Perforin/granzyme-induced apoptosis is the main pathway used by cytotoxic lymphocytes to kill virus-infected and transformed cells. The main function of the FAS–FAS ligand (FASL) pathway is to maintain homeostasis of the lymphoid compartment.
Studies in gene-disrupted mice indicate that perforin is vital for cytotoxic effector function. Perforin-deficient mice are abnormally susceptible to many viruses and other intracellular pathogens, they fail to reject certain types of allograft and they are highly susceptible to spontaneous B-cell lymphomas as they age.
In certain situations, perforin deficiency is associated with reduced immunopathology. For example, non-obese diabetic (NOD) mice that lack perforin develop islet inflammation, but are protected from autoimmune destruction of their pancreatic β-cells and have a reduced incidence of diabetes compared with wild-type NOD mice.
The molecular functions of perforin are not well understood. Perforin has an indispensable role in delivering granzyme B and other granule toxins to the cytosol of target cells. However, the hypothesis that perforin pores allow the passive diffusion of granzymes into cells is not consistent with recent experimental data, and this model is probably an over-simplification.
Recently, a subset of patients with the rare immunodeficiency disease familial haemophagocytic lymphohistiocytosis (FHL) were found to have missense and/or nonsense mutations in both of their perforin alleles. The catalogue of deduced perforin mutations should enable researchers to define in more detail the basis of the normal molecular functions of perforin.
Granzymes trigger several apoptotic pathways in target cells; some of them are dependent on caspases, but others can kill cells when caspases are inhibited, for example by viral proteins that block intrinsic apoptotic pathways.
Deficiency of an individual granzyme might be associated with focal immune deficits — for example, the marked susceptibility of granzyme-A- or -B-deficient mice and, particularly, granzyme A and B double-deficient mice to the poxvirus ectromelia. The same animals do not have increased susceptibility to related poxviruses.
Perforin protects mice against the development of spontaneous B-cell lymphoma and carcinogen-induced sarcoma. It remains to be determined whether surveillance against other cancers, particularly epithelial malignancies, is affected in perforin-deficient mice.
In addition to determining whether a virus is cleared or persists, the effector pathway that is selected by killer lymphocytes (perforin/granzyme, FASL or interferon-γ) can influence immunopathology, often in an organ-specific manner.
Many mechanisms of immune escape that operate at the level of reduced antigen presentation have been described for virus-infected and malignant cells. It is now becoming increasingly clear that alternative mechanisms can operate at the level of inhibiting effector-cell function.
Perforin/granzyme-induced apoptosis is the main pathway used by cytotoxic lymphocytes to eliminate virus-infected or transformed cells. Studies in gene-disrupted mice indicate that perforin is vital for cytotoxic effector function; it has an indispensable, but undefined, role in granzyme-mediated apoptosis. Despite its vital importance, the molecular and cellular functions of perforin and the basis of perforin and granzyme synergy remain poorly understood. The purpose of this review is to evaluate critically recent findings on cytotoxic granule-mediated cell death and to assess the functional significance of postulated cell-death pathways in appropriate pathophysiological contexts, including virus infection and susceptibility to experimental or spontaneous tumorigenesis.
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We thank all of the present and past staff and students of the Cancer Immunology Laboratory at the Peter MacCallum Cancer Institute and the Austin Research Institute (pre-2000), particularly K. Browne, E. Cretney, V. Sutton, S. Street, J. Kelly and K. Thia. Our studies described and cited in this review have been supported by funding from the National Health and Medical Research Council (NHMRC), Australia, The Anti-Cancer Council, Victoria, and the Wellcome Trust, UK, between 1990 and the present. Both J.A.T. and M.J.S. are past recipients of Wellcome Trust Senior Research Fellowships, and current Principal Research Fellows of the NHMRC. We also thank K. Thia for help with preparing the illustrations for this paper.
A common form of cell death, also known as 'intrinsic' or 'programmed' cell death. Many physiological and developmental stimuli cause apoptosis, and themechanism is used frequently to delete unwanted, superfluous or potentially harmful cells, such as those undergoing transformation. Apoptosis involves cell shrinkage, condensation of chromatin in the periphery of the nucleus, cell-membrane blebbing and DNA fragmentation into multiples of about 180 base pairs. Eventually, the cell breaks up into many membrane-bound 'apoptotic bodies', which are phagocytosed by neighbouring cells.
A serine protease inhibitor. Serpins are a large family of intracellular and extracellular protease inhibitors, with many diverse functions. Some serpins show 'cross-class inhibition' and are, therefore, effective inhibitors of other protease families, such as the cysteine proteases. PI9 is a recently described serpin that is synthesized by cytotoxic T lymphocytes (CTLs). It is postulated to neutralize granzyme B molecules that are misdirected to the cytosol, thereby protecting the CTL from accidental suicide. Many viruses encode serpins that block caspases, the enzymes that are responsible for apoptotic death.
- GRAFT-VERSUS-HOST DISEASE
(GVHD). Tissue damage in a recipient of allogeneic transplanted tissue (usually a bone-marrow transplant) that results from the activity of donor cytotoxic T lymphocytes that recognize the recipient's tissue as foreign. GVHD varies markedly in severity, but can be life threatening in severe cases. Typically, damage to the skin and gut mucosa leads to clinical manifestations.
A common form of cell death that frequently results from toxic injury, hypoxia or stress. Necrosis involves cell swelling, dysregulation of plasma-membrane ion and water fluxes, mitochondrial swelling and the eventual release of cell contents into the interstitium. This form of cell death is usually accompanied by inflammation. Cells that are exposed to high concentrations of purified perforin usually die by osmotic lysis, which is a form of necrotic death.
A designation that is used to describe subsets of CD8+ cytotoxic T lyphocytes. TC1 cells typically secrete IFN-γ and GM-CSF and have strong cytotoxic capacity, whereas TC2 cells secrete IL-4 and IL-10 and are less effective killers.
A form of cell killing in which one of a group of similar cells kills another member or members of the group.
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Trapani, J., Smyth, M. Functional significance of the perforin/granzyme cell death pathway. Nat Rev Immunol 2, 735–747 (2002). https://doi.org/10.1038/nri911
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