An estimate of the minimal frequency of cytolytic T lymphocyte effector cells generated in allogeneic reactions

Engers, Howard; Fitch, Frank W.

doi:10.1016/0022-1759(79)90159-5

Cited by 10 publications

(5 citation statements)

References 15 publications

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“…(a) To screen the microwells of a cloning plate (6.4-ram diameter; Costar 3596; Costar, Data Packaging, Cambridge, Mass.) for specific cytolytic activity, such as was done to select for V4, a microcytotoxicity assay using 500 target cells was employed (18). (b) The conventional cytotoxicity assay using 5,000 target cells is essentially as previously described (19), except that centrifugation for 60 see at 200 g preceded the 3.5-h incubation at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Lyt-2-/Lyt 3- variants of a cloned cytolytic T cell line lack an antigen receptor functional in cytolysis.

Dialynas¹,

Loken²,

Glasebrook³

et al. 1981

The Journal of Experimental Medicine

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Detailed analysis of cytolytic T lymphocytes (CTL)1 at the molecular level has been hampered by the cellular heterogeneity intrinsic to most model systems studied. The development of cloned lines (1, 2) of CTL facilitates both biochemical (3) and genetic (this report) analyses of function.CTL have been characterized by a distinctive cell surface antigenic profile that includes Thy-1, Lyt-2, and Lyt-3 (4). Several recent studies using alloantisera indicate that antibodies reactive with either Lyt-2 or Lyt-3, or with the product of a closelinked locus, are able to block CTL activity in the absence of complement (5, 6). Monoclonal antibodies directed against either Lyt-2 (7, 8) or Lyt-3 (J. A. Ledbetter and L. A. Herzenberg, personal communication) also have been found to block CTL activity in the absence of complement. These data imply a role for Lyt-2 and Lyt-3 in cytolysis, but they cannot formally exclude that a molecule required for cytolytic activity merely is located near Lyt-2 and Lyt-3 on the cell surface. Antibody to Thy-1 does not block CTL activity in the absence of complement (7-9).To investigate the role of Lyt-2 and Thy-1 in cytolysis from an independent approach, we have generated, and analyzed the phenotypes of, variants of an especially lytic cloned CTL line (designated L3) which specifically lack either Lyt-2 or Thy-1. An analysis of these variants indicates that neither Lyt-2 nor Lyt-3 is responsible for the lethal hit, but suggests that Lyt-2 and/or Lyt-3 are required for an antigen receptor functional in cytolysis. The data also suggest that the expression of Lyt-3 on the cell surface is not independent of the expression of Lyt-2. Finally the data indicate that Thy-1 plays no role in cytolysis.

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Section: Methodsmentioning

confidence: 99%

Lyt-2-/Lyt 3- variants of a cloned cytolytic T cell line lack an antigen receptor functional in cytolysis.

Dialynas¹,

Loken²,

Glasebrook³

et al. 1981

The Journal of Experimental Medicine

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“…T-cell clones were derived from unidirectional secondary C57BL/6 anti-DBA/2 or A/J anti-DBA/2 MLC as described (11). Clones were assayed for cytolytic activity by testing for ability to lyse P-815 (H-2d) mastocyte target cells, initially in a 51Cr-release microcytotoxicity assay (12) and then in a 3-hr 51Cr-release cytotoxicity assay (13). Clones showing no cytolytic activity were tested for ability to amplify cytolytic activity levels of Tc clones as described (11).…”

Section: Methodsmentioning

confidence: 99%

Cell surface polypeptides of murine T-cell clones expressing cytolytic or amplifier activity.

Sarmiento¹,

Glasebrook²,

Fitch³

1980

Proc. Natl. Acad. Sci. U.S.A.

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“…However, it is not essential to use this sequence of lymphocyte stimulation to derive functional T cell clones since we have been able to obtain such clones after primary stimulation with alloantigen in vitro. Growing cells were evident 6 to 9 days after initiation of the cloning culture at which time individual wells were screened for cytolytic activity using a sensitive micro ^^Cr-release assay (Engers & Fitch 1979). Cells were then expanded and maintained by weekly passage with irradiated alloantigen and TCGF present in secondary MLC supernatant fluids (MLC-TCGF)*.…”

Section: Procedures For Deriving T Cell Clonesmentioning

confidence: 99%

Murine T Lymphocyte Clones with Distinct Immunological Functions

Glasebrook

Sarmiento

Loken

et al. 1981

Immunological Reviews

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127

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An estimate of the minimal frequency of cytolytic T lymphocyte effector cells generated in allogeneic reactions

Cited by 10 publications

References 15 publications

Lyt-2-/Lyt 3- variants of a cloned cytolytic T cell line lack an antigen receptor functional in cytolysis.

Lyt-2-/Lyt 3- variants of a cloned cytolytic T cell line lack an antigen receptor functional in cytolysis.

Cell surface polypeptides of murine T-cell clones expressing cytolytic or amplifier activity.

Murine T Lymphocyte Clones with Distinct Immunological Functions

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