RESUMO
BACKGROUND: This study investigated clinical and laboratory characteristics of human bocavirus type 1 (HBoV1)-plastic bronchiolitis (PB), Mycoplasma pneumoniae (MP)-associated plastic bronchitis (PB) and MP-NPB in children, highlighting inflammation, coagulation, and bronchoscopic needs. METHODS: Data on preschool children with PB during HBoV1 or MP infection were collected, comparing MP-PB to severe Mycoplasma pneumoniae pneumonia. RESULT: Compared with the MP-PB group, the HBoV1-PB group, with younger children, had significantly milder clinical symptoms but higher WBC counts (p = .028). The MP-PB group exhibited notably elevated Fibrinogen (p = .045) and d-dimer levels (p < .001). When contrasting the MP-PB with the MP-NPB group, children in MP-PB group still had higher levels of d-dimer and increased inflammatory indicators such as C-reactive protein, procalcitonin, lactate dehydrogenase, and interleukin-6, which were significantly elevated compared with the MP-NPB group. MP-PB showed a higher prevalence of plastic bronchial casts in lower lobes (p = .016) and a dominance of neutrophils in BALF cytology. Additionally, children in the MP-PB group tended to undergo a greater number of bronchoscopies. CONCLUSION: This study identifies key differences in plastic bronchitis in children due to HBoV1 and MP, highlighting HBoV1's milder inflammation in younger kids and MP's link to severe inflammatory and coagulation responses, guiding clinical diagnosis and treatment.
Assuntos
Bronquite , Mycoplasma pneumoniae , Pneumonia por Mycoplasma , Humanos , Pré-Escolar , Masculino , Feminino , Bronquite/microbiologia , Bronquite/diagnóstico , Bronquite/virologia , Pneumonia por Mycoplasma/sangue , Pneumonia por Mycoplasma/imunologia , Lactente , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/complicações , Infecções por Parvoviridae/diagnóstico , Bocavirus Humano , Bronquiolite/virologia , Bronquiolite/microbiologia , Criança , Líquido da Lavagem Broncoalveolar/virologia , Líquido da Lavagem Broncoalveolar/microbiologia , Produtos de Degradação da Fibrina e do Fibrinogênio/análise , Produtos de Degradação da Fibrina e do Fibrinogênio/metabolismo , Proteína C-Reativa/análiseRESUMO
BACKGROUND: Community-acquired pneumonia often stems from the macrolide-resistant strain of Mycoplasma pneumoniae, yet no effective vaccine exists against it. METHODS: This study proposes a vaccine-immunoinformatics strategy for Mycoplasma pneumoniae and other pathogenic microbes. Specifically, dominant B and T cell epitopes of the Mycoplasma pneumoniae P30 adhesion protein were identified through immunoinformatics method. The vaccine sequence was then constructed by coupling with CTLA-4 extracellular region, a novel molecular adjuvant for antigen-presenting cells. Subsequently, the vaccine's physicochemical properties, antigenicity, and allergenicity were verified. Molecular dynamics modeling was employed to confirm interaction with TLR-2, TLR-4, B7-1, and B7-2. Finally, the vaccine underwent in silico cloning for expression. RESULTS: The vaccine exhibited both antigenicity and non-allergenicity. Molecular dynamics simulation, post-docking with TLR-2, TLR-4, B7-1, and B7-2, demonstrated stable interaction between the vaccine and these molecules. In silico cloning confirmed effective expression of the vaccine gene in insect baculovirus vectors. CONCLUSION: This vaccine-immunoinformatics approach holds promise for the development of vaccines against Mycoplasma pneumoniae and other pathogenic non-viral and non-bacterial microbes.
Assuntos
Vacinas Bacterianas , Antígeno CTLA-4 , Biologia Computacional , Epitopos de Linfócito B , Epitopos de Linfócito T , Mycoplasma pneumoniae , Pneumonia por Mycoplasma , Mycoplasma pneumoniae/imunologia , Mycoplasma pneumoniae/genética , Epitopos de Linfócito T/imunologia , Epitopos de Linfócito T/genética , Vacinas Bacterianas/imunologia , Vacinas Bacterianas/genética , Epitopos de Linfócito B/imunologia , Epitopos de Linfócito B/genética , Humanos , Biologia Computacional/métodos , Pneumonia por Mycoplasma/prevenção & controle , Pneumonia por Mycoplasma/imunologia , Antígeno CTLA-4/imunologia , Simulação de Dinâmica Molecular , Simulação de Acoplamento Molecular , Receptor 2 Toll-Like/imunologia , ImunoinformáticaRESUMO
A non-pathogenic Mycoplasma pneumoniae-based chassis is leading the development of live biotherapeutic products (LBPs) for respiratory diseases. However, reports connecting Guillain-Barré syndrome (GBS) cases to prior M. pneumoniae infections represent a concern for exploiting such a chassis. Galactolipids, especially galactocerebroside (GalCer), are considered the most likely M. pneumoniae antigens triggering autoimmune responses associated with GBS development. In this work, we generated different strains lacking genes involved in galactolipids biosynthesis. Glycolipid profiling of the strains demonstrated that some mutants show a complete lack of galactolipids. Cross-reactivity assays with sera from GBS patients with prior M. pneumoniae infection showed that certain engineered strains exhibit reduced antibody recognition. However, correlation analyses of these results with the glycolipid profile of the engineered strains suggest that other factors different from GalCer contribute to sera recognition, including total ceramide levels, dihexosylceramide (DHCer), and diglycosyldiacylglycerol (DGDAG). Finally, we discuss the best candidate strains as potential GBS-free Mycoplasma chassis.
Assuntos
Glicolipídeos , Síndrome de Guillain-Barré , Mycoplasma pneumoniae , Síndrome de Guillain-Barré/microbiologia , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/imunologia , Humanos , Glicolipídeos/metabolismo , Galactosilceramidas , Reações Cruzadas , Pneumonia por Mycoplasma/microbiologia , Pneumonia por Mycoplasma/imunologia , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologiaRESUMO
BACKGROUND: Mycoplasma pneumoniae is a leading cause of community-acquired respiratory infections. Infantile Feire Kechuan Oral Solution (IFKOS) is effective for treatment of M. pneumoniae infection. The aim of this study was to explore the potential mechanism of IFKOS against M. pneumoniae infection in basal epithelial human lung adenocarcinoma A549 cells. METHODS: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to determine the effects of IFKOS on the viability of A549 cells infected with M. pneumoniae. Optical microscopy was used to observe cell morphology and a Muse cell analyzer was used to assess apoptosis and the cell cycle phase. Enzyme-linked immunosorbent assays were employed to assess the expression levels of interleukin (IL)-4, IL-6, IL-8, IL-17, tumor necrosis factor (TNF)-α, interferon (IFN)-α, and IFN-γ. RESULTS: Under certain conditions, M. pneumoniae infection reduced the viability and inhibited the proliferation of A549 cells, promoted early apoptosis, and arrested cells in the G0/G1 phase, thus shortening the S and G2/M phases (all p < 0.05). M. pneumoniae also upregulated expression of IL-8 and TNF-α and downregulated that of IL-6 (p < 0.05), which switched the immune balance of Th1/Th2 to Th1 cells. IFKOS (5.531 mg/mL) improved the viability and proliferation of M. pneumoniae-infected A549 cells, mitigated early apoptosis, and reversed cell cycle arrest in the G0/G1 phase, thereby extending the S and G2/M phases (all, p < 0.05). IFKOS downregulated expression of IL-8 and TNF-α and upregulated that of IL-6 (p < 0.01), thereby reversing the immune imbalance of Th1/Th2. Secretion of IL-4, IL-17, IFN-α, and IFN-γ was not observed. CONCLUSION: IFKOS played a protective role in the regulation of cell viability, apoptosis, the cell cycle, and Th1/Th2 immune imbalance induced by M. pneumoniae infection and conveyed an anti-inflammatory effect in A549 cells.
Assuntos
Antibacterianos/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Mycoplasma pneumoniae/efeitos dos fármacos , Pneumonia por Mycoplasma/tratamento farmacológico , Células A549 , Anti-Inflamatórios/farmacologia , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Humanos , Pneumonia por Mycoplasma/imunologia , Pneumonia por Mycoplasma/microbiologia , Células Th1/imunologia , Células Th2/imunologiaRESUMO
Mycoplasma pneumoniae (Mp) residing extracellularly in the respiratory tract is the primary cause of bacterial community-acquired pneumonia in humans. However, the detailed pathological mechanism of Mp infection, especially inflammation in the lung, remains unclear. This study examined the role of the neutrophils in the inflammation of Mp-induced pneumonia in mice and the mechanism of neutrophil infiltration into the lungs in the Mp-induced pneumonia. We observed massive infiltration of neutrophils in the bronchoalveolar lavage fluid (BALF) and lung injury after the Mp challenge. The neutrophils were shown to contribute to lung injury in Mp pneumonia but were not involved in eliminating Mp, suggesting that neutrophils are detrimental to the host in Mp pneumonia. Mp also induced the production of inflammatory cytokines and chemokines in the BALF in a toll-like receptor 2 (TLR2)-dependent manner. Particularly, both interleukin (IL)-1α and IL-12 p40 played a crucial role in neutrophil infiltration into the BALF in a coordinated manner. Both IL-1α and IL-12 p40 were released from the alveolar macrophages depending on the TLR2 and reactive oxygen species. In addition, the community-acquired respiratory distress syndrome (CARDS) toxin from Mp were found to induce neutrophil infiltration into BALF in a TLR2-independent and IL-1α-dependent manner. Collectively, the TLR2-dependent production of both IL-1α and IL-12 p40, and CARDS toxin have been elucidated to play an important role in neutrophil infiltration into the lungs subsequently leading to the lung injury upon Mp infection in mice. These data will aid in the development of therapeutics and vaccines for Mp pneumonia. IMPORTANCE Although Mp-induced pneumonia is usually a self-limiting disease, refractory life-threatening pneumonia is often induced. In addition, the development of alternative therapeutic strategies for Mp is expected because of the emergence of antibiotic-resistant Mp. However, the lack of knowledge regarding the pathogenesis of Mp-induced pneumonia, especially inflammation upon the Mp infection, makes it tedious to design novel therapeutics and vaccines. For example, although neutrophil infiltration is widely recognized as one of the characteristics of Mp-induced pneumonia, the precise role of neutrophils in the aggravation of Mp pneumonia remains unclear. This study showed that the infiltration of neutrophils in the lungs is detrimental to the host in Mp-induced pneumonia in mice. Furthermore, the TLR2-dependent IL-1α and IL-12 p40 production, and CARDS toxin play important roles in neutrophil infiltration into the lung, following lung injury. Our findings apply to the rational design of novel therapeutics and vaccines against Mp.
Assuntos
Proteínas de Bactérias/imunologia , Toxinas Bacterianas/metabolismo , Interleucina-12/metabolismo , Interleucina-1alfa/metabolismo , Lesão Pulmonar/imunologia , Neutrófilos/imunologia , Pneumonia por Mycoplasma/imunologia , Receptor 2 Toll-Like/metabolismo , Animais , Líquido da Lavagem Broncoalveolar , Inflamação/imunologia , Inflamação/patologia , Pulmão , Lesão Pulmonar/patologia , Macrófagos Alveolares/imunologia , Camundongos , Mycoplasma pneumoniae/imunologia , Infiltração de Neutrófilos , Espécies Reativas de Oxigênio , Síndrome do Desconforto Respiratório/imunologia , Receptor 2 Toll-Like/genéticaRESUMO
BACKGROUND: Mycoplasma pneumoniae (M. pneumoniae) is implicated in several immune-mediated extrapulmonary manifestations, including reactive arthritis. Recently, increased total serum IgE were reported in children developing M. pneumoniae-related extrapulmonary diseases (MpEPDs). Here, we aimed at analyzing these aspects in children affected with rheumatic disorders and, in detail, Juvenile Idiopathic Arthritis (JIA). METHODS: M. pneumoniae serology (IgG and IgM) and total serum IgE were concomitantly analyzed in 139 pediatric patients diagnosed with: JIA (Group 1, n = 85), or any rheumatic disease other than JIA (Group 2, n = 27), or non-inflammatory endocrinological disorders (Group 3, n = 27). RESULTS: Overall, 19.4% M. pneumoniae seroprevalence was observed in this hospitalized pediatric population, without signicant differences among the three groups. No significant differences in total serum IgE levels were noted among these groups; however, a second analysis excluding children with very high (and clearly abnormal) IgE levels suggested that JIA patients and, in detail, those with oligopolyarticular forms may have higher serum IgE concentrations. This relative difference among groups in serum IgE level seems to be more pronounced in M. pneumoniae seropositive children. CONCLUSIONS: M. pneumoniae infection should be actively sought in children developing immune-mediated diseases, including patients affected with JIA and, especially, in oligopolyarticular forms. There is some evidence that total serum IgE levels may tend to be increased in patients with oligopolyarticular JIA subtype and especially in those resulting as M. pneumoniae seropositive. However, further and focused research is needed to confirm these preliminary results and to clarify the relation between M. pneumoniae infection, atopic status, and immune-mediated arthritis.
Assuntos
Anticorpos Antibacterianos/sangue , Artrite Juvenil/microbiologia , Imunoglobulina E/sangue , Mycoplasma pneumoniae/isolamento & purificação , Pneumonia por Mycoplasma/epidemiologia , Anticorpos Antibacterianos/imunologia , Artrite Juvenil/sangue , Artrite Juvenil/imunologia , Criança , Pré-Escolar , Estudos Transversais , Feminino , Humanos , Imunoglobulina E/imunologia , Lactente , Recém-Nascido , Masculino , Mycoplasma pneumoniae/imunologia , Pneumonia por Mycoplasma/sangue , Pneumonia por Mycoplasma/imunologia , Pneumonia por Mycoplasma/microbiologia , Estudos SoroepidemiológicosRESUMO
Pneumonia caused by Mycoplasma pneumoniae (M. pneumoniae) is a major cause of communityacquired pneumonia in children. In some cases, M. pneumoniae pneumonia (MPP) can develop into refractory MPP (RMPP), which shows no clinical or radiological response to macrolides, and can progress to severe and complicated pneumonia. However, the pathogenesis of RMPP remains poorly understood. The present study aimed to identify target genes that could be used as biomarkers for the clinical diagnosis of earlystage RMPP through highthroughput sequencing technology. The differences in long noncoding (lnc)RNAs, mRNAs and circular (circ)RNAs were examined between wholeblood samples from two patients with nonrefractory MPP (NRMPP), two patients with RMPP and three healthy children using ribosomal (r)RNAdepleted RNAsequencing techniques and an integrated mRNA/circRNA analysis. A total of 17 lncRNAs (four upregulated and 13 downregulated), 18 mRNAs (six upregulated and 12 downregulated) and 24 circRNAs (12 upregulated and 12 downregulated) were the most significantly differentially expressed (P<0.05) between the NRMPP and RMPP groups. Upon functional analysis, the significantly differentially expressed genes encoded by the targeting mRNAs (prostaglandinendoperoxide synthase 2, IL8 and foslike antigen 1) were screened and identified to be enriched in the 'IL17 signaling pathway'. Furthermore, the key circRNAs in the NRMPP and RMPP comparative groups were primarily enriched in 'herpes simplex virus 1 infection', 'viral carcinogenesis' and 'RNA transport'. In the present study, a comprehensive analysis of the differences between the NRMPP and RMPP cases was performed based on rRNAdepleted RNAsequencing techniques, and the selected genes and circRNAs may be closely associated with the complex pathogenesis of RMPP.
Assuntos
Antibacterianos/farmacologia , Infecções Comunitárias Adquiridas/tratamento farmacológico , Farmacorresistência Bacteriana/genética , Mycoplasma pneumoniae/patogenicidade , Pneumonia por Mycoplasma/tratamento farmacológico , Antibacterianos/uso terapêutico , Biomarcadores/análise , Criança , Pré-Escolar , Infecções Comunitárias Adquiridas/genética , Infecções Comunitárias Adquiridas/imunologia , Infecções Comunitárias Adquiridas/microbiologia , Feminino , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/imunologia , Humanos , Lactente , Macrolídeos/farmacologia , Macrolídeos/uso terapêutico , Masculino , Pneumonia por Mycoplasma/genética , Pneumonia por Mycoplasma/imunologia , Pneumonia por Mycoplasma/microbiologia , RNA Circular/análise , RNA Longo não Codificante/análise , RNA Mensageiro/análise , RNA Ribossômico , Análise de Sequência de RNA/métodosRESUMO
Mycoplasma pneumoniae-induced pneumonia (MPP) is a common cause of community-acquired respiratory tract infections, increasing risk of morbidity and mortality, in children. However, diagnosing early-stage MPP is difficult owing to the lack of good diagnostic methods. Here, we examined the protein profile of bronchoalveolar lavage fluid (BALF) and found that S100A8/A9 was highly expressed. Enzyme-linked immunosorbent assays used to assess protein levels in serum samples indicated that S100A8/A9 concentrations were also increased in serum obtained from children with MPP, with no change in S100A8/A9 levels in children with viral or bacterial pneumonia. In vitro, S100A8/A9 treatment significantly increased apoptosis in a human alveolar basal epithelial cell line (A549 cells). Bioinformatics analyses indicated that up-regulated S100A8/A9 proteins participated in the interleukin (IL)-17 signaling pathway. The origin of the increased S100A8/A9 was investigated in A549 cells and in neutrophils obtained from children with MPP. Treatment of neutrophils, but not of A549 cells, with IL-17A released S100A8/A9 into the culture medium. In summary, we demonstrated that S100A8/A9, possibly released from neutrophils, is a new potential biomarker for the clinical diagnosis of children MPP and involved in the development of this disease through enhancing apoptosis of alveolar basal epithelial cells.
Assuntos
Células Epiteliais Alveolares/metabolismo , Apoptose , Calgranulina A/metabolismo , Calgranulina B/metabolismo , Interleucina-17/farmacologia , Mycoplasma pneumoniae/patogenicidade , Neutrófilos/efeitos dos fármacos , Comunicação Parácrina , Pneumonia por Mycoplasma/metabolismo , Células A549 , Células Epiteliais Alveolares/imunologia , Células Epiteliais Alveolares/microbiologia , Células Epiteliais Alveolares/patologia , Biomarcadores/metabolismo , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Interações Hospedeiro-Patógeno , Humanos , Lactente , Masculino , Mycoplasma pneumoniae/imunologia , Neutrófilos/imunologia , Neutrófilos/metabolismo , Neutrófilos/microbiologia , Pneumonia por Mycoplasma/imunologia , Pneumonia por Mycoplasma/microbiologia , Pneumonia por Mycoplasma/patologia , Transdução de SinaisRESUMO
OBJECTIVE: Mycoplasma pneumoniae (M. pneumoniae), an extracellular pathogen lacking a cell wall, causes respiratory infection in adults and children and has been implicated in asthma exacerbation; immunoglobulin (Ig) E may be involved in these exacerbations. Specific IgM and IgG immune response to M. pneumoniae has been reported, but less is known about IgE M. pneumoniae antibody (Ab) responses in asthma. Previous studies in our laboratory demonstrated that asthmatic children have increased IgM M. pneumoniae levels, but not IgE. Thus, we sought to investigate whether past M. pneumoniae infection triggers production of M. pneumoniae-specific IgE Abs in adult subjects with/without asthma. METHODS: M. pneumoniae- IgE and -IgM Ab responses were studied in adult asthmatic (N=22) and non-asthmatic (N=22) subjects (ELISA). Data are reported as antibody index. Threshold detection levels: IgE, IgM: 0.2, 0.9, respectively. RESULTS: M. pneumoniae-IgE Ab levels were low and below the threshold of detection in both asthmatic and non-asthmatics (0.002±0.008 vs. 0.02±0.03; P=0.021). However, specific-IgM levels were slightly higher in non-asthmatics compared with asthmatics (0.96±0.37 vs. 0.79±0.31; P=0.054). M. pneumoniae-IgM Ab positivity was similar in both groups (P=1.0). CONCLUSION: IgM M. pneumoniae Abs may play an important role in non-asthma and persist for months after acute infection. IgE M. pneumoniae Abs may play a less important role in both groups.
Assuntos
Anticorpos Antibacterianos/sangue , Asma/imunologia , Imunoglobulina E/imunologia , Imunoglobulina M/sangue , Mycoplasma pneumoniae/imunologia , Pneumonia por Mycoplasma/imunologia , Adulto , Anticorpos Antibacterianos/imunologia , Asma/sangue , Asma/complicações , Asma/epidemiologia , Estudos de Casos e Controles , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , New York/epidemiologia , Pneumonia por Mycoplasma/sangue , Pneumonia por Mycoplasma/epidemiologia , Pneumonia por Mycoplasma/etiologia , PrognósticoRESUMO
MicroRNAs play a key role in Mannan-binding lectin-mediated resistance to Mycoplasma ovipneumoniae pneumonia, by regulating the translation of mRNAs of target genes, thereby regulating the immune response. Additionally, TRAF6 is a key molecule in Toll-like receptor signal transduction, which mediates inflammation and apoptosis signaling pathways and is widely involved in inflammation and immune response. While the molecular regulation mechanism has not been reported. In this study, we screened differentially expressed miRNAs and genes of Anti-infection for M. pneumonia on Sheep, through relevant bioinformatics analysis. Further, the effect of differential expression of NF-κB signaling pathway related genes on the molecular mechanism of M. pneumonia was detected. We used miRNA-mRNA integrated analysed, the target gene TRAF6 of miR-509-5p was selected. TRAF6 dual luciferase reporter vector was co-transfected into HEK 293T cells and primary sheep respiratory mucosal epithelial cells to detect changes in luciferase activity. qRT-PCR was used to analyze the effect of miR-509-5p on the expression and regulation of TRAF6 and other genes related to the NF-κB signaling pathway. The result confirmed that TRAF6 was a target gene of miR-509-5p. Compared with miR-509-5p-NC group, the luciferase activity of miR-509-5p group was significantly down-regulated (P < 0.01). Further, in sheep respiratory mucosal epithelial cells, miR-509-5p mimic could significantly down-regulate the fold change value of TRAF6 (P < 0.01). On the contrary, miR-509-5p-inhibitor up-regulated the fold change value of TRAF6 (P < 0.05). Interestingly, the expression levels of other genes were different. Among them, miR-509-5p mimic significantly up-regulated TLR4 and IRAK4 (P < 0.05), significantly down-regulated TAK1 (P < 0.05) and NF-κB (P < 0.01). miR-509-5p-inhibitor significantly up-regulated NF-κB (P < 0.05) and TAK1 (P < 0.01). miR-509-5p targets TRAF6 to affect the expression of downstream genes, which negatively regulates the NF-κB pathway, thereby affecting the inflammatory response.
Assuntos
MicroRNAs/metabolismo , NF-kappa B/metabolismo , Pneumonia por Mycoplasma/veterinária , Doenças dos Ovinos/microbiologia , Animais , Células Cultivadas , Células Epiteliais , Regulação da Expressão Gênica , Células HEK293 , Humanos , MicroRNAs/genética , NF-kappa B/genética , Pneumonia por Mycoplasma/imunologia , Pneumonia por Mycoplasma/metabolismo , Mucosa Respiratória/citologia , Ovinos , Doenças dos Ovinos/imunologia , Doenças dos Ovinos/metabolismo , Transdução de Sinais , Fator 6 Associado a Receptor de TNF/genética , Fator 6 Associado a Receptor de TNF/metabolismoRESUMO
Mycoplasma pneumoniae is the most common pathogen of community-acquired pneumonia in humans. Due to its high rates of antibiotic resistance, vaccination has become the best method to control the dissemination of M. pneumoniae. The recombinant carboxyl terminus of the P1 (P1C) protein is an immunodominant antigen, but it has negative effects such as poor stability and lower purity. In the current study, T-B epitopes of the P1C protein were predicted according to bioinformatics analysis and assessed for efficacy in peptide vaccination. BALB/c mice were subcutaneously inoculated with the T-B epitope peptides four times and then infected with M. pneumoniae through the respiratory tract. The results showed that the T-B epitope peptides of the P1C protein (P1C103-117, P1C155-169, P1C224-238 and P1C244-258) induced strong antigen-specific serum antibody responses and cellular immune responses with high levels of serum IgG, IgA antibodies and Th1-biased (IFN-γ and IL-2) cytokines. Immunization with T-B epitope peptides significantly reduced the M. pneumoniae burden and the degree of inflammation in the challenged mice. Furthermore, the levels of IFN-γ and TNF-α in the supernatants of lung homogenates were observably reduced compared to those in the PBS group. Overall, our findings demonstrate that T-B epitopes (P1C103-117, P1C155-169, P1C224-238 and P1C244-258) play significant roles in the P1C protein and can be used to induce powerful humoral and cellular immune responses to provide significant protection against M. pneumoniae pulmonary infection, which provides new insight into the design of potential multiepitope vaccines to prevent host infection by M. pneumoniae.
Assuntos
Epitopos de Linfócito B/imunologia , Epitopos de Linfócito T/imunologia , Mycoplasma pneumoniae/imunologia , Peptídeos/imunologia , Pneumonia por Mycoplasma/imunologia , Animais , Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/imunologia , Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/imunologia , Modelos Animais de Doenças , Camundongos , Camundongos Endogâmicos BALB C , Pneumonia por Mycoplasma/microbiologia , Pneumonia por Mycoplasma/prevenção & controle , Infecções Respiratórias/imunologia , Infecções Respiratórias/microbiologia , Infecções Respiratórias/prevenção & controleRESUMO
Mannose-binding lectin (MBL) glycoproteins in blood can selectively recognise lectins on the surface of bacteria, and play an important role in natural immunity. Micro RNAs (miRNAs) are key molecules that regulate gene expression at the post-transcriptional level in vivo, and their pathways are specific and effective. Previous studies indicate that small RNAs such as miRNAs perform regulatory roles in immunology. Herein, we investigated differential expression of miRNAs during MBL protein immunotherapy in sheep following treatment with different MBL genotypes (resistant and susceptible), and identified miRNAs linked to different target genes and pathways. RNA was extracted from liver tissue of resistant and susceptible sheep, miRNAs were identified by high-throughput sequencing, and differentially expressed miRNAs were analysed by SOAP to predict target genes and biological pathways. Results: Some miRNAs (oar-mir-143, oar-mir-10b, oar-mir-382, oar-mir-432 and oar-mir-379) were up-regulated, while others were down-regulated. GPATCH3 and DNAJC5 were predicted target genes of oar-mir-379, DMRT1 and GATA4 were linked to oar-mir-382, and oar-mir-432 was associated with STAT2, DMRT1 and ATG16L1. Identification of miRNAs differentially expressed in resistant and susceptible sheep may expand our understanding of miRNAs in immune regulation, and the role of MBL in innate immunity.
Assuntos
Lectinas de Ligação a Manose/genética , MicroRNAs/fisiologia , Pneumonia por Mycoplasma/veterinária , Doenças dos Ovinos/genética , Animais , Resistência à Doença/genética , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala/veterinária , MicroRNAs/genética , MicroRNAs/metabolismo , Pneumonia por Mycoplasma/genética , Pneumonia por Mycoplasma/imunologia , Fator de Transcrição STAT2/metabolismo , Ovinos , Doenças dos Ovinos/imunologiaRESUMO
BACKGROUND: Mycoplasma pneumoniae is one of the major pathogens causing community-acquired pneumonia in children. Although usually self-limited, Mycoplasma pneumoniae pneumonia (MPP) may lead to complicated morbidity that can even be life-threatening. Upon MPP infection, alveolar macrophage becomes attracted and activated and will induce subsequent cytokine and chemokine reaction. Refractory Mycoplasma pneumoniae pneumonia (RMPP) is manifested by clinical or radiological deterioration despite proper antibiotic therapy. RMPP is characterized with excessive inflammation and may need subsequent glucocorticoid treatment. AIM: The aim of this study was to investigate the change of plasma chemokines in non-refractory Mycoplasma pneumoniae pneumonia (NRMPP) and RMPP before and after antibiotic or methylprednisolone treatment. METHOD: A total of 42 children with MPP were enrolled in this study. Plasma specimens were collected at admission and one to two weeks after antibiotic or methylprednisolone treatment with declined fever. Plasma specimens were then indicated to chemokines detection. RESULTS: Mycoplasma pneumoniae pneumonia altered the chemokine profile through the observation of decreased plasma M1 related chemokines (CCL2, CCL8 and CXCL10) and increased M2 related chemokines (CCL17 and CCL22) after treatment.When the patients were divided into RMPP and NRMPP groups and the chemokines before treatment were compared, the RMPP group showed higher CXCL10 but lower CCL3 and CCL11 than the NRMPP group. CONCLUSION: Unique changes in macrophage related chemokines is observed in the course of MPP infection. NRMPP and RMPP infection in children showed distinct manifestation in chemokine profiles.
Assuntos
Quimiocinas/sangue , Quimiocinas/imunologia , Mycoplasma pneumoniae/imunologia , Pneumonia por Mycoplasma/complicações , Pneumonia por Mycoplasma/imunologia , Adolescente , Quimiocinas/classificação , Criança , Pré-Escolar , Infecções Comunitárias Adquiridas/complicações , Infecções Comunitárias Adquiridas/imunologia , Infecções Comunitárias Adquiridas/microbiologia , Feminino , Humanos , Masculino , Pneumonia por Mycoplasma/epidemiologiaRESUMO
BACKGROUND: Although Mycoplasma pneumoniae (MP) infection has been implicated in the pathogenesis of allergic diseases, the mechanism of this trigger remains unknown. We explored the mechanism for how MP infection could tilt the balance between regulatory T cells (Tregs) and Th17 cells. METHODS: We analyzed the frequency, phenotype, and function of Tregs in patients at the different stages of MP and various virus infections over a period of more than 1 year. We examined the effect of monocytes to elucidate signals that can regulate the balance between Treg and Th17 cells. RESULTS: The functional activity of Tregs was profoundly impaired during the acute stage of MP as well as viral infections. Upon resolution, however, the Treg function remained impaired even 1 year after MP infection. In the resolution stage, the impaired Treg function was associated with an increase in interleukin (IL) 17A+ Tregs and Th17 cells. Development of Th17 cells was dependent on the "aberrant" proinflammatory monocytes (pMOs), characterized by potent ability to produce IL-6 in a Toll-like receptor 2-dependent manner. CONCLUSIONS: Depending on the prevalence of the pMOs, Tregs and Th17 cells could mutually regulate the number and function of the other. The pMOs/IL-6 could be crucial therapeutic targets against MP-induced allergic diseases.
Assuntos
Monócitos/imunologia , Pneumonia por Mycoplasma , Linfócitos T Reguladores , Células Th17 , Humanos , Interleucina-6/imunologia , Pneumonia por Mycoplasma/imunologia , Linfócitos T Reguladores/imunologia , Células Th17/imunologiaRESUMO
Mycoplasma pneumoniae is a bacterial human pathogen that causes primary atypical pneumonia. M. pneumoniae motility and infectivity are mediated by the immunodominant proteins P1 and P40/P90, which form a transmembrane adhesion complex. Here we report the structure of P1, determined by X-ray crystallography and cryo-electron microscopy, and the X-ray structure of P40/P90. Contrary to what had been suggested, the binding site for sialic acid was found in P40/P90 and not in P1. Genetic and clinical variability concentrates on the N-terminal domain surfaces of P1 and P40/P90. Polyclonal antibodies generated against the mostly conserved C-terminal domain of P1 inhibited adhesion of M. pneumoniae, and serology assays with sera from infected patients were positive when tested against this C-terminal domain. P40/P90 also showed strong reactivity against human infected sera. The architectural elements determined for P1 and P40/P90 open new possibilities in vaccine development against M. pneumoniae infections.
Assuntos
Adesinas Bacterianas/imunologia , Aderência Bacteriana/imunologia , Mycoplasma pneumoniae/imunologia , Pneumonia por Mycoplasma/imunologia , Adesinas Bacterianas/isolamento & purificação , Adesinas Bacterianas/ultraestrutura , Microscopia Crioeletrônica , Cristalografia por Raios X , Mycoplasma pneumoniae/isolamento & purificação , Mycoplasma pneumoniae/patogenicidade , Pneumonia por Mycoplasma/sangue , Pneumonia por Mycoplasma/microbiologia , Domínios Proteicos/imunologiaAssuntos
COVID-19/diagnóstico , Coinfecção/diagnóstico , Mycoplasma pneumoniae/isolamento & purificação , Pneumonia por Mycoplasma/diagnóstico , SARS-CoV-2/isolamento & purificação , Urticária/diagnóstico , Anticorpos Antibacterianos/imunologia , Anticorpos Antibacterianos/isolamento & purificação , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/isolamento & purificação , COVID-19/sangue , COVID-19/imunologia , COVID-19/virologia , Teste Sorológico para COVID-19 , Criança , Coinfecção/sangue , Coinfecção/imunologia , Coinfecção/microbiologia , Ensaio de Imunoadsorção Enzimática , Reações Falso-Positivas , Humanos , Imunoglobulina M/imunologia , Imunoglobulina M/isolamento & purificação , Masculino , Pessoa de Meia-Idade , Mycoplasma pneumoniae/imunologia , Pneumonia por Mycoplasma/sangue , Pneumonia por Mycoplasma/imunologia , Pneumonia por Mycoplasma/microbiologia , SARS-CoV-2/imunologia , Pele/irrigação sanguínea , Pele/microbiologia , Urticária/sangue , Urticária/imunologia , Urticária/microbiologiaRESUMO
AIMS: The study aimed to investigate whether IL-23 is amplified in monocyte subsets of MP pneumonia and to determine its relevant pathway. MATERIALS AND METHODS: We firstly analyze the IL-23p19 expression in monocyte subgroups in MP pneumonia patients and healthy controls subjects by using flow cytometry. Then, we also analyzed the percentage of IL-17+γδT cells and Th17 cells in patients with MP pneumonia and controls subjects. At the same time, the relation between IL-23 and IL-17 were also assessed. Furthermore, we constructed the recombinant community-acquired respiratory distress syndrome (CARDS) toxin and intend to stimulate peripheral blood mononuclear cells and RAW264.7 cells in vitro. IL-23p19 was detected by flow cytometry and the mRNA levels were measured by real-time PCR. Finally, TLR4 pathway was also investigated by TAK242 inhibitor. KEY FINDINGS: It turned out that the expression of IL-23p19 was increased in CD14brightCD16+ monocyte of MP pneumonia patients than controls subjects. The patients with MP pneumonia had significantly higher the percentage of IL-17+γδT cells and Th17 cells than controls subjects. Interestingly, the levels of IL-23 were positively related to IL-17 in MP pneumonia patients. CD16+ monocytes and RAW264.7 cells, respectively can be induced by CARDS toxin to secrete IL-23 by TLR4 pathway in vitro. SIGNIFICANCE: These results indicated that IL-23-IL-17+γδT/Th17 axis may play a role in the pathogenesis of MP pneumonia, whereas IL-23 derived from CD16+ monocytes was expanded in MP pneumonia by TLR4 pathway.
Assuntos
Interleucina-17/imunologia , Interleucina-23/imunologia , Mycoplasma pneumoniae/imunologia , Pneumonia por Mycoplasma/imunologia , Receptores de IgG/imunologia , Receptor 4 Toll-Like/imunologia , Animais , Criança , Pré-Escolar , Feminino , Proteínas Ligadas por GPI/imunologia , Humanos , Masculino , Camundongos , Monócitos/imunologia , Monócitos/patologia , Pneumonia por Mycoplasma/patologia , Células RAW 264.7 , Transdução de SinaisRESUMO
miR-143-3p is correlated with inflammatory pain responses, such as hsa-miR-143-3p expression reduction in fibromyalgia. The present study aimed to explore the effects of miR-143-3p and Toll-like receptor (TLR) 4/myeloid differentiation factor 88 (MyD88)/NF-κB signaling pathway on pulmonary inflammatory factors levels and alveolar epithelial cell apoptosis in mycoplasmal pneumonia mice. Twenty mice were selected as normal group. The 120 successfully modeled Mycoplasma pneumoniae (MP) infection mice were randomly divided into model group (without any treatment), negative control (NC) group (injected with NC mimic), miR-143-3p mimic group (injected with miR-143-3p mimic), miR-143-3p inhibitor group (injected with miR-143-3p inhibitor), TAK-242 group (treatment with TAK-242), and miR-143-3p inhibitor + TAK-242 group (treatment with miR-143-3p inhibitor + TAK-242). Compared with model group, model mice had up-regulated miR-143-3p expression and decreased MyD88 and p-NF-κB p50 protein expressions (all P<0.05); Model mice treated with miR-143-3p mimic and TAK-242 had reduced interleukin (IL)-2 and tumor necrosis factor (TNF)-α contents and protein expressions of MyD88, p-NF-κB p50, increased IL-10 content, fewer alveolar epithelial cell apoptosis, lower Bax expression and higher Bcl-2 expression (all P<0.05); however, mice with miR-143-3p inhibitor treatment showed opposite trends in terms of above indicators. The exacerbation of mycoplasmal pneumonia caused by miR-143-3p inhibitor was partly improved by miR-143-3p inhibitor + TAK-242 combination treatment (all P<0.05). Therefore, up-regulation of miR-143-3p expression may ameliorate pulmonary inflammatory factors levels and reduce alveolar epithelial cell apoptosis in mycoplasmal pneumonia mice by inhibiting TLR4/MyD88/NF-κB signaling pathway.
Assuntos
Mediadores da Inflamação/metabolismo , MicroRNAs/metabolismo , Pneumonia por Mycoplasma/imunologia , Transdução de Sinais/genética , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Apoptose/imunologia , Modelos Animais de Doenças , Células Epiteliais/imunologia , Células Epiteliais/patologia , Humanos , Masculino , Camundongos , MicroRNAs/agonistas , MicroRNAs/antagonistas & inibidores , Mycoplasma pneumoniae/imunologia , Fator 88 de Diferenciação Mieloide/metabolismo , Subunidade p50 de NF-kappa B/metabolismo , Pneumonia por Mycoplasma/tratamento farmacológico , Pneumonia por Mycoplasma/microbiologia , Pneumonia por Mycoplasma/patologia , Alvéolos Pulmonares/citologia , Alvéolos Pulmonares/imunologia , Alvéolos Pulmonares/patologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia , Sulfonamidas/administração & dosagem , Receptor 4 Toll-Like/metabolismo , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/imunologiaRESUMO
BACKGROUND: Mycoplasma pneumoniae (M. pneumoniae) infection may progress to bronchiolitis obliterans (BO), with an underlying chronic inflammatory process. The aim of this study was to investigate the cytokine profiles in pulmonary T-lymphocytes and their associations with lung function in patients with BO following M. pneumoniae infection. METHODS: Bronchoalveolar lavage (BAL) samples were obtained from 10 controls and 18 children with M. pneumoniae-associated BO. We analyzed the BAL T cells for interferon (IFN)-γ, interleukin (IL)-4, IL-9, IL-17, CD25, and Foxp3 by intercellular flow cytometry. The associations with T-cell subpopulations and lung function parameters were determined. RESULTS: In BAL fluid, significantly increased proportions of T-helper 1 (Th1), Th17, and Tc1 cells were found in M. pneumoniae-associated BO patients when compared with controls. The percentages of Th17 cells showed correlations with forced expiratory volume in 1 second % predicted value (r = -0585; P < .05) and forced expiratory flow at 25% to 75% (FEF25%-75% ) % predicted value (r = -.618; P < .01). Higher proportions of Tc1 (r = -.488; P < .05) and Tc17 (r = -.542; P < .05) were significantly correlated with a reduced FEF25%-75% % predicted value in M. pneumoniae-associated BO patients. CONCLUSIONS: Our comprehensive cytokines analysis of BAL T cells revealed correlations of IL-17-producing and IFN-γ-producing T cells with lung function, suggesting that increased T-cell subpopulations may play a role in M. pneumoniae-associated BO progression.
Assuntos
Bronquiolite Obliterante/imunologia , Líquido da Lavagem Broncoalveolar/imunologia , Citocinas/imunologia , Pneumonia por Mycoplasma/imunologia , Linfócitos T/imunologia , Bronquiolite Obliterante/etiologia , Bronquiolite Obliterante/fisiopatologia , Líquido da Lavagem Broncoalveolar/citologia , Criança , Feminino , Volume Expiratório Forçado , Humanos , Pulmão/fisiopatologia , Masculino , Mycoplasma pneumoniae , Pneumonia por Mycoplasma/complicações , Pneumonia por Mycoplasma/fisiopatologiaRESUMO
BACKGROUND: An experiment was conducted to reveal why the Argali hybrid sheep are susceptible to Mycoplasma ovipneumoniae infection, the causative agent of mycoplasma ovipneumonia, a chronic respiratory disease that is harmful to the sheep industry. RESULTS: After nine Argali hybrid sheep, divided into three groups, were experimentally infected with an M. ovipneumoniae strain at 0, 4 and 14 days, transcriptome profiling of lung tissues was performed by deep RNA sequencing, using the Illumina platform. Analysis of differentially expressed genes was performed to determine concomitant gene-specific temporal patterns of mRNA expression in the lungs after M. ovipneumoniae infection. 156 differentially expressed genes (44 up-regulated, 112 down-regulated) were found when comparing transcriptomic data at 4 and 0 days post-infection, and 367 (35 up-regulated, 332 down-regulated) when comparing 14 versus 0 days post-infection. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that the differentially expressed genes at 4 and 14 versus 0 days post-infection were enriched in 109 and 150 pathways, respectively, and the Primary immunodeficiency pathway was considered most closely related to MO infection (p < .01). Hyper-IgM syndrome was identified based on the B-cell Immunodeficiency signaling pathway from differentially expressed genes related to M. ovipneumoniae infection. Gene Ontology analysis showed that differentially expressed genes in different groups were enriched for 497 and 928 terms, where those most closely related to M. ovipneumoniae infection are ciliated motor damage (p < .01). CONCLUSIONS: The situation that ciliary movement is significantly inhibited and B cells in immunodeficiency are possibly the most important reason why Argali hybrid sheep are susceptible to MO.