Isolation and characterization of a serine esterase from cytolytic T cell granules

Young, John Ding‐E; Leong, Lauren Gee; Liu, Chau‐Ching; Damiano, Angela; Wall, Doris A.; Cohn, Zanvil A.

doi:10.1016/0092-8674(86)90441-1

Cited by 168 publications

(56 citation statements)

References 69 publications

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“…An indication of the proportion is suggested by the amount of a serine esterase (termed BLT-serine esterase after the synthetic substrate it cleaves) released into the culture medium when CTLs attack target cells (33) . Because the enzyme seems to be located in the same cytoplasmic granules that contain perforin (33)(34)(35), the amount of secreted enzyme probably approximates the number of toxic granules that are released. Secreted perforin, in contrast, is not detectable in the culture medium, perhaps because of a strong propensity to bind to cell membranes .…”

Section: Inhibition Of Granule-mediated Lysis Of Red Blood Cells By Cmentioning

confidence: 99%

Resistance of cytotoxic T lymphocytes to the lytic effects of their toxic granules.

Verret

Firmenich

Kranz

et al. 1987

The Journal of Experimental Medicine

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A cytotoxic T lymphocyte (CTL) characteristically kills target cells one after the other by releasing toxic granules that contain one or more cytolytic components. To determine how CTLs avoid destroying themselves when they release granules and lyse target cells, 7 murine CD8+ CTL cell lines were compared with 19 other cell lines for susceptibility to lysis by the isolated toxic granules. Murine CD8+ CTLs were clearly the most resistant cells: granules did not lyse them even after they were exposed to azide, cyanide, and 2-deoxyglucose, conditions that were found to enhance the susceptibility of all the other cells tested, including other T cells. Thus, resistance of CD8+ CTLs to cytotoxic granules appears to be independent of cellular ATP. To reconcile these findings with other observations that, under some circumstances, CTLs can be lysed by other CTLs, we suggest a model in which a CTL releases only a limited proportion of its toxic granules at each antigen-specific encounter with a target cell; the amount released is sufficient to kill most target cells but to leave the CTL undamaged and with enough granules to attack other target cells.

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Section: Inhibition Of Granule-mediated Lysis Of Red Blood Cells By Cmentioning

confidence: 99%

Resistance of cytotoxic T lymphocytes to the lytic effects of their toxic granules.

Verret

Firmenich

Kranz

et al. 1987

The Journal of Experimental Medicine

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“…In the mouse, seven granzymes named A to lymphocyte (CTL), natural killer (NK) and lymphokine activated G have been identified (10)(11)(12)(13)(14)(15) and the respective cDNAs have killer (LAK) cell-mediated target cell lysis, the molecular been isolated (16)(17)(18)(19)(20)(21)(22). In human CTLs, only the granzymes A mechanism responsible for the 'lethal hit' delivery is still poorly and B have been characterized at the protein and gene level understood.…”

Section: Introductionmentioning

confidence: 99%

Structure and evolutionary origin of the human granzyme H gene

Haddad¹,

Jenne

Tschopp

et al. 1991

Int Immunol

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Among the molecules proposed to be involved in cytotoxic T lymphocyte (CTL), natural killer (NK) and lymphokine activated killer (LAK) cell-mediated lysis are the granzymes, a family of serine proteases stored in the cytoplasmic granules of CTLs, NK and LAK cells. In addition to the granzymes A and B, a third member of this family has been cloned in man and designated granzyme H. We present the complete gene sequence including the 5' promoter region and demonstrate that the granzyme H sequence represents a functional gene expressed in activated T cells. Granzyme H shows the highest degree (>54%) of amino acid sequence homology with granzyme B and cathepsin G and, like these genes, consists of five exons separated by introns at equivalent positions. The evolutionary history of granzyme H has been analyzed by reconstructing an evolutionary tree for granzyme sequences. We provide evidence that interlocus recombination between the ancestral genes of granzyme B and granzyme H occurred about 21 million years ago, leading to a replacement of exon 3, intron 3 and part of exon 4 in human granzyme H by human granzyme B sequences. Our results suggest that the ancestral gene of granzyme H is more closely related to cathepsin G and granzyme B than to the murine granzymes C to G. Thus, granzyme H does not represent a human counterpart of the known murine granzymes A to G. It diverged from cathepsin G before mammalian radiation and should, therefore, exist in other mammalian lineages as well.

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“…Subcellular Fractionation and Affinity Purification. Fractionation of mast cells using a discontinuous Percoll gradient and microassays for protein, proteoglycan, hemolysis, and enzymes were done exactly as described (26,27). The Percoll was pelleted by high-speed centrifugation (26), after which granule fractions (the equivalent of 5 x 108 cells per ml) were resuspended in relaxation buffer.…”

mentioning

confidence: 99%

Identification, purification, and characterization of a mast cell-associated cytolytic factor related to tumor necrosis factor.

Young

Liu

Butler

et al. 1987

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

214

109

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The role of mast cells and mast-cell-derived factors in natural cytotoxic reactions was investigated. Cultured and freshly isolated murine mast cells are shown to be cytotoxic to WEHI-164 and YAC-1 targets in 18-hr viability assays but not in 4-hr assays. Here, we describe a cytotoxic factor in murine mast cells that is immunologically related to tumor necrosis factor (TNF). This TNF-like factor lyses WEHI-164 cells with a slow time course requiring 16-20 hr for the lytic reaction to complete. Antibodies specific for human and murine TNF and human lymphotoxin partially block mast cell lysis of WEHI-164 cells. These antibodies react on immunoblots with one major mast cell protein band of 50 kDa. Immunoblot analysis shows this factor in cloned and uncloned cultured mouse mast cells and in mature "connective tissue-type" mast cells freshly purified from rat or mouse peritoneal cavities. The amount of this factor is greatly enhanced in cells that have been stimulated with a combination of phorbol ester/concanavalin A or bacterial lipopolysaccharide. Subcellular fractionation analysis of mast cells with Percoll gradients reveals two pools of TNF-related cytotoxic activity that are associated with free cytosolic material and granule fractions. In contrast to cytotoxic T lymphocytes and natural killer cells, granuleenriched fractions of mast cells do not contain any hemolytic activity. The localization of the TNF-like molecule in mast cell granules may play a strategical role in the rapid delivery of this mediator to the target cell membrane following cell surface stimulation and degranulation.

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Isolation and characterization of a serine esterase from cytolytic T cell granules

Cited by 168 publications

References 69 publications

Resistance of cytotoxic T lymphocytes to the lytic effects of their toxic granules.

Resistance of cytotoxic T lymphocytes to the lytic effects of their toxic granules.

Structure and evolutionary origin of the human granzyme H gene

Identification, purification, and characterization of a mast cell-associated cytolytic factor related to tumor necrosis factor.

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