Background: Infections can aggravate the course of neurodegenerative diseases including amyotrophic lateral sclerosis (ALS). Mutations in the anti-oxidant enzyme Cu, Zn superoxide dismutase (EC 1.15.1.1, SODI) are associated with familial ALS. Streptococcus pneumoniae, the most frequent respiratory pathogen, causes damage by the action of the cholesterol-binding virulence factor pneumolysin and by stimulation of the innate immune system, particularly via Toll-like-receptor 2. Methods: SH-SY5Y neuroblastoma cells transfected with the G93A mutant of SODI typical for familial ALS (G93A- SODI) and SH-SY5Y neuroblastoma cells transfected with wildtype SODI were both exposed to pneumolysin and in co-cultures with cultured human macrophages treated with the Toll like receptor 2 agonist N-palmitoyl-S-[2,3-bis(palmitoyloxy)-(2RS)-propyl]-[R]-cysteinyl-[S]-seryl-[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]-lysine x 3 HCI (Pam(3)CSK(4)). Cell viability and apoptotic cell death were compared morphologically and by in-situ tailing. With the help of the WST-I test, cell viability was quantified, and by measurement of neuron-specific enolase in the culture supernatant neuronal damage in co-cultures was investigated. Intracellular calcium levels were measured by fluorescence analysis using fura-2 AM. Results: SH-SY5Y neuroblastoma cells transfected with the G93A mutant of SODI typical for familial ALS (G93A- SODI) were more vulnerable to the neurotoxic action of pneumolysin and to the attack of monocytes stimulated by Pam(3)CSK(4) than SH-SY5Y cells transfected with wild-type human SODI. The enhanced pneumolysin toxicity in G93A- SODI neuronal cells depended on the inability of these cells to cope with an increased calcium influx caused by pores formed by pneumolysin. This inability was caused by an impaired capacity of the mitochondria to remove cytoplasmic calcium. Treatment of G93A- SODI SH-SY5Y neuroblastoma cells with the antioxidant N-acetylcysteine reduced the toxicity of pneumolysin. Conclusion: The particular vulnerability of G93A-SODI neuronal cells to hemolysins and inflammation may be partly responsible for the clinical deterioration of ALS patients during infections. These findings link infection and motor neuron disease and suggest early treatment of respiratory infections in ALS patients.

Goos, M., Zech, W.D., Jaiswal, M.K., Balakrishnan, S., Ebert, S., Mitchell, T., et al. (2007). Expression of a Cu, Zn superoxide dismutase typical for familial amyotrophic lateral sclerosis increases the vulnerability of neuroblastoma cells to infectious injury. BMC INFECTIOUS DISEASES, 7, 131 [10.1186/1471-2334-7-131].

Expression of a Cu, Zn superoxide dismutase typical for familial amyotrophic lateral sclerosis increases the vulnerability of neuroblastoma cells to infectious injury

CARRI', MARIA TERESA;
2007

Abstract

Background: Infections can aggravate the course of neurodegenerative diseases including amyotrophic lateral sclerosis (ALS). Mutations in the anti-oxidant enzyme Cu, Zn superoxide dismutase (EC 1.15.1.1, SODI) are associated with familial ALS. Streptococcus pneumoniae, the most frequent respiratory pathogen, causes damage by the action of the cholesterol-binding virulence factor pneumolysin and by stimulation of the innate immune system, particularly via Toll-like-receptor 2. Methods: SH-SY5Y neuroblastoma cells transfected with the G93A mutant of SODI typical for familial ALS (G93A- SODI) and SH-SY5Y neuroblastoma cells transfected with wildtype SODI were both exposed to pneumolysin and in co-cultures with cultured human macrophages treated with the Toll like receptor 2 agonist N-palmitoyl-S-[2,3-bis(palmitoyloxy)-(2RS)-propyl]-[R]-cysteinyl-[S]-seryl-[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]-lysine x 3 HCI (Pam(3)CSK(4)). Cell viability and apoptotic cell death were compared morphologically and by in-situ tailing. With the help of the WST-I test, cell viability was quantified, and by measurement of neuron-specific enolase in the culture supernatant neuronal damage in co-cultures was investigated. Intracellular calcium levels were measured by fluorescence analysis using fura-2 AM. Results: SH-SY5Y neuroblastoma cells transfected with the G93A mutant of SODI typical for familial ALS (G93A- SODI) were more vulnerable to the neurotoxic action of pneumolysin and to the attack of monocytes stimulated by Pam(3)CSK(4) than SH-SY5Y cells transfected with wild-type human SODI. The enhanced pneumolysin toxicity in G93A- SODI neuronal cells depended on the inability of these cells to cope with an increased calcium influx caused by pores formed by pneumolysin. This inability was caused by an impaired capacity of the mitochondria to remove cytoplasmic calcium. Treatment of G93A- SODI SH-SY5Y neuroblastoma cells with the antioxidant N-acetylcysteine reduced the toxicity of pneumolysin. Conclusion: The particular vulnerability of G93A-SODI neuronal cells to hemolysins and inflammation may be partly responsible for the clinical deterioration of ALS patients during infections. These findings link infection and motor neuron disease and suggest early treatment of respiratory infections in ALS patients.
Pubblicato
Rilevanza internazionale
Articolo
Sì, ma tipo non specificato
Settore BIO/11
English
Con Impact Factor ISI
acetylcysteine; adenine; calcium; copper zinc superoxide dismutase; cysteine; fura 2 acetoxymethyl ester; guanine; hemolysin; lysine; neuron specific enolase; pneumolysin; serine; toll like receptor 2; antioxidant; bacterial protein; caspase 3; CD68 antigen, human; differentiation antigen; leukocyte antigen; Pam(3)CSK(4) peptide; peptide; plY protein, Streptococcus pneumoniae; streptolysin; superoxide dismutase; TLR2 protein, human; amyotrophic lateral sclerosis; apoptosis; article; calcium cell level; calcium transport; cell stimulation; cell structure; cell viability; central nervous system infection; coculture; controlled study; cytoplasm; disorders of mitochondrial functions; enzyme analysis; fluorescence analysis; genetic transfection; human; human cell; inflammation; macrophage; monocyte; nerve cell lesion; neuroblastoma cell; protein expression; respiratory tract infection; wild type; cell culture; cell survival; cytology; drug antagonism; drug effect; enzymology; genetics; immunohistochemistry; pathology; tumor cell line; acetylcysteine; amyotrophic lateral sclerosis; antigens, CD; differentiation, myelomonocytic; antioxidants; apoptosis; bacterial proteins; calcium; caspase 3; cell line, tumor; cell survival; cells, cultured; coculture techniques; humans; immunohistochemistry; macrophages; monocytes; mutation; neuroblastoma; peptides; streptolysins; superoxide dismutase; toll-like receptor 2; transfection
Goos, M., Zech, W.D., Jaiswal, M.K., Balakrishnan, S., Ebert, S., Mitchell, T., et al. (2007). Expression of a Cu, Zn superoxide dismutase typical for familial amyotrophic lateral sclerosis increases the vulnerability of neuroblastoma cells to infectious injury. BMC INFECTIOUS DISEASES, 7, 131 [10.1186/1471-2334-7-131].
Goos, M; Zech, W; Jaiswal, M; Balakrishnan, S; Ebert, S; Mitchell, T; Carri', Mt; Keller, B; Nau, R
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2108/8037
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