Evaluation of cell replication by bovine T cells in polyclonally activated cultures using carboxyfluorescein succinimidyl ester (CFSE) loading and flow cytometric analysis

Sathiyaseelan, Thillainayagam; Baldwin, Cynthia L.

doi:10.1053/rvsc.2000.0429

Cited by 30 publications

(17 citation statements)

References 15 publications

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“…We observed active cell division among leukocytes that had migrated through an infected endothelial membrane by observing the modal decrease in fluorescence of CFSE-labelled cells over time [10,21,25]. We did not measure proliferation after migration through quiescent or TNFastimulated endothelia, but, given the activation status of the transmigrating leukocytes under these circumstances, division probably occurs similarly.…”

Section: Discussionmentioning

confidence: 99%

In vitro infection of aortic endothelial cells by caprine arthritis encephalitis virus enhances in vitro transmigration of peripheral blood leukocytes and modulates their phenotypic expression

et al. 2003

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-Infection of goats by caprine arthritis-encephalitis virus (CAEV) provides a convenient example of the infiltration of various tissues by leukocytes following a natural lentiviral infection. This event is important in determining organ susceptibility and local immunity. Caprine vascular endothelial cells are susceptible to infection by CAEV in vitro, so we have investigated the consequences of this infection on the transmigration of uninfected leukocytes in an in vitro model. After in vitro infection by CAEV or stimulation by TNFa, the endothelial cells allowed the passage of tenfold more leukocytes from uninfected donors than did the uninfected endothelial cells. The transmigrating leukocytes were enriched in CD8 + lymphocytes, and the leukocytes appeared to have been activated during transmigration, as demonstrated by their expression of IL2R, MHC class II antigens and gamma-delta T-lymphocyte markers. CD4 + , CD8 + and B-lymphocytes all proliferated in culture after transmigration. These results suggest that any possible infection or specific stimulation of endothelia in an infected animal could profoundly influence the choice of target organs and could activate the cells involved in local mucosal immune responses. caev / endothelial cell / leukocyte / transmigration / in vitro

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Section: Discussionmentioning

confidence: 99%

In vitro infection of aortic endothelial cells by caprine arthritis encephalitis virus enhances in vitro transmigration of peripheral blood leukocytes and modulates their phenotypic expression

et al. 2003

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“…Lymphocytes were loaded with 2.5 mM carboxyfluorescein succinimidyl ester according to the manufacturer's protocol and as described by Sathiyaseelan & Baldwin (2000). Lymphocytes were placed either in culture with increasing concentrations of bovine SPP1 or in coculture with luteal cells containing increasing concentrations of bovine SPP1 (SigmaAldrich) and T-cell proliferation was analyzed after 24 h using Guava EasyCyte Plus (Millipore).…”

Section: T Lymphocyte Proliferationmentioning

confidence: 99%

Expression and regulation of secreted phosphoprotein 1 in the bovine corpus luteum and effects on T lymphocyte chemotaxis

Poole¹,

Ndiaye²,

Pate³

2013

Reproduction

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Secreted phosphoprotein 1 (SPP1) in the bovine corpus luteum (CL) regulates cell function during the transitional periods of luteinization and luteal regression. The objectives were to i) characterize SPP1 expression in the CL throughout the estrous cycle, ii) determine factors that regulate SPP1 expression in luteal cells, and iii) examine the role of SPP1 in lymphocyte chemotaxis, proliferation, and function. SPP1 mRNA was greater in fully functional (d10) CL and late cycle (d18) CL compared with developing (d4) CL. Additionally, SPP1 mRNA increased within 1 h and remained elevated 4 and 8 h following induction of luteolysis with prostaglandin (PG)F 2a . Expression of the SPP1 receptor, b 3 integrin, was not different throughout the estrous cycle but decreased following induction of luteolysis. Expression of CD44 increased during the estrous cycle but did not change during luteal regression. In cultured luteal cells, SPP1 mRNA was upregulated by PGF 2a and/or tumor necrosis factor a. Western blots revealed the presence of both full-length SPP1 and multiple cleavage products in cultured luteal cells and luteal tissue. Depletion of endogenous SPP1 did not hinder luteal cell-induced lymphocyte proliferation or lymphocyte phenotype but did inhibit lymphocyte migration toward luteal cells. Based on these data, it is concluded that SPP1 is initially activated to establish and maintain cellular interactions between steroidogenic and nonsteroidogenic cells during the development of the CL. Upon induction of luteolysis, SPP1 serves as a signaling molecule to recruit or activate immune cells to facilitate luteal regression and tissue degradation.

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“…Upon thawing, dead cells and debris were removed by centrifugation on Ficoll density gradient (Eurobio) prior to use in stimulation assays. Loading of cells with carboxyfluorescein diacetate succinimidyl ester (CFSE) (Invitrogen, Cergy-Pontoise, France) was performed as described previously [22] with minor modifications. Briefly, cells were resuspended at 2 × 10 7 cells/mL in HBSS containing CFSE at a final concentration of 0.5 μM.…”

Section: Preparation Of Cells From Lymph Nodes and Cfse Stainingmentioning

confidence: 99%

Analysis of cellular responses toMycoplasma mycoidessubsp.mycoidessmall colony biotype associated with control of contagious bovine pleuropneumonia

Totté

Rodrigues

YaYa³

et al. 2007

Vet. Res.

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-A better understanding of protective immune memory against contagious bovine pleuropneumonia (CBPP) is needed in order to facilitate the development of safer vaccines based on selected components of the pathogen. For this purpose, cells collected from lymph nodes draining the lungs of Mycoplasma mycoides subsp. mycoides small colony biotype (MmmSC)-infected cattle were stimulated with the pathogen in vitro and evaluated concurrently for proliferation (CFSE based method), expression of activation, memory markers and cytokine production. Direct evidence is presented for a major contribution of CD4 + T cells to the vigorous proliferative and T1 biased cytokine recall responses observed in cattle that have recovered from infection but not in animals developing the acute form of the disease. Two different phenotypes of MmmSC-specific memory CD4 were observed based on CD62L expression and proliferative capacities. Furthermore, recall proliferation of B cells also occurred but was strictly dependent on the presence of CD4. The information provided in this study will facilitate the search for MmmSC antigens that have potential for the development of subunit vaccines against CBPP.contagious bovine pleuropneumonia / Mycoplasma mycoides subsp. mycoides SC / vaccine / memory / CD4

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Evaluation of cell replication by bovine T cells in polyclonally activated cultures using carboxyfluorescein succinimidyl ester (CFSE) loading and flow cytometric analysis

Cited by 30 publications

References 15 publications

In vitro infection of aortic endothelial cells by caprine arthritis encephalitis virus enhances in vitro transmigration of peripheral blood leukocytes and modulates their phenotypic expression

In vitro infection of aortic endothelial cells by caprine arthritis encephalitis virus enhances in vitro transmigration of peripheral blood leukocytes and modulates their phenotypic expression

Expression and regulation of secreted phosphoprotein 1 in the bovine corpus luteum and effects on T lymphocyte chemotaxis

Analysis of cellular responses toMycoplasma mycoidessubsp.mycoidessmall colony biotype associated with control of contagious bovine pleuropneumonia

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