Clonal Growth of T Cells in vitro: Preliminary Attempts to a Quantitative Approach

Watanabe, Takeshi; Fathman, C. Garrison; Coutinho, António

doi:10.1111/j.1600-065x.1977.tb00233.x

Cited by 52 publications

(29 citation statements)

References 31 publications

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“…Unlike polyclonal Con A stimulation which is known to reverse with time (30), SCIF added at 3-day intervals induced exponential growth of alloantigen primed Ly2 .3 + T cells over a 3-4 wk period . Consequently, to avoid a crowding effect the cells had to be subcultured each 4-6th day .…”

Section: Discussionmentioning

confidence: 98%

T-T-cell interactions during the vitro cytotoxic allograft responses. I. Soluble products from activated Lyl+ T cells trigger autonomously antigen-primed Ly23+ T cells to cell proliferation and cytolytic activity.

Wagner¹,

Röllinghoff²

1978

The Journal of Experimental Medicine

308

103

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Secondary murine cytotoxic T lymphocyte responses from alloantigen-primed T cells can be induced in vitro by apparently unrelated regimens, such as addition of either concanavalin A (Con A), conditioned medium from Con A stimulated lymphocyte cultures, conditioned medium from secondary mixed lymphocyte cultures (MLC), or stimulator cells sharing only the I-region with the stimulating cells used for primary sensitization. We now report that upon polyclonal (Con A), or antigen-specific (MLC) stimulation, Lyl+ T cells release a factor, which in turn triggers alloantigen primed Ly23+ T cells to proliferation and cytolytic activity. The secondary cytotoxic T lymphocyte inducing factor (SCIF) is produced within 24 h. For its production, an intact protein metabolism, not DNA metabolism, is required. Once induced, the functional activity of SCIF is nonspecific and not H-2 restricted. SCIF allows exponential growth and long-term propagation of cytolytic Ly23+ T cells with specificity to alloantigens used for primary sensitization. SCIF induced activation of alloantigen primed Ly23+ T cells does not require the presence of alloantigens. The results therefore reveal a process by which Lyl+ T-cell-derived nonspecific factor(s) induce autonomously Ly23+ T-cell-mediated, antigen-specific, cytotoxic T lymphocyte responses.

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Section: Discussionmentioning

confidence: 98%

T-T-cell interactions during the vitro cytotoxic allograft responses. I. Soluble products from activated Lyl+ T cells trigger autonomously antigen-primed Ly23+ T cells to cell proliferation and cytolytic activity.

Wagner¹,

Röllinghoff²

1978

The Journal of Experimental Medicine

308

103

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“…The cultures were set up in 1 ml vol in 3-cm plastic Petri dishes and were activated with lipepelysaccharide (LPS) (100 ~ag/ml culture) or native dextran (5-10 mg/ culture) and the response tested at day 2 against SRC as well as SRC coupled with stearoyl dextran or FITC. Low cell density cultures contained 3 × 105 cells/ml in 2 ml medium (10). The cultures were supplemented with 5% human A serum and 2 mercaptoethanol at a final concentration of 5 × 10 -5 M. The mitogens were used at the same concentration as above and the response was tested 4 days later.…”

Section: Preparation Of Lymphocytesmentioning

confidence: 99%

Immunological unresponsiveness to native dextran B512 in young animals of dextran high responder strains is due to lack of Ig receptors expression. Evidence for a nonrandom expression of V-genes.

Fernández¹,

Möller²

1978

The Journal of Experimental Medicine

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Young mice of dextran high responder strains were found to be complete nonresponders to the alpha-1-6 epitope of dextran during 30-40 days after birth. They also failed to respond to thymus-dependent dextran-protein conjugates. Cells from young and adult mice were activated equally well to polyclonal antibody synthesis by the polyclonal B-cell-activating property of dextran. There was no age difference in the immune response to haptens conjugated to dextran, indicating that dextran can function as an efficient carrier also in young mice. Unresponsiveness could not be attributed to suppressor T cells or to a suppressive environment in young animals, as shown by transfer experiments, in which living or irradiated cells from young and adult mice were admixed in various ways and transferred to irradiated recipients of different ages. Cells from young mice did not affect response of adult cells (and the reverse), nor did the age of the irradiated recipient influence the response. When lymphocytes from young and adult mice were polyclonally activated in vitro by lipopolysaccharide, only cells from young mice failed to synthesize antibodies against the alpha-1-6 epitope of dextran, although they produced antibodies of all other specificities tested for. It was concluded that young animals fail to express immunoglobulins directed against the alpha-1-6 epitope during the first 30-40 days after birth. Since the mice possess the VH gene coding for antibodies against this particular epitope, it was concluded that the timing of V gene expression is regulated during development, possibly at the V-C gene translocation level.

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“…Indeed, there is evidence that elderly humans have fewer lymphocytes responsive to phytohemagglutinin [Inkeles et al, 1977] and to SK/SD [Sohnle et al, 1982], In stud ies of lymphocyte proliferation, this defi ciency could be overcome by repeated cell divisions to increase the number of re sponding cells. Since the mitotic process is time-consuming, the peak of lymphocyte proliferative responses involving fewer ini tially responsive cells would be delayed [ Watanabe et al, 1977], However, as pre viously demonstrated for LMIF [Rocklin, 1973] and as demonstrated in our LMIF system , lymphokine production does not require the participa tion of dividing cells. Therefore, another process such as recruitment of uncommitted cells must be involved to explain the de layed peak of lymphokine production in the elderly subjects.…”

Section: Discussionmentioning

confidence: 63%

Kinetics of Lymphokine Production by Lymphocytes from Elderly Humans

Sohnle¹,

Collins-Lech²,

Huhta³

1983

Gerontology

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Production in vitro of the lymphokine leukocyte migration inhibition factor (LMIF) was studied for elderly as compared to young human subjects over an 8-day period of culture. When stimulated by either the antigen streptokinase/streptodornase (SK/SD) or the mitogen concanavalin A (Con A), the peak of LMIF production was retarded in cultures of lymphocytes from elderly subjects. Absolute peak values and numbers of positive responses were diminished in this group when the cells were stimulated with SK/SD but not with Con A. However, if LMIF production during the first 48-hour period was examined, the responses to Con A were also diminished in the elderly group. Retardation of lymphokine production by lymphocytes from elderly humans may be relevant to the assessment of immunologic competence in this group as well as to potential adverse effects on in vivo reactions of cell-mediated immunity.

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Clonal Growth of T Cells in vitro: Preliminary Attempts to a Quantitative Approach

Cited by 52 publications

References 31 publications

T-T-cell interactions during the vitro cytotoxic allograft responses. I. Soluble products from activated Lyl+ T cells trigger autonomously antigen-primed Ly23+ T cells to cell proliferation and cytolytic activity.

T-T-cell interactions during the vitro cytotoxic allograft responses. I. Soluble products from activated Lyl+ T cells trigger autonomously antigen-primed Ly23+ T cells to cell proliferation and cytolytic activity.

Immunological unresponsiveness to native dextran B512 in young animals of dextran high responder strains is due to lack of Ig receptors expression. Evidence for a nonrandom expression of V-genes.

Kinetics of Lymphokine Production by Lymphocytes from Elderly Humans

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