluric conditions, macrophages can be seen surrounding renal calcium oxalate (CaOx) crystal deposits. We hypothesize that macrophages play a role in degrading and destroying these deposits and investigated inflammatory response and phagocytic mechanisms when macrophages are exposed to human kidney stones and inorganic crystals.
Materials and Methods
Human monocytes were differentiated into resting, fully-differentiated macrophages by treating with recombinant human M-CSF or GM-CSF for 6 days. After confirming phenotype by flow cytometry, macrophages were exposed for 20 hours to fragments of sterile human CaOx stones or CaOx crystals. Crystal uptake was determined, and supernatant cytokine and chemokine profiles were analyzed using antibody arrays. qRT-PCR was used to validate mRNA profile expression.
Results
Under direct-vision fluorescent microscopy, activated human macrophages were noted to surround both stone fragments and synthesized crystals and destroy them in a step-by-step process that involved clathrin-mediated endocytosis and phagocytosis. An inflammatory cascade was released by macrophages, including chemokines CCL2, CCL3, interleukin-1 receptor antagonist (IL-1ra), complement component C5/C5a and IL-8. The response patterns to stone and crystal material was dependent on macrophage phenotype and activation status.
Conclusions
In our in vitro study, macrophages differentiated with M-CSF displayed a greater ability to phagocytize crystal deposits than those treated with GM-CSF. Following clathrin-mediated endocytosis, macrophages released a number of cytokines crucial for inflammatory immune response, suggesting that tissue macrophages play an important role in preventing kidney stone disease by removing and digesting interstitial renal crystal deposits.
“…Nanoparticles have many diverse potential applications, and a large number of nanoparticle groups have been developed including: viral carriers3, organic cationic compounds4, recombinant proteins5 and inorganic nanoparticles6. In all cases the selection of a suitable nanotransporter is crucial for successful application for specific target cells7.…”
Here we report a powerful method that facilitates the transport of biologically active materials across the cell wall barrier in plant cells. Positively charged delaminated layered double hydroxide lactate nanosheets (LDH-lactate-NS) with a 0.5‒2 nm thickness and 30‒60 nm diameter exhibit a high adsorptive capacity for negatively charged biomolecules, including fluorescent dyes such as tetramethyl rhodamine isothiocyanate (TRITC), fluorescein isothiocyanate isomer I(FITC) and DNA molecules, forming neutral LDH-nanosheet conjugates. These neutral conjugates can shuttle the bound fluorescent dye into the cytosol of intact plant cell very efficiently. Furthermore, typical inhibitors of endocytosis and low temperature incubation did not prevent LDH-lactate-NS internalization, suggesting that LDH-lactate-NS penetrated the plasma membrane via non-endocytic pathways, which will widen the applicability to a variety of plant cells. Moreover, the absence of unwanted side effects in our cytological studies, and the nuclear localization of ssDNA-FITC suggest that nano-LDHs have potential application as a novel gene carrier to plants.
“…The production of a range of different cytokines and co‐stimulatory molecules was measured, with mean results shown as black squares and 95% confidence intervals as bars. The red triangles indicate the response predicted mathematically; it is clear that these lie within the confidence intervals in the majority of cases. Reproduced with permission from Williams et al [159]. Copyright R ockefeller U niversity P ress 2014.…”
Section: Inorganic Adjuvant Systemsmentioning
confidence: 94%
“…A series of layered double hydroxide materials was prepared, and the response of human dendritic cells in co‐culture or of mice upon OVA vaccination quantified. The physicochemical properties of the layered double hydroxides were then considered to be causative of the immune response, and structure/property relationships were elucidated. Reproduced with permission from Williams et al [159]. Copyright R ockefeller U niversity P ress 2014.…”
Objectives To review the state of the art and assess future potential in the use of inorganic particulates as vaccine adjuvants. Key findings An adjuvant is an entity added to a vaccine formulation to ensure that robust immunity to the antigen is inculcated. The inclusion of an adjuvant is typically vital for the efficacy of vaccines using inactivated organisms, subunit and DNA antigens. With increasing research efforts being focused on subunit and DNA antigens because of their improved safety profiles, the development of appropriate adjuvants is becoming ever more crucial. Despite this, very few adjuvants are licensed for use in humans (four by the FDA, five by the European Medicines Agency). The most widely used adjuvant, alum, has been used for nearly 90 years, yet its mechanism of action remains poorly understood. In addition, while alum produces a powerful antibody Th2 response, it does not provoke the cellular immune response required for the elimination of intracellular infections or cancers. New adjuvants are therefore needed, and inorganic systems have attracted much attention in this regard. Summary In this review, the inorganic adjuvants currently in use are considered, and the efforts made to date to understand their mechanisms of action are summarised. We then move on to survey the literature on inorganic particulate adjuvants, focusing on the most interesting recent developments in this area and their future potential.
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