Characterization of lymphocyte subpopulations in sheep by rosette formation, adherence to nylon wool and mitogen responsiveness

Outteridge, P.M.; Licence, S. T.; Binns, R. M.

doi:10.1016/0165-2427(81)90034-9

Cited by 24 publications

(8 citation statements)

References 16 publications

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“…There are few reports describing the lymphocyte subpopulations in normal sheep (14)(15)(16)(17)(18). We found that the rosettes formed with SRBC and ovine PBL are fragile but that fixation with glutaraldehyde allowed the identification of a higher percentage of rosette-forming ovine T cetls than reported previously (14).…”

Section: Discussioncontrasting

confidence: 53%

Lymphocyte subpopulations in sheep during the early stage of experimental infection with bovine leukaemia virus

Wh²,

Kf³

1989

Immunol Cell Biol

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Summary Stieep were experiraentalty infected with bovine leukaemia virus (BLV) by the inoculation of PBL from leukaemic sheep. Antibodies to viral structural proteins were detected at from 2 to 6 weeks after inoculation. At seroconversion, all sheep had a marked increase in the number of circulating lymphocytes, due essentially to an increase in the number of B cells. The number of circulating B cells then decreased but remained higher than pre-infection levels. A second increase in this population preceded the development of a B cell lymphoblastic leukaemia. Generalized lymphosarcoma was diagnosed at necropsy of all sheep. Variation between individual sheep in the time from infection to the development of tumours allowed two clearly delineated groups of nine sheep to be compared. A study of changes in the B cell and T cell populations during the first 16 weeks of infection suggested that the initial response to infection influences the subsequent rate of leukaemogenesis. At seroconversion the number of circulating B cells was significantly higher in group 1 (1016±l-51X10'/l)than in group2(6-47±276X10'/l). Group 1 sheep became leukaemic at 20-50 weeks after infection, whereas group 2 sheep did not do so until 70-95 weeks after infection.

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Section: Discussioncontrasting

confidence: 53%

Lymphocyte subpopulations in sheep during the early stage of experimental infection with bovine leukaemia virus

Wh²,

Kf³

1989

Immunol Cell Biol

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“…Furthermore, this study underlines the validity and value of our use of the calculated elution pattern of each subpopulation, as an index of subpopulation adherence, a calculation which is inherently subject to vigorous backchecking because of the cellular 'balance sheet' involved. Since the initial description [9], the method has proved itself in this series and in studies of pig lymphocytes with a weak Fc receptor [4] and sheep lymphocyte subpopu lations [14], Technical parameters of the method have been discussed thoroughly elsewhere [9] but two fur ther points are noteworthy. Confirmation that sub population elution is not random was proved by our observation that 96% of NAD cells from a first co lumn loaded on a second column eluted in the NAD fraction.…”

Section: Discussionmentioning

confidence: 99%

“…Studies were made of: slg4 lymphocytes by immunofluorescence, Fc+ lympho cytes by rosette formation with bovine RBC sensitized with hyper immune pig antibody, E rosette-forming lymphocytes by rosette formation with SRBC in dextran (DS*) orsaline (S*) and Null cells using the formula Null% = 100 -(sIg*°/o + DS*%). E rosettes were separated preparatively by centrifugation at -400 g, on density lay ers (12% FicollTriosil) for I h at 4 0C and the lymphocytes released by ammonium chloride-Tris (ACT) lysis, exactly as for sheep lym phocytes [ 14]. Lymphocytes were separated on nylon wool columns for tissue culture or for analysis of subpopulation nylon-adherence as detailed previously for pig [4,9] and sheep [14] lymphocytes.…”

Section: Methodsmentioning

confidence: 99%

“…E rosettes were separated preparatively by centrifugation at -400 g, on density lay ers (12% FicollTriosil) for I h at 4 0C and the lymphocytes released by ammonium chloride-Tris (ACT) lysis, exactly as for sheep lym phocytes [ 14]. Lymphocytes were separated on nylon wool columns for tissue culture or for analysis of subpopulation nylon-adherence as detailed previously for pig [4,9] and sheep [14] lymphocytes. Techniques for tissue culture, tritiated thymidine incorporation, cell harvesting, and scintillation counting in the in vitro mitogen studies were identical to those described for sheep lymphocytes [ 14].…”

Section: Methodsmentioning

confidence: 99%

“…Lymphocytes were separated on nylon wool columns for tissue culture or for analysis of subpopulation nylon-adherence as detailed previously for pig [4,9] and sheep [14] lymphocytes. Techniques for tissue culture, tritiated thymidine incorporation, cell harvesting, and scintillation counting in the in vitro mitogen studies were identical to those described for sheep lymphocytes [ 14].…”

Section: Methodsmentioning

confidence: 99%

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Subpopulations of Pig Blood E-Rosette-Forming Lymphocytes and Thymus-Dependent Null Cells: Separation by Nylon Wool Columns, Rosette Formation and Macrophage-Dependent Mitogen and Antigen Responsiveness

Outteridge¹,

Binns²,

Licence³

1982

Int Arch Allergy Immunol

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Pig peripheral blood lymphocyte subpopulations were characterized by their adherence to nylon wool. Thus whilst B cells, defined by surface Ig and Fc receptor, were predominantly adherent, thymus-dependent Null cells and the T cells forming rosettes with sheep red blood cells in dextran (DS⁺) were predominantly non-adherent. There was a direct relationship between the size of the non-adherent fraction and the proportion of the total DS⁺ cells which were in the non-adherent fraction. Using nylon wool and/or DS rosette separation, lymphocytes from normal and BCG sensitized animals were stimulated in vitro with phytohemagglutinin (PHA), concanavalin A (Con A) and tuberculin (PPD). The cells responding to PHA and Con A (T cell mitogens in other species) were DS⁺ lymphocytes. Non-rosetting, adherent (B cells) and non-adherent (thymus dependent Null cells) lymphocytes were unresponsive, even in the presence of homologous alveolar macrophages at levels which greatly enhanced the PPD response. Cells responding to PPD were present in the DS⁺ and DS^––, adherent and non-adherent fractions but their response was apparently controlled by macrophage dependence and suppression. Techniques are described for the preparation and storage in liquid nitrogen of alveolar macrophages resulting in high numbers of viable cells, capable of helping in vitro responses, even after 3 years of storage.

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An immunological classification of ovine lymphomas

Dixon

Moriarty

Johnstone

1984

Journal of Comparative Pathology

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Characterization of lymphocyte subpopulations in sheep by rosette formation, adherence to nylon wool and mitogen responsiveness

Cited by 24 publications

References 16 publications

Lymphocyte subpopulations in sheep during the early stage of experimental infection with bovine leukaemia virus

Lymphocyte subpopulations in sheep during the early stage of experimental infection with bovine leukaemia virus

Subpopulations of Pig Blood E-Rosette-Forming Lymphocytes and Thymus-Dependent Null Cells: Separation by Nylon Wool Columns, Rosette Formation and Macrophage-Dependent Mitogen and Antigen Responsiveness

An immunological classification of ovine lymphomas

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