A Rapid and Simple Method to Obtain Canine Peripheral Blood-Derived Macrophages

Goto‐Koshino, Yuko; Ohno, Koichi; Nakajima, Mayumi; Mochizuki, Hiroyasu; Kanemoto, Hideyuki; Tsujimoto, Hajime

doi:10.1292/jvms.10-0470

Cited by 22 publications

(7 citation statements)

References 22 publications

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“…In our study, canine MDMs were negative for MHCII in both the control and ATRA-treated samples. Canine MDMs have been either positive [ 30 ], or nearly negative [ 31 ], for MHCII in previous reports, with expression likely depending on differentiation conditions that varied between studies. Our differentiation conditions were similar to those reported by Hartley et al [ 25 ], although their MDM phenotype was not reported.…”

Section: Discussionmentioning

confidence: 99%

The Effect of Arginase on Canine T-Lymphocyte Functions and its Modulation by All-Trans Retinoid Acid (ATRA) in Canine Monocyte-Derived Macrophages

Soileau

Quick

Moeller

et al. 2022

Veterinary Sciences

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Immunosuppressive myeloid cells in the tumor microenvironment play a major role in suppressing tumor immunity via the production of arginase, IL-10, and others. The objectives of this study were to determine the ability of all-trans retinoic acid (ATRA) to decrease the expression of arginase and other soluble mediators by canine monocyte-derived macrophages (MDMs) and to determine the inhibitory activity of arginase on canine T-lymphocytes. The immunomodulatory ability of ATRA (2 µM) on canine MDMs was evaluated via reverse transcription quantitative PCR (RT-qPCR), flow cytometry, arginase activity assay, and enzyme-linked immunoassay (ELISA). Arginase effects on T-lymphocyte phenotype and proliferation were then evaluated by flow cytometry. ATRA consistently decreased MDM expression of IL6, TGFB1, NOS2, ARG1, and CIITA transcripts, by approximately 2–4-fold, although this did not reach statistical significance for ARG1 or CIITA. Furthermore, arginase activity was decreased in ATRA-treated MDMs while the MDM phenotype remained unchanged. Arginase decreased the expression of granzyme B on CD8+ T-lymphocytes and inhibited CD4+ and CD8+ T-lymphocyte proliferation. These findings suggested that ATRA could inhibit canine MDM production of soluble inflammatory/immunosuppressive mediators. These data also revealed that arginase decreased canine T-lymphocyte proliferation and granzyme B expression. Further studies are needed to determine whether ATRA could reverse the immunosuppressive effects of myeloid cells on canine T-lymphocytes in vivo.

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Section: Discussionmentioning

confidence: 99%

The Effect of Arginase on Canine T-Lymphocyte Functions and its Modulation by All-Trans Retinoid Acid (ATRA) in Canine Monocyte-Derived Macrophages

Soileau

Quick

Moeller

et al. 2022

Veterinary Sciences

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“…Monocytes from peripheral blood were isolated as previously described for canine monocytes [ 28 ] with some modifications. Briefly, the gradient centrifugation steps occurred at 450× g without break at deceleration and the subsequent washes to remove platelets were performed at 200× g for a total of three washes.…”

Section: Methodsmentioning

confidence: 99%

Host Gene Expression of Macrophages in Response to Feline Coronavirus Infection

Drechsler

Vasconcelos

Griggs

et al. 2020

Cells

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Feline coronavirus is a highly contagious virus potentially resulting in feline infectious peritonitis (FIP), while the pathogenesis of FIP remains not well understood, particularly in the events leading to the disease. A predominant theory is that the pathogenic FIPV arises from a mutation, so that it could replicate not only in enterocytes of the intestines but also in monocytes, subsequently systemically transporting the virus. The immune status and genetics of affected cats certainly play an important role in the pathogenesis. Considering the importance of genetics and host immune responses in viral infections, the goal of this study was to elucidate host gene expression in macrophages using RNA sequencing. Macrophages from healthy male cats infected with FIPV 79-1146 ex vivo displayed a differential host gene expression. Despite the virus uptake, aligned viral reads did not increase from 2 to 17 h. The overlap of host gene expression among macrophages from different cats was limited, even though viral transcripts were detected in the cells. Interestingly, some of the downregulated genes in all macrophages were involved in immune signaling, while some upregulated genes common for all cats were found to be inhibiting immune activation. Our results highlight individual host responses playing an important role, consistent with the fact that few cats develop feline infectious peritonitis despite a common presence of enteric FCoV.

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“…PBMCs were incubated overnight at 37°C in a culture medium (RPMI-1640, Wako, Tokyo, Japan) to isolate the lymphocytes as floating cells. The adhered cells were collected as monocytes [12]. Total mRNA was extracted from lymphocytes and monocytes using a commercially available kit (RNAspin Mini RNA Isolation Kit; GE Healthcare, Buckinghamshire, England), according to the manufacturer’s instructions.…”

Section: Methodsmentioning

confidence: 99%

Sphingosine-1-phosphate (S1P) signaling regulates the production of intestinal IgA and its potential role in the pathogenesis of canine inflammatory bowel disease

Nakazawa

Maeda

Yokoyama

et al. 2019

The Journal of Veterinary Medical Science

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Inflammatory bowel disease (IBD) is a common gastrointestinal disease in dogs. Decreased production of intestinal immunoglobulin A (IgA) has been suggested as a possible pathogenesis in a subset of canine IBD; however, the underlying cause remains unclear. Sphingosine-1-phosphate (S1P) is a lipid mediator that regulates intestinal IgA production by controlling lymphocyte trafficking in mice. The objectives of this study were to clarify the role of S1P in IgA production in dogs and to evaluate the expression of S1P-related molecules in dogs with IBD. First, an S1P receptor antagonist was administrated to five healthy dogs. The S1P receptor antagonist significantly decreased the IgA concentration in sera and feces but did not affect the IgG concentration. Moreover, the immunoreactivity of intestinal IgA was significantly decreased by S1P signal blockade. These results indicate that S1P signaling specifically regulates the intestinal IgA production in dogs. Subsequently, the intestinal S1P concentration and the expression of S1P-related molecules were measured in dogs with IBD and healthy dogs. The intestinal concentration of S1P was significantly lower in dogs with IBD than in healthy dogs. In addition, the gene expression levels of S1P receptor (S1P1) and S1P synthase (SK1) were significantly lower in dogs with IBD than in healthy dogs. Taken together, these observations suggest that decreased S1P production, likely caused by a lower expression of S1P synthetase, leads to attenuation of S1P/S1P1 signaling pathway and the production of intestinal IgA in dogs with IBD.

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A Rapid and Simple Method to Obtain Canine Peripheral Blood-Derived Macrophages

Cited by 22 publications

References 22 publications

The Effect of Arginase on Canine T-Lymphocyte Functions and its Modulation by All-Trans Retinoid Acid (ATRA) in Canine Monocyte-Derived Macrophages

The Effect of Arginase on Canine T-Lymphocyte Functions and its Modulation by All-Trans Retinoid Acid (ATRA) in Canine Monocyte-Derived Macrophages

Host Gene Expression of Macrophages in Response to Feline Coronavirus Infection

Sphingosine-1-phosphate (S1P) signaling regulates the production of intestinal IgA and its potential role in the pathogenesis of canine inflammatory bowel disease

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