Production and characterization of monoclonal antibodies against cathepsin B and cathepsin B-Like proteins of Naegleria fowleri

Seong, Gi-Sang; Sohn, Hae-Jin; Kang, Hyung Suk; Seo, Ga-Eun; Kim, Jong-Hyun; Shin, Ho‐Joon

doi:10.1016/j.exppara.2017.09.004

Cited by 12 publications

(13 citation statements)

References 32 publications

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“…Therefore, the pathogenicity of N. fowleri is not affected by high temperature as N. lovaniensis is . Hence, both contact‐dependent mechanism (food cups), as well as contact‐independent mechanism (secretion of various proteins), are significantly involved in evasion of the host immune response by N. fowleri …”

Section: Naegleria Fowleri and Host Immune Responsementioning

confidence: 99%

“…The study indicated that incubation of N. fowleri trophozoites with ebselen results in loss of the nuclear membrane, in the cytoplasmic membrane alteration and appearance of electron‐dense granules while incubation with BAY 11‐7082 and BAY 11‐7085 cause cytoplasmic and nuclear membranes disruption, in the cytoplasm presence of large lipid droplets and appearance of chromatin residues and several vesicles, has been observed. These amoebicidal compounds that are permeable to the BBB are considered as potential new drugs for the treatment of N. fowleri …”

Section: Managementmentioning

confidence: 99%

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Naegleria fowleri: Sources of infection, pathophysiology, diagnosis, and management; a review

Jahangeer

Mahmood

Munir

et al. 2019

Clin Exp Pharma Physio

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Naegleria fowleri, a thermophilic flagellate amoeba known as a “brain‐eating” amoeba, is the aetiological agent of a perilous and devastating waterborne disease known as primary amoebic meningoencephalitis (PAM), both in humans as well as in animals. PAM is a rare but fatal disease affecting young adults all around the world, particularly in the developed world but recently reported from developing countries, with 95%–99% mortality rate. Swimmers and divers are at high risk of PAM as the warm water is the most propitious environment adapted by N. fowleri to cause this infection. Infective amoeba in the trophozoite phase enter the victim's body through the nose, crossing the cribriform plate to reach the human brain and cause severe destruction of the central nervous system (CNS). The brain damage leads to brain haemorrhage and death occurs within 3–7 days in undiagnosed cases and maltreated cases. Though the exact pathogenesis of N. fowleri is still not known, it has exhibited two primary mechanisms, contact‐independent (brain damage through different proteins) and contact‐dependent (brain damage through surface structures food cups), that predominantly contribute to the pathogen invading the host CNS. For the management of this life‐threatening infection different treatment regimens have been applied but still the survival rate is only 5% which is ascribed to its misdiagnosis, as the PAM symptoms closely resembled bacterial meningitis. The main objectives of this review article are to compile data to explore the sources and routes of N. fowleri infection, its association in causing PAM along with its pathophysiology; latest techniques used for accurate diagnosis, management options along with challenges for Pakistan to control this drastic disorder.

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Section: Naegleria Fowleri and Host Immune Responsementioning

confidence: 99%

Section: Managementmentioning

confidence: 99%

Naegleria fowleri: Sources of infection, pathophysiology, diagnosis, and management; a review

Jahangeer

Mahmood

Munir

et al. 2019

Clin Exp Pharma Physio

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“…Recently, the NfCPB and NfCPB-L proteins of Naegleria fowleri were observed on cell membranes, especially on psuedopodia and food-cup structures which were associated with cytotoxicity and phagocytosis (Seong et al, 2017).…”

Section: Discussionmentioning

confidence: 99%

“…In order to analyze the purified MBP and evaluate the monoclonal antibody to react the purified MBP, SDS-PAGE and western blot were performed as described by Laemmli (Laemmli, 1970;Seong et al, 2017). The samples were mixed SDS-PAGE buffer containing 2-mercaptoethanol, boiled for 5 min, and loaded on to 12% polyacrylamide gels.…”

Section: Sodium Dodecyl Sulfate-polyacrylamide Gel Electrophoresis (Smentioning

confidence: 99%

Production of a monoclonal antibody against a mannose-binding protein of Acanthamoeba culbertsoni and its localization

Kang

Park

Shin

et al. 2018

Experimental Parasitology

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Amoebae from the genus Acanthamoeba are facultative pathogens of humans and other animals. In humans they most frequently infect the eye causing a sight threatening infection known as Acanthamoeba keratitis (AK), and also cause an often fatal encephalitis (GAE). A mannose-binding protein (MBP) has been identified as being important for Acanthamoeba infection especially in AK. This lectin has previously been characterized from Acanthamoeba castellanii as consisting of multiple 130 kDa subunits. MBP expression correlates with pathogenic potential and is expressed in a number of Acanthamoeba species. Here we report the purification of a similar lectin from Acanthamoeba culbertsoni and the production of a monoclonal antibody to it. The A. culbertsoni MBP was isolated by affinity chromatography using α-D-mannose agarose and has an apparent molecular weight of 83 kDa. The monoclonal antibody is an IgM that is useful in both western blots and immunofluorescence. We expect that this antibody will be useful in the study of the pathology of A. culbertsoni and in its identification in clinical samples.

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“…A. castellanii (ATCC NO. 50492), In addition, briefly, pathogenic N. fowleri (Carter Nf69 strain, ATCC 30215) was axenically cultured at 37℃ in Nelson's medium (Seong et al, 2017). CHO cells were cultured with Earle's minimum essential medium (EMEM) containing 10% fetal bovine serum at 37℃ in a 5% CO 2 incubator (Kang et al, 2005).…”

mentioning

confidence: 99%

Protective Effects of a Monoclonal Antibody to a Mannose-Binding Protein ofAcanthamoeba culbertsoni

Park

Kang

Jung

2018

BSL

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Acanthamoeba culbertsoni is the causative agent of granulomatous amoebic encephalitis (GAE), a condition that predominantly occurs in immunocompromised individuals and which is typically fatal. A mannose-binding protein (MBP) among lectins was shown to have strong A. castellanii pathogenic potential when correlated with major virulence proteins. In this study, protective effects were analyzed using the monoclonal antibody to A. culbertsoni MBP by quantification and were also compared with other free-living amoebae. For the amoebial cytotoxicity to the target cell, amoeba trophozoites were incubated with Chinese hamster ovary (CHO) cells. For the protective effects of antibodies, amoebae were preincubated with them for 4 h and then added to the target cells. After 24 h, the supernatants were collected and examined for host cell cytotoxicity by measuring lactate dehydrogenase (LDH) release. The cytotoxicity of A. culbertsoni to the CHO cells showed about 87.4%. When the monoclonal antibody was pre-incubated with A. culbertsoni, the amoebial cytotoxicity was remarkably decreased as shown at LDH release (1.858 absorbance), which was represented with about 49.9%. Taken together, it suggested that the monoclonal antibody against MBP be important to inhibit the cytotoxicity of A. culbertsoni trophozoites to the target cell. The antibody will be applied into an in vivo functional analysis, which would help to develop therapeutics.

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Production and characterization of monoclonal antibodies against cathepsin B and cathepsin B-Like proteins of Naegleria fowleri

Cited by 12 publications

References 32 publications

Naegleria fowleri: Sources of infection, pathophysiology, diagnosis, and management; a review

Naegleria fowleri: Sources of infection, pathophysiology, diagnosis, and management; a review

Production of a monoclonal antibody against a mannose-binding protein of Acanthamoeba culbertsoni and its localization

Protective Effects of a Monoclonal Antibody to a Mannose-Binding Protein ofAcanthamoeba culbertsoni

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