Separation of phagocytic leukocytes from the peripheral blood of chickens

Mills, Jennifer N.; Wilcox, G. E.

doi:10.1080/03079459308418925

Cited by 10 publications

(4 citation statements)

References 22 publications

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“…To the best of our knowledge, the distribution of cells in the buffy coat of selected reptile and nonpoultry avian species has never been studied in detail. Still, our data are in accordance with a classical report‐based separation of cells by Percoll gradients . This study by Mills and Wilcox with chicken blood reported that thrombocytes appear admixed with lymphocytes in lighter fractions (which corresponds to the upper buffy coat parts), whereas denser fractions contained mostly heterophils.…”

Section: Discussionsupporting

confidence: 93%

“…Still, our data are in accordance with a classical report‐based separation of cells by Percoll gradients . This study by Mills and Wilcox with chicken blood reported that thrombocytes appear admixed with lymphocytes in lighter fractions (which corresponds to the upper buffy coat parts), whereas denser fractions contained mostly heterophils. These cells were also present among nucleated erythrocytes, and to a lesser extent in lighter fractions of lymphocytes.…”

Section: Discussionsupporting

confidence: 93%

“…These cells were also present among nucleated erythrocytes, and to a lesser extent in lighter fractions of lymphocytes. Notably, these authors reported that immature chicken erythrocytes had the same density as granulocytes as we highlighted here.…”

Section: Discussionsupporting

confidence: 76%

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Unraveling avian and reptilian hematology: An optical and electron microscopic study of the buffy coat

Pinto¹,

Lopes

Malhão

et al. 2018

Veterinary Clinical Pathol

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Section: Discussionsupporting

confidence: 93%

Section: Discussionsupporting

confidence: 93%

See 1 more Smart Citation

Unraveling avian and reptilian hematology: An optical and electron microscopic study of the buffy coat

Pinto¹,

Lopes

Malhão

et al. 2018

Veterinary Clinical Pathol

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“…Blood was obtained from the brachial vein in an heparinized syringe, afterwards it was diluted using an equal volume of Hank's balanced saline solution (HBSS) and homogenized. Blood was carefully deposited in a gradient with 70 and 80% of Percoll (Pharmacia) and centrifuged to 1150 g for 30 min at room temperature as has been previously reported (Mills & Wilcox, 1993;Desmidt et al ., 1996). Cells were washed twice with HBSS by centrifugation (1150 g for 10 min) and the pellet was suspended in RPMI medium containing 10% fetal bovine serum and 10 UI/ml penicillin, and 10 mg/ml streptomycin (complete RPMI medium).…”

Section: Proliferation Assaysmentioning

confidence: 99%

Comparison of the immune responses against Salmonella enterica serovar Gallinarum infection between naked neck chickens and a commercial chicken line

Alvarez

Ledesma²,

Téllez³

et al. 2003

Avian Pathology

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The immune responses of indigenous naked neck (NaNa and Nana) and normally feathered (nana) chickens against a Salmonella Gallinarum (SG) infection were evaluated and compared with those of a commercial line (B-380). Groups of 28-day-old chickens (NaNa, Nana, nana, and B-380) were immunized orally and subcutaneously with 50 microg of SG antigens. Control non-immunized animals were inoculated with sterile saline solution. All chickens were challenged with 1 LD(50) of SG and mortality was recorded daily for 20 days. Antibodies to SG were measured in sera before immunization, before the challenge, 10 days after the challenge, and at sacrifice. Peripheral blood lymphocyte proliferation assays were performed using concanavalin A and SG antigens. Results showed that non-immunized Nana chickens exhibited the best natural resistance to Salmonella infection, since only 30% of them died. In contrast, all control B-380 chickens died by the 13th day. Immunization with SG induced immunity in chickens of all genotypes. Indigenous naked neck and normally feathered chickens showed a higher survival rate when compared with B-380 chickens. Immunized Nana chickens showed the highest antibody titres (P<0.05) as well as the highest thymidine incorporation in peripheral blood lymphocytes stimulated with con A or SG antigens (P<0.05). The results show that Nana chickens are the most resistant to SG infection and the best responders to vaccination with SG antigens.

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Characterisation of the green turtle’s leukocyte subpopulations by flow cytometry and evaluation of their phagocytic activity

Tenería

Franco-Noguez²,

González-Ballesteros

et al. 2014

Vet Res Commun

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Phagocytosis is a fundamental aspect of innate immunity that is conserved across many species making it a potentially useful health-assessment tool for wildlife. In non-mammalian vertebrates, heterophils, monocytes, macrophages, melanomacrophages, and thrombocytes all have phagocytic properties. Recently, B lymphocytes from fish, amphibians, and aquatic turtles have also showed phagocytic capacity. Phagocytes can be studied by flow cytometry; however, the use of this tool is complicated in reptiles partly because nucleated erythrocytes complicate the procedure. We separated green turtle leukocytes by density gradient centrifugation and identified subpopulations by flow cytometry and confocal microscopy. Additionally, we assessed their ability to phagocytize Fluorspheres and Ovoalbumin-Alexa. We found that heterophils and lymphocytes but not monocytes could be easily identified by flow cytometry. While heterophils from adults and juvenile turtles were equally able to phagocytize fluorspheres, adults had significantly more phagocytic ability for OVA-Alexa. Lymphocytes had a mild phagocytic activity with fluorospheres (27-38 %; 39-45 %) and OVA-Alexa (35-46 %; 14-22 %) in juvenile and adult green turtles, respectively. Confocal microscopy confirmed phagocytosis of fluorospheres in both heterophils and lymphocytes. This provides the first evidence that green turtle lymphocytes have phagocytic activity and that this assay could potentially be useful to measure one aspect of innate immunity in this species.

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Separation of phagocytic leukocytes from the peripheral blood of chickens

Cited by 10 publications

References 22 publications

Unraveling avian and reptilian hematology: An optical and electron microscopic study of the buffy coat

Unraveling avian and reptilian hematology: An optical and electron microscopic study of the buffy coat

Comparison of the immune responses against Salmonella enterica serovar Gallinarum infection between naked neck chickens and a commercial chicken line

Characterisation of the green turtle’s leukocyte subpopulations by flow cytometry and evaluation of their phagocytic activity

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