1.Species Identification of Coagulase Negative Staphylococci by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism of Heat Shock Protein 60 Gene.
Eun Jee OH ; Jung Do JANG ; Yeon Joon PARK ; Sun Moo KIM ; Byung Kee KIM
Korean Journal of Clinical Microbiology 2000;3(1):36-42
BACKGROUND: An accurate and rapid method for specise identification of coagulase negative staphylococci(CNS) has been increasingly necessary for the clinical significance and planning the management of patients with staphylococcal infections. Recently, it has been reported that there is a highly conserved area on their 60KDa heat shock protein(HSP60) gene sequences between the interspecies of CNS and it can be amplified by a set of universal degenerate primer. This led us our attention to focus on whether the PCR-based RFLP method using Mse / restriction enzyme could be a useful tool for the species identification of CNS. METHODS: In the present study, we performed PCR-based RFLP analysis using a set of degenerate primers covering HSP60 and Mse / restriction enzyme on the reference strains and 25 clinical isolates(10 of S. epidermidis, 10 of S. haemolyticus, 4 of S. lugdunensis and 1 of S. warneri) which were previously identified by the API-STAPH, Vitek GPI card and/or with conventional biochemical test. RESULT: All the seven reference strains revealed that each strain has a distinct electrophoresed band patterns with combination of different number (up to 8) and size of fragments. And these distinct band patterns showed remarkable concordance with the seven reference strains and 25 clinical isolates. CONCLUSION: These result strongly suggest that the PCR-RFLP method using degenerate primers covering the HSP60 gene and Mse / digestion enzyme offer a convenient and accurate tool for species-specific identification of CNS.
Chaperonin 60*
;
Coagulase*
;
Digestion
;
Heat-Shock Proteins*
;
Hot Temperature*
;
Humans
;
Polymerase Chain Reaction
;
Polymorphism, Restriction Fragment Length
;
Shock
;
Staphylococcal Infections
2.Heat shock proteins of the hyperthermophilic archaea.
Huayou CHEN ; Chunxia ZHANG ; Xiaoke MA ; Yi ZHANG
Chinese Journal of Biotechnology 2008;24(12):2011-2021
As thermostable enzymes and organisms are much more needed, researches on heat shock proteins(HSPs) of hyperthermophilic archaea have drawn more concerns. HSPs from hyperthermophilic archaea are concise only with HSP60, sHSP, prefoldin and AAA+proteins, but without HSP100s, HSP90s, HSP70 (DnaK), HSP40 (DnaJ) and GrpE which are common in mesophilic or thermophilic archaea. Accordingly, studies on the structure, function and operation mechanism of these four groups are much more important and meaningful. This review focuses on the recent progress in the researchs on the structure, function, operation mechanism and cooperation of the HSPs from hyperthermophilic archaea. The problems and obfuscations in these HSPs are analyzed, and farther research direction and key points are put out.
Archaea
;
classification
;
metabolism
;
Archaeal Proteins
;
metabolism
;
Chaperonin 60
;
metabolism
;
Heat-Shock Proteins
;
genetics
;
metabolism
;
Molecular Chaperones
;
metabolism
3.Polymorphisms of hsp 60 gene in Chinese Han people.
Jing WANG ; Xiao-bo YANG ; Yun BAI ; Qi JIANG ; Yue-feng HE ; Yong-wen CHEN ; Mei-an HE ; Tang-chun WU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2006;24(8):471-474
OBJECTIVETo identify the single nucleotide polymorphisms (SNPs) in the regulatory and coding regions of heat shock protein 60 gene and search for its genetic makers in Chinese Han people.
METHODSThe 5' flank region, parts of the exons and introns of hsp60 gene were resequenced to identify the SNPs in Chinese Han people, and then the sequenced results to the Japanese, European and African's data in National Center for Biotechnology Information (NCBI) and HapMap databases were compared.
RESULTSOne novel SNP was identified in exon 2 resulting in synonymous variant and the G allele frequency was 0.025. There were 11 reported SNPs in the sequenced region. The minor allele frequencies of rs1116734, rs3749095, rs1050347, rs8539 were 0.51, 0.30, 0.29, 0.49. The heterozygosity of the other 7 SNPs was 0. The distributions of rs1116734, rs1050347, rs8539, rs3749095 in Chinese Han people were similar to the Japanese's. The hsp60 rs3749095 which was not found in Japanese people was a high-frequency SNP in Chinese Han people; the distribution of rs8539 in Chinese Han people was quite different from European and African's (P < 0.01).
CONCLUSIONThe SNPs of hsp60 in Chinese Han people are different from the other peoples. The SNPs of hsp60 gene rs1116734, rs3749095, rs1050347, rs8539 are very common in Chinese Han people and might be used for candidate genetic markers of hsp60 gene.
Adolescent ; Adult ; Alleles ; Chaperonin 60 ; genetics ; China ; ethnology ; Exons ; genetics ; Female ; Gene Frequency ; Humans ; Male ; Polymorphism, Single Nucleotide
4.Immune Responses Induced by HSP60 DNA Vaccine against Toxoplasma gondii Infection in Kunming Mice
Zhong Yuan LI ; Jing LU ; Nian Zhang ZHANG ; Jia CHEN ; Xing Quan ZHU
The Korean Journal of Parasitology 2018;56(3):237-245
Toxoplasma gondii can infect all the vertebrates including human, and leads to serious toxoplasmosis and considerable veterinary problems. T. gondii heat shock protein 60 (HSP60) is associated with the activation of antigen presenting cells by inducing initial immune responses and releasing inflammatory cytokines. It might be a potential DNA vaccine candidate for this parasite. A pVAX-HSP60 DNA vaccine was constructed and immune responses was evaluated in Kunming mice in this study. Our data indicated that the innate and adaptive immune responses was elicited by successive immunizations with pVAX-HSP60 DNA, showing apparent increases of CD3e+CD4+ and CD3e+CD8a+ T cells in spleen tissues of the HSP60 DNA-immunized mice (24.70±1.23% and 10.90±0.89%, P < 0.05) and higher levels of specific antibodies in sera. Furthermore, the survival period of the immunized mice (10.53±4.78 day) were significantly prolonged during the acute T. gondii infection. Decrease of brain cysts was significant in the experimental group during the chronic infection (P < 0.01). Taken together, TgHSP60 DNA can be as a vaccine candidate to prevent the acute and chronic T. gondii infections.
Animals
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Antibodies
;
Antigen-Presenting Cells
;
Brain
;
Chaperonin 60
;
Cytokines
;
DNA
;
Humans
;
Immunization
;
Mice
;
Parasites
;
Spleen
;
T-Lymphocytes
;
Toxoplasma
;
Toxoplasmosis
;
Vertebrates
5.Changes of CK-MB and HSP 60 in electrical-injuried rats.
Hui-tong LIU ; Gao-wen FU ; Ze ZHAO ; Su-zhen DING ; Qiao-feng WANG ; Lei CHEN ; Ya-nan XIE ; Zhen-yuan WANG
Journal of Forensic Medicine 2012;28(5):333-336
OBJECTIVE:
To investigate the changes of creatine kinase-MB (CK-MB) and heat shock protein 60 (HSP 60) in rats without electric marks after electric injury, to identify the relationship of the CK-MB, HSP 60 and the time of electric injuries, and to evaluate the damage to cells after electric injury.
METHODS:
The animal model of electric injury without electric marks was established by alternating current (voltage 110 V). Automatic biochemistry analyzer was used to detect the serum CK-MB and immunohistochemical staining technology was used to analyze the tissues of myocardium and left lobe of liver.
RESULTS:
The amount of serum CK-MB was increased when the rats were injuried, and reached the peak at 30min. Then the amount of CK-MB began to decrease and showed a slight downward trend in 3-5 h after electric injury, and leveled off at 6 h. Immunohistochemistry staining also showed the changes of HSP 60 of rats' myocardial cells and hepatic cells regularly after electric injury.
CONCLUSION
The regular changes of serum CK-MB and tissular HSP 60 in rats can be used to diagnosis electric injury and assess the injury of internal organs after the electric injury without electric marks.
Animals
;
Chaperonin 60/metabolism*
;
Creatine Kinase, MB Form/metabolism*
;
Electric Injuries/complications*
;
Immunohistochemistry
;
Liver/pathology*
;
Myocardium/pathology*
;
Rats
6.Heat Shock Protein 60 and 27 expressions on the trophoblast of preterm birth.
Suk Young KIM ; Eun Joo CHUNG ; Hyun Myong OH ; Jung Min KIM ; Deul Lae MIN ; You Jin HWANG
Korean Journal of Obstetrics and Gynecology 2008;51(9):957-964
OBJECTIVE: To compare the distribution and expression of hsp60 and hsp27 in placental trophoblast between preterm and term placenta and to observe hsp immune response in relation to the pathogenesis of preterm birth. METHODS: 22 cases of preterm trophoblast, between 24 weeks and 36 weeks gestation, which were developed spontaneous onset or less than 24 hours after rupture of membrane were obtained. And aged-matched, 22 cases of normal term trophoblast, as control were also obtained after informed consent from each patient. The protein extraction form trophoblast was stained by immunohistochemical methods and was measured by the assay of Western blots. And the density of band using Image-writer were taken and statistical assay were performed as significance <0.05. RESULTS: The expressions of hsp60 and hsp27 in trophoblast of preterm and term placenta were identified by immunohistochemical staining method. The hsp60 had significantly higher expression in trophoblast of preterm birth than in that of term birth (P<0.001) and the hsp27 also had significantly higher expression in trophoblast of preterm birth than in that of term birth (P<0.02) CONCLUSION: The higher expression of hsp60 and hsp27 in trophoblast of preterm birth might be suggested the development of immune response to occur preterm labor Further study are necessary to determine the exact actions of hsp60 and27 in trophoblast and to understand the immune mechanism of preterm birth.
Blotting, Western
;
Chaperonin 60
;
Female
;
Heat-Shock Proteins
;
Hot Temperature
;
Humans
;
Informed Consent
;
Membranes
;
Obstetric Labor, Premature
;
Placenta
;
Pregnancy
;
Premature Birth
;
Rupture
;
Term Birth
;
Trophoblasts
7.Expression of Heat Shock Protein 60 kDa Is Upregulated in Cervical Cancer.
You Jin HWANG ; Soon Pyo LEE ; Suk Young KIM ; Young Hwan CHOI ; Min Ji KIM ; Choong Ho LEE ; Joo Young LEE ; Dae Young KIM
Yonsei Medical Journal 2009;50(3):399-406
PURPOSE: Cervical cancer caused by the human papilloma virus (HPV) continues to be the cause of yearly death among women. However, it is a curable disease when diagnosed at an early stage. Recently, several researches have reported that heat shock protein (HSP) 60, a chaperone protein of molecular weight of 60 kDa, is involved in carcinogenesis and apoptosis. In order to evaluate the prognostic significance of HSP60 in cervical cancer, we examined differences in the HSP60 expression between cervical cancer and normal tissues in women. MATERIALS AND METHODS: Tissue samples were collected from 20 cervical cancer patients and 20 normal controls. HSP60 expression of cervical cancer and normal tissues were verified by the 2D gel proteomics, semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot analyses. RESULTS: In 2D proteomic analysis, an increase of HSP60 expression was detected in cervical cancer tissues and confirmed by Western blot analysis (p < 0.05). However, messenger RNA (mRNA) levels of HSP60 did not display any significant differences between cervical cancer and normal tissues. CONCLUSION: These results suggest that HSP60 may be involved in the development of cervical cancer and have profound biological and prognostic significance.
Adult
;
Aged
;
Blotting, Western
;
Chaperonin 60/*metabolism
;
Electrophoresis, Gel, Two-Dimensional
;
Female
;
Humans
;
Male
;
Middle Aged
;
Reverse Transcriptase Polymerase Chain Reaction
;
Uterine Cervical Neoplasms/*metabolism
8.Prevalence of Anaplasma and Bartonella spp. in Ticks Collected from Korean Water Deer (Hydropotes inermis argyropus).
Jun Gu KANG ; Sungjin KO ; Heung Chul KIM ; Sung Tae CHONG ; Terry A KLEIN ; Jeong Byoung CHAE ; Yong Sun JO ; Kyoung Seong CHOI ; Do Hyeon YU ; Bae Keun PARK ; Jinho PARK ; Joon Seok CHAE
The Korean Journal of Parasitology 2016;54(1):87-91
Deer serve as reservoirs of tick-borne pathogens that impact on medical and veterinary health worldwide. In the Republic of Korea, the population of Korean water deer (KWD, Hydropotes inermis argyropus) has greatly increased from 1982 to 2011, in part, as a result of reforestation programs established following the Korean War when much of the land was barren of trees. Eighty seven Haemaphysalis flava, 228 Haemaphysalis longicornis, 8 Ixodes nipponensis, and 40 Ixodes persulcatus (21 larvae, 114 nymphs, and 228 adults) were collected from 27 out of 70 KWD. A total of 89/363 ticks (266 pools, 24.5% minimum infection rate) and 5 (1.4%) fed ticks were positive for Anaplasma phagocytophilum using nested PCR targeting the 16S rRNA and groEL genes, respectively. The 16S rRNA gene fragment sequences of 88/89 (98.9%) of positive samples for A. phagocytophilum corresponded to previously described gene sequences from KWD spleen tissues. The 16S rRNA gene fragment sequences of 20/363 (5.5%) of the ticks were positive for A. bovis and were identical to previously reported sequences. Using the ITS specific nested PCR, 11/363 (3.0%) of the ticks were positive for Bartonella spp. This is the first report of Anaplasma and Bartonella spp. detected in ticks collected from KWD, suggesting that ticks are vectors of Anaplasma and Bartonella spp. between reservoir hosts in natural surroundings.
Anaplasma/genetics/*physiology
;
Animals
;
Arachnid Vectors/microbiology
;
Bartonella/genetics/*physiology
;
Chaperonin 60/genetics
;
Deer/parasitology
;
Disease Reservoirs/veterinary
;
RNA, Ribosomal, 16S/genetics
;
Republic of Korea/epidemiology
;
Ticks/*microbiology
9.Identification of mono- or poly-specific monoclonal antibody to Porphyromonas gingivalis heat-shock protein 60.
Jeomil CHOI ; Sang Yull LEE ; Koanhoi KIM ; Bong Kyu CHOI ; Myung Jin KIM
Journal of Periodontal & Implant Science 2011;41(2):54-59
PURPOSE: The aim of this study was to define the immunoreactive specificity of Porphyromonas gingivalis (P. gingivalis) heat shock protein (HSP) 60 in periodontitis and atherosclerosis. METHODS: In an attempt to define the cross-reactive bacterial heat-shock protein with human self-antigen at molecular level, we have introduced a novel strategy for cloning hybridoma producing anti-P. gingivalis HSP 60 which is polyreactive to bacterial HSPs or to the human homolog. RESULTS: Five cross-reactive clones were obtained which recognized the #19 peptide (TLVVNRLRGSLKICAVKAPG) among 37 synthetic peptides (20-mer, 5 amino acids overlapping) spanning the whole molecule of P. gingivalis HSP 60. We have also established three anti-P. gingivalis HSP 60 monoclonal antibodies demonstrating mono-specificity. These clones recognized the #29 peptide (TVPGGGTTYIRAIAALEGLK). CONCLUSIONS: Peptide #19 and #29 of P. gingivalis HSP 60 might be important immunoreactive epitopes in the immunopathogenic mechanism of bacterial antigen-triggered autoimmune diseases.
Amino Acids
;
Antibodies
;
Antibodies, Monoclonal
;
Autoimmune Diseases
;
Chaperonin 60
;
Clone Cells
;
Cloning, Organism
;
Epitopes
;
Heat-Shock Proteins
;
Humans
;
Hybridomas
;
Peptides
;
Periodontitis
;
Porphyromonas
;
Porphyromonas gingivalis
;
Sensitivity and Specificity
10.Studies of heat shock protein 60 and heat shock protein 70 in oral lichen planus.
Chinese Journal of Stomatology 2003;38(4):275-278
OBJECTIVETo study the role of heat shock protein (HSP) 60 and HSP70 in the pathogenesis of oral lichen planus (OLP).
METHODSThe immunohistochemical SP methods were used to detected the expression of HSP60 and HSP70 in 62 cases of oral lichen planus, 10 cases of normal oral mucosa, 21 cases of chronic discoid lupus erythematosus (DLE), 10 cases of benign lymphoadenosis of mucosa, 46 cases of leukoplakia. RT-PCR was used to evaluate the expression of HSP60 mRNA and HSP70 mRNA in 12 cases of OLP and 5 cases of normal oral mucosa.
RESULTSHSP60 was expressed at much higher levels in OLP than other groups. The expression of HSP70 decreased in epithelial cells in erosive lichen planus group. HSP60 mRNA and HSP70 mRNA were expressed at high level in OLP.
CONCLUSIONHSP60 and HSP70 play an important role in pathogenesis of OLP.
Chaperonin 60 ; biosynthesis ; genetics ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; Humans ; Lichen Planus, Oral ; metabolism ; pathology ; Mouth Mucosa ; metabolism ; RNA, Messenger ; biosynthesis