scholarly journals The murine cytomegalovirus MCK-2 facilitates in vivo infection transfer from dendritic cells to salivary gland acinar cells.

2021 ◽  
Author(s):  
Jiawei Ma ◽  
Kimberley Bruce ◽  
Philip G. Stevenson ◽  
Helen E. Farrell
Keyword(s):  
Dendritic Cells ◽  
Salivary Gland ◽  
Epithelial Cells ◽  
Salivary Glands ◽  
Immune Evasion ◽  
Acinar Cells ◽  
Lung Infection ◽  
Human Cmv

The cytomegaloviruses (CMVs) spread systemically via myeloid cells and demonstrate a broad tissue tropism. Human CMV (HCMV) UL128 encodes a component of the virion pentameric complex (PC) that is important for entry into epithelial cells and cell-cell spread in vitro . It possesses N-terminal amino acid sequences similar to C-C chemokines. While the species-specificity of HCMV precludes confirmation of UL128 function in vivo , UL128-like counterparts in experimental animals have demonstrated a role for salivary gland infection. How they achieve this has not been defined, although effects on monocyte tropism and immune evasion have been proposed. By tracking infected cells following lung infection, we show that although the UL128-like protein in mouse CMV (MCMV; designated MCK-2), facilitated entry into lung macrophages, it was dispensable for subsequent viremia mediated by CD11c + dendritic cells (DC) and extravasation to the salivary glands. Notably, MCK-2 was important for transfer of MCMV infection from DC to salivary gland acinar epithelial cells. Acinar cell infection of MCMVs deleted of MCK-2 was not rescued by T-cell depletion, arguing against an immune evasion mechanism for MCK-2 in the salivary glands. In contrast to lung infection, peritoneal MCMV inoculation yields a mixed monocyte/DC viremia. In this setting, MCK-2 again promoted DC-dependent infection of salivary gland acinar cells, but it was not required for monocyte-dependent spread to the lung. Thus, the action of MCK-2 in MCMV spread was specific to DC-acinar cell interaction. IMPORTANCE Cytomegaloviruses (CMVs) establish myeloid cell-associated viremias and persistent shedding from the salivary glands. In vitro studies with human CMV (HCMV) have implicated HCMV UL128 in epithelial tropism, but its role in vivo is unknown. Here we analysed how a murine CMV (CMV) protein with similar physical properties - designated MCK-2 - contributes to host colonization. We demonstrate that MCK-2 is dispensable for the initial systemic spread from primary infection sites, but within the salivary gland facilitates transfer of infection from dendritic cells (DC) to epithelial acinar cells. Virus transfer from extravasated monocytes to the lungs did not require MCK-2, indicating a tissue-specific effect. These results provide new information about how persistent viral tropism determinants operate in vivo .

scholarly journals Retrovirus-mediated Gene Transfer into Rat Salivary Gland Cells In Vitro and In Vivo

1997 ◽  
Vol 45 (11) ◽  
pp. 1533-1545 ◽  
Author(s):  
Tibor Barka ◽  
Hendrika M. van der Noen
Keyword(s):  
Enzyme Activity ◽  
Salivary Gland ◽  
Gene Transfer ◽  
Cell Line ◽  
Submandibular Gland ◽  
Acinar Cells ◽  
Retroviral Vector ◽  
Placental Alkaline Phosphatase

A retroviral vector DAP that encodes the human placental alkaline phosphatase (PLAP) and the neomycin-resistant gene was used to transduce the salivary gland-derived cell line A5 in vitro and acinar cells in rat submandibular gland in vivo. Expression of the transduced PLAP gene was established by histochemical staining for heat-resistant AP and by determination of enzyme activity. From the in vitro experiments, we concluded that the salivary gland-derived cell line A5 can be infected by the retroviral vector DAP. In the transduced cells the viral long terminal repeat (LTR) promoter was effective, and the cells expressed heat-stable PLAP which was localized mostly in the plasma membrane and could be released by treatment with bromelain or phosphatidyinositol-specific phospholipase C. A5-DAP cells secreted PLAP into the medium. Clones of A5-DAP cells expressed various levels of the enzyme. The level of enzyme activity in different clones was unrelated to growth rate. Retrograde ductal injection of the viral vector into the duct of the submandibular gland of rats resulted in integration and long-term expression of PLAP gene in acinar cells. Expression of PLAP was seen up to 25 days, the limit of the observation period. To facilitate integration of the viral DNA, cell division of acinar cells was induced by administration of the β-adrenergic agonist isoproterenol before administration of the virus. PLAP was secreted into submandibular saliva. The data support the notion that salivary glands are suitable targets for gene transfer in vivo by a retroviral vector. (J Histochem Cytochem 45:1533–1545, 1997)

scholarly journals Laminin-1 Peptides Conjugated to Fibrin Hydrogels Promote Salivary Gland Regeneration in Irradiated Mouse Submandibular Glands

2021 ◽  
Vol 9 ◽  
Author(s):  
Kihoon Nam ◽  
Harim T. dos Santos ◽  
Frank Maslow ◽  
Bryan G. Trump ◽  
Pedro Lei ◽  
...  
Keyword(s):  
Radiation Therapy ◽  
Salivary Gland ◽  
Salivary Glands ◽  
Tissue Regeneration ◽  
Secretory Function ◽  
Gland Morphogenesis ◽  
Salivary Gland Regeneration ◽  
Laminin 1

Previous studies demonstrated that salivary gland morphogenesis and differentiation are enhanced by modification of fibrin hydrogels chemically conjugated to Laminin-1 peptides. Specifically, Laminin-1 peptides (A99: CGGALRGDN-amide and YIGSR: CGGADPGYIGSRGAA-amide) chemically conjugated to fibrin promoted formation of newly organized salivary epithelium both in vitro (e.g., using organoids) and in vivo (e.g., in a wounded mouse model). While these studies were successful, the model’s usefulness for inducing regenerative patterns after radiation therapy remains unknown. Therefore, the goal of the current study was to determine whether transdermal injection with the Laminin-1 peptides A99 and YIGSR chemically conjugated to fibrin hydrogels promotes tissue regeneration in irradiated salivary glands. Results indicate that A99 and YIGSR chemically conjugated to fibrin hydrogels promote formation of functional salivary tissue when transdermally injected to irradiated salivary glands. In contrast, when left untreated, irradiated salivary glands display a loss in structure and functionality. Together, these studies indicate that fibrin hydrogel-based implantable scaffolds containing Laminin-1 peptides promote secretory function of irradiated salivary glands.

scholarly journals Rat aquaporin-5 4.3-kb 5′-flanking region differentially regulates expression in salivary gland and lung in vivo

2008 ◽  
Vol 295 (1) ◽  
pp. C111-C120 ◽  
Author(s):  
Beiyun Zhou ◽  
David K. Ann ◽  
Per Flodby ◽  
Parviz Minoo ◽  
Janice M. Liebler ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Salivary Glands ◽  
Transgene Expression ◽  
Fluorescent Protein ◽  
Acinar Cells ◽  
Regulatory Elements ◽  
Egfp Expression ◽  
Type I ◽  
Aquaporin 5

We previously cloned a 4.3-kb genomic fragment encompassing 5′-flanking regulatory elements of rat aquaporin-5 ( Aqp5) that demonstrated preferential transcriptional activity in lung and salivary cells in vitro. To investigate the ability of Aqp5 regulatory elements to direct transgene expression in vivo, transgenic (TG) mice and rats were generated in which the 4.3-kb Aqp5 fragment directed the expression of enhanced green fluorescent protein (EGFP). RT-PCR revealed relative promoter specificity for the lung and salivary glands in TG mice. Immunofluorescence microscopy showed strong EGFP expression in salivary acinar cells but not in lung type I (AT1) cells, both known sites of endogenous AQP5 expression. Similar results were obtained in TG rats generated by lentiviral transgenesis. EGFP mRNA was detected in both salivary glands and lung. Robust EGFP fluorescence was observed in frozen sections of the rat salivary gland but not in the lung or other tested tissues. The percentage of EGFP-positive acinar cells was increased in parotid and submandibular glands of TG rats receiving a chronic injection of the β-adrenergic receptor agonist isoproterenol. EGFP-positive cells in the lung that were also reactive with the AT1-cell specific monoclonal antibody VIIIB2 were identified by flow cytometry. These findings demonstrate that the 4.3-kb Aqp5 promoter/enhancer directs strong cell-specific transgene expression in salivary gland and low-level AT1 cell-specific expression in the lung. While these Aqp5 regulatory elements should be useful for functional studies in salivary glands, additional upstream or intronic cis-active elements are likely required for robust expression in the lung.

scholarly journals CpG Oligodeoxynucleotides Facilitate Delivery of Whole Inactivated H9N2 Influenza Virus via Transepithelial Dendrites of Dendritic Cells in Nasal Mucosa

2015 ◽  
Vol 89 (11) ◽  
pp. 5904-5918 ◽  
Author(s):  
Tao Qin ◽  
Yinyan Yin ◽  
Qinghua Yu ◽  
Lulu Huang ◽  
Xiaoqing Wang ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Influenza Virus ◽  
Epithelial Cells ◽  
Immune Responses ◽  
Nasal Mucosa ◽  
Nasal Epithelial Cells ◽  
H9n2 Influenza Virus ◽  
Cpg Oligodeoxynucleotides

ABSTRACTThe spread of the low-pathogenicity avian H9N2 influenza virus has seriously increased the risk of a new influenza pandemic. Although whole inactivated virus (WIV) vaccine via intranasal pathway is the effective method of blocking virus transmission, the mucosal barrier seems to be a major factor hampering its development. CpG oligodeoxynucleotides, a known adjuvant, can target downstream dendritic cells (DCs) and effectively enhance the mucosal and systemic immune responses. However, the ability of CpGs to assist H9N2 WIV in transepithelial transport remains unknown. Here,in vitroandin vivo, we showed that CpGs provided assistance for H9N2 WIV in recruiting DCs to the nasal epithelial cells (ECs) and forming transepithelial dendrites (TEDs) to capture luminal viruses. CD103+DCs participated in this process. Chemokine CCL20 from nasal ECs played a key role in driving DC recruitment and TED formation. Virus-loaded DCs quickly migrated into the draining cervical lymph nodes (CLNs) for antigen presentation. In addition, the competence of CpGs was independent of direct epithelial transport via the transcellular or paracellular pathway. Taken together, our data demonstrated that CpGs enhanced the transport of H9N2 WIV via TEDs of nasal DCs, which might be a novel mechanism for optimal adaptive immune responses.IMPORTANCEThis paper demonstrates by both anin vivoand anin vitrococulture model that CpG oligodeoxynucleotides, known as an adjuvant generally targeting downstream immune responses, also are crucial for the transport of H9N2 WIV across nasal epithelial cells (ECs) via the uptake of transepithelial dendrites (TEDs). Our results prove for the first time to our knowledge that the immune-potentiating mechanism of CpGs is based on strengthening the transepithelial uptake of H9N2 WIV in nasal mucosa. These findings provide a fresh perspective for further improvement of intranasal influenza vaccines, which are urgently needed in the face of the potential threat of H9N2 influenza.

scholarly journals Regenerating Salivary Glands in the Microenvironment of Induced Pluripotent Stem Cells

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Hitomi Ono ◽  
Aya Obana ◽  
Yu Usami ◽  
Manabu Sakai ◽  
Kanji Nohara ◽  
...  
Keyword(s):  
Stem Cells ◽  
Salivary Gland ◽  
Salivary Glands ◽  
Ips Cells ◽  
Initial Attempt ◽  
Cell Niche ◽  
Salivary Gland Regeneration ◽  
Induced Pluripotent

This report describes our initial attempt to regenerate salivary glands using induced pluripotent stem (iPS) cellsin vivoandin vitro. Glandular tissues that were similar to the adult submandibular glands (SMGs) and sublingual glands could be partially produced by the transplantation of iPS cells into mouse salivary glands. However, the tumorigenicity of iPS cells has not been resolved yet. It is well known that stem cells affect their microenvironment, known as a stem cell niche. We focused on the niche and the interaction between iPS cells and salivary gland cells in our study on salivary gland regeneration. Coculture of embryonic SMG cells and iPS cells have better-developed epithelial structures and fewer undifferentiated specific markers than monoculture of embryonic SMG cellsin vitro. These results suggest that iPS cells have a potential ability to accelerate differentiation for salivary gland development and regeneration.

The value of the twinkling artefact for the diagnosis of sialolithiasis of the large salivary glands

2017 ◽  
Vol 132 (2) ◽  
pp. 162-167 ◽  
Author(s):  
G Pabst ◽  
K Strobel ◽  
J Zehnder
Keyword(s):  
Salivary Gland ◽  
Salivary Glands ◽  
Doppler Ultrasound ◽  
Model Test ◽  
Power Doppler ◽  
Intensity Level ◽  
Colour Doppler

AbstractObjective:The imaging of stones in the salivary glands and ducts poses a challenge, even to experienced ultrasound examiners. This study investigated whether the ‘twinkling artefact’, which occurs at internal calcific foci during Doppler ultrasound examinations, is useful for detecting salivary gland stones.Methods:In a model test, 20 salivary stones were analysedin vitro, via Doppler ultrasound, with regard to their representability and the triggering of the twinkling artefact. In a follow-up study, 28 patients with sialolithiasis and food-related large salivary gland swellings were examined, using both power and colour Doppler modes, with regard to the twinkling artefact. All ultrasound examinations were performed by an experienced examiner and retrospectively graded by two experienced sonographers.Results:All stones could reliably be detected using the twinkling artefact in the model test. Twenty-seven of 28 salivary stones (96 per cent) also showed twinklingin vivo, during patient assessment. The power Doppler mode showed a significantly higher intensity level of twinkling than the colour Doppler mode (p< 0.0001).Conclusion:The twinkling artefact is a very reliable sign for the diagnosis of sialolithiasis. Power Doppler is superior to colour Doppler for detection of the twinkling artefact.

Three-dimensional Growth of Renal Epithelial Cells in Vitro: A Tool in Toxicity Testing

1993 ◽  
Vol 21 (2) ◽  
pp. 191-195 ◽  
Author(s):  
Knut-Jan Andersen ◽  
Erik Ilsø Christensen ◽  
Hogne Vik
Keyword(s):  
Epithelial Cells ◽  
Three Dimensional ◽  
Toxicity Testing ◽  
Renal Tissue ◽  
Epithelial Cell Line ◽  
Multicellular Spheroids ◽  
Renal Epithelial Cell ◽  
Renal Epithelial Cells

The tissue culture of multicellular spheroids from the renal epithelial cell line LLC-PK1 (proximal tubule) is described. This represents a biological system of intermediate complexity between renal tissue in vivo and simple monolayer cultures. The multicellular structures, which show many similarities to kidney tubules in vivo, including a vectorial water transport, should prove useful for studying the potential nephrotoxicity of drugs and chemicals in vitro. In addition, the propagation of renal epithelial cells as multicellular spheroids in serum-free culture may provide information on the release of specific biological parameters, which may be suppressed or masked in serum-supplemented media.

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