Different methods of cell quantification can lead to different results: a comparison of digital methods using a pilot study of dendritic cells in HIV-positive patients

Fernandes, D-T; Heerden, W-F van; Ribeiro, A-C; Brandão, Thaís Bianca; Mello, E-S; Rivera, César; Heerden, M-B van; Gondak, Rogério; Santos‐Silva, Alan Roger; Vargas, Pablo Agustı́n; Ma, Le

doi:10.4317/medoral.23472

Cited by 4 publications

(4 citation statements)

References 29 publications

(47 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Depending on the state of maturation, DCs play an important role in the immune response. 22 Many different subtypes of immature DCs (CD1a+, CD207+, and S100+) and mature DCs (CD83+ and CD208+) have been described, 23,24,25,26 making it difficult to compare the studies. In the present study, we used the CD1a and CD83 markers for immature and mature DCs, respectively, since they are recognized and well-accepted DCs maturation markers in formalin fixed paraffin-embedded human tissues.…”

Section: Discussionmentioning

confidence: 99%

Impact of smoking on dendritic cells in patients with oral squamous cell carcinoma

et al. 2021

View full text Add to dashboard Cite

Smoking has been shown to alter innate and adaptive immune responses and is directly associated with the onset of oral squamous cell carcinoma (OSCC). The purpose of this study was to evaluate the effect of cigarette smoke exposure on dendritic cells (DCs) from OSCC patients. CD1a and CD83 antibodies were used to identify immature and mature DCs, respectively, by immunohistochemistry in OSCC samples of 24 smokers and 24 non-smokers. Density of DCs was calculated in intra and peritumoral areas. Clinical and microscopic findings were reviewed and analyzed for all patients. Smokers with OSCC had a lower density of intra and peritumoral DCs when compared to non-smokers. Tumors classified as moderately/poorly differentiated had lower peritumoral CD1a+ DCs than well-differentiated tumors (p < 0.001). Smoking contributed to a depletion of immature and mature DCs in the OSCC.

show abstract

Section: Discussionmentioning

confidence: 99%

Impact of smoking on dendritic cells in patients with oral squamous cell carcinoma

et al. 2021

View full text Add to dashboard Cite

show abstract

“…The putative mature cod piscidin is rich in isoleucine (27.3%) and histidine (22.7%) and it contains the two conserved histidines that define the piscidin group of antimicrobial peptides (Figure 2). This result shows that piscidins are an ancient family of host defense peptides not restricted to higher teleosts, since the cod, G. morhua, is a basal teleost of the Paracanthopterygii superorder [51][52][53]. Most piscidin genes were identified to have a date code for a precursor comprising a 22-residue signal peptide, a mature (active) peptide of 22-25 residues and a variable C-terminal region.…”

Section: Phylogenetic Inference Of Piscidin Genesmentioning

confidence: 92%

Expression of the Antimicrobial Peptide Piscidin 1 and Neuropeptides in Fish Gill and Skin: A Potential Participation in Neuro-Immune Interaction

et al. 2022

View full text Add to dashboard Cite

Antimicrobial peptides (AMPs) are found widespread in nature and possess antimicrobial and immunomodulatory activities. Due to their multifunctional properties, these peptides are a focus of growing body of interest and have been characterized in several fish species. Due to their similarities in amino-acid composition and amphipathic design, it has been suggested that neuropeptides may be directly involved in the innate immune response against pathogen intruders. In this review, we report the molecular characterization of the fish-specific AMP piscidin1, the production of an antibody raised against this peptide and the immunohistochemical identification of this peptide and enkephalins in the neuroepithelial cells (NECs) in the gill of several teleost fish species living in different habitats. In spite of the abundant literature on Piscidin1, the biological role of this peptide in fish visceral organs remains poorly explored, as well as the role of the neuropeptides in neuroimmune interaction in fish. The NECs, by their role as sensors of hypoxia changes in the external environments, in combination with their endocrine nature and secretion of immunomodulatory substances would influence various types of immune cells that contain piscidin, such as mast cells and eosinophils, both showing interaction with the nervous system. The discovery of piscidins in the gill and skin, their diversity and their role in the regulation of immune response will lead to better selection of these immunomodulatory molecules as drug targets to retain antimicrobial barrier function and for aquaculture therapy in the future.

show abstract

“…MNPs have pleiomorphic shapes with long dendrites, they are difficult to identify using standard digital image analysis algorithms [11]. To assess MNP density, we and others have previously used pixel counts, which can accurately identify changes in staining patterns [10,23,28,29]. Since correlation between pixel counts and nuclear counts varies widely depending on the antibody used [10], pixel-based approaches cannot provide accurate information on cell numbers.…”

Section: Automated Cell Identification Of Cells In Immunohistochemicalmentioning

confidence: 99%

MNPmApp: An image analysis tool to quantify mononuclear phagocyte distribution in mucosal tissues

et al. 2022

View full text Add to dashboard Cite

Mononuclear phagocytes (MNPs) such as dendritic cells and macrophages perform key sentinel functions in mucosal tissues and are responsible for inducing and maintaining adaptive immune responses to mucosal pathogens. Positioning of MNPs at the mucosal epithelial interface facilitates their access to luminally-derived antigens and may regulate MNP function through soluble mediators or surface receptor interactions. Therefore, accurately quantifying the distribution of MNPs within mucosal tissues as well as their spatial relationship with other cells is important to infer functional cellular interactions in health and disease. In this study, we developed and validated a MATLAB-based tissue cytometry platform, termed "MNP mapping application" (MNPmApp), that performs high throughput analyses of MNP density and distribution in the gastrointestinal mucosa based on digital multicolor fluorescence microscopy images and that integrates a Monte Carlo modeling feature to assess randomness of MNP distribution.MNPmApp identified MNPs in tissue sections of the human gastric mucosa with a specificity of 98.3 ± 1.6% and a sensitivity of 76.4 ± 15.1%. Monte Carlo modeling revealed that mean MNP-MNP distances were significantly lower than anticipated based on random cell placement, whereas MNP-epithelial distances did not significantly differ from those of randomly placed cells.Interestingly, H. pylori infection had no significant impact on MNP density or distribution with regards to MNP-epithelial distances or MNP-MNP distances in gastric tissue. Overall, our analysis demonstrates that MNPmApp is a useful tool for unbiased quantitation of MNPs and their distribution at mucosal sites.

show abstract

Different methods of cell quantification can lead to different results: a comparison of digital methods using a pilot study of dendritic cells in HIV-positive patients

Cited by 4 publications

References 29 publications

Impact of smoking on dendritic cells in patients with oral squamous cell carcinoma

Impact of smoking on dendritic cells in patients with oral squamous cell carcinoma

Expression of the Antimicrobial Peptide Piscidin 1 and Neuropeptides in Fish Gill and Skin: A Potential Participation in Neuro-Immune Interaction

MNPmApp: An image analysis tool to quantify mononuclear phagocyte distribution in mucosal tissues

Contact Info

Product

Resources

About