1.Pro- and anti-inflammatory cytokine expression and histopathological characteristics in canine brain with traumatic brain injury.
Chi Ho YU ; Ji Young YHEE ; Jong Hyuk KIM ; Keum Soon IM ; Na Hyun KIM ; Dong In JUNG ; Hee Chun LEE ; Seung Ki CHON ; Jung Hyang SUR
Journal of Veterinary Science 2011;12(3):299-301
We analyzed the expression level and cellular localization of pro- and anti-inflammatory cytokines and histopathologically characterized canine traumatic brain injury (TBI). Canine TBI brains revealed subarachnoid and cerebral cortical hemorrhage, neutrophilic infiltration, neuronal necrosis, astrocytosis, and vasogenic edema. Immunohistochemical evaluations suggested that both pro-inflammatory cytokines [interleukin (IL)-1beta, IL-6, and tumor necrosis factor-alpha] and anti-inflammatory cytokines [IL-10 and transforming growth factor-beta (TGF-beta)] were highly expressed in neurons and neutrophils. In particular, the highest magnitude of expression was identified for IL-1beta and TGF-beta. This data helps describe the pathologic characteristics of canine TBI, and may help in the design of potential therapeutic approaches to control secondary damage by inflammatory cytokines.
Animals
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Brain/*immunology/*pathology
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Brain Injuries/immunology/*pathology/*veterinary
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Dogs
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Humans
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Interleukin-10/immunology/metabolism
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Interleukin-1beta/immunology/metabolism
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Interleukin-6/immunology/metabolism
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Transforming Growth Factor beta/immunology/metabolism
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Tumor Necrosis Factor-alpha/immunology/metabolism
2.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
;
immunology
;
metabolism
;
pathology
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Glioma
;
immunology
;
metabolism
;
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
;
immunology
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Rabbits
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Recombinant Fusion Proteins
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biosynthesis
;
immunology
3.Encephalic lymphomatoid granulomatosis.
Li YU ; Zhen-Xi LIU ; Sha XIAO ; Cheng-Yi LUO
Chinese Journal of Pathology 2005;34(2):121-122
4.Role of Brain Inflammation in Epileptogenesis.
Yonsei Medical Journal 2008;49(1):1-18
Inflammation is known to participate in the mediation of a growing number of acute and chronic neurological disorders. Even so, the involvement of inflammation in the pathogenesis of epilepsy and seizure-induced brain damage has only recently been appreciated. Inflammatory processes, including activation of microglia and astrocytes and production of proinflammatory cytokines and related molecules, have been described in human epilepsy patients as well as in experimental models of epilepsy. For many decades, a functional role for brain inflammation has been implied by the effective use of anti-inflammatory treatments, such as steroids, in treating intractable pediatric epilepsy of diverse causes. Conversely, common pediatric infectious or autoimmune diseases are often accompanied by seizures during the course of illness. In addition, genetic susceptibility to inflammation correlated with an increased risk of epilepsy. Mounting evidence thus supports the hypothesis that inflammation may contribute to epileptogenesis and cause neuronal injury in epilepsy. We provide an overview of the current knowledge that implicates brain inflammation as a common predisposing factor in epilepsy, particularly childhood epilepsy.
Animals
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Blood-Brain Barrier
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Chronic Disease
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Encephalitis/genetics/immunology/metabolism/*pathology
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Epilepsy/immunology/metabolism/*pathology/therapy
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Gene Expression Regulation
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Humans
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Nervous System Diseases/immunology/pathology
5.Screening serum response special antibodies of U251 cell line from surface display phage antibody library.
Min YU ; De-Yong TAN ; Wei QIAN ; Jian-Hua LAI ; Gui-Lin SUN
Chinese Journal of Biotechnology 2004;20(3):356-360
U251 cell is a sensitive cell line to serum, which stops at G0 phase of cell cycle in no-serum medium, and recovers growth when the serum is added into no-serum medium. The cell can express corresponding proteins in different phase of cell cycle. Therefore it is very signification for the study of cell cycle regulation mechanism that explores these proteins. In this paper, the mouse antibody phage display library was added into the bottle in which the serum starvation U251 cells had been cultured, and the special antibody phages were absorbed. Then the absorbed antibody phages were amplified by adding E. coli TG1 and helper phage M13K07. Amplified antibody phages were added into bottle in which the serum cultured cell after serum starvation (follow named as serum recovered cells) were incubated, so that the cell absorbed the no-special antibody phages for the serum starvation cell and the special antibody phages were in supernatant. The remaining no-special antibody phages in the supernatant were discarded by repeating above program 3-4 times. The pure special antibody phages were gotten, and amplified by adding the host cell E. coli TG1 and helper phage M13K07. Then the host bacterium infected special antibody phage was spread on the plate medium with ampicillin, and the monoclonal antibody phages were gotten. Using same as above program, the monoclonal antibody phages absorbed specially for serum recovered U251 cells were obtained when the serum recovered cells instead of serum starvation cells and serum starvation cells instead of serum recovered cells. In this study, ninety-six positive monoclonal antibody phages that absorbed specially the serum starvation cells and eighty-two positive monoclonal antibody phages that absorbed specially the serum recovered cells were obtained. By using cell immunochemistry assay, two special signification antibodies were obtained. one (No.11) was the strong response in serum starvation cells, the other (No.2) was the strong response in serum recovered cells. The antibody No.2 had the distinctive response to the serum recovered cells in different incubation time (15min, 30min, 1h, 2h, 4h, 8h, 12h and 48h) after serum starvation. The results showed that No.2 antibody would be useful to research the factors of cell cycle regulation and apply to tumor diagnosis.
Antibodies, Neoplasm
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genetics
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immunology
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isolation & purification
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Bacteriophages
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genetics
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Brain Neoplasms
;
immunology
;
pathology
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Cell Line, Tumor
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Escherichia coli
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genetics
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metabolism
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Glioma
;
immunology
;
pathology
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Humans
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Peptide Library
6.Magnetic resonance imaging characteristics of spinal cord lesions and brain abnormalities in Chinese patients with neuromyelitis optica.
Lei WU ; Yang YANG ; De-hui HUANG ; Hui XIAO ; Wei-ping WU
Journal of Southern Medical University 2010;30(12):2733-2737
OBJECTIVETo analyze the magnetic resonance imaging (MRI) characteristics of neuromyelitis optica (NMO) in Chinese patients.
METHODSWe retrospectively reviewed the MRI films of 61 patients with NMO (including 57 female and 4 male patients) admitted in our department.
RESULTSOf these patients, 39 (79.6%) showed positivity for serum aquaporin-4 (AQP4) antibody. On MRI, 18 patients showed involvement of the cervical cord alone, 27 had both cervical and thoracic segment involvement, and 16 displayed thoracic segment involvement. The lesions appeared linear (9 cases), diffuse (23 cases), or both (29 cases), mostly located axially with occasional lateral distribution. Thirty-nine of the 61 patients (63.9%) had brain abnormalities, 31 presented with supratentorial lesions (mostly in the juxtacortical, subcortical, deep white matter and lateral ventricle-adjacent regions, n=27), 15 showed infratentorial lesions (mostly in the preiaqueduct-fourth ventricular-central canal, n=13), and 7 had supra- and infratentorial lesions simultaneously.
CONCLUSIONNMO has complex MRI presentation, and linear lesions in the spinal cord and preiaqueduct-fourth ventricular-central canal lesions, where AQP4 is high expressed, can be characteristic for NMO. MRI and AQP4 antibody detection are suggested for suspected cases for early diagnosis.
Adult ; Aquaporin 4 ; immunology ; Brain ; pathology ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Neuromyelitis Optica ; pathology ; Retrospective Studies ; Spinal Cord ; pathology
7.Relationship between cellular immune response and apoptosis of the brain neurons after brain trauma in rats.
Ke-Jia CHEN ; Wei LI ; Zhi-Gang WANG
Journal of Southern Medical University 2009;29(3):497-499
OBJECTIVETo investigate the relation between CD4(+) and CD8(+) T lymphocyte infiltration and apoptosis of the neurons in the local traumatic brain tissue after brain trauma in rats.
METHODSIn rat models of brain trauma, the changes in the number of CD4(+) and CD8(+) T lymphocytes and the apoptosis of neurons in the local traumatic brain tissue were observed by immunohistochemistry at different time points after brain trauma.
RESULTSTwenty-four hours after brain trauma, a significant increase in the number of CD4(+) and CD8(+) T lymphocytes occurred in the injured brain tissue, both reaching the highest levels on day 10, at the point of which the number of CD4(+) cells increased by about 15 folds and that of CD8(+) cells by about 20 folds compared with the control groups. The CD4(+) and CD8(+) T lymphocytes both began to decrease 30 days after the injury. A similar pattern of alterations was found in the apoptosis of neurons in the local brain tissue. Correlation analysis demonstrated a close positive correlation between the changes in CD4(+) and CD8(+) lymphocyte numbers and the number apoptotic neurons in the injured brain tissue.
CONCLUSIONSBrain trauma induces obvious increases in CD4(+) and CD8(+) T lymphocytes and enhanced cellular immune response in the injured brain tissue to mediate neuronal apoptosis and further exacerbate the brain tissue injuries.
Animals ; Apoptosis ; immunology ; Brain Injuries ; immunology ; pathology ; CD4-Positive T-Lymphocytes ; cytology ; immunology ; CD8-Positive T-Lymphocytes ; cytology ; immunology ; Female ; Immunity, Cellular ; Male ; Neurons ; immunology ; pathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley
8.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
9.Immunopathological Changes in the Brain of Immunosuppressed Mice Experimentally Infected with Toxocara canis.
Mohamed M EID ; Samy I EL-KOWRANY ; Ahmad A OTHMAN ; Dina I El GENDY ; Eman M SAIED
The Korean Journal of Parasitology 2015;53(1):51-58
Toxocariasis is a soil-transmitted helminthozoonosis due to infection of humans by larvae of Toxocara canis. The disease could produce cognitive and behavioral disturbances especially in children. Meanwhile, in our modern era, the incidence of immunosuppression has been progressively increasing due to increased incidence of malignancy as well as increased use of immunosuppressive agents. The present study aimed at comparing some of the pathological and immunological alterations in the brain of normal and immunosuppressed mice experimentally infected with T. canis. Therefore, 180 Swiss albino mice were divided into 4 groups including normal (control) group, immunocompetent T. canis-infected group, immunosuppressed group (control), and immunosuppressed infected group. Infected mice were subjected to larval counts in the brain, and the brains from all mice were assessed for histopathological changes, astrogliosis, and IL-5 mRNA expression levels in brain tissues. The results showed that under immunosuppression, there were significant increase in brain larval counts, significant enhancement of reactive gliosis, and significant reduction in IL-5 mRNA expression. All these changes were maximal in the chronic stage of infection. In conclusion, the immunopathological alterations in the brains of infected animals were progressive over time, and were exaggerated under the effect of immunosuppression as did the intensity of cerebral infection.
Animals
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Brain/*pathology
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Disease Models, Animal
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Female
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Gene Expression Profiling
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Histocytochemistry
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*Immunocompromised Host
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Immunohistochemistry
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Interleukin-5/genetics
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Male
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Mice
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Parasite Load
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Toxocara canis/*immunology
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Toxocariasis/*immunology/*pathology
10.Immunopathological Changes in the Brain of Immunosuppressed Mice Experimentally Infected with Toxocara canis.
Mohamed M EID ; Samy I EL-KOWRANY ; Ahmad A OTHMAN ; Dina I El GENDY ; Eman M SAIED
The Korean Journal of Parasitology 2015;53(1):51-58
Toxocariasis is a soil-transmitted helminthozoonosis due to infection of humans by larvae of Toxocara canis. The disease could produce cognitive and behavioral disturbances especially in children. Meanwhile, in our modern era, the incidence of immunosuppression has been progressively increasing due to increased incidence of malignancy as well as increased use of immunosuppressive agents. The present study aimed at comparing some of the pathological and immunological alterations in the brain of normal and immunosuppressed mice experimentally infected with T. canis. Therefore, 180 Swiss albino mice were divided into 4 groups including normal (control) group, immunocompetent T. canis-infected group, immunosuppressed group (control), and immunosuppressed infected group. Infected mice were subjected to larval counts in the brain, and the brains from all mice were assessed for histopathological changes, astrogliosis, and IL-5 mRNA expression levels in brain tissues. The results showed that under immunosuppression, there were significant increase in brain larval counts, significant enhancement of reactive gliosis, and significant reduction in IL-5 mRNA expression. All these changes were maximal in the chronic stage of infection. In conclusion, the immunopathological alterations in the brains of infected animals were progressive over time, and were exaggerated under the effect of immunosuppression as did the intensity of cerebral infection.
Animals
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Brain/*pathology
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Disease Models, Animal
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Female
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Gene Expression Profiling
;
Histocytochemistry
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*Immunocompromised Host
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Immunohistochemistry
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Interleukin-5/genetics
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Male
;
Mice
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Parasite Load
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Toxocara canis/*immunology
;
Toxocariasis/*immunology/*pathology