Combination therapy with transductive anti-death FNK protein and FK506 ameliorates brain damage with focal transient ischemia in rat

Katsura, Ken‐ichiro; Takahashi, Kumiko; Asoh, Sadamitsu; Watanabe, Megumi; Sakurazawa, Makoto; Ohsawa, Ikuroh; Mori, Takashi; Igarashi, Hironaka; Ohkubo, Seiji; Katayama, Yasuo; Ohta, Shigeo

doi:10.1111/j.0022-3042.2008.05360.x

Cited by 10 publications

(7 citation statements)

References 60 publications

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“…Therapeutic proteins and peptides have been delivered to several cell types or tissues such as brain (19,29,32,36) muscle (5,8,49,50), heart (3,15), eye lens epithelial cells (22), and into living organisms (43). The recombinant proteins are considered for use in cell transplantation as well (33,35).…”

Section: Discussionmentioning

confidence: 99%

Endotoxin Deactivation by Transient Acidification

Ribeiro

Klein

et al. 2010

Cell Transplant

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Recombinant proteins are an important tool for research and therapeutic applications. Therapeutic proteins have been delivered to several cell types and tissues and might be used to improve the outcome of the cell transplantation. Recombinant proteins are propagated in bacteria, which will contaminate them with the lypopolysacharide-endotoxin found in the outer bacterial membrane. Endotoxin could interfere with in vitro biological assays and is the major pathological factor, which must be removed or inactivated before in vivo administration. Here we describe a one-step protocol in which the endotoxin activity on recombinant proteins is remarkably reduced by transient exposure to acidic conditions. Maximum endotoxin deactivation occurs at acidic pH below their respective isoelectric point (pI). This method does not require additional protein purification or separation of the protein from the endotoxin fraction. The endotoxin level was measured both in vitro and in vivo. For in vitro assessment we have utilized Limulus Amebocyte Lysate method for in vivo the pyrogenic test. We have tested the above-mentioned method with 5 different recombinant proteins including a monoclonal antibody clone 5c8 against CD154 produced by hybridomas. More than 99% of endotoxin was deactivated in all of the proteins, the recovery of the protein after deactivation varied between maximum 72.9 and minimum 46.8%. The anti CD154 clone 5c8 activity remained unchanged as verified by the measurement of binding capability to activated lymphocytes. Furthermore, the effectiveness of this method was not significantly altered by urea, commonly used in protein purification. This procedure provides a simple and cost-efficient way to reduce the endotoxin activity in antibodies and recombinant proteins.

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Section: Discussionmentioning

confidence: 99%

Endotoxin Deactivation by Transient Acidification

Ribeiro

Klein

et al. 2010

Cell Transplant

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“…Transient focal cerebral ischemia was induced using the intraluminal filament occlusion model in male Sprague‐Dawley rats weighing 300–370 g (Katsura et al. 2008).…”

Section: Methodsmentioning

confidence: 99%

“…Next, we defined ischemic penumbra as the region that was rescued by reperfusion, and the ischemic core as the region that was not rescued by reperfusion as described previously (Memezawa et al. 1992; Katsura et al. 2008).…”

Section: Methodsmentioning

confidence: 99%

Biochemical and histopathological alterations in TAR DNA‐binding protein‐43 after acute ischemic stroke in rats

Kanazawa

Kakita

Igarashi

et al. 2011

Journal of Neurochemistry

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J. Neurochem. (2011) 116, 957–965. Abstract Nuclear factor TAR DNA‐binding protein‐43 (TDP‐43) is considered to play roles in pathogenesis of human neurodegenerative diseases, so‐called TDP‐43 proteinopathy, via its proteolytic cleavage, abnormal phosphorylation, subcellular redistribution, and insolubilization generating TDP‐43‐positive neuronal intracellular inclusions. The purpose of this study was to elucidate biochemical and histopathological alternations in TDP‐43 specific to acute ischemic stroke. Adult male rats were subjected to a 90‐min middle cerebral artery occlusion. We examined the proteolytic cleavage, phosphorylation, subcellular localization, and solubility of TDP‐43 by immunoblottings and histopathological examinations using the ischemic and sham‐operated cortex. The level of full‐length TDP‐43 (43 kDa) decreased and that of the 25‐kDa C‐terminal fragment increased after acute ischemic stroke, which can be explained by proteolytic cleavage of TDP‐43. Cytoplasmic redistribution and altered nuclear distribution of TDP‐43 was observed after acute ischemic stroke, whereas abnormal phosphorylation and insolubilization of TDP‐43 as well as formation of intracellular inclusions were not observed. Ischemic neurons with the cytoplasmic redistribution of TDP‐43 expressed ubiquitin and activated caspase 3 and were terminal deoxynucleotidyl transferase‐mediated uridine 5′‐triphosphate‐biotin nick end labeling‐positive. In conclusion, biochemical and histopathological alterations in TDP‐43 were identified in rats after acute ischemic stroke, although there was very less similarity between TDP‐43 alterations observed in acute ischemic stroke and those observed in TDP‐43 proteinopathy.

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“…It is the first mutant with a gain-of-function phenotype among the mammalian anti-apoptotic factors and shows anti-necrotic as well as anti-apoptotic activity [ 183 ]. By fusing it with the human immunodeficiency virus type 1/trans-activator of transcription (HIV-TAT) protein transduction domain, it can be transduced into neuronal cells rapidly [ 184 ]. The protein transduction domain fused FNK (PTD-FNK) protein has been shown to be significantly protective against rat brain focal ischemia, even when administered 3 h after ischemia [ 184 ].…”

Section: Protein Therapymentioning

confidence: 99%

Combination of Therapeutic Hypothermia and Other Neuroprotective Strategies after An Ischemic Cerebral Insult

Goossens¹,

Hachimi-Idrissi

2014

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Abrupt deprivation of substrates to neuronal tissue triggers a number of pathological events (the “ischemic cascade”) that lead to cell death. As this is a process of delayed neuronal cell death and not an instantaneous event, several pharmacological and non-pharmacological strategies have been developed to attenuate or block this cascade. The most promising neuroprotectant so far is therapeutic hypothermia and its beneficial effects have inspired researchers to further improve its protective benefit by combining it with other neuroprotective agents. This review provides an overview of all neuroprotective strategies that have been combined with therapeutic hypothermia in rodent models of focal cerebral ischemia. A distinction is made between drugs interrupting only one event of the ischemic cascade from those mitigating different pathways and having multimodal effects. Also the combination of therapeutic hypothermia with hemicraniectomy, gene therapy and protein therapy is briefly discussed. Furthermore, those combinations that have been studied in a clinical setting are also reviewed.

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Combination therapy with transductive anti-death FNK protein and FK506 ameliorates brain damage with focal transient ischemia in rat

Cited by 10 publications

References 60 publications

Endotoxin Deactivation by Transient Acidification

Endotoxin Deactivation by Transient Acidification

Biochemical and histopathological alterations in TAR DNA‐binding protein‐43 after acute ischemic stroke in rats

Combination of Therapeutic Hypothermia and Other Neuroprotective Strategies after An Ischemic Cerebral Insult

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