scholarly journals Probiotic microbes induce different resonses from bovine bronchial alveolar lavage cells in vitro

2019 ◽  
Author(s):  
Susan D. Eicher ◽  
Carol Chitko-McKown ◽  
Keith A. Bryan
Keyword(s):  
Fluorescence Intensity ◽  
Oxidative Burst ◽  
Mean Fluorescence Intensity ◽  
Lung Lavage ◽  
Control Treatment ◽  
Upper Respiratory Tract ◽  
E Coli ◽  
Leukocyte Function ◽  
Probiotic Microbes

Abstract Objective Probiotics are fed to improve enteric health, and they may also affect respiratory immunity through their exposure to the upper respiratory tract upon ingestion. However, their effect on the respiratory system is not known. Our aim was to determine how probiotics affect functions and markers of bronchoalveolar lung lavage cells (BAL) isolated from lungs of calves at slaughter. Results Treatments consisted of ten probiotic species and one control treatment. Probiotics and BAL were incubated 1:1 for 2 h at 37° C and 5% CO2. The cell surface markers measured included CD14, CD205, and CD18, and E. coli bioparticles were used to measure phagocytosis and oxidative burst. Differences were considered significant at P ≤ 0.05 and were noted for percent cells fluorescing and mean fluorescence intensity for CD14 and CD205. Additionally, oxidative burst was different as measured by both percentage of cells fluorescing and mean fluorescence intensity, and phagocytosis differed among species as measured by mean fluorescence intensity. Overall, probiotic species differed in their ability to suppress or increase leukocyte function showing that probiotic bacteria differentially modulate BAL.

scholarly journals Variation in the response of bovine alveolar lavage cells to diverse species of probiotic bacteria.

2020 ◽  
Author(s):  
Susan D. Eicher ◽  
Carol Chitko-McKown ◽  
Keith A. Bryan
Keyword(s):  
Fluorescence Intensity ◽  
Oxidative Burst ◽  
Mean Fluorescence Intensity ◽  
Lung Lavage ◽  
Probiotic Bacteria ◽  
Control Treatment ◽  
Upper Respiratory Tract ◽  
E Coli ◽  
Leukocyte Function ◽  
Upper Respiratory

Abstract Objective Probiotics are fed to improve enteric health, and they may also affect respiratory immunity through their exposure to the upper respiratory tract upon ingestion. However, their effect on the respiratory system is not known. Our aim was to determine how probiotics affect functions and markers of bronchoalveolar lung lavage cells (BAL) isolated from lungs of calves at slaughter. Results Treatments consisted of ten probiotic species and one control treatment. Probiotics and BAL were incubated 1:1 for 2 h at 37° C and 5% CO2. The cell surface markers measured included CD14, CD205, and CD18, and E. coli bioparticles were used to measure phagocytosis and oxidative burst. Differences were considered significant at P ≤ 0.05 and were noted for percent cells fluorescing and mean fluorescence intensity for CD14 and CD205. Additionally, oxidative burst was different as measured by both percentage of cells fluorescing and mean fluorescence intensity, and phagocytosis differed among species as measured by mean fluorescence intensity. Overall, probiotic species differed in their ability to suppress or increase leukocyte function showing that probiotic bacteria differentially modulate BAL.

scholarly journals VapC-1 of Nontypeable Haemophilus influenzae Is a Ribonuclease

2007 ◽  
Vol 189 (14) ◽  
pp. 5041-5048 ◽  
Author(s):  
Dayle A. Daines ◽  
Mack H. Wu ◽  
Sarah Y. Yuan
Keyword(s):  
Haemophilus Influenzae ◽  
Growth Inhibition ◽  
Gene Pair ◽  
Gene Families ◽  
Upper Respiratory Tract ◽  
Nontypeable Haemophilus Influenzae ◽  
E Coli ◽  
Conserved Gene

ABSTRACT Nontypeable Haemophilus influenzae (NTHi) organisms are obligate parasites of the human upper respiratory tract that can exist as commensals or pathogens. Toxin-antitoxin (TA) loci are highly conserved gene pairs that encode both a toxin and antitoxin moiety. Seven TA gene families have been identified to date, and NTHi carries two alleles of the vapBC family. Here, we have characterized the function of one of the NTHi alleles, vapBC-1. The gene pair is transcribed as an operon in two NTHi clinical isolates, and promoter fusions display an inverse relationship to culture density. The antitoxin VapB-1 forms homomultimers both in vitro and in vivo. The expression of the toxin VapC-1 conferred growth inhibition to an Escherichia coli expression strain and was successfully purified only when cloned in tandem with its cognate antitoxin. Using total RNA isolated from both E. coli and NTHi, we show for the first time that VapC-1 is an RNase that is active on free RNA but does not degrade DNA in vitro. Preincubation of the purified toxin and antitoxin together results in the formation of a protein complex that abrogates the activity of the toxin. We conclude that the NTHi vapBC-1 gene pair functions as a classical TA locus and that the induction of VapC-1 RNase activity leads to growth inhibition via the mechanism of mRNA cleavage.

scholarly journals In vitro observation: the GFP-E. coli adhering to porcine erythrocytes can be removed by porcine alveolar macrophages

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6439 ◽  
Author(s):  
Wei Yin ◽  
Chun Wang ◽  
Kuohai Fan ◽  
Na Sun ◽  
Yaogui Sun ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Confocal Microscopy ◽  
Fluorescence Intensity ◽  
Alveolar Macrophages ◽  
Control Group ◽  
Laser Confocal Microscopy ◽  
Functional Protein ◽  
Blank Control Group ◽  
E Coli

Although the activation of pathogen phagocytosis via complement system has been studied, erythrocyte-phagocyte interactions in pigs are not clearly understood. Therefore, we sought to investigate the ability of porcine erythrocytes to clear immune complexes (ICs) by using laser confocal microscopy and flow cytometry to observe the immune adhesion of porcine erythrocytes to fluorescent bacilli and the immune presentation process of transferring fluorescent bacilli to macrophages. Isolated porcine alveolar macrophages (PAMs) had uniform morphology and size, and a survival rate of 97.2%. The phagocytosis rate was 98.8%. After WT E. coli was labeled with Fluorescein Isothiocyanate (FITC), the bacteria showed a bright green fluorescence, and the labeling rate was 92.3%. When laser confocal microscopy was utilized to observe the co-incubation system of porcine erythrocytes, PAM, and fluorescent E. coli, the fluorescence intensity of bacilli decreased with increasing observation time and even disappeared. Flow Cytometry examination showed that the average fluorescence intensity of PAMs co-incubated with porcine erythrocytes adhered to WT-E. coli-FITC, was significantly higher than that of normal PAMs. Furthermore, when porcine erythrocytes adhered to WT E. coli were incubated with PAMs, the surface mean fluorescence intensity of porcine erythrocytes was significantly higher than that of the blank control group. This shows that PAMs can competitively bind to the oposinized E. coli adhered to the surface of porcine erythrocytes, and these oposinized pathogens can enter macrophages by the process of phagocytosis, which promoting the internalization of ICs or pathogens. During this process, the physical morphology of porcine erythrocytes was not damaged, but the levels of its main functional protein CR1-like were reduced.

Antioxidant supplementation enhances bacterial peritonitis in mice by inhibiting phagocytosis

2014 ◽  
Vol 63 (3) ◽  
pp. 355-366 ◽  
Author(s):  
Manish Goswami ◽  
Deepak Sharma ◽  
Nazir M. Khan ◽  
Rahul Checker ◽  
Santosh Kumar Sandur ◽  
...  
Keyword(s):  
Oxidative Burst ◽  
Bacterial Load ◽  
Neutrophil Infiltration ◽  
Peritoneal Macrophages ◽  
Antioxidant Supplementation ◽  
E Coli ◽  
Bacterial Peritonitis ◽  
Therapeutic Advantage ◽  
Tnf Α

Antioxidants are known to exhibit numerous health benefits including anti-ageing, anti-apoptotic and immuno-stimulatory effects. However, we present the data showing counterproductive effects of therapeutically relevant antioxidants on bacterial clearance by the immune system in a murine peritonitic model. The antioxidants ascorbic acid, glutathione and N-acetylcysteine augmented morbidity and mortality in mice carrying Eshcerichia coli-induced acute bacterial peritonitis. Treatment of peritonitic mice with antioxidants significantly increased their bacterial load in the range of 0.3–2 logs. Antioxidant administration to peritonitic mice resulted in decreased numbers of macrophages, B-cells and dendritic cells at the primary site of infection and increased neutrophil infiltration. Serum TNF-α levels were also decreased in antioxidant-treated peritonitic mice. In vitro experiments showed that antioxidants reduced the phagocytic efficacy of peritoneal macrophages by ~60–75 % and also decreased E. coli-induced oxidative burst in macrophages cells. Taken together, our data indicate that the antioxidants increased the severity of peritonitis by decreasing the phagocytic efficiency, oxidative burst, and TNF-α production, and increasing neutrophil infiltration. Based on these results, we propose that antioxidant supplementation during the course of bacterial infection is not recommended as it could be detrimental for the host. In addition, the present study underlines the importance of timing and context of antioxidant administration rather than indiscriminate usage to gain the best possible therapeutic advantage of these redox compounds.

scholarly journals CHITOSAN INHIBITION TEST AGAINST E. coli AND DIGESTIBILITY OF THE RATION IN THE IN-VITRO METHOD

2020 ◽  
Vol 11 (2) ◽  
pp. 230
Author(s):  
Eli Sahara ◽  
Sofia Sandi ◽  
Fitra Yosi
Keyword(s):  
Escherichia Coli ◽  
Crude Protein ◽  
Dry Matter ◽  
Control Treatment ◽  
Clear Zone ◽  
Bacterial Colony ◽  
E Coli ◽  
Chicken Feces ◽  
Inhibitory Power

Diarrhea and vomiting are often caused by E coli bacteria. E coli bacteria has a strain of Shigatoxigenic Escherichia coli (STEC), producing Shiga poisons or poisons such as Shiga (verotoxin) which are harmful and pollute nature. This strain of the E coli bacterium has a detrimental effect because it excludes one or both types of Shiga Like Toxin -1 (Stx -1) and Shiga Like Toxin-2 (Stx-2) toxins. This bacterial infection has the potential as a zoonotic agent because it has been found in feces and sheep meat, feces and beef meat, chicken feces and human feces. If this bacterial colony inceases in the digestive tract of poultry it will disturb the productivity of the livestock.  Therefore it must be watched out and studied more deeply. The objectives of the study are 1) to see the inhibitory power of chitosan on the growth and development of E coli bacteria in vitro 2) the test of digestibility of dry matter (BK) and crude protein (PK) ration in vitro. The treatments given in this test are: R0 = control (without chitosan), R1 = 0.5% chitosan, R2 = 1% chitosan, R3 = 1.5% chitosan, R4 = 2% chitosan, R5 = 2.5% chitosan. The parameters measured were 1) inhibition of chitosan against E. coli growth based on clear zone diameter 2) digestibility of dry matter (BK) and crude protein (PK) ration in vitro. The results showed that the higher level of chitosan administration showed greater inhibition, which was indicated by the greater diameter of the clear zone caused. The provision of 2.5% chitosan shows medium inhibition that is has a range of 10-14 mm. The addition of a dose of 1.5% chitosan in the ration was able to increase the digestibility of dry matter by 7.86% and the digestibility of crude protein 11.20% higher than the control treatment (without chitosan). The conclusion of this study is that chitosan can inhibit the growth of E coli and improve the digestibility of dry matter (BK) and crude protein (PK) for the better.AbstrakPenyakit diare dan muntah-muntah sering disebabkan oleh bakteri E coli.  Bakteri E coli memiliki strain Shigatoxigenic Escherichia coli (STEC), menghasilkan racun Shiga atau  racun seperti Shiga (verotoxin) yang  berbahaya dan mencemari  alam. Strain dari bakteri E coli  ini mempunyai efek merugikan karena mengeluarkan salah satu atau kedua jenis toxin Shiga Like Toxin -1 (Stx -1) maupun Shiga Like Toxin-2 (Stx-2).  Infeksi bakteri ini berpotensi sebagai agen zoonosis karena sudah pernah ditemukan pada feses dan daging domba, feses dan daging sapi serta feses ayam dan feses manusia.  Jika koloni bakteri ini tinggi dalam saluran pencernaan unggas akan mengganggu produktivitas ternak tersebut.  Oleh sebab itu harus diwaspadai dan dikaji lebih mendalam. Tujuan penelitian adalah 1) melihat daya hambat kitosan terhadap pertumbuhan dan perkembangan bakteri E coli secara in vitro 2) menguji kecernaan bahan kering (BK) dan protein kasar (PK) ransum secara in vitro. Perlakuan yang diberikan dalam pengujian ini adalah: R0 = kontrol (tanpa kitosan), R1 = 0,5% kitosan, R2 = 1 % kitosan, R3 = 1,5% kitosan, R4 = 2% kitosan, R5 = 2,5% kitosan.   Parameter yang diukur adalah 1) daya hambat kitosan terhadap pertumbuhan E. coli berdasarkan diameter zona bening (in vitro) 2) kecernaan bahan kering (BK) dan protein kasar (PK) ransum secara in vitro. Hasil penelitian menunjukkan bahwa semakin tinggi level pemberian kitosan menunjukkan daya hambat yang semakin besar yang ditandai oleh semakin besarnya diameter zona bening yang ditimbulkan. Pemberian 2,5% kitosan menunjukkan daya hambat sedang yaitu memiliki range 10 - 14 mm. Penambahan dosis 1,5% kitosan dalam ransum, mampu meningkatkan kecernaan bahan kering 7,86% dan kecernaan protein kasar 11,20% lebih tinggi dari perlakuan kontrol (tanpa kitosan). Kesimpulan penelitian ini adalah bahwa kitosan mampu menghambat pertumbuhan E coli dan meningkatkan kecernaan bahan kering (BK) serta protein kasar (PK) menjadi lebih baik.Kata kunci: Kitosan, daya hambat, E. coli, kecernaan ransum, in vitro

Expression of the cell adhesion molecules on leukocytes that demarginate during acute maximal exercise

1999 ◽  
Vol 86 (3) ◽  
pp. 970-976 ◽  
Author(s):  
Stephan F. van Eeden ◽  
John Granton ◽  
Jennifer M. Hards ◽  
Barbara Moore ◽  
James C. Hogg
Keyword(s):  
Fluorescence Intensity ◽  
Mean Fluorescence Intensity ◽  
Maximal Exercise ◽  
Flow Cytometric Analysis ◽  
Cycle Ergometer ◽  
Phenotypic Characteristics ◽  
Flow Cytometric ◽  
The Mean ◽  
Low Levels

The pulmonary vascular bed is an important reservoir for the marginated pool of leukocytes that can be mobilized by exercise or catecholamines. This study was designed to determine the phenotypic characteristics of leukocytes that are mobilized into the circulation during exercise. Twenty healthy volunteers performed incremental exercise to exhaustion [maximal O2consumption (V˙o2 max)] on a cycle ergometer. Blood was collected at baseline, at 3-min intervals during exercise, atV˙o2 max, and 30 min after exercise. Total white cell, polymorphonuclear leukocyte (PMN), and lymphocyte counts increased with exercise toV˙o2 max( P < 0.05). Flow cytometric analysis showed that the mean fluorescence intensity of L-selectin on PMN (from 14.9 ± 1 at baseline to 9.5 ± 1.6 atV˙o2 max, P < 0.05) and lymphocytes (from 11.7 ± 1.2 at baseline to 8 ± 0.8 atV˙o2 max, P < 0.05) decreased with exercise. Mean fluorescence intensity of CD11b on PMN increased with exercise (from 10.2 ± 0.6 at baseline to 25 ± 2.5 atV˙o2 max, P < 0.002) but remained unchanged on lymphocytes. Myeloperoxidase levels in PMN did not change with exercise. In vitro studies showed that neither catecholamines nor plasma collected atV˙o2 maxduring exercise changed leukocyte L-selectin or CD11b levels. We conclude that PMN released from the marginated pool during exercise express low levels of L-selectin and high levels of CD11b.

scholarly journals Effects of calcitriol on oxidative burst, phagocytic function, and leukocyte cytokine production in shelter dogs

2020 ◽  
Author(s):  
Jared Jaffey ◽  
Mariah Bessette ◽  
Zenan Tao ◽  
Nancy Bradley-Siemens ◽  
Melissa Thompson
Keyword(s):  
Vitamin D ◽  
Oxidative Burst ◽  
Whole Blood ◽  
Vitamin D Supplementation ◽  
Phagocytic Function ◽  
E Coli ◽  
Tnf Α ◽  
Shelter Dogs ◽  
Necrosis Factor

Abstract Background The active metabolite of vitamin D, calcitriol, has been shown across many different species to augment innate immune responses and dampen aberrant proinflammatory cytokine production. Community acquired infections are common in shelters and consume limited shelter resources, impact adoption rates, and can result in unnecessary euthanasia. Prophylactic oral vitamin D supplementation decreases the incidence and severity of upper and lower respiratory tract infections in humans. Before a clinical trial investigating the clinical benefit of oral vitamin D supplementation in shelter dogs can be pursued, an in vitro study evaluating the immunomodulatory effects of calcitriol in blood from shelter dogs is warranted. Therefore, the objective of this study was to determine if incubation of whole blood obtained from apparently healthy dogs housed in a shelter for ≥ 7 days with calcitriol would alter granulocyte/monocyte (GM) oxidative burst and phagocytic function as well as pathogen-associated molecular pattern (PAMP)-stimulated leukocyte production of tumor necrosis factor (TNF)-α, and interleukin (IL)-6, and IL-10. Results Ten dogs housed in a shelter for ≥ 7 days were enrolled in a prospective cohort study. Whole blood from these dogs was incubated with calcitriol (10-7 M) or ethanol (control) for 24 h. Subsequent to this incubation phagocytosis of opsonized-Escherichia coli (E. coli) and E. coli-induced oxidative burst were evaluated via flow cytometry. In addition, leukocyte production of TNF-α, IL-6, and IL-10 were measured using a canine-specific multiplex assay. Calcitriol significantly decreased leukocyte TNF-α production (p = 0.009) and increased IL-10 production (p = 0.002). Tumor necrosis factor-α-to-IL-10 ratio was significantly decreased with calcitriol (p = 0.017), while IL-6 production as well as GM oxidative burst and phagocytic function were not significantly affected. Conclusions These data indicate that calcitriol attenuates proinflammatory immune responses without affecting GM oxidative burst or phagocytic function in vitro in whole blood obtained from apparently healthy shelter dogs.

Reduction of Escherichia coli O157:H7 and Salmonella on Laboratory-Inoculated Alfalfa Seed with Commercial Citrus-Related Products†

2003 ◽  
Vol 66 (7) ◽  
pp. 1158-1165 ◽  
Author(s):  
WILLIAM F. FETT ◽  
PETER H. COOKE
Keyword(s):  
Escherichia Coli ◽  
Tap Water ◽  
The United States ◽  
Control Treatment ◽  
In Vitro Assays ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Alfalfa Seed ◽  
Alfalfa Sprouts

Alfalfa sprouts contaminated with the bacterial pathogens Escherichia coli O157:H7 and Salmonella have been the source of numerous outbreaks of foodborne illness in the United States and in other countries. The seed used for sprouting appears to be the primary source of these pathogens. The aim of this study was to determine whether the efficacy of commercial citrus-related products for sanitizing sprouting seed is similar to that of high levels of chlorine. Five products (Citrex, Pangermex, Citricidal, Citrobio, and Environné) were tested at concentrations of up to 20,000 ppm in sterile tap water and compared with buffered chlorine (at 16,000 ppm). Alfalfa seeds were inoculated with four-strain cocktails of Salmonella and E. coli O157:H7 to give final initial concentrations of ca. 9.0 and 7.0 CFU/g, respectively. Treatments (10 min) with Citrex, Pangermex, and Citricidal at 20,000 ppm and chlorine at 16,000 ppm produced similar log reductions for alfalfa seed inoculated with four-strain cocktails of E. coli O157:H7 and Salmonella (3.42 to 3.46 log CFU/g and 3.56 to 3.74 log CFU/g, respectively), and all four treatments were significantly (P &lt; 0.05) more effective than the control treatment (a buffer wash). Citrobio at 20,000 ppm was as effective as the other three products and chlorine against Salmonella but not against E. coli O157:H7. Environné was not more effective (producing reductions of 2.2 to 2.9 log CFU/g) than the control treatment (which produced reductions of 2.1 to 2.3 log CFU/g) against either pathogen. None of the treatments reduced seed germination. In vitro assays, as well as transmission electron microscopy, confirmed the antibacterial nature of the products that were effective against the two pathogens and indicated that they were bactericidal. When used at 20,000 ppm, the effective citrus-related products may be viable alternatives to chlorine for the sanitization of sprouting seed pending regulatory approval.

CD20 levels determine the in vitro susceptibility to rituximab and complement of B-cell chronic lymphocytic leukemia: further regulation by CD55 and CD59

Blood ◽  
2001 ◽  
Vol 98 (12) ◽  
pp. 3383-3389 ◽  
Author(s):  
Josée Golay ◽  
Manuela Lazzari ◽  
Valeria Facchinetti ◽  
Sergio Bernasconi ◽  
Gianmaria Borleri ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Fluorescence Intensity ◽  
Mean Fluorescence Intensity ◽  
Lymphocytic Leukemia ◽  
In Vitro Susceptibility ◽  
Prolymphocytic Leukemia ◽  
Complement Dependent Cytotoxicity ◽  
Cell Chronic Lymphocytic Leukemia

Abstract Complement-dependent cytotoxicity is thought to be an important mechanism of action of the anti-CD20 monoclonal antibody rituximab. This study investigates the sensitivity of freshly isolated cells obtained from 33 patients with B-cell chronic lymphocytic leukemia (B-CLL), 5 patients with prolymphocytic leukemia (PLL), and 6 patients with mantle cell lymphoma (MCL) to be lysed by rituximab and complement in vitro. The results showed that in B-CLL and PLL, the levels of CD20, measured by standard immunofluorescence or using calibrated beads, correlated linearly with the lytic response (coefficient greater than or equal to 0.9;P &lt; .0001). Furthermore, the correlation remained highly significant when the 6 patients with MCL were included in the analysis (coefficient 0.91; P &lt; .0001), which suggests that CD20 levels primarily determine lysis regardless of diagnostic group. The role of the complement inhibitors CD46, CD55, and CD59 was also investigated. All B-CLL and PLL cells expressed these molecules, but at different levels. CD46 was relatively weak on all samples (mean fluorescence intensity less than 100), whereas CD55 and CD59 showed variability of expression (mean fluorescence intensity 20-1200 and 20-250, respectively). Although CD55 and CD59 levels did not permit prediction of complement susceptibility, the functional block of these inhibitors demonstrated that they play an important role in regulating complement-dependent cytotoxicity. Thus, lysis of poorly responding B-CLL samples was increased 5- to 6-fold after blocking both CD55 and CD59, whereas that of high responders was essentially complete in the presence of a single blocking antibody. These data demonstrate that CD20, CD55, and CD59 are important factors determining the in vitro response to rituximab and complement and indicate potential strategies to improve the clinical response to this biologic therapy.

scholarly journals Effects of calcitriol on oxidative burst, phagocytic function, and leukocyte cytokine production in shelter dogs

2020 ◽  
Author(s):  
Jared Jaffey ◽  
Mariah Bessette ◽  
Zenan Tao ◽  
Nancy Bradley-Siemens ◽  
Melissa Thompson
Keyword(s):  
Vitamin D ◽  
Oxidative Burst ◽  
Whole Blood ◽  
Vitamin D Supplementation ◽  
Phagocytic Function ◽  
E Coli ◽  
Tnf Α ◽  
Shelter Dogs ◽  
Necrosis Factor

Abstract Background The active metabolite of vitamin D, calcitriol, has been shown across many different species to augment innate immune responses and dampen aberrant proinflammatory cytokine production. Community acquired infections are common in shelters and consume limited shelter resources, impact adoption rates, and can result in unnecessary euthanasia. Prophylactic oral vitamin D supplementation decreases the incidence and severity of upper and lower respiratory tract infections in humans. Before a clinical trial investigating the clinical benefit of oral vitamin D supplementation in shelter dogs can be pursued, an in vitro study evaluating the immunomodulatory effects of calcitriol in blood from shelter dogs is warranted. Therefore, the objective of this study was to determine if incubation of whole blood obtained from apparently healthy dogs housed in a shelter for ≥ 7 days with calcitriol would alter granulocyte/monocyte (GM) oxidative burst and phagocytic function as well as pathogen-associated molecular pattern (PAMP)-stimulated leukocyte production of tumor necrosis factor (TNF)-α, and interleukin (IL)-6, and IL-10. Results Ten dogs housed in a shelter for ≥ 7 days were enrolled in a prospective cohort study. Whole blood from these dogs was incubated with calcitriol (10-7 M) or diluent (control) for 24 h. Subsequent to this incubation phagocytosis of opsonized-Escherichia coli (E. coli) and E. coli-induced oxidative burst were evaluated via flow cytometry. In addition, leukocyte production of TNF-α, IL-6, and IL-10 were measured using a canine-specific multiplex assay. Calcitriol significantly decreased leukocyte TNF-α production (p = 0.009) and increased IL-10 production (p = 0.002). Tumor necrosis factor-α-to-IL-10 ratio was significantly decreased with calcitriol (p = 0.017), while IL-6 production as well as GM oxidative burst and phagocytic function were not significantly affected. Conclusions These data indicate that calcitriol attenuates proinflammatory immune responses without affecting GM oxidative burst or phagocytic function in vitro in whole blood obtained from apparently healthy shelter dogs.

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