1.Clinical Characteristics of Autoimmune Disease with Dual Seropositive Antibodies of Leucine-rich Glioma Inactivated 1 and Contactin-associated Protein 2.
Li Ling DONG ; Hong Zhi GUAN ; Yan HUANG ; Hong Lin HAO ; Jing Wen NIU ; Qing LIU ; Qiang LU ; Dan XU ; Jun Yi ZHANG ; Li Xin ZHOU ; Li Ri JIN ; Hai Tao REN ; Yi Cheng ZHU ; Bin PENG ; Li Ying CUI ; Xiang Qin ZHOU
Acta Academiae Medicinae Sinicae 2019;41(3):344-350
Objective To explore the clinical characteristics of autoimmune disease with dual seropositive antibodies of leucine-rich glioma inactivated 1(LGI1)and contactin-associated protein 2(Caspr2).Methods The clinical data of seven patients with dual seropositive LGI1 and Caspr2 antibodies who were admitted to the Neurology Department of Peking Union Medical College Hospital from July 2014 to December 2017 were retrospectively analyzed.Results Central,peripheral and autonomic nervous systems were all involved in the seven cases;100%(7/7)presented with insomnia,myokymia,neuropahic pain and hyperhydrosis;71%(5/7)showed memory decline or psychiatric and behavioral symptoms;57%(4/7)had urinary hesitation or constipation;and 43%(3/7)had seizure.Electromyography showed 100%(6/6) of the patients had prolonged afterdischarges following normal M waves and/or abnormal spontaneous firing.Electroencephalography revealed slow waves or basic rhythm slowing in 71%(5/7)of patients.Electrocardiography showed sinus tachycardia,axis deviation,and prolonged QT intervals in 71%(5/7)of patients.One patient died from arrhythmia before immunotherapy.One died from pulmonary infection after immunotherapy.Improvement with immunotherapy was documented in the other five cases.No relapse was noted during the 1-2-year follow-up.Conclusions Autoimmune disease with dual seropositive antibodies of LGI1 and Caspr2 can diffusely affect the central,peripheral,and autonomic nervous systems.The possibility of this disease should be considered in patients with acute and subacute onset of neuropsychiatric symptoms,especially in patients with accompanying insomnia,myokymia,and hyperhydrosis.
Autoantibodies
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blood
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Autoimmune Diseases
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immunology
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Humans
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Membrane Proteins
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immunology
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Nerve Tissue Proteins
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immunology
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Proteins
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immunology
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Retrospective Studies
2.Immune plexins and semaphorins: old proteins, new immune functions.
Kelly RONEY ; Eda HOLL ; Jenny TING
Protein & Cell 2013;4(1):17-26
Plexins and semaphorins are a large family of proteins that are involved in cell movement and response. The importance of plexins and semaphorins has been emphasized by their discovery in many organ systems including the nervous (Nkyimbeng-Takwi and Chapoval, 2011; McCormick and Leipzig, 2012; Yaron and Sprinzak, 2012), epithelial (Miao et al., 1999; Fujii et al., 2002), and immune systems (Takamatsu and Kumanogoh, 2012) as well as diverse cell processes including angiogenesis (Serini et al., 2009; Sakurai et al., 2012), embryogenesis (Perala et al., 2012), and cancer (Potiron et al., 2009; Micucci et al., 2010). Plexins and semaphorins are transmembrane proteins that share a conserved extracellular semaphorin domain (Hota and Buck, 2012). The plexins and semaphorins are divided into four and eight subfamilies respectively based on their structural homology. Semaphorins are relatively small proteins containing the extracellular semaphorin domain and short intracellular tails. Plexins contain the semaphorin domain and long intracellular tails (Hota and Buck, 2012). The majority of plexin and semaphorin research has focused on the nervous system, particularly the developing nervous system, where these proteins are found to mediate many common neuronal cell processes including cell movement, cytoskeletal rearrangement, and signal transduction (Choi et al., 2008; Takamatsu et al., 2010). Their roles in the immune system are the focus of this review.
Animals
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Cell Adhesion Molecules
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immunology
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metabolism
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Humans
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Immunity
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Nerve Tissue Proteins
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immunology
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metabolism
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Semaphorins
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immunology
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metabolism
3.Advances in research on the target antigens of antisperm antibodies.
National Journal of Andrology 2004;10(6):458-464
Antisperm antibodies can lead to immunological infertility. Further research on the target antigens of antisperm antibodies may help to discover the causal relationship of antisperm antibodies to infertility. This paper summarizes the structure and function of the six target antigens of antisperm antibodies found recently, so as to discover the causal relationship of the antibodies to infertility and provide a basis for screening a vaccine for immunological contraception.
Amyloid beta-Protein Precursor
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chemistry
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immunology
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Antibodies
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immunology
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Antigen-Antibody Reactions
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Cytoskeletal Proteins
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Glycoproteins
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chemistry
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immunology
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Humans
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Infertility, Male
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etiology
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Male
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Nerve Tissue Proteins
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chemistry
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immunology
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Nuclear Pore Complex Proteins
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Nuclear Proteins
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Spermatozoa
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immunology
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Zyxin
4.Prokaryotic expression, purification of human LINGO-1(aa76-319) and preparation of its polyclonal antibody.
Jun LV ; Xin LU ; Xiao-Dan JIANG ; Chang-Chen HU ; Ying-Qian CAI ; Mou-Xuan DU ; Yu-Xi ZOU ; Ling-Sha QIN
Journal of Southern Medical University 2009;29(11):2175-2178
OBJECTIVETo express and purify the fusion protein of extracellular domain of human Ig domain-containing, neurite outgrowth inhibitor (Nogo) receptor-interacting protein-1 (LINGO-1(aa76-319)) in prokaryotic cells and prepare the rabbit anti-LINGO-1 polyclonal antibody (pAb).
METHODSThe 732 bp DNA sequence of hLINGO-1(aa76-319) was obtained from pCMV-SPORT6 by PCR and inserted into pET30a(+) plasmid to construct the prokaryotic expression plasmid pET30a(+)-hLINGO-1(aa76-319), which was subsequently transformed into E.coli. The target fusion protein was expressed with IPTG induction and purified by Ni(2+)-NTA affinity chromatography column. The antiserum against hLINGO-1(aa76-319) was obtained from the rabbits immunized with hLINGO-1(aa76-319), and the titer of the pAb was determined using enzyme linked immunosorbent assay (ELISA) and its specificity identified using Western blotting.
RESULTSThe prokaryotic expression plasmid pET30a(+)-hLINGO-1(aa76-319) was constructed successfully. Efficient expression of the target fusion protein was achieved with IPTG induction at the optimal concentration of 0.4 mmol/L and culture temperature at 37 degrees celsius; for 2.5 h. The hLINGO-1(aa76-319) fusion protein was effectively expressed in E.coli as inclusion bodies, and the soluble protein was obtained through denaturation and refolding procedures, and the purified fusion protein showed a purity above 90%. The titer of the anti-hLINGO-1(aa76-319) pAb obtained by immunizing the rabbits with the purified protein reached 1:1.6x10(6), and Western blotting confirmed its good specificity.
CONCLUSIONThe fusion protein hLINGO-1(aa76-319) with high purity has been obtained and the anti-hLINGO-1(aa76-319) pAb obtained shows a high titer and good specificity, which provide important experimental basis for further functional investigation of LINGO-1.
Animals ; Antibodies ; immunology ; isolation & purification ; Antibody Specificity ; Escherichia coli ; genetics ; metabolism ; Humans ; Immune Sera ; immunology ; Membrane Proteins ; biosynthesis ; genetics ; immunology ; Nerve Tissue Proteins ; biosynthesis ; genetics ; immunology ; Plasmids ; genetics ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology
5.Preparation of anti-LRRC4 polyclonal antibody and its application in constructing expression profile of human gliomas with different pathological grades.
Dan LI ; Ming-hua WU ; Qiong CHEN ; He HUANG ; Chen HUANG ; Wei-song LI ; Xiao-ling LI ; Gui-yuan LI
Journal of Central South University(Medical Sciences) 2007;32(3):373-379
OBJECTIVE:
To prepare anti-LRRC4 polyclonal antibody and analyze the correlation between the expression of LRRC4 and pathological grades of gliomas in rabbits.
METHODS:
Appropriate protein sequence with good hydrophilicity and antigenicity was chosen by analyzing with DS Gene 1.1 software. The corresponding nucleic acid sequence amplified by PCR was used to construct a recombinant pGEX-4T-2/276 bp. E.coli JM109 transformed with the recombinant was induced by IPTG to express GST-fusion protein, and the fusion protein expressed as insoluble inclusion bodies. Then the purified inclusion body was used to immunize rabbits. Once the titer of antiserum reached 1:10(8) by indirect ELISA, the serum was collected and purified. The expression-profile of LRRC4 in embryonic tissues and gliomas with various pathological grades were obtained by western blot and immunohistochemistry with the anti-LRRC4 polyclonal antibody.
RESULTS:
The highly specific anti-LRRC4 polyclonal antibody whose titer reached 1:10(8) was prepared. The relatively specific expression of LRRC4 was detected in the normal brain, but reduced expression or loss of expression in gliomas was also noticed by immunohistochemistry, and there was a correlation between the expression level of lrrc4 and the pathological grade of gliomas.
CONCLUSION
The anti-LRRC4 polyclonal antibody with high titer and specificity has been obtained. A correlation between the expression level of LRRC4 and the pathological grade of gliomas is detected, which lays the foundation for advanced research of LRRC4.
Animals
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Antibodies, Monoclonal
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immunology
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Antibody Specificity
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immunology
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Blotting, Western
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Brain
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metabolism
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pathology
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Brain Neoplasms
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immunology
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metabolism
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pathology
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Glioma
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immunology
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metabolism
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pathology
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Immunohistochemistry
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Male
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Nerve Tissue Proteins
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biosynthesis
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genetics
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immunology
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Rabbits
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Recombinant Fusion Proteins
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biosynthesis
;
immunology
6.Comparison between CD271 and CD133 used for immunomagnetic positive sorting enriching in mesenchymal stem cells from bone marrow.
Wei LIN ; Xue-Mei TANG ; Yuan KONG ; Hui WANG ; Kai-Yan LIU
Journal of Experimental Hematology 2008;16(2):333-338
This study was aimed to find a better method to isolate and enrich mesenchymal stem cells (MSCs)from bone marrow between CD271 (low affinity nerve growth factor receptor, LNGFR) and CD133 used for immunomagnetic positive selections through comparison of characteristics of MSCs isolated by these two agents. CD271+ and CD133+ cells were isolated from bone marrow and their colony forming unit-fibroblast (CFU-F) efficiency and proliferative capacity were assessed. Cell surface phenotype, adipogenic and osteogenic inductions were also assayed on the cells (after passage 3) isolated by both methods. The results showed that the purities of immunomagnetically selected CD271+ and CD133+ cells were (89.50+/-0.98)% and (88.03+/-3.06)% respectively. The CFU-F median frequency of CD271+ cells was 3 times as high as that of CD133+ cells, no CFU-F was observed in CD271- cells, while a few CFU-F was found in the CD133- cells. Phenotype of cells (after passage 3) isolated by the two methods was same, that is CD34-, CD14-, CD45-, CD90+, CD29+, CD44+, CD105+, CD73+. CD271+ cells possessed faster proliferation and stronger osteogenic and adipogenic differentiation potential than that of CD133+ cells. It is concluded that as compared with CD133 positive selection, CD271 positive selection is a better method for isolating and enriching mesenchymal stem cells from bone marrow.
AC133 Antigen
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Antigens, CD
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immunology
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metabolism
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Bone Marrow Cells
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cytology
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Glycoproteins
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immunology
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metabolism
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Humans
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Immunomagnetic Separation
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methods
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Mesenchymal Stromal Cells
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cytology
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Nerve Tissue Proteins
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immunology
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metabolism
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Peptides
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immunology
;
metabolism
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Receptors, Nerve Growth Factor
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immunology
;
metabolism
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Stem Cells
7.Identification and characterization of a novel HBV large protein binding protein: CDK5RAP3.
Xue-li GONG ; Ben LI ; Jian-long ZHANG ; Jin-qian ZHANG ; Jun CHENG
Chinese Journal of Hepatology 2010;18(5):381-382
Carrier Proteins
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genetics
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metabolism
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Cell Line, Tumor
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Gene Library
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Hepatitis B Surface Antigens
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immunology
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metabolism
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Hepatitis B virus
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genetics
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immunology
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Humans
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Immunoprecipitation
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Intracellular Signaling Peptides and Proteins
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metabolism
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Nerve Tissue Proteins
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metabolism
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Viral Proteins
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immunology
;
metabolism
8.In Vivo Effects of Adipose-Derived Stem Cells in Inducing Neuronal Regeneration in Sprague-Dawley Rats Undergoing Nerve Defect Bridged with Polycaprolactone Nanotubes.
Dong Yeon KIM ; Yong Seong CHOI ; Sung Eun KIM ; Jung Ho LEE ; Sue Min KIM ; Young Jin KIM ; Jong Won RHIE ; Young Joon JUN
Journal of Korean Medical Science 2014;29(Suppl 3):S183-S192
There have been many attempts for regeneration of peripheral nerve injury. In this study, we examined the in vivo effects of non-differentiated and neuronal differentiated adipose-derived stem cells (ADSCs) in inducing the neuronal regeneration in the Sprague-Dawley (SD) rats undergoing nerve defect bridged with the PCL nanotubes. Then, we performed immunohistochemical and histopathologic examinations, as well as the electromyography, in three groups: the control group (14 sciatic nerves transplanted with the PCL nanotube scaffold), the experimental group I (14 sciatic nerves with the non-differentiated ADSCs at a density of 7x105 cells/0.1 mL) and the experimental group II (14 sciatic nerves with the neuronal differentiated ADSCs at 7x105 cells/0.1 mL). Six weeks postoperatively, the degree of the neuronal induction and that of immunoreactivity to nestin, MAP-2 and GFAP was significantly higher in the experimental group I and II as compared with the control group. In addition, the nerve conduction velocity (NCV) was significantly higher in the experimental group I and II as compared with the control group (P=0.021 and P=0.020, respectively). On the other hand, there was no significant difference in the NCV between the two experimental groups (P>0.05). Thus, our results will contribute to treating patients with peripheral nerve defects using PCL nanotubes with ADSCs.
Adipose Tissue/cytology
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Animals
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Cell Differentiation
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Electromyography
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Male
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Nanotubes
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*Nerve Regeneration
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Nerve Tissue Proteins/immunology
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Nestin/immunology
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Neural Conduction/physiology
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Peripheral Nerve Injuries/*surgery
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Phosphoprotein Phosphatases/immunology
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Polyesters/*therapeutic use
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Rats
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Rats, Sprague-Dawley
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Sciatic Nerve/injuries/surgery
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Stem Cell Transplantation/*methods
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Stem Cells/*cytology
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Tissue Engineering/methods
9.Relationship between anti-myelin basic protein antibody and myelinoclasis in rat brain stem after brain trauma.
Wei LI ; Shan-Cheng CHEN ; Zhi-Gang WANG ; Xiu-Bao SONG ; Yu-Ping WANG ; Mei ZHANG
Journal of Southern Medical University 2008;28(6):1028-1030
OBJECTIVETo investigate the relations between anti-myelin basic protein antibody (anti-MBP) variation and myelinoclasis in the brain stem following brain trauma.
METHODSIn rat models of brain trauma, MBP content and anti-MBP titer in the blood were measured using enzyme-linked immunosorbent assay (ELISA) at different time points after brain trauma, and the degree of myelinoclasis in the brain stem slices was assessed with osmic acid staining.
RESULTSEarly after brain trauma, MBP content in the blood increased followed by significant reduction 10 days later. Four days after the trauma, anti-MBP titer was markedly increased, accompanied by obvious exacerbation of myelinoclasis in the brain stem, both reaching the highest levels on day 10, at the point of which anti-MBP titer increased by 4 folds and the number of myelinoclasis by 10 folds compared with the control group. Anti-MBP titer and brain stem myelinolysis both lowered 30 days later. Correlation analysis showed an intimate positive correlation between anti-MBP titer and the degree of myelinoclasis.
CONCLUSIONAfter brain trauma, MBP is released as a specific antigen into the blood to stimulate the immune system for anti-MBP production, and the antibody is intimately related to the brain stem myelinoclasis.
Animals ; Antibodies ; metabolism ; Brain Injuries ; complications ; Brain Stem ; immunology ; pathology ; Demyelinating Autoimmune Diseases, CNS ; etiology ; immunology ; Female ; Male ; Myelin Basic Protein ; Nerve Tissue Proteins ; blood ; immunology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Transcription Factors ; blood ; immunology
10.Transformation of antimicrobial peptide fusion gene of cecropin B and rabbit NP-1 to Houttuynia cordata.
Yan DONG ; Ying ZHANG ; Lang YI ; Huili LAI ; Yaming ZHANG ; Lian ZHOU ; Peixun WANG
China Journal of Chinese Materia Medica 2010;35(13):1660-1665
OBJECTIVETo transform the antimicrobial peptide fusion gene of cecropin B and rabbit NP-1(CN) into Houttuynia cordata to improve its antimicrobic capability.
METHODThe fusion gene of CN designed and synthesized artificially was recombined with expression vector pBI121. The recombined vector was transformed to Agrobacterium tumefaciens LBA4404, by which CN gene was transformed to the explants of H. cordata. The transgenic regeneration plantlets were selected by kanamycin and rapid screening PCR. The transgenic plants were identified by PCR-Southern of genomic DNA and RT-PCR. The disease resistances were detected by antibacterial zone trail of leaf extracts to E. coli K12 and infection by Rhizoctonia solani.
RESULTGene of interesting CN was inserted into genomic DNA and expressed in transformed H, cordata, whose resistance to E. coli K12 and Rh. solani was stronger than that of the non-transformed control.
CONCLUSIONThe fusion gene CN can improve antimicrobic capability of transformed H. cordata.
Animals ; Anti-Bacterial Agents ; immunology ; pharmacology ; C-Reactive Protein ; genetics ; metabolism ; pharmacology ; Houttuynia ; genetics ; immunology ; microbiology ; Immunity, Innate ; Insect Proteins ; genetics ; immunology ; pharmacology ; Nerve Tissue Proteins ; genetics ; metabolism ; pharmacology ; Plant Diseases ; immunology ; microbiology ; Plants, Genetically Modified ; genetics ; immunology ; microbiology ; Rabbits ; Recombinant Fusion Proteins ; genetics ; immunology ; pharmacology ; Rhizoctonia ; physiology ; Transformation, Genetic