3.Identification of Outer Membrane Vesicles Derived from Orientia tsutsugamushi.
Sun Myoung LEE ; Hea Yoon KWON ; Jae Hyong IM ; Ji Hyeon BAEK ; Jae Seung KANG ; Jin Soo LEE
Journal of Korean Medical Science 2015;30(7):866-870
Orientia tsutsugamushi, a causative pathogen of Scrub typhus, is a gram-negative intracellular bacterium. Outer membrane vesicles (OMVs) are produced from the membrane of bacteria and play many roles related to the survival of the pathogen. However, there have been no reports confirming whether O. tsutsugamushi indeed produce OMVs. O. tsutsugamushi boryong was cultured in ECV-304 cells for the purification of OMVs. Western blot analysis and immunoenrichment using anti-O. tsutsugamushi monoclonal antibody and electron microscopy were employed for identification and characterization of OMVs. We confirm the presence of OMVs derived from O. tsutsugamushi, and also found that those OMVs contain a major surface antigen of 56-kDa protein and variant immunogenic antigens.
Antibodies, Monoclonal/*immunology
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Antigens, Bacterial/*immunology
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Antigens, Surface/*immunology
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Cell Line
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Cell Membrane/immunology
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Humans
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Microscopy, Electron
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Orientia tsutsugamushi/*immunology/metabolism
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Scrub Typhus/diagnosis/microbiology
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Secretory Vesicles/*immunology
4.Detection of anti-Sp17 antibodies in infertile patients' serum and its clinical significance.
Chun-hua ZHANG ; Fang-qiu LI ; Ai-long YANG ; Wei SUN ; Jia-wen MIAO
National Journal of Andrology 2007;13(1):27-29
OBJECTIVETo test anti-Sp17 antibodies in the serum of AsAb positive infertile patients, to investigate the proportion of anti-Spl7 antibodies in AsAb and their potential application to the serologic diagnosis of immune infertility and immunocontraception.
METHODSWith human recombinant Sp17 as the antigen, the ELISA method was used to detect the positive rate, antibody titre and content of anti-Sp17 antibodies in the AsAb positive serum.
RESULTSThe positive rate of anti-Sp17 antibodies in the AsAb positive serum was 56.5%, with no significant difference in the gender aspect. The percentage of anti-Sp17 antibodies in AsAb was (10.09 +/-7.45) %, with statistical significance (P <0.05).
CONCLUSIONSp17 is an important sperm antigen. Anti-Sp17 antibodies in the serum can be taken as auxiliary diagnostic index of infertility, and Sp17 is shown to be a potential candidate immunocontraception vaccine.
Adult ; Antigens, Surface ; immunology ; Autoantibodies ; blood ; Carrier Proteins ; immunology ; Contraception, Immunologic ; Female ; Humans ; Infertility, Male ; blood ; immunology ; Male ; Spermatozoa ; immunology
8.Advances in sperm membrane antigens.
National Journal of Andrology 2003;9(4):292-295
A series of sperm membrane antigens, the important molecular markers associated with the program of spermatogenesis and maturation of sperm, which also are for attachment of the physiological mechanism of the sperm-egg interaction and fusion and the pathological changes in the infertility, were reviewed. Some traditional crucial sperm membrane proteins, such as FA-1, PH-20, fertilin, have been cloned and sequenced, and some novel gene which encode new proteins associated with the sperm membrane also have been screened out successively. All of these provided a basement for lucubration of these proteins and the research of the contraception vaccine.
Animals
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Antigens, Surface
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genetics
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immunology
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Cell Membrane
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immunology
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Cloning, Molecular
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Humans
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Male
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Mice
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Spermatozoa
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immunology
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Vaccines, Contraceptive
9.Extramedullary infiltration of acute monocytic leukemia/monoblastic sarcoma: a clinicopathologic and immunophenotype analysis of 5 cases.
Zhuo ZUO ; Wei-Ping LU ; Jian-Bo YU ; Ji-Man LI ; Dian-Ying LIAO
Chinese Journal of Pathology 2008;37(1):27-30
OBJECTIVETo study the clinicopathologic features, diagnosis and differential diagnosis of extramedullary infiltration of acute monocytic leukemia/monoblastic sarcoma.
METHODSFive cases of extramedullary infiltration of acute monocytic leukemia/monoblastic sarcoma were selected from 102 cases of myeloid sarcoma diagnosed during the period from 1990 to 2006. The clinicopathologic findings and followup data were retrospectively analyzed. Immunohistochemical study was also carried out with SP method.
RESULTSAmong the 5 cases studied, 3 were males and 2 were females, including 2 children and 3 adults. Generalized lymphadenopathy was found in 4 patients and skin lesions were observed in 2 patients. The tumor cells in all cases were positive for CD68 (KP1), CD68 (PGM1), lysozyme and CD45. They were negative for MPO, CD15, CD163, TdT, CD117, T and B cell markers. The Ki-67 index ranged from 40% to 80%. Follow-up data were available in all the 5 patients. Four of the 5 patients died of the disease, with the average survival time being 6.25 months.
CONCLUSIONSMonoblastic sarcoma is a rare disease with poor prognosis. It is almost impossible to distinguish monoblastic sarcoma from granulocytic sarcoma and other types of small round cell tumors on the basis of morphologic examination alone. Immunohistochemistry is mandatory for a correct diagnosis.
Adult ; Antigens, CD ; immunology ; Antigens, Differentiation, Myelomonocytic ; immunology ; Child ; Diagnosis, Differential ; Female ; Humans ; Immunohistochemistry ; methods ; Immunophenotyping ; Leukemia, Monocytic, Acute ; immunology ; pathology ; Leukocyte Common Antigens ; Lewis X Antigen ; immunology ; Male ; Receptors, Cell Surface ; immunology ; Sarcoma ; immunology ; pathology ; Sarcoma, Myeloid ; immunology ; pathology
10.Reactivity of a novel monoclonal antibody ZCH-2B8a on normal hematopoietic cells and malignant cell lines and its significance.
Yong-Min TANG ; Li GUO ; Shi-Long YANG ; Hong-Qiang SHEN ; Bai-Qin QIAN ; Yi ZHANG ; Hai-Zhong ZHANG
Journal of Experimental Hematology 2006;14(5):990-994
ZCH-2B8a (IgG2a) is a novel monoclonal antibody (McAb) generated in laboratory of Children Hospital of Medical College, Zhejiang University recently using human myeloblastic leukemia cell line KG1a as immunogen. This antibody has been submitted to the 8th International Workshop and Conference on Human Leukocyte Differentiation Antigens (HLDA8) and the results showed that the antibody recognized an unknown molecule on the surface of some blood cells. The aim of this study was to investigate the reactivity of this antibody on normal blood cells and malignant cell lines and to explore its possible application in clinical practice. The multi-parameter flow cytometry was used to analyze the expression pattern of 2B8a antigen in triplicate on normal blood components including T cells, B cells, natural killers (NK), neutrophils, monocytes, dendritic cells (DC), red blood cells (RBC), platelets (Plt), hematopoietic stem/progenitor cells derived from either bone marrow or G-CSF mobilized peripheral blood CD34(+) cells and malignant cell lines including 14 hematopoietic, 5 neuroblastoma, 1 colon cancer and 1 amniotic epithelium cell lines. The amount of positive cells > or = 20% was considered as positivity. The results showed that 2B8a antibody reacted to 3/3 specimens of blood B cells with a positive rate of 26.29% and 2/3 specimens of monocytes with an average positive rate of 59.84%. 2B8a was weakly reactive to neutrophils (23.72%) and negative for T cells, NK, DC, RBC and Plt. The antibody reacted to all 3 marrow CD34(+) cells with an average positive rate of 39.33% while it was negative for G-CSF-mobilized CD34(+) peripheral blood stem/progenitor cells (PBSC, 1.25%). Cell line analysis showed that the antibody notably reacted to three out of 4 cell lines (Raji, SMS-SB, Nalm-6 and Nall-1) with the positive rates of 98.78%, 98.61%, 94.93% respectively and weakly to one of them with 5.68% in B lineage cell lines and monoblastic cell line (U937, 67.78%) while it was only weakly positive or negative for other myeloid leukemia cell lines including Meg01 (33.40%), HL-60 (29.70%), K562 (28.19%), KG1a (16.23%) and HEL92.1.7 (8.02%). Among 4 T lineage leukemia, 5 neuroblastoma and 1 colon cancer cell lines tested, only Molt-3 was found weakly positive (31.40%) for 2B8a, while the remaining 3 T cell lines (Molt4, JM and CCRF-CEM), 5 neuroblastoma cell lines (LA-N1, KCNR, BE, SK-N-SH, SK-N-AS) and the colon cancer cell line (HR8348) tested were negative. An amniotic epithelium cell line (FL) was showed positive for the antibody (45.03%). It is concluded that 2B8a antibody primarily reacts to B lineage and monocytic lineage cells which may bear the diagnostic and therapeutic applications among different types of hematopoietic malignancies.
Antibodies, Monoclonal
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biosynthesis
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immunology
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Antibody Specificity
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Antigen-Antibody Reactions
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Antigens, Neoplasm
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analysis
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immunology
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Antigens, Surface
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immunology
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B-Lymphocytes
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cytology
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immunology
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HLA Antigens
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immunology
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Hematopoietic Stem Cells
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cytology
;
immunology
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Hematopoietic System
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cytology
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Humans
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Leukemia, Myeloid, Acute
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immunology
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pathology
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Tumor Cells, Cultured