ABSTRACT
BACKGROUND: There are a million ragpickers in India who gather and trade recyclable municipal solid wastes materials for a living. The objective of this study was to examine whether their occupation adversely affects their immunity. METHODS: Seventy-four women ragpickers (median age, 30 years) and 65 age-matched control housemaids were enrolled. Flow cytometry was used to measure leukocyte subsets, and leukocyte expressions of Fcγ receptor I (CD64), FcγRIII (CD16), complement receptor 1 (CD35) and CR3 (CD11b/CD18), and CD14. Serum total immunoglobulin-E was estimated with enzyme-linked immunosorbent assay. RESULTS: Compared with the controls, ragpickers had significantly (p < 0.0001) higher levels of CD8+T-cytotoxic, CD16+CD56+natural killer, and CD4+CD45RO+memory T-cells, but depleted levels of CD19+B-cells. The percentage of CD4+T-helper-cells was lower than the control group (p < 0.0001), but their absolute number was relatively unchanged (p = 0.42) due to 11% higher lymphocyte counts in ragpickers. In ragpickers, the percentages of CD14+CD16+intermediate and CD14dim CD16+nonclassical monocyte subsets were elevated with a decline in CD14+CD16-classical monocytes. The expressions of CD64, CD16, CD35, and CD11b/CD18 on both monocytes and neutrophils, and CD14 on monocytes were significantly higher in ragpickers. In addition, ragpickers had 2.7-times more serum immunoglobulin-E than the controls (p < 0.0001). After controlling potential confounders, the profession of ragpicking was positively associated with the changes. CONCLUSION: Ragpicking is associated with alterations in both innate (neutrophils, monocytes, and natural killer cell numbers and expression of complement and Fcγ receptors) and adaptive immunity (numbers of circulating B cells, helper, cytotoxic, and memory T cells).
Subject(s)
Female , Humans , Adaptive Immunity , B-Lymphocytes , Complement System Proteins , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , India , Killer Cells, Natural , Leukocytes , Lymphocyte Count , Lymphocytes , Memory , Monocytes , Neutrophils , Occupations , Receptors, Complement , Solid Waste , T-LymphocytesABSTRACT
O Sistema Complemento (SC) desempenha papel importante no controle de infecções atuando na eliminação do patógeno e na regulação da resposta imune. Contudo, uma ativação desregulada do SC gera efeitos deletérios ao hospedeiro, contribuindo para a patogênese de diversas doenças, como na Dengue. Entretanto, o envolvimento do SCna infecção pelo vírus Dengue (DENV) ainda tem vários aspectos a serem investigados. Assim, avaliamos a contribuição do SC na infecção in vitro de monócitos pelo DENV; o perfil de expressão dos Receptores de Complemento (CR) CR1, CR2,CR3, CR4, CD46, CD55 e CD59, e de moléculas de ativação nos monócitos e linfócitos T circulantes de pacientes infectados pelos DENV-1,-2 ou -4 por citometria de fluxo.Dosamos os níveis de SC5b-9 e citocinas em pacientes por ELISA. Avaliamos ainda, a contribuição da ativação do SC na permeabilidade e viabilidade endotelial, utilizando modelo in vitro de células endoteliais (CEs) pela medida da resistência elétrica transendotelial (TEER) e liberação de LDH sobrenadante de culturas. Por fim investigamos a interação DENV-2 com componentes purificados do SC por eletroforesedas proteínas. Como achados principais, observamos diminuição na frequência de monócitos CD14+ expressando CR3, CR4 e CD59 em pacientes comparado aos controles saudáveis. De forma interessante, o bloqueio do CR3 levou à diminuição em cerca de 30 por cento da infecção in vitro pelo DENV-2 em monócitos, sem alterar o fenótipo de ativação ou a ativação da caspase-1 destas células. No entanto, com o bloqueio deCR3, detectamos diminuição na produção de TNF-alfa e IFN-alfa. Não observamos diferença significativa na frequência de linfócitos T expressando CR3, CD46, CD55 eCD59 de em pacientes-Dengue-4...
Subject(s)
Humans , Complement Activation , Dengue/diagnosis , Dengue/epidemiology , Receptors, Complement , Fluorescent Antibody Technique, IndirectABSTRACT
PURPOSE: Although the polymorphisms of erythrocyte complement receptor type 1 (CR1) in patients with malaria have been extensively studied, a question of whether the polymorphisms of CR1 are associated with severe malaria remains controversial. Furthermore, no study has examined the association of CR1 polymorphisms with malaria in Chinese population. Therefore, we investigated the relationship of CR1 gene polymorphism and malaria in Chinese population. MATERIALS AND METHODS: We analyzed polymorphisms of CR1 gene rs2274567 G/A, rs4844600 G/A, and rs2296160 C/T in 509 patients with malaria and 503 controls, using the Taqman genotyping assay and PCR-direct sequencing. RESULTS: There were no significant differences in the genotype, allele and haplotype frequencies of CR1 gene rs2274567 G/A, rs4844600 G/A, and rs2296160 C/T polymorphisms between patients with malaria and controls. Furthermore, there was no association of polymorphisms in the CR1 gene with the severity of malaria in Chinese population. CONCLUSION: These findings suggest that CR1 gene rs2274567 G/A, rs4844600 G/A, and rs2296160 C/T polymorphisms may not be involved in susceptibility to malaria in Chinese population.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Alleles , Asian People , Case-Control Studies , China , Erythrocytes/parasitology , Genetic Predisposition to Disease , Genotype , Haplotypes , Malaria/ethnology , Polymorphism, Single Nucleotide/genetics , Promoter Regions, Genetic/genetics , Receptors, Complement/blood , Taq PolymeraseABSTRACT
<p><b>OBJECTIVE</b>To study the expression of C3aR and C5aR in trichloroethylene-sensitized mouse liver injury and discuss the pathogenesis of Dermatitis Medicamentosa-like of TCE (DMLT).</p><p><b>METHODS</b>6∼8 w female BALB/c mouse were randomly divided into blank control group, solvent control group and TCE treatment group. TCE was given to the mouse for sensitization at 1th, 4th, 7th, 10th day and challenge at 17th day and 19th day. Before killing mouse, liver weight and body weight were recorded. The livers were separated at 24 h, 48 h, 72 h and 7 d after challenge. And the liver sections were used for immunofluorescence stain and RT-PCR to detect the expression levels of C3aR and C5aR.</p><p><b>RESULTS</b>Microscopic examination showed no significant change in liver structure or organization in TCE non-sensitized group, while liver cell oedema, cell necrosis and inflammatory cell infiltration were clearly observed in TCE-sensitized groups. The expression levels of C3aR and C5aR in 24 h, 48 h, 72 h and 7 d TCE-sensitized groups were significant higher than blank control group, solvent control group and related TCE non-sensitized groups (P < 0.05).</p><p><b>CONCLUSION</b>Complement activation was involved in TCE-induced liver injury and C3aR and C5aR might play essential role in the process.</p>
Subject(s)
Animals , Female , Mice , Chemical and Drug Induced Liver Injury , Dermatitis, Occupational , Edema , Liver , Mice, Inbred BALB C , Receptor, Anaphylatoxin C5a , Metabolism , Receptors, Complement , Metabolism , Solvents , Toxicity , Trichloroethylene , ToxicityABSTRACT
OBJECTIVE@#To determine whether the requirements for sialic acid varies and whether several types of silaic acid independent receptors utilized for invasion mechanisms of fresh filed isolates collected around Nanay river basin, Iquitos.@*METHODS@#The field isolates were cultured as described previously by Jensen and Trager and MR4 protocol with little modifications. The erythrocytes preparation and subsequent enzyme treatment was done as described previously by Sharma. with little modification. Invasion assay was performed as described previously by Sharma et al with little modification.@*RESULTS@#The Nanay river basin isolates showed five types of invasion mechanisms or types of receptors-ligand interactions. Here we observed that an equal numbers of neuraminidase sensitive and resistant invasion receptor-ligand interaction profiles as the most common receptor-ligand invasion profiles. Neuraminidase resistance trypsin sensitive chymotrypsin sensitive (NM(R)T(S)CT(S)) invasion of receptor-ligand interaction profile was found in seven isolates, Five field isolates and one reference strain showed neuraminidase sensitive, trypsin sensitive and chymotrypsin resistant (NM(S)T(S)CT(R)) invasion of receptor-ligand interactions, six isolates including one reference strains dd2 showed neuraminidase sensitive, trypsin and chymotrypsin resistance (NM(S)T(R)CT(R)) indicating its dependence on sialic acids and independence of trypsin and chymotrypsin sensitive proteins. Four isolates showed neuraminidase sensitive, trypsin sensitive and chymotrypsin sensitive (NM(S)T(S)CT(S)) invasion of receptor-ligand interactions, seven isolates were neuraminidase resistant, trypsin sensitive and chymotrypsin resistance (NM(R)T(S)CT(R)) invasion of receptor-ligand interactions, indicating its dependence on trypsin sensitive proteins.@*CONCLUSIONS@#The Nanay river basin isolates showed five types of invasion mechanisms or types of receptors-ligand interactions. A full understanding of theses invasion mechanisms may allow the development of novel prophylactic and therapeutic strategies that block erythrocyte receptor-ligand invasion mechanisms.
Subject(s)
Antigens, Protozoan , Metabolism , Chymotrypsin , Erythrocytes , Allergy and Immunology , Metabolism , Parasitology , Hemagglutination Tests , Membrane Glycoproteins , Metabolism , N-Acetylneuraminic Acid , Metabolism , Neuraminidase , Peru , Plasmodium falciparum , Metabolism , Virulence , Protozoan Proteins , Metabolism , Receptors, Cell Surface , Metabolism , Receptors, Complement , Metabolism , Receptors, Immunologic , Metabolism , Sialic Acid Binding Ig-like Lectin 1 , Trypsin , Virulence , Virulence Factors , MetabolismABSTRACT
The complement system is a key component of innate immunity. More than 45 genes encoding the proteins of complement components or their isotypes and subunits, receptors, and regulators have been discovered. These genes are distributed throughout different chromosomes, with 19 genes comprising three significant complement gene clusters in the human genome. Genetic deficiency of any early component of the classical pathway (C1q, C1r/s, C2, C4, and C3) is associated with autoimmune diseases due to the failure of clearance of immune complexes (IC) and apoptotic materials, and the impairment of normal humoral response. Deficiencies of mannan-binding lectin (MBL) and the early components of the alternative (factor D, properdin) and terminal pathways (from C3 onward components: C5, C6, C7, C8, C9) increase susceptibility to infections and their recurrence. While the association of MBL deficiency with a number of autoimmune and infectious disorders has been well established, the effects of the deficiency of other lectin pathway components (ficolins, MASPs) have been less extensively investigated due to our incomplete knowledge of the genetic background of such deficiencies and the functional activity of those components. For complement regulators and receptors, the consequences of their genetic deficiency vary depending on their specific involvement in the regulatory or signalling steps within the complement cascade and beyond. This article reviews current knowledge and concepts about the genetic load of complement component deficiencies and their association with diseases. An integrative presentation of genetic data with the latest updates provides a background to further investigations of the disease association investigations of the complement system from the perspective of systems biology and systems genetics.
Subject(s)
Animals , Humans , Complement Activation , Genetics , Complement System Proteins , Genetics , HLA Antigens , Genetics , Immunity, Innate , Genetics , Immunologic Deficiency Syndromes , Genetics , Lectins , Metabolism , Multigene Family , Receptors, Complement , GeneticsABSTRACT
O Sistema Complemento (SC) consiste numa rede de proteínas altamente regulada, podendo ser ativado pelas vias alternativa, clássica e da lectina e que convergem na clivagem de C3 e formação do complexo de ataque à membrana (C5b-9). Receptores do complemento (RC), envolvidos na regulação do SC, induzem mecanismos para eliminação de patógenos e a interação das imunidades inata e adaptativa. Estudos vem indicando que uma ativação desregulada do SC contribuiria à gravidade da dengue. A infecção humana pelo vírus Dengue (DENV) é caracterizada por um amplo espectro de sintomas clínicos, variando desde uma febre branda até distúrbios hemodinâmicos, podendo evoluir para choque hipovolêmico e morte. O objetivo principal do trabalho foi investigar o envolvimento do SC na infecção natural pelo DENV, através: (i) da avaliação quanto a expressão dos RC CR1/CD35, CR2/CD21, CR4/CD11c e CD59 nos monócitos CD14+ e linfócitos T CD4+ e TCD8+ por citometria de fluxo, (ii) dosagem de quantidades plasmáticas de SC5b-9 e das citocinas/quimiocinas TNF-alfa, IFN-gamma, CCL5, CCL2 por ELISA e por fim, (iii) avaliação da influência dos componentes do SC na alteração da permeabilidade de células endoteliais (CEs) pelo ensaio de citotoxidade pela enzima LDH. Neste estudo foram incluídos 66 pacientes com infecção pelo DENV-1 ou -2 e 12 indivíduos saudáveis. Os pacientes-DENV foram agrupados segundo a nova classificação da OMS: febre do dengue sem sinais de alarme (FD Sem SA), FD com sinais de alarme (FD Com SA) e dengue grave (G). Nossos resultados demonstraram diminuição significativa na frequência de monócitos CD14+ expressando CR1/CD35, CR4/CD11c e CD59 nos pacientes-DENV comparado aos controles. Além disso, diminuição na frequência de células T CD4+ e CD8+ expressando CR1/CD35 e CR2/CD21 nos pacientes-DENV comparado aos controles. Não observamos diferenças quanto a frequência de nonócitos CD14+ expressando CR2/CD21 ou de linfócitos T CD4+ e CD8+ expressando CR4/CD11c ou CD59 entre os grupos. A quantificação plasmática das citocinas/quimiocinas revelou: (i) maiores quantidades de TNF-alfa nos pacientes FD Sem SA e FD Com SA, (ii) quantidades aumentadas de CCL2 e (iii) diminuídas de CCL5 comparados aos controles saudáveis. Por fim, (iv) a quantidade de IFN-gamma não foi diferente entre grupos de pacientes e controles. Análises de correlação entre as quantidades de citocinas e expressão dos RC revelaram que TNF-alfa foi diretamente correlacionada com a frequência de monócitos CD14+ expressando de CD59 e CR4/CD11c, enquanto que CCL2 foi inversamente correlacionada com a frequência de linfócitos TCD8+ expressando CD59Quanto a dosagem do SC5b-9, quantidades plasmáticas desta molécula foram mais elevadas nos pacientes graves e que apresentaram manifestações hemorrágicas e extravasamento plasmático comparado aqueles sem estes sintomas. A adição de plasma de pacientes com quantidades elevadas de SC5b-9 promoveu maior lise de CEs in vitro, comparado aos plasmas de pacientes com quantidades menores de SC5b-9. Desta forma, nós concluímos que na infecção pelo DENV ocorre uma modulação dos RC nos monócitos e linfócitos T de pacientes, o que poderia levar a uma alteração funcional destas células. Além disso, TNF-alfa e CCL2 poderiam estar modulando a expressão de alguns dos RC nas células. Por fim, quantidades elevadas de SC5b-9, associada a ativação do SC, estaria associada as manifestações clínicas graves e na citotoxidade de CEs. Como perspectivas, serão avaliadas as alterações funcionais das células imunes e o grau de contribuição dos componentes do SC na patogênese da dengue. Desta forma, nossos dados levam a concluir que componentes do SC contribuem à patogênese da doença.
Subject(s)
Complement Membrane Attack Complex , Dengue Virus , Dengue/epidemiology , Receptors, Complement , Virulence , Dengue/historyABSTRACT
<p><b>OBJECTIVE</b>Different from primary membranous nephropathy, hepatitis B virus associated membranous nephropathy (HBV-MN) shows lower deposits of membrane attack complex (C5b-9) in glomerular subepithelium. The causes of relatively low complement activation in this disease remain unclear. The aim of this study was to investigate the influence of hepatitis B x protein (HBx) on the expression of CD59 and Crry in mouse podocytes.</p><p><b>METHOD</b>Cultured mouse podocytes were divided into adenovirus vector hepatitis B virus X gene (Ad-HBx) transfected group (Ad-X group), blank podocytes group (B group) and adenovirus vector transfected group (Ad group). CD59 and Crry mRNA expression were assayed by semiquantitative RT-PCR. CD59 and Crry expression were tested by flow cytometry. The effect of HBx on complement activation was evaluated with MTT method. And then, the effects of P38MAPK, PI-3K and ERK1/2 pathway inhibitors (SB203580, LY294002, U0126) and DMSO on CD59 and Crry expression were respectively detected by flow cytometry.</p><p><b>RESULT</b>Proteins CD59 and Crry expression rates (%) in group B, Ad group and Ad-X group were 17.71 ± 3.81, 18.29 ± 3.36 and 45.7 ± 9.01; 18 ± 2.31, 21.78 ± 2.01 and 47.45 ± 9.95, respectively. Compared with group B, CD59 and Crry expression in group Ad was not significantly different (P values for both > 0.05), but CD59 and Crry protein expression in Ad-X group was significantly higher than that in groups B and Ad (P values for both < 0.005); CD59 and Crry gene expression in group Ad was not significantly different from that in group B (P values for both > 0.05). However, CD59 and Crry gene expression of Ad-X group was significantly higher than that in groups B and Ad (P values for both < 0.05). Flow cytometry detected CD59 protein expression rates (%) were 17.35 ± 1.24, 46.19 ± 9.77, 43.03 ± 6.83 and 40.04 ± 6.39 and Crry protein expression rates (%) were 18.14 ± 3.56, 31.95 ± 1.68, 31.95 ± 1.69 and 37.14 ± 3.92 after SB203580, LY294002, U0126 and DMSO were added to Ad-X group respectively. P38 pathway inhibition resulted in significantly lower CD59 and Crry expression than Ad-X group (P values for all < 0.005), but PI-3K, ERK1/2 pathway inhibitors and DMSO had no significant effect on the expression of CD59 and Crry (P values for all > 0.05). The inhibition rates of cell lysis were significantly higher in Ad-X group than in groups B and Ad at each serum dilution point (P values for all < 0.05), while groups B and Ad had no significant difference in cell viability.</p><p><b>CONCLUSION</b>HBx can up-regulate CD59 and Crry expression in podocytes through activating P38 pathway, resulting in decreased complement activation, which may facilitate latent HBV infection in podocytes and play a role in development of hepatitis B virus associated glomerulonephritis (HBV-GN).</p>
Subject(s)
Animals , Mice , CD59 Antigens , Metabolism , Cells, Cultured , Podocytes , Allergy and Immunology , Metabolism , Receptors, Complement , Metabolism , Trans-Activators , Metabolism , p38 Mitogen-Activated Protein Kinases , MetabolismABSTRACT
Astragalus heteropolysaccharides (AHPS) is obtained from the dried roots of Astragalus membranaceus (Fisch.) Bunge var. mongholious (Bunge) Hsiao. In the present study, we observed its effects on erythrocyte immune adherence function in mice with adjuvant-induced arthritis (AA). The mice were treated intragastrically with AHPS of 1 000, 500, and 250 mg x kg(-1) x d(-1) separately and treated with tripterygium glycosides (TG) of 60 mg x kg(-1) x d(-1) as positive control. The number of complement receptor type 1 (CR1) on erythrocyte, the concentration of circulating immune complex (CIC) in serum and the amount of immune complex (IC) deposition in synovium of knee joint were determined by flow cytometry, polyethylene glycol (PEG-6000) precipitation and ponceau S (P-S) staining and fluorescent immunohistochemistry respectively. The pathological change of knee joint was evaluated by histological section. The results showed that both AHPS and TG improved significantly the primary and secondary local or systemic symptoms of the mice with AA and reduced the synovium hyperplasia, inflammatory cell infiltrate, pannus and cartilage demolish of knee joint, and AHPS of 1 000, 500, and 250 mg x kg(-1) x d(-1) could significantly increase the number of CR1 on erythrocyte, improve the elimination of CIC in the peripheral blood and reduce the deposition of IC in joint synovium in a dose-dependent manner (P < 0.01 or P < 0.05). The results indicate that one of the therapeutic effective mechanisms of AHPS on mice with AA could be to increase gene expression of CR1 of mice with AA.
Subject(s)
Animals , Male , Mice , Antigen-Antibody Complex , Blood , Metabolism , Arthritis, Experimental , Metabolism , Pathology , Astragalus Plant , Chemistry , Dose-Response Relationship, Drug , Erythrocytes , Allergy and Immunology , Knee Joint , Pathology , Plant Roots , Chemistry , Plants, Medicinal , Chemistry , Polysaccharides , Pharmacology , Random Allocation , Receptors, Complement , Blood , Synovial Membrane , Allergy and ImmunologyABSTRACT
C1q nephropathy is a proliferative glomerulopathy with extensive mesangial deposition of C1q. A three-year old boy presented with a nephrotic-range proteinuria during an acute phase of Epstein-Barr virus (EBV) infection, and he had a family history of Dent's disease. The renal biopsy findings were compatible with C1q nephropathy. However, EBV in situ hybridization was negative. The CLCN5 gene analysis revealed an R637X hemizygous mutation, which was the same as that detected in his maternal cousin, the proband of the family. The causal relationship between EBV infection and C1q nephropathy remains to be determined. Moreover, the effects of underlying Dent's disease in the process of C1q nephropathy has to be considered.
Subject(s)
Child, Preschool , Humans , Male , Biopsy , Epstein-Barr Virus Infections/metabolism , Glomerulonephritis/pathology , In Situ Hybridization , Infectious Mononucleosis/complications , Kidney Diseases/complications , Kidney Tubules/pathology , Membrane Glycoproteins/chemistry , Mutation , Nephrosis , Proteinuria/complications , Receptors, Complement/chemistry , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To investigate the role of sulfated tyrosine in regulating the activity of tyrosylprotein sulfotransferases (TPST) 1 and TPST2.</p><p><b>METHODS</b>Constructs of TPST 1 and TPST2 were amplified by polymerase chain reaction (PCR), then fused into immunoglobulin G1 Fc region. All the variants in which sulfated tyrosines were mutated to phenylalanine were made by the PCR-based Quick Change method and confirmed by sequencing the entire reading frame. Small hairpin RNA (shRNA) constructs-targeting nucleotides 259-275 of TPST1 and nucleotides 73-94 of TPST2 were generated and subcloned into pBluescript. Human embryonic kidney (HEK) 293T cells were transfected with these plasmids. One day later, cells were split: one part was labeled with 35S-cysteine and methionine or 35S-Na2SO3 overnight, the second part was used for 125I labeled binding experiment, and the third part was retained for binding and flow cytometry.</p><p><b>RESULTS</b>Tyrosines at position 326 of TPST1 and position 325 of TPST2 were sulfated posttranslationally. Tyrosine sulfation of TPSTs was effectively inhibited by sulfation inhibitors, including specific shRNAs and non-specific NaCIO3. shRNAs reduced the sulfation of C3a receptor and C5a receptor, and partially blocked the binding of these two receptors to their respective ligands.</p><p><b>CONCLUSIONS</b>The activities of TPSTs were regulated by tyrosine sulfation. Inhibition of sulfotyrosine decreases the binding ability of C3a receptor and C5a receptor to their respective ligands.</p>
Subject(s)
Humans , Cell Line , Complement C3a , Metabolism , Complement C5a , Metabolism , Protein Binding , Protein Processing, Post-Translational , Receptor, Anaphylatoxin C5a , Metabolism , Receptors, Complement , Metabolism , Sulfotransferases , Genetics , Metabolism , Transfection , Tyrosine , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the protective effects of recombinant SCR15-18 domain of human soluble complement receptor type 1 (sCR1-SCR-15-18) in rats underwent myocardial ischemia and reperfusion (I/R).</p><p><b>METHODS</b>Sprague-Dawley rats were randomly divided into three groups (n = 12 each group): sham (SO); 30 min ischemia/3h reperfusion (I/R) and I/R plus sCR1-SCR15-18 (15 mg/kg before I/R, sCR1). Serum LDH, CK and cardiac myeloperoxidase (MPO) activity were measured. Infarct size, myocardial histopathological changes and myocardial C3c were also compared among groups.</p><p><b>RESULTS</b>Infarct size [(16.1 +/- 3.3)% vs. (22.9 +/- 3.0)%, infarct zone/left ventricular mass, P < 0. 05] and CK [(2532.5 +/- 597.1) U/L vs. (3400.9 +/- 534.9) U/L, P < 0. 05] and LDH [(5436.2 +/- 611.3) U/L vs. (6572.0 +/- 476.3) U/L, P < 0. 05] as well as MPO activity in infarct zone [(0.81 +/- 0.14) U/g vs. (1.12 +/- 0.13) U/g, P < 0.05] were significantly decreased post sCR1 compared to I/R group. sCR1 also significantly attenuated histological myocardial injury and reduced the deposition of C3c in infarct zone.</p><p><b>CONCLUSION</b>sCR1-SCR15-18 protein exerts cardioprotective effects in this rat I/R model.</p>
Subject(s)
Animals , Humans , Rats , Creatine Kinase , Metabolism , Disease Models, Animal , L-Lactate Dehydrogenase , Metabolism , Myocardial Reperfusion Injury , Metabolism , Pathology , Peroxidase , Metabolism , Rats, Sprague-Dawley , Receptors, Complement , Therapeutic Uses , Receptors, Complement 3b , Metabolism , Recombinant Proteins , Therapeutic UsesABSTRACT
<p><b>OBJECTIVE</b>To determine the effects of recombinant soluble complement receptor type I (sCR1) on the immune inflammatory reaction in acute spinal cord injury tissue of rats and its protective effects.</p><p><b>METHODS</b>SD rat models of acute spinal cord injury were prepared by modified Allen's method. The motor function of the rat lower extremities in sCR1 group and normal saline (NS) group was evaluated by the tiltboard experiment at 12 h, 1 d, 3 d, 7 d, and 14 d. The neutrophil infiltration and C3c positive expression were observed. The myeloperoxidase activity was assessed in the injury tissue at 12 h, 1 d, 3 d, 7 d, and 14 d after injury in the two groups.</p><p><b>RESULTS</b>The motor function of rat in sCR1 group at 3 d, 7 d, and 14 d was obviously better than that in NS group (P<0.01, P<0.01, P<0.01). C3c positive expression in sCR1 group at each time point after injury was obviously less than that in NS group (P<0.01). The myeloperoxidase activity in sCR1 group at each time point after injury was obviously less than that in NS group (P<0.01).</p><p><b>CONCLUSIONS</b>Recombinant soluble complement receptor type I (sCR1) can lessen the immune inflammatory reaction in acute spinal cord injury tissue and relieve secondary spinal cord injury by inhibiting the activation of the complement system.</p>
Subject(s)
Animals , Rats , Disease Models, Animal , Immunohistochemistry , Inflammation , Peroxidase , Random Allocation , Rats, Sprague-Dawley , Receptors, Complement , Therapeutic Uses , Recombinant Proteins , Therapeutic Uses , Spinal Cord Injuries , Drug Therapy , PathologyABSTRACT
To evaluate the role of erythrocyte CR1 and L-selectin in the pathogenesis of SLE and try to find out any correlation between them with some clinical and laboratory associated data. Thirty patients diagnosed as SLE according to ACR criteria with mean age 23.6 +/- 6.9 in addition to 10 apparently healthy normal [age and sex matched] subjects as a control group were included in this study. There were higher values of erythrocyte CR1 percentage in controls more than patients [p<0.05], while this could not be found as regards level of L-selectin [p>0.05]. A significant association was found between erythrocyte CR1 percentage with fever and arthralgia [p<0.05], but this could not be found as regards the other clinical findings. There was a significant association between the level of L-selectin with arthralgia and cardiac affection [p<0.05], but this was not found with other clinical findings. Significant correlations were found between erythrocyte CR1 percentage and serum creatinine level, urinary pus cells ESR and platelets counts [p<0.05], while no correlations were found between L-selectin level and other laboratory findings. No correlations were found between erythrocyte CR1 percentage and L-selectin level with disease onset, duration or disease activity score. Both erythrocytes CR1 and L-selectin [to a lesser extent] have a crucial role in the pathogenesis of SLE as regards joint and kidney affection through IC mediated vasculitis
Subject(s)
Humans , Male , Female , Receptors, Complement , Erythrocytes , L-Selectin , Kidney Function Tests , Disease ProgressionABSTRACT
<p><b>OBJECTIVE</b>To study the changes of genomic density polymorphism, quantitative expression and the adhesion activity of complement receptor type 1 (ECR1) on erythrocytes in patients with chronic hepatitis.</p><p><b>METHODS</b>Polymerase chain reaction (PCR) and Hind restriction enzyme digestion, the quantitative assay of ECR1 and the activity of erythrocytes immune adhesion test were applied.</p><p><b>RESULTS</b>The spot mutation rate (25.0%-30.3%) of ECR1 density gene in patients with chronic hepatitis was not significantly different from that of healthy individuals (28.0%). The amount of ECR1 in patients with chronic hepatitis, except for the diseases with normal liver function, was significantly lower than that of healthy individuals (t=9.87,P<0.000 1). The quantitative expression of ECR1 in decompensated cirrhosis was obviously lower than that of compensated cirrhosis (t=2.21,P<0.05).</p><p><b>CONCLUSIONS</b>Defective expression of ECR1 in chronic hepatitis B may be acquired through central and/or peripheral mechanisms. It is very important to study the quantitative expression in the patients with chronic hepatitis.</p>
Subject(s)
Humans , Erythrocytes , Allergy and Immunology , Metabolism , Hepatitis B, Chronic , Genetics , Liver Cirrhosis , Genetics , Polymerase Chain Reaction , Polymorphism, Genetic , Receptors, Complement , Genetics , Metabolism , Tissue AdhesionsSubject(s)
Humans , Male , Female , Acute Disease , Chronic Disease , Receptors, Complement/blood , Bronchoalveolar LavageABSTRACT
<p><b>OBJECTIVE</b>To explore the postburn adhesion properties of polymorphonuclear leukocyte (PMN) onto pulmonary vascular endothelial cells (PVEC) in burn patients with acute lung injury (ALI), so as to determine the role of C5a on PVEC-PMN adhesion.</p><p><b>METHODS</b>Microtubule sucking technique was employed to determine the PVEC-PMN adhesion. The myeloperoxidase (MPO) was also assayed to reflect the magnitude of PVEC-PMN adhesion.</p><p><b>RESULTS</b>The magnitude of PVEC-PMN adhesion increased and the adhesion force increased along with an increase in rh-C5a concentration. Simultaneously, the MPO activity was increased, which could be inhibited by anti-C5aR McAb in a concentration 1:104.</p><p><b>CONCLUSION</b>Both C5a and C5aR participated in PVEC-PMN adhesion, which might be important in the pathogenesis of ALI.</p>
Subject(s)
Humans , Acute Disease , Antibodies, Monoclonal , Pharmacology , Antigens, CD , Allergy and Immunology , Burns , Blood , Cell Adhesion , Cells, Cultured , Complement C5a , Pharmacology , Dose-Response Relationship, Drug , Endothelium, Vascular , Cell Biology , Fetus , Lung , Lung Diseases , Neutrophils , Cell Biology , Peroxidase , Metabolism , Receptor, Anaphylatoxin C5a , Receptors, Complement , Allergy and ImmunologyABSTRACT
Complement Receptor 1 (CR1) is a polymorphic glycoprotein expressed on erythrocytes, leukocytes and glomerular podocytes and has a major role in immune complex processing. In addition, it regulates the complement cascade activation by preventing formation of classical and alternative pathway convertases and by acting as a cofactor for Factor I mediated cleavage of C3. In this study, we have examined the expression of erythrocyte CR1 (E-CR1) and glomerular CR1 (G-CR1) in different kinds of nephropathies using ELISA and immunofluorescence microscopy to understand their role in immune complex (IC) mediated renal diseases. E-CR1 was significantly reduced in all categories of lupus nephritis in comparison to normal subjects and non-IC renal diseases. However, other IC mediated diseases like IgA nephropathy and membranoproliferative glomerulonephritis had normal E-CR1 levels. G-CR1 showed distinct differences between IC and non-IC mediated diseases. G-CR1 was virtually absent in lupus kidneys. In other IC mediated diseases, there was a correlation of G-CR1 expression to the IC and complement fragment deposition. G-CR1 serves as a useful diagnostic marker for IC mediated diseases while E-CR1 is useful as a prognostic marker to monitor the course of disease after the treatment has initiated.
Subject(s)
Erythrocytes/chemistry , Female , Follow-Up Studies , Humans , Immune Complex Diseases/diagnosis , India , Kidney Diseases/diagnosis , Kidney Glomerulus , Male , Methods , Prognosis , Receptors, Complement/analysis , Receptors, Complement 3b/analysisABSTRACT
A un siglo de su descubrimiento por Bordet, se trata de poner un poco de orden en los mecanismos de activación del complemento a través de su hasta ahora conocidas rutas de activación clásica o de C1 y la vía alterna o de la properdina. Se hace además referencia a otras vías de activación descritas más recientemente como la activación iniciada por la lectina de unión a la manosa (MBL). Se destaca también la actividad de los componentes inhibidores o controladores, que frenan la actividad del sistema, evitando la producción de daños por la formación y liberación de péptidos con potente acción biológica derivados del mismo, tal el caso de las anafilatoxinas. Se hace además referencia a la presencia de receptores para el complemento, ubicados en la membrana de diversas células del sistema inmunológico, responsables de muchas de las principales actividades del sistema, como la fagocitosis de microorganismos a través de la unión de receptores para C3b (principal opsonina) sobre la membrana de las células fagocitarias
Subject(s)
Humans , Complement System Proteins/physiology , Self-Evaluation Programs , Complement Activation/immunology , Complement Inactivating Agents/immunology , Complement Membrane Attack Complex/immunology , Complement System Proteins/immunology , Receptors, Complement/immunology , Complement Pathway, Alternative/immunology , Complement Pathway, Classical/immunologyABSTRACT
PURPOSE: HL-60 is a promyelocytic cell line. Fc receptors and complement receptor 3 (CR3) play important role in the protective response of granulocytes and monocytes against microbial infection. We quantified the expression of Fc I, Fc II, Fc III, and CD11b/CD18 during differentiation using HL-60 cells by N-N-dimethylformamide (DMF). Functional studies, such as phagocytic activity, respiratory burst and ADCC, were also performed. METHODS: HL-60 cells were induced to differentiate by adding 0.8% DMF. On the 4th and 7th day after stimulation as well as before stimulation, the cells were analyzed for phenotypic and functional differentiaton. Phenotypic analysis was performed by flow cytometry after staining the cells with PE-conjugated anti-human CD64, CD32, CD16, CD11b, CD18, and isotype controls. And the measured fluorescent intensity was transformed into Molecules of Equivalent Soluble Fluorochromes (MESF). Phagocytic activity was also measured by flow cytometry after incubation with fluorochrome-conjugated beads. Respiratory burst was measured by chemiluminescence assay of cells incubated with luminol after stimulation with PMA. ADCC was measured by hemoglobin release assay. RESULTS: The expression of CD11b, CD18 and CD64 on HL-60 cells markedly increased on the 4th day and slightly decreased on the 7th day. Expression of CD32 was already induced before differentiation induction and slightly increased by DMF. CD16 was not expressed during differentiation. In phagocytic assay, the phagocytic cell fraction increased by stimulation on 4th and 7th day. Chemiluminescence showed the DMF increased the respiratory burst of HL-60 cells on the 4th and 7th day. In ADCC, DMF increased the target cell lysis continuously. CONCLUSION: HL-60 cells which were differentiated with DMF for are good models for studying opsonophagocytic assay of immunized sera.