Properties of memory T lymphocytes isolated from the mixed leukocyte reaction.

Inaba, Kayo; Koide, Sumil; Steinman, Ralph M.

doi:10.1073/pnas.82.22.7686

Cited by 35 publications

(26 citation statements)

References 19 publications

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“…Lymphoblasts were sensitized to lectin (3 ~,g/ml Con A, Pharmacia Fine Chemicals, Piscataway, N J), alloantigens, or soluble proteins (50 ~g/ml of either KLH [Calbiochem-Behring, San Diego, CA] or OVA [Sigma Chemical Co., St. Louis, MO]). The procedure was as described (4)(5)(6), and the cell yields are summarized in Table I. Briefly, 5 x 104 spleen DC were cultured with 5 × 106 T cells in 1 ml medium (RPMI-1640 supplemented with 10% FCS 20,,g/ml gentamicin, and 5 x l0 -~ M 2-ME) in 16-mm-diam culture wells.…”

Section: Methodsmentioning

confidence: 99%

Accessory cell-T lymphocyte interactions. Antigen-dependent and -independent clustering.

Inaba¹,

Steinman²

1986

The Journal of Experimental Medicine

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181

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Previous work documented the capacity of dendritic cells (DC) to stimulate primary immune responses and to physically cluster with the responding lymphocytes. Rapid cell-cell aggregation assays were used here to study the interaction of DC and other types of APC with T lymphocytes. Graded doses of APC were sedimented with T cells that had been primed to alloantigens, soluble proteins, or lectin, and then labeled with carboxyfluorescein diacetate. The number of clustered T cells was measured after 10 min at 4 or 37 degrees C. At 4 degrees, binding was antigen-dependent and included greater than 50% of the added T cells. Clustering was mediated by all types of APC tested, including DC, macrophages, B lymphocytes, and fresh Langerhans cells, although DC were the most effective. Specificity was evident in the findings that alloreactive T lymphoblasts bound to allogeneic but not syngeneic APC; KLH- and OVA-reactive T cells bound to syngeneic APC in the presence of specific protein: and Con A blasts needed lectin to cluster. A 30 min pretreatment with chloroquine, a drug known to inhibit APC activity, markedly blocked the specific binding of alloreactive and protein-specific T blasts at 4 degrees C. Since Lyt-2- alloreactive blasts should specifically recognize Ia, presentation of Ia seems to be altered by chloroquine. Binding assays at 37 degrees C gave similar results to those performed at 4 degrees C, with one exception. When DC were used as APC, striking antigen-independent clustering occurred. DC could efficiently cluster primed T cells in the absence of alloantigen, soluble protein, or lectin. We suggest that antigen-independent binding contributes to the distinctive capacity of DC to prime T cells in the afferent limb of the immune response, whereas antigen-dependent binding between other APC and sensitized lymphocytes is critical in the efferent limb.

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Section: Methodsmentioning

confidence: 99%

Accessory cell-T lymphocyte interactions. Antigen-dependent and -independent clustering.

Inaba¹,

Steinman²

1986

The Journal of Experimental Medicine

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181

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“…Memory T cells were prepared by resting the blasts for three or more days in the absence of exogenous IL-2 or alloantigen (19) . The blasts and memory populations were >70% allospecific, as assessed in rapid DC-T binding assays (28 Mixtures of accessory-cell depleted B and T lymphocytes were obtained by passing spleen suspensions over Sephadex G10 (17) .…”

Section: T Cells and T Cell Proliferationmentioning

confidence: 99%

Interleukin 1 enhances T-dependent immune responses by amplifying the function of dendritic cells.

Koide¹,

Inaba²,

Steinman³

1987

The Journal of Experimental Medicine

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192

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The function of exogenous murine recombinant IL-1 alpha as a T lymphocyte-activating molecule was examined. IL-1 did not induce IL-2 release or responsiveness in purified T cells regardless of their state of activation: unprimed lymphocytes, freshly sensitized lymphocytes, or memory cells derived from the blasts. Nor did IL-1 synergize with mitogens, or with antigens, to stimulate proliferation. For example the combinations of IL-1 plus Ia+ peritoneal macrophages, or IL-1 plus Con A, were less than 5% as effective in triggering T cell growth as a low dose (1%) of dendritic cells. However, when IL-1 was added at the onset of culture, the response to limiting doses of dendritic cells was increased 3- to 10-fold in several systems: the syngeneic and allogeneic MLR, Con A- and periodate-induced polyclonal mitogenesis, and T-dependent antibody formation against foreign red cells. The amplifying effect of IL-1 could be obtained if the dendritic cells but not the responding lymphocytes were exposed to IL-1 before use as accessory cells. Optimal activation of dendritic cells required a dose of 5 U/ml (50 pM) and 18 h of exposure, and was not due to carryover of IL-1 into the lymphocyte culture. IL-2, IL-3, and cachectin/TNF did not amplify dendritic cell function, while IFN-gamma diminished it. The enhanced function of IL-1-treated dendritic cells was due to an enhanced clustering with helper T lymphocytes in the first day of the MLR response. Therefore IL-1 does not seem to act as an activating factor for most peripheral T lymphocytes. Instead, IL-1 enhances the function of accessory dendritic cells and represents the first molecule that has been shown to enhance the immune response at this critical level.

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“…Although the clusters contain < 10% of the available T cells, these immunocompetent units are responsible for almost all the interleukins and growth factors produced in the cultures (3,8,14) and physical blocking of cluster formation by interposing an artificial membrane abrogates the observed immunological responses (15). In all instances the responsible cell is an Lyt-2-helper T cell (4,5); but murine DC can also cluster with and activate cytotoxic T cells (1 6).…”

Section: Introductionmentioning

confidence: 99%

“…generate active helper cells from unprimed and memory lymphocytes, and cause B cells to secrete antibodies (2)(3)(4)(5). Cultured spleen cells depleted of DC failed to develop anti-TNP cytotoxic T cells while the addition of a small number of DC restored the antigen dependent response (6).…”

Section: Introductionmentioning

confidence: 99%

Dendritic cell-lymphocyte clusters that form spontaneously in rheumatoid arthritis synovial effusions differ from clusters formed in human mixed leukocyte reactions.

Tsai¹,

Zvaifler²

1988

J. Clin. Invest.

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Lymphocytes cluster about dendritic cells (DC) spontaneously in 48 h cultures of rheumatoid arthritis synovial fluid (RA SF) mononuclear cells and in peripheral blood autologous or allogeneic mixed leukocyte reactions. In the latter case, the clusters are predominantly CD4+ T cells (T4/T8 > 5) and with time progress into blastic cells that express IL-2 (Tac) and/or transferrin (T9) receptors. In contrast, the clusters in RA SF cultures have a T4/T8 ratio of < 1 and a majority of the T8 cells coexpress the Leu 7 marker. T cells in these clusters remain inert and with time the clusters disintegrate. Addition of IL-1, IL-2, or IFN-gamma alone or in combination had no effect on RA

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Properties of memory T lymphocytes isolated from the mixed leukocyte reaction.

Cited by 35 publications

References 19 publications

Accessory cell-T lymphocyte interactions. Antigen-dependent and -independent clustering.

Accessory cell-T lymphocyte interactions. Antigen-dependent and -independent clustering.

Interleukin 1 enhances T-dependent immune responses by amplifying the function of dendritic cells.

Dendritic cell-lymphocyte clusters that form spontaneously in rheumatoid arthritis synovial effusions differ from clusters formed in human mixed leukocyte reactions.

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