Ten serum samples from the patient with bullous pemphigoid with the baseruent mernbrane zone autoantibody titers of 320 or greater were tested, by the method of in vitro complement immunofluorescence, for their ability to fix Factor B and properdin in addition to Clq, C4 and C3. Five samples yielded positive C3 and properdin staining reaciions while four samples demonstrated positive Factor B stainings. All ten samples yieled positive C3, C4 and Clq staining reactions, Heat inactivation or treatment of the complement source with EDTA, MG2-EGTA abolished both C3, properdin and Factor B staining in all ten cases. This result suggest that pemphigoid antibody will fix properdin and Factor B in addition to Clq, C4 and C3, a phenomenon explained by assembly of the C3b amplification mechanism following activation of the classical pathway of complement system.
Complement Factor B* ; Complement System Proteins ; Edetic Acid ; Fluorescent Antibody Technique ; Hot Temperature ; Humans ; Pemphigoid, Bullous* ; Properdin*

The complement system is part of the innate immune response and as such defends against invading pathogens, removes immune complexes and damaged self-cells, aids organ regeneration, confers neuroprotection, and engages with the adaptive immune response via T and B cells. Complement activation can either benefit or harm the host organism; thus, the complement system must maintain a balance between activation on foreign or modified self surfaces and inhibition on intact host cells. Complement regulators are essential for maintaining this balance and are classified as soluble regulators, such as factor H, and membrane-bound regulators. Defective complement regulators can damage the host cell and result in the accumulation of immunological debris. Moreover, defective regulators are associated with several autoimmune diseases such as atypical hemolytic uremic syndrome, dense deposit disease, age-related macular degeneration, and systemic lupus erythematosus. Therefore, understanding the molecular mechanisms by which the complement system is regulated is important for the development of novel therapies for complement-associated diseases.
Adaptive Immunity ; Antigen-Antibody Complex ; Autoimmune Diseases ; Autoimmunity ; B-Lymphocytes ; Complement Activation ; Complement Factor H ; Complement System Proteins* ; Glomerulonephritis, Membranoproliferative ; Hemolytic-Uremic Syndrome ; Immunity, Innate ; Lupus Erythematosus, Systemic ; Macular Degeneration ; Physiology* ; Regeneration

OBJECTIVE: To explore the effects of (resolving stasis, promoting collateral circulation) moxibustion on learning and memory ability and the expressions of brain derived neurotrophic factor (BDNF) and tyrosine kinase B (TrkB) in the rats of vascular dementia (VD) in the microenvironment of neurovascular niche. METHODS: Using 2-vessel occlusion (2-VO), the VD rat models were duplicated. The neural stem cells (NSCs) labeled with lentiviral vector-mediated enhanced green fluorescent protein (EGFP) were co-cultured with endothelial progenitor cells (EPCs) to structure the NSCs + EPCs implant. The implant was transplanted into the lateral ventricle of VD rats and the VD rat models with neurovascular niche were established. In No.1 experiment, the successful-modeled rats were divided into 3 groups, i.e. a NSCs + EPCs moxibustion group, a NSCs + EPCs blank group and a model group, 12 rats in each one. No any treatment was provided in the model group and the NSCs + EPCs blank group. The moxibustion therapy was adopted in the NSCs + EPCs moxibustion group, in which, the suspending moxibustion technique was applied to "Baihui" (GV 20), "Dazhui" (GV 14) and "Shenting" (GV 24), 20 min at each acupoint. The treatment was given once every day and a 14-day treatment was as one course. Totally, 3 courses of treatment were required. At the end of treatment, Morris water maze experiment was adopted to determine the learning and memory ability of the rats in each group. In the No.2 experiment, the model rats were divided into 3 groups, a NSCs + EPCs moxibustion group, a NSCs + EPCs blank group and a model group, 18 rats in each one. In each group, according to the durations of treatment, 3 subgroups were divided and 6 rats in each one. The intervention method was same as the No.1 experiment. Additionally, after corresponding treatment course, using perfusion, the brains were collected in each subgroup and the slices were frozen. BDNF/TrkB expressions were observed in the immunofluorescence test. RESULTS: After treatment, in the NSCs + EPCs moxibustion group, the escape incubation was reduced, the time of the first running-cross platform was shortened and the frequency of running-cross platform increased as compared with the model group and the NSCs + EPCs blank group (<0.01, <0.05). The protein expressions were increased in tendency among the 3 courses of treatment in the NSCs + EPCs moxibustion group, indicating the significant differences (all <0.05), in which, the increase of the protein expressions in the NSCs + EPCs moxibustion group was better than the NSCs + EPCs blank group (<0.05, <0.01). CONCLUSION The moxibustion therapy is the effective approach to VD in clinical treatment. This therapy up-regulates the BDNF/TrkB protein expressions in the microenvironment of neurovascular niche, co-modulates NSCs-EPCs coupling mechanism, promotes nerve neogenesis and repairs the injured nerve.
Animals ; Brain-Derived Neurotrophic Factor ; metabolism ; Complement Factor B ; Dementia, Vascular ; metabolism ; Drugs, Chinese Herbal ; Hippocampus ; Moxibustion ; Protein-Tyrosine Kinases ; metabolism ; Rats ; Rats, Sprague-Dawley

The purine antimetabolite 6-Mercaptopurine (6-MP) has been in clinical use for over 30 years and is still a widely used agent in the treatment of childhood acute lymphoblastic leukemia. The bioavailibility, clinical efficacy and toxicity of 6-MP administered orally for maintenance therapy of children with acute lymphoblastic leukemia are highly variable in many studies, as well as at differnt times in same patient. there are many factors affecting the bioavailibility of 6-MP. The most notably factor being that concomitantly administered drugs and foods might contribute to a decrease in the bioavailibity of this drug. In our sociocultural environment milk is a major constituent of child's foods. Cow's milk contains a high concentration of xanthine oxidase, which could potentially transform 6-TM into 6-thioxanthine (6-TX) and 6-thiouric acid (6-TUA) which have no more therapeutic effects. In this study, we evaluated the effect of various milk products on the bioavailability of 6-MP. Incubation at 37degrees C for 30 min raw or pasteurized milk resulted in transformation of a large quantity of clinically relevant concentration of 6-MP into 6-TUA. The concomitant adminstration of folic acid and allopurinol has markedly inhibitory effect on the 6-MP destroying activity of milk at clinically relevant concentrations. These observations may help to optimize modalities of administration of 6-MP for the treartment of patients with childhood leukemia.
6-Mercaptopurine* ; Allopurinol ; Biological Availability* ; Child ; Complement Factor B ; Folic Acid ; Humans ; Leukemia ; Milk* ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Xanthine Oxidase

PURPOSE: T cell-mediated immune responses, and particularly activation of polyfunctional T cells that simultaneously produce multiple cytokines, are necessary for the control of Mycobacterium tuberculosis. In the present study, we examined if DNA immunization of Mycobacterium tuberculosis resuscitation-promoting factor B (RpfB) elicits polyfunctional T cell responses in mice. MATERIALS AND METHODS: C57BL/6 mice were immunized intramuscularly three times, at 3-week intervals, with RpfB-expressing plasmid DNA. For comparison, protein immunization was performed with recombinant RpfB in control mice. After immunization, RpfB-specific T cell responses were assessed by interferon-gamma (IFN-gamma) enzyme-linked immunosorbent spot assay and intracellular cytokine staining (ICS), and T cell polyfunctionality was assessed from the ICS data. RESULTS: RpfB DNA immunization induced not only humoral immune responses, but also CD8+ and CD4+ T cell responses. Immunodominant T-cell epitopes were identified within RpfB by assays with overlapping peptides. RpfB DNA immunization elicited a polyfunctional CD8+ T cell response that was dominated by a functional phenotype of IFN-gamma+/TNF-alpha+/IL-2-/CD107a+. CONCLUSION: RpfB DNA immunization elicits polyfunctional CD8+ T cell responses, suggesting that RpfB DNA immunization might induce protective immunity against tuberculosis.
Animals ; Complement Factor B* ; Cytokines ; DNA* ; Epitopes, T-Lymphocyte ; Immunity, Humoral ; Immunization* ; Interferon-gamma ; Mice ; Mycobacterium tuberculosis* ; Peptides ; Phenotype ; Plasmids ; T-Lymphocytes ; Tuberculosis ; Vaccines, DNA

Patients should be treated with dignity and respect toward the end of their lives, being freed from unnecessary and painful life-sustaining therapy in hospitals. In Korea, the quality of endof-life (EOL) care has been variable, a major factor being the physicians' perception to the care. A firm consensus of EOL care decision-making has not yet explicitly stated in Korean law and ethics until recently. However, movements to make a law of so-called "the death with dignity act" are presently making its way to the National Assembly, initiated by a law case that allowed the hospital to withdraw mechanical ventilator support per request by the patients' family of a permanently vegetative patient. Socially agreed guidelines for EOL care can facilitate clinical decision process and communication between health service provider and the patient or his/her family. At the same time, EOL care should be individualized also in the same line of guideline to meet patient' and patient' family wish regarding the withdrawal of life-sustaining therapy. The painful EOL care experience of the loved one remains in the memory of the relatives who live on. Physicians should identify, document, respect, and act on behalf of the hospitalized patients' needs, priorities, and preference for EOL care. It has been advocated that competent patients can express their right of self-determination on EOL care through advance directives in Western countries. Advance directives are considered as a tool to facilitate EOL decision making. However, there are barriers to adopt the advance directives as a legitimate tool for an EOL decision making in Korea. For one thing, the reality of death and dying is rarely discussed in our society. In addition, the discussion about EOL care with chronically and critically ill patients has been considered as a taboo in the hospitals. In spite of these difficulties, physicians could do better EOL care by the open communication with patients or with their surrogates. Through the communication, physician should set a goal how to manage the EOL patient. The set goal should be shared among the caregivers to achieve the maximum benefit of the patient. The lack of open discussion with patient prior to EOL care results in inappropriate protraction of a patient's dying process. In summary, physicians, who know the clinical significance of delivering treatments to EOL patients, should play a central role in assisting patients' and their families' to make the best decision on EOL care. Moreover, the concerted actions to improve EOL care in our society among general public, professionals, stakeholders for EOL care, and governmental organizations are required to address ongoing social requests, although a policy or a guideline is made in this time.
Advance Directives ; Caregivers ; Complement Factor B ; Consensus ; Critical Illness ; Decision Making ; Ethics, Medical ; Health Services ; Humans ; Jurisprudence ; Korea ; Love ; Memory ; Physician's Role ; Right to Die ; Taboo ; Ventilators, Mechanical

<b>BACKGROUNDb>The present study was undertaken to replicate the associations of representative polymorphisms in three genes (complement factor H (CFH), complement factor B (BF) and HtrA serine peptidase 1 (HTRA1)) with exudative age-related macular degeneration (AMD) in a Han Chinese population, and to test if the modifiable environmental factors affect AMD susceptibility associated with different type of genotype in these genes.

<b>METHODSb>An age, gender and ethnicity matched case-control study was conducted to genotype the representative single neucleotide polymorphisms (SNPs) loci including rs1061170 and rs1410996 in CFH, rs641153 and rs4151667 in BF and rs11200638 in HTRA1 gene in 144 exudative AMD patients and 126 normal controls using PCR-RFLP and direct resequencing. The demographic characteristics and behavioral risk factors were also recorded. Allelic and genotypic associations for individual SNP and joint associations with two loci were performed. The gene-gene and gene-environment interactions were analyzed using multivariate non-conditional Logistic regression analysis.

<b>RESULTSb>The C risk allele frequencies for CFH Y402H (rs1061170) in cases and controls were 12.5% and 5.4% respectively, which were much lower than those in Caucasians (P < 0.001). Compared with TT homozygous genotype, the CT heterozygous genotype was positively associated with AMD with odds ratio (OR) of 3.23 (1.36 - 5.07). However, the population attributable risk (PAR) of C allele was only 3.3% (1.4% - 4.3%). rs1410996 was also associated with AMD independent of Y402H. The ORs of exudative AMD for individuals carrying one copy risk allele and two copy risk alleles were 2.57 (1.21 - 5.45) and 4.76 (2.15 - 10.55) respectively, with correspondent PARs of 28.3% (2.0% - 40.5%) and 38.2% (21.8% - 45.4%). rs11200638 in HTRA1 was another susceptible locus for AMD and the risk homozygotes were significantly susceptible for exudutive AMD (OR = 3.98, 1.88 - 8.43) with PAR of 38.9% (24.3% - 45.8%). Education status and cigarette smoking were also related to exudative AMD. After controlling for environmental risk factors, CFH and HTRA1 SNPs were independently associated with exudative AMD, with OR of 3.50 (1.45 - 8.45) for CT genotype in Y402H, 3.34 (1.33 - 8.36) for GG genotype in rs1410996 and 3.85 (1.58 - 9.42) for AA genotype in rs11200638 respectively. The interaction analysis between gene and environmental factors showed that smoking synergistically increased susceptibility of AMD for heterozygotes of rs1410996, with OR(interaction) of 7.33 (P(interaction) = 0.029).

<b>CONCLUSIONSb>In a Han Chinese population, CFH and HTRA1 polymorphisms appear to be independently and possibly additively hereditary contributors to exudative AMD. Y402H polymorphism conferred a significant but relatively lower contribution in Chinese than in Caucasians with a low frequency of risk allele. The gene-environment interaction may be a best way to encourage those with a high genetic risk to prevent AMD by avoiding modifiable factors until there is effective treatment for AMD.

Aged ; Asian Continental Ancestry Group ; Complement Factor B ; genetics ; Complement Factor H ; genetics ; Female ; Gene Frequency ; genetics ; Genetic Predisposition to Disease ; genetics ; Genotype ; High-Temperature Requirement A Serine Peptidase 1 ; Humans ; Macular Degeneration ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; genetics ; Risk Factors ; Serine Endopeptidases ; genetics ; Smoking ; adverse effects

Collagen induced arthritis (CIA)is an animal model that in many ways resembles rheumatoid arthritis (RA). CIA can be induced in susceptible animals by immunization with type II collagen (CII). Like RA,CIA is characterized by intense joint inflammation and destruction.On histological examination,there is synovitis accompanied by erosion of cartilage and subchondral bone. Autoanti-bodies to CII initiate joint inflammation by binding to articular cartilage,forming antigen-antibody complexes locally and activating hemolytic complement. Susceptibility to CIA in mice is linked to the expression of specific class II MHC Molecules,which dictate the T cell determinants on CII,and therefore,the subsets of T cells that can be activated by CII.In addition to activation of B cells reactive to CII,the T cells stimulate monocytes/macrophages.These cells amplify the inflammatory cascade by secretion of proinflammatory monokines, including TNF-alpha and IL-1,leading to the production of other proinflammatory proteins,including matrix metalloproteinases (MMPs).The importance of CIA lies in its possible relationship to arthritis in humans.Progress in understanding CIA has contributed to the development of new therapies for RA.In addition,it has been found that mice with human HLA-DR1,DR4 and HLA-DQ8 transgenes,which have been demonstrated to be the susceptibility markers for RA, confer susceptibility to CIA.These observations coupled with the finding of T cells and B cells reactive with CII in the inflamed joints of RA patients establish the potential role of CII autoimmunity in the pathogenesis of RA.
Animals ; Antigen-Antibody Complex ; Arthritis ; Arthritis, Experimental* ; Arthritis, Rheumatoid* ; Autoimmunity* ; B-Lymphocytes ; Cartilage ; Collagen ; Collagen Type II* ; Complement System Proteins ; Humans ; Immunization ; Inflammation ; Joints ; Matrix Metalloproteinases ; Mice ; Models, Animal ; Monokines ; Synovitis ; T-Lymphocytes ; Tumor Necrosis Factor-alpha

The prognosis of myasthenia gravis (MG) has improved dramatically due to advances in critical-care medicine and symptomatic treatments. Its immunopathogenesis is fundamentally a T-cell-dependent autoimmune process resulting from loss of tolerance toward self-antigens in the thymus. Thymectomy is based on this immunological background. For MG patients who are inadequately controlled with sufficient symptomatic treatment or fail to achieve remission after thymectomy, remission is usually achieved through the addition of other immunotherapies. These immunotherapies can be classified into two groups: rapid induction and long-term maintenance. Rapid induction therapy includes intravenous immunoglobulin (IVIg) and plasma exchange (PE). These produce improvement within a few days after initiation, and so are useful for acute exacerbation including myasthenic crisis or in the perioperative period. High-dose prednisone has been more universally preferred for remission induction, but it acts more slowly than IVIg and PE, commonly only after a delay of several weeks. Slow tapering of steroids after a high-dose pulse offers a method of maintaining the state of remission. However, because of significant side effects, other immunosuppressants (ISs) are frequently added as "steroid-sparing agents". The currently available ISs exert their immunosuppressive effects by three mechanisms: 1) blocking the synthesis of DNA and RNA, 2) inhibiting T-cell activation and 3) depleting the B-cell population. In addition, newer drugs including antisense molecule, tumor necrosis factor alpha receptor blocker and complement inhibitors are currently under investigation to confirm their effectiveness. Until now, the treatment of MG has been based primarily on experience rather than gold-standard evidence from randomized controlled trials. It is hoped that well-organized studies and newer experimental trials will lead to improved treatments.
Autoantigens ; B-Lymphocytes ; Complement System Proteins ; DNA ; Humans ; Immunoglobulins ; Immunoglobulins, Intravenous ; Immunosuppressive Agents ; Immunotherapy ; Myasthenia Gravis ; Perioperative Period ; Plasma Exchange ; Prednisone ; Prognosis ; Remission Induction ; RNA ; Steroids ; T-Lymphocytes ; Thymectomy ; Thymus Gland ; Tumor Necrosis Factor-alpha

Rheumatoid arthritis is a chronic inflammatory disease that primarily affects joint synovium. Although its etiology has yet to be identified, the underlying mechanism of joint inflammation is understood as autoimmune process. The inflamed synovium is thickened due to synovial hyperplasia and infiltrating mononuclear cells such as T and B lymphocytes, macrophages, and plasma cells. Key inflammatory cytokines TNF-alpha and IL-1, as shown by the significant therapeutic effect of their blockade, are mainly secreted by macrophages whereas IL-17 is secreted by a newly recognized subset of T cells (Th17 cells) and induces TNF-alpha and IL-1 production by adjacent macrophages, synoviocytes, and chondrocytes. IL-17 has also been shown to induce RANKL from osteoblasts, thereby indicating that this cytokine plays as an upstream molecule that regulates inflammation and osteoclastogenesis. More importantly, IL-17 has been shown to convert acute inflammation into chronic inflammation and when combined with already important cytokines, more marked inflammation occurs. The role of B cells as antigen presenting cells are now being recognized based on the therapeutic effect of rituximab, a B cell inhibitor, in rheumatoid arthritis. Great attention has been turned to anti-citrullinated peptide antibodies because they form immune complex and contribute to inflammation by activating complement system. Recently, clinical trials showed therapeutic efficacy of tocilizumab, monoclonal antibody against IL-6 receptor, suggesting relevant involvement of IL-6 in disease process of rheumatoid arthritis. Thus, various cellular and molecular players seem to interact within rheumatoid synovium to perpetuate inflammation. Further studies are needed to explore the exact mechanisms of development and maintenance of inflammation in rheumatoid arthritis.
Antibodies ; Antibodies, Monoclonal, Humanized ; Antibodies, Monoclonal, Murine-Derived ; Antigen-Antibody Complex ; Antigen-Presenting Cells ; Arthritis, Rheumatoid ; B-Lymphocytes ; Chondrocytes ; Complement System Proteins ; Cytokines ; Hyperplasia ; Inflammation ; Interleukin-1 ; Interleukin-17 ; Interleukin-6 ; Joints ; Macrophages ; Osteoblasts ; Plasma Cells ; Receptors, Interleukin-6 ; Synovial Membrane ; T-Lymphocytes ; Tumor Necrosis Factor-alpha ; Rituximab