RESUMO
The filamentous fungus Aspergillus oryzae, also known as koji mold, has been used for centuries in the production of fermented foods in East Asia. A. oryzae fermentation can produce enzymes and metabolites with various bioactivities. In this study, we investigated whether A. oryzae fermentation extract (AOFE) has any effect on Mycoplasma pneumoniae (Mp) pneumonia. We performed solid-state fermentation of A. oryzae and obtained the ethanol extract. AOFE was analyzed by HPLC, and the major component was identified to be kojic acid. In vitro, AOFE suppressed Mp growth and invasion into A549 lung epithelial cells as determined by the gentamicin protection assay. AOFE treatment also suppressed Mp-stimulated production of tumor necrosis factor (TNF)-α and interleukin (IL)-6 at mRNA and protein levels in murine MH-S alveolar macrophages. In a mouse model of Mp pneumonia, Mp infection induced a marked pulmonary infiltration of neutrophils, which was significantly reduced in mice pre-treated orally with AOFE. AOFE administration also suppressed the production of proinflammatory cytokines and chemokines in the lungs. Collectively, our results show that AOFE has the potential to be developed into a preventive/therapeutic agent for Mp pneumonia.
Assuntos
Aspergillus oryzae , Pneumonia por Mycoplasma , Animais , Camundongos , Mycoplasma pneumoniae/metabolismo , Fermentação , Pneumonia por Mycoplasma/tratamento farmacológico , Pneumonia por Mycoplasma/microbiologia , Pneumonia por Mycoplasma/patologia , Inflamação/microbiologia , Interleucina-6/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
OBJECTIVE: This study aimed to investigate the role of p50-associated cyclooxygenase- (COX-2) extragenic RNA (PACER) on the inflammation of airway epithelium caused by Mycoplasma pneumoniae (MP) infection. METHODS: A549 cells and MP strain were cultured respectively. The expressions of PACER, IL-8, TNF-α and COX-2 in MP-infected cells were detected by qRT-PCR, the concentration of IL-8 and TNF-α in the supernatant of the cells were detected by ELISA, and the expression of COX-2 protein in the cells was detected by western-blot. After knockdown of PACER, the expression of IL-8, TNF-α and COX-2 in MP infected cells were observed. The activity of NF-κB in cells was detected by fluorescence reporter assay, and the interaction between PACER and NF-κB was verified by RNA immunoprecipitation. RESULTS: First, we observed that PACER was upregulated in MP infected A549 cells. Knockdown of PACER suppressed the production of inflammatory cytokines as well as the expression of COX-2 in A549 cells after MP infection. By performing luciferase reporter assay, we found PACER knockdown inhibited NF-κB activation induced by MP. Furthermore, RNA immunoprecipitation showed that PACER could physically bind to NF-κB p50 in MP-treated A549 cells. CONCLUSION: Collectively, our data demonstrated that attenuation of PACER reduces the inflammatory response of MP-infected epithelial cells via regulating NF-κB.
Assuntos
Mycoplasma pneumoniae , NF-kappa B , Pneumonia por Mycoplasma , RNA Longo não Codificante , Células A549 , Humanos , Inflamação/genética , Inflamação/metabolismo , Inflamação/microbiologia , NF-kappa B/metabolismo , Pneumonia por Mycoplasma/genética , Pneumonia por Mycoplasma/metabolismo , Pneumonia por Mycoplasma/microbiologia , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
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-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
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
The secretory function of airway epithelial cells is important in the pathogenesis of Mycoplasma pneumoniae pneumonia (MPP). To investigate the regulatory function of NKILA (nuclear factor-κB (NF-κB) interacting long noncoding RNA (lncRNA)) in MPP, we first detected NKILA as well as the concentration of interleukin 8 (IL-8) and tumor necrosis factor-α (TNF-α) in bronchoalveolar lavage fluid of children with MPP. Then, NKILA was knocked down in epithelial cells to investigate its effect on their secretory function. The results suggested that NKILA was downregulated in children with MPP, while IL-8 and TNF-α levels increased. Knockdown of NKILA in vitro promoted the inflammatory effects of Mycoplasma pneumoniae (MP) in epithelial A549 and BEAS-2B cells. Knockdown of NKILA promoted inhibitor of κBα (IκBα) phosphorylation and degradation, and NF-κB p65 nuclear translocation. Furthermore, RNA immunoprecipitation showed that NKILA could physically bind to IκBα in MP-treated A549 cells. Collectively, our data demonstrated that attenuation of NKILA enhances the effects of MP-stimulated secretory functions of epithelial cells via regulation of NF-κB signaling.
Assuntos
Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Pulmão/patologia , Mycoplasma pneumoniae/fisiologia , NF-kappa B/metabolismo , RNA Longo não Codificante/metabolismo , Células A549 , Líquido da Lavagem Broncoalveolar , Pré-Escolar , Citocinas/genética , Citocinas/metabolismo , Regulação para Baixo/genética , Células Epiteliais/patologia , Feminino , Humanos , Mediadores da Inflamação/metabolismo , Masculino , Inibidor de NF-kappaB alfa/metabolismo , Fosforilação , Pneumonia por Mycoplasma/genética , Pneumonia por Mycoplasma/microbiologia , Ligação Proteica , Transporte Proteico , RNA Longo não Codificante/genética , Fator de Transcrição RelA/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
Mycoplasma pneumoniae pneumonia (MPP) is a type of pneumonia induced by M. pneumoniae (MP) infection. The present study investigated the effect of long noncoding RNA growth arrestspecific 5 (GAS5) in MPP and the underlying molecular mechanism of this. The expression of GAS5, microRNA2223p, (miR2223p) and tissue inhibitor of metalloproteinases3 (TIMP3) in MPP was investigated using reverse transcriptionquantitative PCR. Lipidassociated membrane protein (LAMP)induced THP1 cells were used to model MPP. The viability of LAMPinduced THP1 cells was analyzed using an MTT assay. Expression levels of interleukin (IL)1ß, IL6 and tumor necrosis factorα (TNFα) proinflammatory cytokines, and the antiinflammatory cytokine heme oxygenase1 (HO1) in LAMPinduced THP1 cells were measured by ELISA. A dualluciferase reporter assay assessed the associations among GAS5, miR2223p and TIMP3. The expression of GAS5 and TIMP3 was downregulated in MPP. Expression of miR2223p was upregulated. GAS5overexpression increased the viability of LAMPinduced THP1 cells. GAS5 upregulation decreased the levels of IL1ß, IL6, TNFα and HO1 levels in LAMPinduced THP1 cells. GAS5 directly interacted with miR2223p. TIMP3 was a target of miR2223p. miR2223p upregulation or TIMP3knockdown reversed the promotion effect on cell viability as well as the inhibitory effect on inflammation caused by GAS5overexpression in LAMPinduced THP1 cells. GAS5overexpression increased the viability and decreased the inflammation of LAMPinduced THP1 cells by regulating the miR2223p/TIMP3 axis. These results demonstrated a potential therapeutic target for MPP treatment.
Assuntos
Inflamação/genética , MicroRNAs/genética , Pneumonia por Mycoplasma/genética , RNA Longo não Codificante/genética , Inibidor Tecidual de Metaloproteinase-3/genética , Apoptose/genética , Citocinas/genética , Regulação da Expressão Gênica/genética , Humanos , Inflamação/microbiologia , Inflamação/patologia , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/patogenicidade , Pneumonia por Mycoplasma/microbiologia , Pneumonia por Mycoplasma/patologiaRESUMO
BACKGROUND: Identifying new molecular diagnostic markers for Mycoplasma Pneumoniae Pneumonia (MPP) has always been an essential topic since MPP cases have increased every year, especially among children. Here, we examined the correlation between serum level of Purinergic receptor P2X7, vitamin A, and 25-hydroxy vitamin D (25(OH)D) and the severity of MPP, aiming to identify molecules that have the potential to become diagnostic markers. METHODS: This study was conducted on 186 cases aged 1-14 (136 MPP and 50 non-MPP patients). Serum levels of Purinergic receptor P2X7, vitamin A, 25(OH)D, and multiple inflammatory and immune factors were measured, compared, and tested for statistical significance. RESULTS: Serum P2X7, tumor necrosis factor-α (TNF-α), and interleukin-1ß (IL-1ß) levels were significantly increased in severe MPP patients, while serum vitamin A, 25(OH)D, IgA, and IgG levels were significantly decreased. CONCLUSION: Our results demonstrated a positive correlation between serum P2X7 level and the severity of MPP, and negative correlations between serum levels of vitamin A and 25(OH)D and the severity of MPP, suggesting that high serum levels of P2X7 and low serum levels of vitamin A and 25(OH)D may indicate relatively severer MPP.
Assuntos
Mycoplasma pneumoniae/fisiologia , Pneumonia por Mycoplasma/sangue , Pneumonia por Mycoplasma/microbiologia , Receptores Purinérgicos P2X7/sangue , Vitamina A/sangue , Vitamina D/análogos & derivados , Adolescente , Criança , Pré-Escolar , Citocinas/sangue , Humanos , Imunoglobulina G/sangue , Lactente , Mediadores da Inflamação/sangue , Modelos Logísticos , Análise Multivariada , Vitamina D/sangueRESUMO
To investigate the relationships between LncRNA NNT-AS1, CRP, PCT and their interactions and the refractory mycoplasma pneumoniae pneumonia (RMPP) in children. Serum levels of LncRNA NNT-AS1 of RMPP and non-RMPP (NRMPP) patients were detected by real-time PCR, and were analyzed together with serum c-reactive protein (CRP) and procalcitonin (PCT). Correlations between LncRNA NNT-AS1 and CRP and PCT were analyzed by Pearson correlation test. The ROC curve was used to analyze the potential of LncRNA NNT-AS1, CRP and PCT as biomarkers for predicting RMPP. Logistic regression crossover model and the Excel compiled by Andersson et al. were used to analyze the interactions among the biomarkers. We found that LncRNA NNT-AS1, CRP and PCT were all highly expressed in patients with RMPP. LncRNA NNT-AS1 could positively correlate with the expressions of CRP and PCT, and jointly promote the occurrence of RMPP. The combined diagnosis of LncRNA NNT-AS1, CRP and PCT could predict the occurrence of RMPP.
Assuntos
Proteína C-Reativa/metabolismo , Pneumonia por Mycoplasma/sangue , Pró-Calcitonina/sangue , RNA Longo não Codificante/sangue , Biomarcadores/sangue , Criança , Pré-Escolar , Feminino , Humanos , Modelos Logísticos , Masculino , Mycoplasma pneumoniae/isolamento & purificação , Mycoplasma pneumoniae/patogenicidade , Pneumonia por Mycoplasma/microbiologiaRESUMO
This study aimed to investigate the effect of erythromycin sequential therapy plus azithromycin on lung function and inflammatory factors in children with severe mycoplasma pneumonia (MP). Ninety-three severe MP children were selected and randomized into azithromycin group, erythromycin group, and combination group, 31 cases in each. The disappearance time of cough, fever, lung rale and X-ray shadow in the combination group were shorter than those in the azithromycin group and erythromycin group. The clinical treatment efficiency of the combination group was higher than that of the azithromycin group. After treatment, FVC, FEV1/FVC and PEF in combination group were higher than before treatment; IL-8, IL-6, CRP in combination group were lower than erythromycin group and azithromycin group. IL-8, IL-6, CRP are negatively correlated with disappearance time of cough, fever, pulmonary rale, X-ray shadow and clinical treatment efficiency; FEV1/FVC is positively correlated with disappearance time of cough and fever, pulmonary rales and X-ray shadow, and clinical treatment efficiency. Sequential erythromycin therapy combined with azithromycin in the treatment of MP can effectively inhibit high inflammatory reactions, control the disease in a timely manner, improve lung function and produce fewer adverse reactions.
Assuntos
Antibacterianos/administração & dosagem , Azitromicina/administração & dosagem , Eritromicina/administração & dosagem , Mediadores da Inflamação/sangue , Pulmão/efeitos dos fármacos , Mycoplasma pneumoniae/patogenicidade , Pneumonia por Mycoplasma/tratamento farmacológico , Antibacterianos/efeitos adversos , Azitromicina/efeitos adversos , Biomarcadores/sangue , Proteína C-Reativa/metabolismo , Estudos de Casos e Controles , Criança , Pré-Escolar , China , Quimioterapia Combinada , Eritromicina/efeitos adversos , Feminino , Interações Hospedeiro-Patógeno , Humanos , Interleucina-6/sangue , Interleucina-8/sangue , Pulmão/metabolismo , Pulmão/microbiologia , Pulmão/fisiopatologia , Masculino , Pneumonia por Mycoplasma/sangue , Pneumonia por Mycoplasma/microbiologia , Pneumonia por Mycoplasma/fisiopatologia , Distribuição Aleatória , Recuperação de Função Fisiológica , Índice de Gravidade de Doença , Fatores de Tempo , Resultado do TratamentoRESUMO
OBJECTIVES: The contribution of intracellular and fastidious bacteria in Cystic fibrosis (CF) pulmonary exacerbations, and progressive lung function decline remains unknown. This project aimed to explore their impact on bacterial microbiota diversity over time in CF children. METHODS: Sixty-one children enrolled in the MUCOVIB multicentre prospective cohort provided 746 samples, mostly nasopharyngeal swabs, throat swabs and sputa which were analysed using culture, specific real-time qPCRs and 16S rRNA amplicon metagenomics. RESULTS: Chlamydia pneumoniae (n = 3) and Mycoplasma pneumoniae (n = 1) were prospectively documented in 6.6% of CF children. Microbiota alpha-diversity in children with a documented C. pneumoniae was highly variable, similarly to children infected with Staphylococcus aureus or Pseudomonas aeruginosa. The transition from routine follow-up visits to pulmonary exacerbation (n = 17) yielded variable changes in diversity indexes with some extreme loss of diversity. CONCLUSIONS: The high rate of C. pneumoniae detection supports the need for regular screenings in CF patients. A minor impact of C. pneumoniae on the microbial community structure was documented. Although detected in a single patient, M. pneumoniae should also be considered as a possible aetiology of lung infection in CF subjects.
Assuntos
Chlamydophila pneumoniae/isolamento & purificação , Fibrose Cística/microbiologia , Microbiota , Mycoplasma pneumoniae/isolamento & purificação , Sistema Respiratório/microbiologia , Biodiversidade , Criança , Pré-Escolar , Infecções por Chlamydophila/microbiologia , Chlamydophila pneumoniae/genética , DNA Bacteriano , Humanos , Metagenômica , Mycoplasma pneumoniae/genética , Pneumonia por Mycoplasma/microbiologia , Estudos Prospectivos , RNA Ribossômico 16S , Escarro/microbiologiaRESUMO
Introduction. Infections with the respiratory pathogen Mycoplasma pneumoniae are often chronic, recurrent and resistant, persisting after antibiotic treatment. M. pneumoniae grown on glass forms protective biofilms, consistent with a role for biofilms in persistence. These biofilms consist of towers of bacteria interspersed with individual adherent cells.Hypothesis/Gap Statement. A tissue culture model for M. pneumoniae biofilms has not been described or evaluated to address whether growth, development and resistance properties are consistent with persistence in the host. Moreover, it is unclear whether the M. pneumoniae cells in the biofilm towers and individual bacterial cells have distinct roles in disease.Aim. We evaluated the properties of biofilms of M. pneumoniae grown on the immortalized human bronchial epithelial cell line BEAS-2B in relation to persistence in the host. We observed nucleation of biofilm towers and the disposition of individual cells in culture, leading to a model of how tower and individual cells contribute to infection and disease.Methodology. With submerged BEAS-2B cells as a substrate, we evaluated growth and development of M. pneumoniae biofilms using scanning electron microscopy and confocal laser scanning microscopy. We characterized resistance to erythromycin and complement using minimum inhibitory concentration assays and quantification of colony forming units. We monitored biofilm tower formation using time-lapse microscopic analysis of host-cell-free M. pneumoniae cultures.Results. Bacteria grown on host cells underwent similar development to those grown without host cells, including tower formation, rounding and incidence of individual cells outside towers. Erythromycin and complement significantly reduced growth of M. pneumoniae. Towers formed exclusively from pre-existing aggregates of bacteria. We discuss a model of the M. pneumoniae biofilm life cycle in which protective towers derive from pre-existing aggregates, and generate individual cytotoxic cells.Conclusion . M. pneumoniae can form protective biofilms in a tissue culture model, implicating biofilms in chronic infections, with aggregates of M. pneumoniae cells being important for establishing infections.
Assuntos
Biofilmes , Brônquios/microbiologia , Mycoplasma pneumoniae/fisiologia , Pneumonia por Mycoplasma/microbiologia , Antibacterianos/farmacologia , Brônquios/ultraestrutura , Linhagem Celular , Células Epiteliais/microbiologia , Células Epiteliais/ultraestrutura , Humanos , Microscopia Eletrônica de Varredura , Mycoplasma pneumoniae/efeitos dos fármacos , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/ultraestruturaRESUMO
There is limited data on serum biomarkers in distinguishing Mycoplasma pneumoniae (MP) from Streptococcus pneumoniae (SP) and viral pneumoniae (VP) etiologies of community-acquired pneumonia (CAP). A retrospective study of inpatients diagnosed with CAP at the First Affiliated Hospital of Dali University (Dali, Yunnan, China) between January 2018 and June 2020 was conducted. The demographic, clinical and laboratory data of the patients with CAP were analyzed. Univariate analyses identified predictors for MP infections. The discriminative power of C-reactive protein (CRP), procalcitonin (PCT), CRP/PCT and CRP/PCT >350 µg/ng was assessed by area under the curve (AUC) of the receiver operating characteristic (ROC) curves. A total of 552 CAP patients, including 247 (44.7%) with MP, 152 (27.6%) with SP and 153 (27.7%) with influenza A and B viruses, were enrolled. When comparing MP with SP, cough and CRP/PCT >350 µg/ng (odds ratio [OR]) 2.88, p < .001) were predictors for MP. CRP/PCT >350 µg/ng had 76% sensitivity and 100% specificity (AUC = 0.89, p < .001, 95% confidence interval [CI]:0.81-0.94) to predict MP infections. Furthermore, similar results were again obtained when comparing MP with VP. CRP/PCT >350 µg/ng present better information (OR: 4.70; AUC = 0.92, p < .001, 87% sensitivity and 100% specificity). In addition, comparing MP and non-MP (SP and VP combined), CRP/PCT >350 µg/ng exhibited excellent performance (AUC = 0.90, 95%CI 0.83-0.95, p < .001, 76% sensitivity and 100% specificity). CRP/PCT ratio may be a potential index to distinguish MP-CAP from non-MP-CAP.
Assuntos
Proteína C-Reativa/metabolismo , Infecções Comunitárias Adquiridas/sangue , Infecções Comunitárias Adquiridas/microbiologia , Hospitalização , Mycoplasma pneumoniae/fisiologia , Pneumonia por Mycoplasma/sangue , Pneumonia por Mycoplasma/microbiologia , Pró-Calcitonina/sangue , Adulto , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-IdadeRESUMO
Herein, we found that serum concentration of superoxide dismutase 3 (SOD3) was significantly reduced in children with mycoplasma pneumonia (MP) infection. To study the roles of SOD3 in inflammatory regulation of MP infection, human A549 type II alveolar epithelial cells were stimulated with 107 CCU/ml of MP to build MP infection in vitro. Secretion of pro-inflammatory cytokine interleukin (IL)-8 and tumor necrosis factor (TNF)-α were measured via enzyme-linked immunosorbent assay (ELISA) to assess the inflammatory response of A549 cells. Levofloxacin (LVFX) was used as an anti-inflammatory drug while recombinant TNF-α was used as an inflammatory promotor in MP-infected cells. Transcriptional activity of nuclear factor (NF)-κB was assessed by detecting protein levels of nuclear NF-κB and cytoplasm NF-κB using Western blot analysis. Our data suggested that the expression of SOD3 mRNA and protein, as well as content of SOD3 in cultured supernatant, were time-dependently inhibited in MP-infected A549 cells. However, lentiviruses-mediated SOD3 overexpression alleviated inflammatory response of MP-infected A549 cells, and prevented the unclear translocation of NF-κB, as evidenced by obviously reducing the production of IL-8 and TNF-α in cell cultured supernatant, as well as decreasing nuclear NF-κB while increasing cytoplasm NF-κB. Inspiringly, SOD3 overexpression induced anti-inflammatory effect and the inactivation of NF-κB was similar to that of 2 lg/ml of LVFX, but reversed by additional TNF-α treatment. Therefore, we can conclude that transcriptional activity of NF-jB was the underlying mechanism, by which SOD3 regulated inflammatory response in MP infection in vitro.
Assuntos
Inflamação/genética , Interleucina-8/genética , Pneumonia por Mycoplasma/genética , Superóxido Dismutase/genética , Fator de Necrose Tumoral alfa/genética , Células A549 , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/genética , Criança , Humanos , Inflamação/tratamento farmacológico , Inflamação/microbiologia , Levofloxacino/farmacologia , Lipopolissacarídeos/farmacologia , Mycoplasma pneumoniae/patogenicidade , Pneumonia por Mycoplasma/microbiologia , Pneumonia por Mycoplasma/patologia , RNA Mensageiro/genéticaRESUMO
BACKGROUND: The prevalence of macrolide-resistant Mycoplasma pneumoniae (MP) pneumonia has been rapidly increased. MP pneumonia is a risk factor for the development of post-infectious bronchiolitis obliterans (PIBO). The aim of the present study was to identify the risk factors for the development of PIBO after MP pneumonia in the era of increasing macrolide resistance of MP. MATERIALS AND METHODS: This retrospective study enrolled 150 children with a mean age of 6.0 years admitted to the hospital due to MP pneumonia between May 2019 and February 2020 at a tertiary hospital. The clinical, radiologic, and laboratory data were obtained using retrospective chart review. RESULTS: Eighteen children (12%) were diagnosed with PIBO after MP pneumonia. PIBO was diagnosed after a mean duration of 100.0 days (range, 6-268 days) from symptom onset. The respiratory virus co-infection (adjusted odds ratio [aOR], 4.069; 95% confidence interval [95% CI], 1.224-13.523), adenovirus co-infection (aOR, 5.607; 95% CI, 1.801-17.454), longer duration between symptom onset and admission (aOR, 1.150; 95% CI, 1.020-1.298), higher levels of serum lactate dehydrogenase (LDH) at the time of admission (aOR, 1.001; 95% CI, 1.000-1.003), and poor response to stepwise treatment increased the risk for development of PIBO after MP pneumonia. However, macrolide resistance of MP was not associated with development of PIBO after MP pneumonia. CONCLUSION: The present study suggests that respiratory virus co-infection, including adenovirus, poor response to the treatment of MP pneumonia, and higher levels of serum LDH, but not macrolide resistance of MP, are risk factors of PIBO after MP pneumonia.
Assuntos
Bronquiolite Obliterante/etiologia , Mycoplasma pneumoniae , Pneumonia por Mycoplasma/complicações , Infecções por Adenoviridae/complicações , Fatores Etários , Antibacterianos , Criança , Pré-Escolar , Coinfecção/complicações , Farmacorresistência Bacteriana , Feminino , Humanos , L-Lactato Desidrogenase/sangue , Macrolídeos , Masculino , Pneumonia por Mycoplasma/microbiologia , Estudos Retrospectivos , Fatores de Risco , Fatores de TempoAssuntos
Anticorpos Antibacterianos/sangue , COVID-19/complicações , Síndrome de Linfonodos Mucocutâneos/complicações , Mycoplasma pneumoniae/patogenicidade , Pneumonia por Mycoplasma/complicações , SARS-CoV-2/patogenicidade , Síndrome Respiratória Aguda Grave/complicações , Adolescente , Western Blotting , COVID-19/diagnóstico , COVID-19/patologia , COVID-19/virologia , Teste para COVID-19 , Criança , Pré-Escolar , Coinfecção , Conjuntivite/diagnóstico , Conjuntivite/fisiopatologia , Edema/diagnóstico , Edema/fisiopatologia , Exantema/diagnóstico , Exantema/fisiopatologia , Feminino , Febre/diagnóstico , Febre/fisiopatologia , Humanos , Lactente , Masculino , Síndrome de Linfonodos Mucocutâneos/diagnóstico , Síndrome de Linfonodos Mucocutâneos/patologia , Síndrome de Linfonodos Mucocutâneos/virologia , Mycoplasma pneumoniae/crescimento & desenvolvimento , Pneumonia por Mycoplasma/diagnóstico , Pneumonia por Mycoplasma/microbiologia , Pneumonia por Mycoplasma/patologia , SARS-CoV-2/crescimento & desenvolvimento , Síndrome Respiratória Aguda Grave/diagnóstico , Síndrome Respiratória Aguda Grave/patologia , Síndrome Respiratória Aguda Grave/virologia , Estomatite/diagnóstico , Estomatite/fisiopatologiaRESUMO
BACKGROUND: To analyze the clinical characteristics of Mycoplasma pneumoniae pneumonia with hypoxia in children, and identify the associated risk factors of hypoxia in MPP. METHODS: A retrospective case-control study was performed on 345 children with Mycoplasma pneumoniae pneumonia (MPP) admitted to our hospital wards from January 2017 to June 2019. They were divided into three groups, namely MPP with hypoxia, refractory Mycoplasma pneumoniae pneumonia (RMPP), and general Mycoplasma pneumoniae pneumonia (GMPP). The clinical features, laboratory findings, imaging, and management were collected and compared in the three groups. RESULTS: The MPP with hypoxia patients (n = 69) had longer disease duration, a higher extra-pulmonary complications rate, and more severe radiological abnormalities (P < 0.05). They also needed more complicated treatments (P < 0.05). Meanwhile, the levels of white blood cell count (WBC), C-reactive protein (CRP), lactic dehydrogenase (LDH), interleukin (IL)-6, ferritin, D-dimer, fibrinogen (FG), alanine aminotransferase (ALT) and the percentage of neutrophils in the MPP with hypoxia group were significantly higher than those in the RMPP group and the GMPP group (P < 0.05). In ROC curve analysis, the percentage of neutrophils, WBC, CRP, LDH, IL-6, ferritin, D-dimer, and ALT were contributed to identify the MPP with hypoxia patients. Multivariate logistic regression analysis revealed that ferritin> 174.15 ng/mL, IL-6 > 25.475 pg/ml, and pleural effusion were significantly associated with the incidence of hypoxia in MPP (P < 0.01). CONCLUSION: MPP with hypoxia patients presented more serious clinical manifestations. Ferritin> 174.15 ng/mL, IL-6 > 25.475 pg/ml and pleural effusion were related risk factors for hypoxia in MPP.
Assuntos
Hipóxia/sangue , Hipóxia/complicações , Mycoplasma pneumoniae , Pneumonia por Mycoplasma/sangue , Pneumonia por Mycoplasma/complicações , Proteína C-Reativa/análise , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Ferritinas/sangue , Produtos de Degradação da Fibrina e do Fibrinogênio/análise , Humanos , Interleucina-6/sangue , Contagem de Leucócitos , Masculino , Neutrófilos/metabolismo , Derrame Pleural , Pneumonia por Mycoplasma/microbiologia , Curva ROC , Estudos Retrospectivos , Fatores de RiscoRESUMO
Human surfactant protein-A2 (hSP-A2) is a component of pulmonary surfactant that plays an important role in the lung's immune system by interacting with viruses, bacteria, and fungi to facilitate pathogen clearance and by downregulating inflammatory responses after an allergic challenge. Genetic variation in SP-A2 at position Gln223Lys is present in up to â¼30% of the population and has been associated with several lung diseases, such as asthma, pulmonary fibrosis, and lung cancer (M. M. Pettigrew, J. F. Gent, Y. Zhu, E. W. Triche, et al., BMC Med Genet 8:15, 2007, https://bmcmedgenet.biomedcentral.com/articles/10.1186/1471-2350-8-15; Y. Wang, P. J. Kuan, C. Zing, J. T. Cronkhite, et al., Am J Hum Genet 84:52-59, 2009, https://www.cell.com/ajhg/fulltext/S0002-9297(08)00595-8). Previous work performed by our group showed differences in levels of SP-A binding to non-live mycoplasma membrane fractions that were dependent on the presence of a lysine (K) or a glutamine (Q) at amino acid position 223 in the carbohydrate region of SP-A2. On the basis of these differences, we have derived 20-amino-acid peptides flanking this region of interest in order to test the ability of each to regulate various immune responses to live Mycoplasma pneumoniae in SP-A knockout mice and RAW 264.7 cells. In both models, the 20-mer containing 223Q significantly decreased both tumor necrosis factor alpha (TNF-α) mRNA levels and protein levels in comparison to the 20-mer containing 223K during M. pneumoniae infection. While neither of the 20-mer peptides (223Q and 223K) had an effect on p38 phosphorylation during M. pneumoniae infection, the 223Q-20mer peptide significantly reduced NF-κB p65 phosphorylation in both models. Taken together, our data suggest that small peptides derived from the lectin domain of SP-A2 that contain the major allelic variant (223Q) maintain activity in reducing TNF-α induction during M. pneumoniae infection.
Assuntos
Anti-Inflamatórios/farmacologia , Interações entre Hospedeiro e Microrganismos/imunologia , Mycoplasma pneumoniae/imunologia , Peptídeos/farmacologia , Pneumonia por Mycoplasma/tratamento farmacológico , Proteína A Associada a Surfactante Pulmonar/imunologia , Fator de Necrose Tumoral alfa/imunologia , Adesinas Bacterianas/genética , Adesinas Bacterianas/imunologia , Animais , Anti-Inflamatórios/síntese química , Modelos Animais de Doenças , Regulação da Expressão Gênica , Interações entre Hospedeiro e Microrganismos/genética , Humanos , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Pulmão/microbiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mycoplasma pneumoniae/efeitos dos fármacos , Mycoplasma pneumoniae/patogenicidade , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Neutrófilos/microbiologia , Peptídeos/síntese química , Pneumonia por Mycoplasma/genética , Pneumonia por Mycoplasma/imunologia , Pneumonia por Mycoplasma/microbiologia , Domínios Proteicos , Proteína A Associada a Surfactante Pulmonar/química , Proteína A Associada a Surfactante Pulmonar/deficiência , Proteína A Associada a Surfactante Pulmonar/genética , Células RAW 264.7 , RNA Mensageiro/genética , RNA Mensageiro/imunologia , Transdução de Sinais , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/imunologia , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/genética , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/imunologiaRESUMO
PURPOSE: To evaluate the natural history and ophthalmologic morbidity of Mycoplasma pneumoniae-induced rash and mucositis (MIRM) and propose a treatment algorithm. DESIGN: Retrospective, interventional case series. METHODS: Retrospective chart review of all MIRM patients examined by the department of ophthalmology at a tertiary children's hospital. Diagnosis was established clinically concomitant with either positive Mycoplasma pneumoniae IgM or PCR testing from January 1, 2010, until December 31, 2019. The main outcome measures were best-corrected visual acuity, long-term ocular sequelae, and duration and type of ophthalmic intervention. RESULTS: There were 15 patients (10 male and 5 female) aged 10.9 ± 4.2 years who had primary episodes of MIRM; of those, 4 had multiple episodes. All patients required topical steroid treatment, 3 required amniotic membrane transplantation, and 1 patient underwent placement of a sutureless biologic corneal badage device. There were no patients who suffered visual loss, but 1 was left with mild symblephara near the lateral canthus in each eye and 2 others had scarring of the eyelid margins and blepharitis. CONCLUSIONS: The ocular morbidity is significantly less in MIRM than in other closely related syndromes such as erythema multiforme, Stevens-Johnson syndrome, and toxic epidermal necrolysis. However, these patients still require close observation and a low threshold for intervention to avoid permanent ophthalmic sequelae and possible blindness.