A simple and rapid flow cytometric method for detection of porcine cell surface markers

Stabel, Thomas J.; Bolin, Steven R.; Pesch, B.A.; Rahner, T. E.

doi:10.1016/s0022-1759(00)00289-1

Cited by 17 publications

(14 citation statements)

References 8 publications

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“…Cells from the TBLNs were phenotyped by flow cytometry, as described previously (38). Commercially available antibodies to the following markers were used: CD3 (2B3C), CD25 (PGBL25A), MHC class II (TH16B), B cell (BB6-11C9), CD4 (PT90A), CD8 (76-2-11), and ␥/␦ T-cell (PGBL22A) (VMRD Inc., Pullman, WA).…”

Section: Methodsmentioning

confidence: 99%

Cytokine Protein Expression Levels in Tracheobronchial Lymph Node Homogenates of Pigs Infected with Pseudorabies Virus

Miller

Zanella

Waters

et al. 2010

Clin Vaccine Immunol

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Section: Methodsmentioning

confidence: 99%

Cytokine Protein Expression Levels in Tracheobronchial Lymph Node Homogenates of Pigs Infected with Pseudorabies Virus

Miller

Zanella

Waters

et al. 2010

Clin Vaccine Immunol

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“…The ratio of lymphocytes added to monocytes, MDMs, and immature MDCs resembled the ratio of lymphocytes to monocytes in the peripheral blood of pigs of the same age, approximately 10:1 (56). The cultures were incubated at 37°C for 2 d in a humidified, 5% CO 2 atmosphere.…”

Section: Cell Culturementioning

confidence: 99%

Effects of Porcine Reproductive and Respiratory Syndrome Virus-Infected Antigen-Presenting Cells on T Cell Activation and Antiviral Cytokine Production

2006

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The ability of porcine reproductive and respiratory syndrome virus (PRRSV) to suppress T cell expression of CD25 (alpha chain of interleukin [IL]-2 receptor), interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha) was determined by flow cytometry in naive porcine T cells in response to mitogen (concanavalin A) and cytokine inducers (phorbol 12-myristate 13-acetate plus ionomycin [PMA/I]). Four PRRSV isolates of varying clinical virulence and three different types of porcine myeloid antigen-presenting cells (APCs) were used. T cells cultured with monocytes infected with virulent PRRSV (VR-2385, SDSU-73, and VR-2332), but not with a vaccine strain (Ingelvac PRRS MLV; Boehringer Ingelheim Vetmedica, St. Joseph, MO), demonstrated significantly reduced CD25 expression (%CD25(+)) and IFN-gamma expression (%IFN-gamma (+)) compared with T cells incubated with uninoculated monocyte cultures. T cells cultured with monocytes infected with all four PRRSV isolates demonstrated significantly reduced %TNF-alpha (+). The significant reduction of %CD25(+), %IFN-gamma (+), and %TNF-alpha (+) was not detected in T cells cultured with monocyte-derived macrophages (MDMs) and immature monocyte-derived dendritic cells (MDCs) infected with any PRRSV isolates. Heat-inactivated PRRSV did not induce significantly reduced T cell responses in any APC cultures. The reduction of T cell response in monocyte cultures was not due to PRRSV-induced T cell death. Gene expression of IL-10 detected by semiquantitative reverse transcriptase-polymerase chain reaction was significantly increased in virulent PRRSV-infected monocyte cultures after PMA/I, but not concanavalin A, stimulation compared with IL-10 gene expression from uninoculated monocyte cultures. Increased IL-10 gene expression contributed to significantly reduced %IFN-gamma (+) and %TNF-alpha (+), but not %CD25(+), as determined by IL-10 neutralization assay. This study reports that PRRSV has the ability to suppress T cell responses. The suppressive ability of PRRSV is associated with viral virulence and is mediated by virus-infected monocytes, but not by virus-infected MDMs and immature MDCs.

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“…Mouse IgC2a and IgC2b, k-isotype standard antibodies (Southern Biotechnology) were used as control antibodies. Whole blood ahquots (100 |iL) were suspended in 100 (iL of cold PBS/azide (0.1%) and incubated in the dark at 4°C for 20 min with pretitered dilutions of the monoclonal antibodies and equal concentration of isotypic control antibodies (Stabel et al, 2000). Erythrocytes were lysed with freshly diluted lysing solution (150 mMNH4Cl, 10 mMNaHCOg, and 1 mMEDTA in distilled water), for 10 min at room temperature.…”

Section: Immune Functionmentioning

confidence: 99%

Immune response and blood chemistry of pigs fed conjugated linoleic acid1

Wiegand

Pompeu

Thiel-Cooper

et al. 2011

Journal of Animal Science

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Immune function (response to concanavalin A, cytokine production, and lymphocyte profiles) and blood chemistry variables were measured in growing-finishing pigs (Yorkshire/Landrace/Duroc dam × Hampshire sire) fed varying percentages of CLA (0, 0.12, 0.25, 0.50, and 1.0%). Blood was collected at 0, 14, 28, 42, and 56 d on feed (DOF). Total white blood cell (WBC) count increased (P < 0.01) linearly to 42 DOF. No differences (P = 0.53) were observed for WBC across CLA treatment. Nitric oxide was greater (P < 0.01) for the 1.0% CLA treatment compared with all other treatments. Flow cytometry using fluorescent labeled monoclonal antibodies to the CD4, CD8, double-positive CD4/CD8, and CD2 surface markers was used to determine lymphocyte subpopulations. Supplementation of CLA had no effect (P = 0.61) on lymphocyte subpopulation cell distribution. Most blood chemistry variables were within the normal metabolic range for pigs. A decrease was observed over DOF for P (P < 0.01) and K (P < 0.05). Additionally, Na and Cl concentrations increased (P < 0.05) from 14 to 28 DOF and decreased over the remainder of the trial. Electrolyte balance was not different (P = 0.38) across CLA treatments and was likely explained by no differences in feed intake among the CLA treatment groups. Blood lipid variables indicated that total cholesterol (P < 0.001), triglycerides (P < 0.001), high-density lipoproteins (P < 0.001), and low-density lipoproteins (P < 0.01) increased as the amount of CLA in the diet increased, but none of the results from these treatments exceeded the normal range of acceptability. These results suggested that CLA was safe when fed to growing-finishing pigs and had little effect on their immune function and blood chemistry variables.

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A simple and rapid flow cytometric method for detection of porcine cell surface markers

Cited by 17 publications

References 8 publications

Cytokine Protein Expression Levels in Tracheobronchial Lymph Node Homogenates of Pigs Infected with Pseudorabies Virus

Cytokine Protein Expression Levels in Tracheobronchial Lymph Node Homogenates of Pigs Infected with Pseudorabies Virus

Effects of Porcine Reproductive and Respiratory Syndrome Virus-Infected Antigen-Presenting Cells on T Cell Activation and Antiviral Cytokine Production

Immune response and blood chemistry of pigs fed conjugated linoleic acid1

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