scholarly journals Detection of Acanthamoeba spp. using carboxylesterase antibody and its usage for diagnosing Acanthamoeba-keratitis

PLoS ONE ◽  
2022 ◽  
Vol 17 (1) ◽  
pp. e0262223
Author(s):  
Min-Jeong Kim ◽  
Ki-Back Chu ◽  
Hae-Ahm Lee ◽  
Fu-Shi Quan ◽  
Hyun-Hee Kong ◽  
...  
Keyword(s):  
Acanthamoeba Keratitis ◽  
Incidence Rates ◽  
Conditioned Media ◽  
Corneal Epithelial ◽  
Corneal Infection ◽  
Cell Lysates ◽  
Diagnostic Potential ◽  
Specific Antibody Production ◽  
Multiple Species ◽  
High Degree

Contact lens usage has contributed to increased incidence rates of Acanthamoeba keratitis (AK), a serious corneal infection that can lead to blindness. Since symptoms associated with AK closely resemble those incurred by bacterial or fungal keratitis, developing a diagnostic method enabling rapid detection with a high degree of Acanthamoeba-specificity would be beneficial. Here, we produced a polyclonal antibody targeting the carboxylesterase (CE) superfamily protein secreted by the pathogenic Acanthamoeba and evaluated its diagnostic potential. Western blot analysis revealed that the CE antibody specifically interacts with the cell lysates and conditioned media of pathogenic Acanthamoeba, which were not observed from the cell lysates and conditioned media of human corneal epithelial (HCE) cells, Fusarium solani, Staphylococcus aureus, and Pseudomonas aeruginosa. High titers of A. castellanii-specific antibody production were confirmed sera of immunized mice via ELISA, and these antibodies were capable of detecting A. castellanii from the cell lysates and their conditioned media. The specificity of the CE antibody was further confirmed on A. castellanii trophozoites and cysts co-cultured with HCE cells, F. solani, S. aureus, and P. aeruginosa using immunocytochemistry. Additionally, the CE antibody produced in this study successfully interacted with 7 different Acanthamoeba species. Our findings demonstrate that the polyclonal CE antibody specifically detects multiple species belong to the genus Acanthamoeba, thus highlighting its potential as AK diagnostic tool.

scholarly journals Identification, Diagnostic Potential, and Natural Expression of Immunodominant Seroreactive Peptides Encoded by FiveMycobacterium tuberculosis-Specific Genomic Regions

2010 ◽  
Vol 18 (3) ◽  
pp. 477-482 ◽  
Author(s):  
Noora Y. Al-Khodari ◽  
Rajaa Al-Attiyah ◽  
Abu S. Mustafa
Keyword(s):  
Healthy Subjects ◽  
Comparative Genomic ◽  
Whole Cell ◽  
Reading Frame ◽  
Cell Lysates ◽  
Antipeptide Antibodies ◽  
Antibody Reactivity ◽  
Diagnostic Potential ◽  
Genomic Regions ◽  
Natural Expression

ABSTRACTComparative genomic studies have identified severalMycobacterium tuberculosis-specific genomic regions of difference (RDs) which are absent in the vaccine strains ofMycobacterium bovisBCG and which may be useful in the specific diagnosis of tuberculosis (TB). In this study, a total of 775 synthetic peptides covering the sequences of 39 open reading frame (ORF) proteins encoded by genes predicted in five RDs ofM. tuberculosis, i.e., RD1, RD4, RD5, RD6, and RD7, were tested by enzyme-linked immunosorbent assays for antibody reactivity with sera from HIV-negative pulmonary TB patients (n= 100) andM. bovisBCG-vaccinated healthy subjects (n= 100). The results identified three immunodominant peptides reactive with TB sera, i.e., amino acids (aa) 346 to 370 of RD1ORF Rv3876, aa 241 to 265 of RD6ORF Rv1508c, and aa 325 to 336 of RD6ORF Rv1516c. These peptides had significantly stronger antibody reactivity with sera from TB patients than with sera from healthy subjects (P< 0.05) and significantly higher rates of positivity with TB sera (positives = 66 to 93%) than sera from healthy subjects (positives = 10 to 28%). Antipeptide antibodies were raised in rabbits after immunization with pools of 11 peptides corresponding to each protein. Probing of culture filtrates and whole-cell lysates ofM. tuberculosiswith antipeptide antibodies suggested the natural expression of Rv1516c in whole-cell lysates ofM. tuberculosis. The results suggest the potential of the identified immunodominant RD peptides in the serodiagnosis of TB.

scholarly journals Host Range Restriction of Epstein-Barr Virus and Related Lymphocryptoviruses

2015 ◽  
Vol 89 (17) ◽  
pp. 9133-9136 ◽  
Author(s):  
Janine Mühe ◽  
Fred Wang
Keyword(s):  
B Cells ◽  
Host Range ◽  
Epstein Barr Virus ◽  
Nonhuman Primates ◽  
Range Restriction ◽  
Barr Virus ◽  
Epstein Barr ◽  
Multiple Species ◽  
High Degree ◽  
Host Range Restriction

Epstein-Barr-related herpesviruses, or lymphocryptoviruses (LCV), naturally infect humans and nonhuman primates (NHP), but their host range is not well characterized. Using LCV and B cells from multiple species of Hominidae and Cercopithecidae, we show that LCV can immortalize B cells from some nonnative species but that growth transformation is restricted to B cells from their own family of hominoids or Old World NHP, suggesting a high degree of LCV adaptation to their natural primate host.

scholarly journals A Case of Medication-Resistant Acanthamoeba Keratitis Treated by Corneal Crosslinking in Turkey

2013 ◽  
Vol 2013 ◽  
pp. 1-4 ◽  
Author(s):  
Goktug Demirci ◽  
Akif Ozdamar
Keyword(s):  
Visual Acuity ◽  
Local Anesthesia ◽  
Symptomatic Relief ◽  
Acanthamoeba Keratitis ◽  
Symptomatic Improvement ◽  
Corneal Epithelial ◽  
Corneal Crosslinking ◽  
Corneal Melting

Purpose. To report a case of medication-resistant acanthamoeba keratitis (AK) treated successfully by corneal crosslinking (CXL).Methods. A 26-year-old male with medication-resistant AK underwent a standard CXL procedure with local anesthesia, followed by central corneal epithelial debridement, application of riboflavin 0.1%, and UV-A irradiation.Results. The patient experienced a dramatic symptomatic improvement within 24 hours. At two months, keratitis was healed with a semitransparent paracentral scar that did not affect visual acuity.Conclusions. Our experience, considered in the context of recent studies, suggests that CXL may be an option for selected patients with medication-resistant AK and corneal melting. CXL allows patients to avoid emergency keratoplasty and experience rapid symptomatic relief.

Elevated Corneal Epithelial Lines in Acanthamoeba Keratitis

1988 ◽  
Vol 106 (9) ◽  
pp. 1202-1206 ◽  
Author(s):  
G. J. Florakis ◽  
R. Folberg ◽  
J. H. Krachmer ◽  
D. T. Tse ◽  
T. J. Roussel ◽  
...  
Keyword(s):  
Acanthamoeba Keratitis ◽  
Corneal Epithelial

scholarly journals Furin Functions as a Nonproteolytic Chaperone for Matrix Metalloproteinase-28: MMP-28 Propeptide Sequence Requirement

2011 ◽  
Vol 2011 ◽  
pp. 1-12 ◽  
Author(s):  
Maria Pavlaki ◽  
Stanley Zucker ◽  
Antoine Dufour ◽  
Nikki Calabrese ◽  
Wadie Bahou ◽  
...  
Keyword(s):  
Cell Migration ◽  
Matrix Metalloproteinase ◽  
Consensus Sequence ◽  
Mechanisms Of Action ◽  
Conditioned Media ◽  
Transfected Cells ◽  
Cell Lysates ◽  
Protein Levels ◽  
In Cells

Although MMP-28 is involved in numerous important physiologic and pathologic conditions, the mechanisms of action of this secreted proteinase is not well understood. We now have demonstrated that furin serves as an intermolecular chaperone for MMP-28 secretion by interacting with the propeptide domain of MMP-28. Employing COS-1 cells transfected with MMP-28 cDNA, protein levels of MMP-28 were quite low in conditioned media as compared to cell lysates. Coexpression of MMP-28 with furin cDNA resulted in markedly enhanced MMP-28 secretion. Contrary to expectation, cleavage of MMP-28 at the furin consensus sequence did not occur and proteolytic inactive furin was equally effective in enhancing MMP-28 secretion. Furin and MMP-28 coimmunoprecipitated and were partially coimmunolocalized in the cytoplasm of transfected cells. Cotransfection with furin cDNA also enhanced MMP-28 induced cell migration. In conclusion, our data provide a novel mechanism for MMP-28 function in cells in which furin serves as an intermolecular chaperone.

scholarly journals Oral Immunization with Acanthamoeba castellanii Mannose-Binding Protein Ameliorates Amoebic Keratitis

2006 ◽  
Vol 74 (12) ◽  
pp. 7032-7034 ◽  
Author(s):  
M. Garate ◽  
H. Alizadeh ◽  
S. Neelam ◽  
J. Y. Niederkorn ◽  
N. Panjwani
Keyword(s):  
Binding Protein ◽  
Immunoglobulin A ◽  
Acanthamoeba Castellanii ◽  
Oral Immunization ◽  
Corneal Epithelial Cells ◽  
Corneal Epithelial ◽  
Corneal Infection ◽  
Mannose Binding Protein ◽  
Mannose Binding ◽  
Amoebic Keratitis

ABSTRACT Acanthamoeba castellanii mannose-binding protein (MBP) mediates adhesion of the amoebae to corneal epithelial cells, a key first step in the pathogenesis of Acanthamoeba keratitis (AK), a devastating corneal infection. In the present study, we demonstrate that oral immunization with recombinant MBP ameliorates AK in a hamster animal model and that this protection is associated with the presence of elevated levels of anti-MBP immunoglobulin A in the tear fluid of the immunized animals.

scholarly journals Aminoglucoside/Imidazole in the Treatment of Acanthamoeba Keratitis

2018 ◽  
Vol 2 (2) ◽  
Keyword(s):  
Visual Acuity ◽  
Acanthamoeba Keratitis ◽  
Final Visual Acuity ◽  
Free Living ◽  
Corneal Infection ◽  
Free Living Amoeba

Acanthamoeba Keratitis (AK) is an infrequent corneal infection caused by free living amoeba, it is frequently misdiagnosed and medically/surgical treated with low or no response in advanced cases. In this paper we present five cases of AK with early diagnoses and good response to aminoglucoside/imidazole treatment and achieving acceptable final visual acuity in each case.

scholarly journals Infectious Keratitis Associated with Contact Lenses

2021 ◽  
Vol 20 (4) ◽  
pp. 140-143
Author(s):  
Kyong Jin Cho
Keyword(s):  
Patient Education ◽  
Contact Lens ◽  
Contact Lenses ◽  
Infectious Keratitis ◽  
Corneal Surface ◽  
Corneal Epithelial Cells ◽  
Myopia Progression ◽  
Corneal Epithelial ◽  
Corneal Infection ◽  
Tear Flow

The use of contact lenses for correcting refraction, suppressing myopia progression, and cosmetic purposes is increasing steadily. Contact lenses have various effects on the corneal surface and corneal infection can occur following obstruction of tear flow, micro-damage to corneal epithelial cells, corneal hypoxia, changes in corneal immunity, and exposure to contaminants. When a patient who used to wear contact lenses presents with keratitis, it is important to distinguish whether it is an infection; if it is an infection, it is important to find the causative strain and promptly treat it appropriately. Since improper lens care is related to infection, appropriate patient education is necessary, and the risk of contact lens infection should be reduced through regular ophthalmic examinations.

Homocysteine rapidly increases matrix metalloproteinase-2 expression and activity in cultured human vascular smooth muscle cells

2005 ◽  
Vol 94 (12) ◽  
pp. 1285-1293 ◽  
Author(s):  
Gabriella Doronzo ◽  
Isabella Russo ◽  
Luigi Mattiello ◽  
Mariella Trovati ◽  
Giovanni Anfossi
Keyword(s):  
Smooth Muscle ◽  
Matrix Metalloproteinase ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells ◽  
Conditioned Media ◽  
Matrix Metalloproteinase 2 ◽  
Cell Lysates ◽  
Increased Risk

SummaryIn this study we aimed to test the hypothesis that in human vascular smooth muscle cells (VSMC) homocysteine influences synthesis and release of matrix metalloproteinase-2 (MMP-2), which is deeply involved in vascular remodeling and atherosclerotic plaque instabilization. Experiments were carried out in cultured human VSMC exposed to 50–500 μmol/l homocysteine after a 24-hour culture with MEM containing 0.1% BSA. Both in supernatants and cell lysates we evaluated MMP-2 activity (gelatin zimography), MMP-2 and TIMP-2 protein synthesis (Western immunoblotting). Homocysteine effects were investigated also after cell exposure to i) specific MEK inhibitor PD98059 (30 μmol/l) to evaluate the involvement of Mitogen-Activated Protein Kinase (MAPK) and ii) specific phosphatidylinositol 3-kinase (PI3-K) inhibitor LY294002 (100 μmol/l) to evaluate the involvement of PI3-K pathway. Gelatin zimography evidenced that MMP-2 activity is increased both in conditioned media and in cell lysates starting from 8-hour incubation with 100 μmol/l homocysteine. Western blot analysis evidenced increased MMP-2 levels in both conditioned media and cell lysates. Cell exposure to PD98059 and LY294002 prevented homocysteine effects on MMP-2 synthesis. Homocysteine, at concentrations associated with increased risk of cardiovascular events, increases MMP-2 activity, synthesis and secretion in VSMC through a mechanism involving the activation of MAPK and PI3-K pathways. These data suggest that homocysteine is directly involved in mechanisms leading to remodelling and instabilization of atherosclerotic plaques.

scholarly journals Autophagy Inhibitors as a Potential Antiamoebic Treatment for Acanthamoeba Keratitis

2015 ◽  
Vol 59 (7) ◽  
pp. 4020-4025 ◽  
Author(s):  
Eun-Kyung Moon ◽  
So-Hee Kim ◽  
Yeonchul Hong ◽  
Dong-Il Chung ◽  
Youn-Kyoung Goo ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Acanthamoeba Keratitis ◽  
New Combination ◽  
Corneal Epithelial Cells ◽  
Content Type ◽  
Corneal Epithelial ◽  
Cytopathic Effects ◽  
Combination Treatments ◽  
Human Corneal Epithelial Cells ◽  
Significant Block

ABSTRACTAcanthamoebacysts are resistant to extreme physical and chemical conditions. Autophagy is an essential pathway for encystation ofAcanthamoebacells. To evaluate the possibility of an autophagicAcanthamoebaencystation mechanism, we evaluated autophagy inhibitors, such as 3-methyladenine (3MA), LY294002, wortmannin, bafilomycin A, and chloroquine. Among these autophagy inhibitors, the use of 3MA and chloroquine showed a significant reduction in the encystation ratio inAcanthamoebacells. Wortmannin also inhibited the formation of mature cysts, while LY294002 and bafilomycin A did not affect the encystation ofAcanthamoebacells. Transmission electron microscopy revealed that 3MA and wortmannin inhibited autophagy formation and that chloroquine interfered with the formation of autolysosomes. Inhibition of autophagy or autolysosome formation resulted in a significant block in the encystation inAcanthamoebacells. Clinical treatment with 0.02% polyhexamethylene biguanide (PHMB) showed high cytopathic effects onAcanthamoebatrophozoites and cysts; however, it also revealed high cytopathic effects on human corneal epithelial cells. In this study, we investigated effects of the combination of a low (0.00125%) concentration of PHMB with each of the autophagy inhibitors 3MA, wortmannin, and chloroquine onAcanthamoebaand human corneal epithelial cells. These new combination treatments showed low cytopathic effects on human corneal cells and high cytopathic effects onAcanthamoebacells. Taken together, these results provide fundamental information for optimizing the treatment ofAcanthamoebakeratitis.

Sign in / Sign up

Export Citation Format

Share Document