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1.
Mol Microbiol ; 120(5): 684-701, 2023 11.
Artículo en Inglés | MEDLINE | ID: mdl-37718557

RESUMEN

Syphilis is a persistent sexually transmitted disease caused by infiltration of the elusive pathogen Treponema pallidum. Despite the prevalence of human polymorphonuclear neutrophils (hPMNs) within cutaneous lesions, which are characteristic of incipient syphilis, their role in T. pallidum infection remains unclear. Tp92 is the only T. pallidum helical outer membrane protein that exhibits structural features similar to those of outer membrane proteins in other gram-negative bacteria. However, the functional mechanism of this protein in immune cells remains unclear. Neutrophils are short-lived cells that undergo innate apoptosis in response to external stimuli that typically influence this process. In this study, we determined that Tp92 impedes the activation of procaspase-3 via the ERK MAPK, PI3K/Akt, and NF-κB signaling pathways, consequently suppressing caspase-3 activity within hPMNs, and thereby preventing hPMNs apoptosis. Furthermore, Tp92 could also modulate hPMNs apoptosis by enhancing the expression of the anti-apoptotic protein Mcl-1, stimulating IL-8 secretion, and preserving the mitochondrial membrane potential. These findings provide valuable insights into the molecular mechanisms underlying T. pallidum infection and suggest potential therapeutic targets for syphilis treatment.


Asunto(s)
FN-kappa B , Sífilis , Humanos , FN-kappa B/metabolismo , Treponema pallidum/genética , Treponema pallidum/metabolismo , Sífilis/metabolismo , Sífilis/microbiología , Sífilis/patología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas de la Membrana/metabolismo , Neutrófilos , Apoptosis
2.
J Infect Dis ; 228(4): 453-463, 2023 08 16.
Artículo en Inglés | MEDLINE | ID: mdl-36961856

RESUMEN

BACKGROUND: Whether polymorphonuclear neutrophils (PMN) exert a protective role upon chlamydial infection by expressing inducible nitric oxide (NO) synthase (iNOS) and producing NO remains unclear. METHODS: This issue was addressed using BALB/c mice infected with Chlamydia psittaci 6BC strain. Methods included flow cytometry, immunofluorescence, qRT-PCR, and western blot. RESULTS: The number of PMN was significantly increased during C. psittaci infection, which was accompanied by increased iNOS expression and NO production in the mouse lungs. PMN were the major source of NO during pulmonary C. psittaci infection and inhibited C. psittaci multiplication in an iNOS/NO-dependent manner. Depletion of PMN aggravated C. psittaci-induced disease and increased C. psittaci burden. Nuclear factor-κB (NF-κB) and STAT1 signaling pathways, but not MAPK signaling pathways, were required for the induction of iNOS expression and NO production in PMN by C. psittaci infection. Thus, our findings highlight the protective role of NO-producing PMN in C. psittaci infection. CONCLUSIONS: NO-producing PMN confer a protective role during pulmonary C. psittaci infection in mice, and thus our study sheds new light on PMN function during Chlamydia infection.


Asunto(s)
Infecciones por Chlamydia , Chlamydophila psittaci , Neumonía , Ratones , Animales , Chlamydophila psittaci/metabolismo , Óxido Nítrico/metabolismo , Neutrófilos/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Pulmón/metabolismo
3.
Microbiol Immunol ; 67(11): 469-479, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-37615441

RESUMEN

Chlamydia psittaci is a zoonotic pathogen found in birds and humans. Macrophages, major components of the innate immune system, can resist chlamydial infections and trigger adaptive immune responses. However, the molecular mechanisms underlying the action of macrophages against C. psittaci infection are not well understood. This study investigated the roles and mechanisms of plasmid-encoded protein CPSIT_p7 of C. psittaci in regulating autophagy in RAW264.7 cells. The results demonstrated that stimulation of RAW264.7 with C. psittaci plasmid protein CPSIT_p7 induced the expressions of the autophagy signaling primary regulators LC3 and Beclin1, which could also significantly induce the phosphorylation levels of ERK, JNK, p38, and Akt. Next, siRNA knockdown of TLR2 resulted in significant downregulation of CPSIT_p7-triggered autophagy in RAW264.7 cells. Moreover, the extracellular regulated protein kinase (ERK) inhibitor PD98059 markedly reduced autophagy in CPSIT_p7-stimulated macrophages. In summary, these results indicated that TLR2 plays an essential role in the induction of autophagy through the ERK signaling pathway in CPSIT_p7-stimulated RAW264.7 cells.


Asunto(s)
Chlamydophila psittaci , Psitacosis , Animales , Humanos , Ratones , Autofagia , Chlamydophila psittaci/genética , Chlamydophila psittaci/metabolismo , Psitacosis/genética , Psitacosis/metabolismo , Células RAW 264.7 , Transducción de Señal , Receptor Toll-Like 2/genética , Receptor Toll-Like 2/metabolismo
4.
Environ Res ; 239(Pt 1): 117266, 2023 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-37775001

RESUMEN

Triple-negative breast cancer (TNBC) has an escalating morbidity and a dismal prognosis. Obesity has been reported to be strongly linked to adverse TNBC outcomes. Exosomes (Exos) transport RNA and proteins between cells and serve as intermediaries for cell-to-cell communication. Accumulated evidence suggests that adipose-secreted circular RNAs (circRNAs) can modulate protein glycosylation in TNBC to facilitate tumor cell outgrowth. Herein, exo-circCRIM1 expression was found to be elevated in TNBC patients with a high body fat percentage. Functional experiments demonstrated that by inhibiting miR-503-5p, exo-circCRIM1 enhanced TNBC evolution and metastasis while activating glycosylation hydrolase OGA. Furthermore, OGA negatively regulates FBP1 by decreasing its protein stability. Moreover, the levels of OGA and FBP1 were positively related to the infiltration level of some immune cells in TNBC. These findings indicate that exo-cirCRIM1 secreted by adipocytes contributes to TNBC progression by inhibiting miR-503-5p and activating the OGA/FBP1 signaling pathway. The findings reveal a novel intercellular signaling pathway mediated by adipose-derived exosomes and suggest that treatment targeting the secreted exosome-circCRIM1 may reverse TNBC progression.


Asunto(s)
Exosomas , MicroARNs , Neoplasias de la Mama Triple Negativas , Humanos , MicroARNs/genética , MicroARNs/metabolismo , Neoplasias de la Mama Triple Negativas/genética , Neoplasias de la Mama Triple Negativas/metabolismo , Neoplasias de la Mama Triple Negativas/patología , Línea Celular Tumoral , Exosomas/metabolismo , Exosomas/patología , Adipocitos/metabolismo , Adipocitos/patología , Proliferación Celular
5.
Int J Mol Sci ; 24(8)2023 Apr 10.
Artículo en Inglés | MEDLINE | ID: mdl-37108176

RESUMEN

Chlamydia psittaci (C. psittaci), a zoonotic pathogen, poses a potential threat to public health security and the development of animal husbandry. Vaccine-based preventative measures for infectious diseases have a promising landscape. DNA vaccines, with many advantages, have become one of the dominant candidate strategies in preventing and controlling the chlamydial infection. Our previous study showed that CPSIT_p7 protein is an effective candidate for a vaccine against C. psittaci. Thus, this study evaluated the protective immunity of pcDNA3.1(+)/CPSIT_p7 against C. psittaci infection in BALB/c mice. We found that pcDNA3.1(+)/CPSIT_p7 can induce strong humoral and cellular immune responses. The IFN-γ and IL-6 levels in the infected lungs of mice immunized with pcDNA3.1(+)/CPSIT_p7 reduced substantially. In addition, the pcDNA3.1(+)/CPSIT_p7 vaccine diminished pulmonary pathological lesions and reduced the C. psittaci load in the lungs of infected mice. It is worth noting that pcDNA3.1(+)/CPSIT_p7 suppressed C. psittaci dissemination in BALB/c mice. In a word, these results demonstrate that the pcDNA3.1(+)/CPSIT_p7 DNA vaccine has good immunogenicity and immunity protection effectiveness against C. psittaci infection in BALB/c mice, especially pulmonary infection, and provides essential practical experience and insights for the development of a DNA vaccine against chlamydial infection.


Asunto(s)
Infecciones por Chlamydia , Chlamydophila psittaci , Psitacosis , Vacunas de ADN , Animales , Ratones , Chlamydophila psittaci/genética , Vacunas de ADN/genética , Ratones Endogámicos BALB C , Proteínas Bacterianas/genética , Vacunas Bacterianas , Psitacosis/prevención & control , Pulmón/patología , Infecciones por Chlamydia/prevención & control , Plásmidos/genética , ADN
6.
Molecules ; 28(20)2023 Oct 13.
Artículo en Inglés | MEDLINE | ID: mdl-37894556

RESUMEN

Mycoplasma pneumoniae (M. pneumoniae) is an atypical bacterial pathogen responsible for community-acquired pneumonia primarily among school-aged children and young adults. Camellia oleifera (C. oleifera) has been used as a medicinal and edible plant in China for centuries, the constituents from which possessed various bioactivities. Notably, flavonoids existing in residues of C. oleifera defatted seeds exhibited significant anti-inflammatory activities. In the present study, we investigated the impact of total flavonoids from C. oleifera (TFCO) seed extract on M. pneumoniae pneumonia. TFCO was obtained using multiple column chromatography methods and identified as kaempferol glycosides via UPLC-HRESIMS. In a M. pneumoniae pneumonia mouse model, TFCO significantly reduced the lung damage, suppressed IL-1ß, IL-6, and TNF-α production, and curbed TLR2 activation triggered by M. pneumoniae. Similarly, in RAW264.7 macrophage cells stimulated by lipid-associated membrane proteins (LAMPs), TFCO suppressed the generation of proinflammatory cytokines and TLR2 expression. Moreover, TFCO diminished the phosphorylation of IκBα, JNK, ERK, p38, and p65 nuclear translocation in vitro. In conclusion, TFCO alleviated M. pneumoniae-induced lung damage via inhibition of TLR2-mediated NF-κB and MAPK pathways, suggesting its potential therapeutic application in M. pneumoniae-triggered lung inflammation.


Asunto(s)
Camellia , Lesión Pulmonar , Neumonía , Animales , Niño , Ratones , Humanos , FN-kappa B/metabolismo , Mycoplasma pneumoniae/metabolismo , Receptor Toll-Like 2/genética , Receptor Toll-Like 2/metabolismo , Flavonoides
7.
Int J Med Microbiol ; 312(4): 151553, 2022 May.
Artículo en Inglés | MEDLINE | ID: mdl-35358795

RESUMEN

BACKGROUND: Neurosyphilis is a serious complication caused by the invasion of the central nervous system by Treponema pallidum subsp. pallidum (T. pallidum). However, the molecular mechanism by which T. pallidum crosses the blood-brain barrier has not been fully elucidated. OBJECTIVES: The primary purpose of this experimental design was to explore the effect of the T. pallidum adhesion protein Tp0751 on the blood-brain barrier and cerebrovascular endothelial cells. METHODS: BEnd3 cells were used to construct a monolayer blood-brain barrier model in vitro. The integrity of blood-brain barrier model was evaluated by a transendothelial cell resistance meter and transmission electron microscope after the stimulation of recombinant protein TP0751. Hoechst 33258 staining and flow cytometry were used to detect the apoptosis rate. Western blotting assay was used to measure the expression of tight junction proteins and apoptosis-related proteins. The enzyme activity detection kit was responsible for detecting the enzyme activities of Caspase 3, Caspase 8 and Caspase 9. The expression of pro-inflammatory cytokines TNF-α, IL-1ß and IL-6 at the transcription and translation levels were detected by qRT-PCR and ELISA, respectively. RESULTS: The results showed that, the tight junction structures between cells showed no obvious fragmentation, but the levels of the tight junction proteins zonula occludens-1 and occludin were reduced by the effects of Tp0751 on bEnd3 cells. In addition, further research demonstrated that after incubation with bEnd3 cells, Tp0751 induced cell apoptosis in a concentration- and time-dependent manner via the caspase 8/caspase 3 pathway. These apoptotic processes may have contributed to the changes in tight junction proteins expression. Furthermore, the Tp0751 protein may be involved in the pathogenic process by which T. pallidum crosses the blood-brain barrier by promoting secretion of the proinflammatory factor interleukin-6. CONCLUSIONS: On the whole, this study is the first to reveal and highlight that Tp0751 may affect the expression of tight junction proteins by inducing apoptosis and promoting the secretion of the inflammatory cytokine IL-6, thus playing a role in the progression of neurosyphilis caused by T. pallidum.


Asunto(s)
Neurosífilis , Treponema pallidum , Apoptosis , Proteínas Bacterianas , Caspasa 3/metabolismo , Caspasa 8/metabolismo , Citocinas/metabolismo , Células Endoteliales , Humanos , Interleucina-6/metabolismo , Neurosífilis/metabolismo , Neurosífilis/microbiología , Proteínas de Uniones Estrechas/metabolismo , Uniones Estrechas/metabolismo , Treponema , Treponema pallidum/metabolismo
8.
Int J Med Microbiol ; 312(8): 151571, 2022 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-36511277

RESUMEN

This study tested the hypothesis that Chlamydia psittaci (C. psittaci) survives and multiplies in human neutrophils by activating P2X7, a nonselective cationic channel receptor expressed constitutively on the surface of these cells. Findings illustrated that P2X7 receptor expression was enhanced in C. psittaci-infected neutrophils. C. psittaci was able to inhibite spontaneous apoptosis of neutrophils through mitochondrial-induced ATP release and IL-8 production. Importantly, inhibiting ATP activation of the P2X7 receptor with AZ10606120 promotes apoptosis, while stimulating P2X7 receptor expression with BzATP delayed spontaneous apoptosis of human neutrophils, suggesting that C. psittaci inhibits apoptosis of human neutrophils by activating P2X7 receptor. This study reveals new insights into the survival advantages of the latent persistent state of C. psittaci and the mechanism by which it evades the innate immune response.


Asunto(s)
Chlamydophila psittaci , Humanos , Adenosina Trifosfato/metabolismo , Adenosina Trifosfato/farmacología , Apoptosis , Chlamydophila psittaci/metabolismo , Expresión Génica , Receptores Purinérgicos P2X7
9.
J Clin Lab Anal ; 36(9): e24635, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35908795

RESUMEN

OBJECTIVE: To discover novel serodiagnostic candidates for the serological diagnosis of syphilis. METHODS: Two recombinant Treponema pallidum proteins Tp0100 and Tp1016 were expressed, purified, and identified by Western Blotting. A total of 600 clinical serum samples were tested with the Tp0100-based ELISA, the Tp1016-based ELISA, and the commercial LICA Syphilis TP kit (ChIVD, Beijing, China). The sensitivities were determined by testing 340 samples from individuals with clinically diagnosed primary, secondary, latent, and tertiary syphilis. The specificities were determined by screening 260 samples from healthy controls and individuals with potentially cross-reactive infections, including leptospirosis, Lyme disease, hepatitis B, tuberculosis, rheumatoid arthritis, systemic lupus erythematosus. Kappa (κ) values were applied to compare the agreement between clinical syphilis diagnosis and the Tp0100-based ELISA, the Tp1016-based ELISA, or the LICA Syphilis TP test. RESULTS: Using clinical syphilis diagnosis as the gold standard, Tp0100 exhibited an overall sensitivity of 95.6% and specificity of 98.1% for testing IgG antibody while Tp1016 demonstrated only an overall sensitivity of 75.0% and specificity of 79.6%. In contrast, the LICA Syphilis TP test revealed an overall sensitivity of 97.6% and specificity of 96.2%. In addition, the overall percent agreement and corresponding κ values were 96.7% (95% CI 95.6%-97.8%) and 0.93 for the Tp0100-based ELISA, 77.0% (95% CI 74.3%-79.7%) and 0.54 for the Tp1016-based ELISA, and 97.0% (95% CI 96.0%-98.0%) and 0.94 for the LICA Syphilis TP test, respectively. CONCLUSION: The recombinant T. pallidum protein Tp0100 shows promise as a novel diagnostic antigen in the serological tests for syphilis.


Asunto(s)
Sífilis , Treponema pallidum , Anticuerpos Antibacterianos , Antígenos Bacterianos , Ensayo de Inmunoadsorción Enzimática , Humanos , Proteínas Recombinantes , Sensibilidad y Especificidad , Pruebas Serológicas , Sífilis/diagnóstico , Serodiagnóstico de la Sífilis , Treponema pallidum/genética
10.
Microb Pathog ; 147: 104256, 2020 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-32416138

RESUMEN

Chlamydia psittaci is the pathogen of psittacosis, and it has emerged as a significant public health threat. Because most infections are easily overlooked, a vaccine is recognized as the best solution to control the spread of C. psittaci. Our previous study showed that Pgp3 protein is efficacious as a subunit vaccine while not the best candidate due to the negative effects. Thus, in this study, we tested the ability of a tandem epitope vaccine candidate designated SP based on Pgp3-dominant epitopes to induce protective immunity against pulmonary chlamydial infection. BALB/c mice were intraperitoneally inoculated with multiepitope peptide antigens followed by intranasal infection with C. psittaci. We found that the multiepitope peptide antigens induced strong humoral and cellular immune responses with high Th1-related (IFN-γ and IL-2) and proinflammatory (IL-6) cytokine levels. Meanwhile, the pathogen burden and inflammatory infiltration were significantly reduced in lungs of SP-immunized mice after chlamydial challenge. In addition, the IFN-γ and IL-6 secretion levels in the infected lungs were substantially reduced. Overall, our findings demonstrate that the peptide vaccine SP plays a significant role with good immunogenicity and protective efficacy against C. psittaci lung infection in BALB/c mice, providing important insights towards understanding the potential of peptide vaccines as new vaccine antigens for inducing protective immunity against chlamydial infection.


Asunto(s)
Infecciones por Chlamydia , Chlamydophila psittaci , Animales , Anticuerpos Antibacterianos , Vacunas Bacterianas , Citocinas , Pulmón , Ratones , Ratones Endogámicos BALB C , Vacunas de Subunidad
11.
Cell Mol Biol Lett ; 25: 38, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32782452

RESUMEN

AIMS: Daxx is a highly conserved nuclear protein with an important role in transcription, apoptosis and other cell processes. We investigated the role of HPV16 E6 in Daxx-induced apoptosis through their interactions in C33A cells. METHODS: The binding of HPV16 E6 and Daxx was confirmed in C33A cells using co-immunoprecipitation and indirect immunofluorescence assays. Quantitative PCR and western blotting were performed to determine the RNA and protein expressions of Daxx, respectively. Automatic cell count and MTT assays were performed to investigate the proliferation of C33A cells. The apoptosis rate of C33A cells was determined via flow cytometry using Annexin V-FITC/PI staining. The relative activity of caspase-8 was tested using ELISA. RESULTS: HPV16 E6 can bind with Daxx and cause its translocation in C33A cells. The transfected HPV16 E6 can cause a decrease in relative quantification for Daxx in Daxx-overexpressing cells. After Daxx transfection, cell proliferation was found to decrease sharply and cell apoptosis to increase sharply. However, when HPV16 E6 was co-transfected with Daxx, this decrease and increase both became gentle. Similarly, HPV16 E6 made the Daxx-induced increase in caspase-8 activity milder. CONCLUSIONS: HPV16 E6 is involved in inhibiting apoptosis through deregulation of Daxx-induced caspase-8 activities.


Asunto(s)
Apoptosis/fisiología , Proteínas Co-Represoras/metabolismo , Chaperonas Moleculares/metabolismo , Proteínas Oncogénicas Virales/metabolismo , Proteínas Represoras/metabolismo , Caspasa 8/metabolismo , Línea Celular Tumoral , Proliferación Celular/fisiología , Humanos , ARN/metabolismo , Transfección/métodos
12.
J Cell Mol Med ; 23(11): 7490-7504, 2019 11.
Artículo en Inglés | MEDLINE | ID: mdl-31493340

RESUMEN

Syphilis is a chronic bacterial infection caused by Treponema pallidum (T pallidum) and the pathogenesis that T pallidum infection induces immunopathological damages in skin and other tissues remains unclear. We have previously reported that recombinant flagellins of T pallidum can elicit IL-6 and IL-8 transcriptions via TLR5 pathway. To identify the domains which induced the pro-inflammatory activity and the importance of the interactions between TLR5 and domains, homology-based modelling and comparative structural analyses revealed that Tpflagellins can combine with TLR5 directly. Deletion mutations showed that the ND1 domain binding to TLR5 is required but not sufficient in TLR5 activation. Moreover, site-directed mutagenesis analysis indicated that the arginine residue (Tpflagellins R89) of the ND1 domain and its adjacent residues (Tpflagellins L93 and E113) constitute a hot spot that elicits IL-6, IL-8 transcriptions and TLR5 activation, and affects the binding of Tpflagellins to TLR5. Taken together, these results give insight into the pathogenesis of T pallidum and may contribute to the future design of Tpflagellins-based therapeutics and syphilis vaccine.


Asunto(s)
Flagelina/genética , Flagelina/metabolismo , Receptor Toll-Like 5/metabolismo , Treponema pallidum/genética , Treponema pallidum/metabolismo , Secuencia de Aminoácidos , Línea Celular , Humanos , Interleucina-6/genética , Interleucina-6/metabolismo , Interleucina-8/genética , Interleucina-8/metabolismo , Unión Proteica/genética , Transducción de Señal/genética , Sífilis/genética , Sífilis/metabolismo , Células THP-1 , Transcripción Genética/genética
13.
J Cell Biochem ; 120(3): 4409-4422, 2019 03.
Artículo en Inglés | MEDLINE | ID: mdl-30260037

RESUMEN

Chlamydia psittaci is an obligate intracellular pathogen with a biphasic developmental life cycle. It is auxotrophic for a variety of essential metabolites and obtains amino acids from eukaryotic host cells. Chlamydia can develop inside host cells within chlamydial inclusions. A pathway secreting proteins from inclusions into the host cellular cytoplasm is the type III secretion system (T3SS). The T3SS is universal among several Gram-negative bacteria. Here, we show that CPSIT_0959 of C. psittaci is expressed midcycle and secreted into the infected cellular cytoplasm via the T3SS. Recombinant CPSIT_0959 possesses cysteine desulfurase and PLP-binding activity, which removes sulfur from cysteine to produce alanine, and helps chlamydial replication. Our study shows that CPSIT_0959 improve the infectivity of offspring elementary bodies and seems to promote the replication by its product. This phenomenon has inhibited by the PLP-dependent enzymes inhibitor. Moreover, CPSIT_0959 increased expression of Bim and tBid, and decreased the mitochondrial membrane potential of host mitochondria to induce apoptosis in the latecycle for release of offspring. These results demonstrate that CPSIT_0959 has cysteine desulfurase and PLP-binding activity and is likely to contribute to apoptosis of the infected cells via a mitochondria-mediated pathway to improve the infectivity of progeny.


Asunto(s)
Proteínas Bacterianas , Liasas de Carbono-Azufre , Chlamydophila psittaci , Psitacosis , Sistemas de Secreción Tipo III , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Liasas de Carbono-Azufre/genética , Liasas de Carbono-Azufre/metabolismo , Chlamydophila psittaci/enzimología , Chlamydophila psittaci/genética , Femenino , Células HeLa , Humanos , Potencial de la Membrana Mitocondrial , Ratones , Ratones Endogámicos BALB C , Mitocondrias/metabolismo , Psitacosis/genética , Psitacosis/metabolismo , Sistemas de Secreción Tipo III/genética , Sistemas de Secreción Tipo III/metabolismo
14.
Appl Microbiol Biotechnol ; 103(2): 941-952, 2019 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-30467705

RESUMEN

Chlamydia psittaci is an obligate intracellular pathogen with a broad host range that can lead to severe infectious disease by transferring from birds to humans. Vaccination has been considered the best way to prevent chlamydial infection; nevertheless, there is currently still no commercially available vaccine that can inhibit the spread of C. psittaci. In previous study, major outer membrane protein (MOMP) of C. psittaci was confirmed to be an appropriate candidate antigen for limiting C. psittaci respiratory infections in a murine model, and plasmid-encoded CPSIT_p6 also has functions similar to those of MOMP in our study. Therefore, according to bioinformatics analysis, we developed a recombinant peptide containing multiple antigenic epitopes from MOMP (24-32, 262-272) and CPSIT_p6 protein (109-119, 173-181) and evaluated the efficacy of peptide immunization. BALB/c mice were inoculated intraperitoneally with the recombinant multi-epitope antigens three times at 2-week intervals and subsequently intranasally infected with C. psittaci. We found that the recombinant multi-epitope antigens induced strong humoral and Th1 cellular immune responses by producing meaningfully high levels of antigen-specific antibodies, interferon-gamma (IFN-γ), or interleukin-2 (IL-2). Vaccination significantly reduced the bacterial burden and the degree of inflammation in the infected lungs and led to lower levels of IFN-γ and IL-6. Furthermore, adoptive transfer of CD4+ splenocytes harvested from the vaccinated mice produced a significantly lower chlamydial load, indicating the importance of the cellular immune response. Therefore, the recombinant multi-epitope antigens may provide the basis for a new peptide-based vaccine against C. psittaci infection.


Asunto(s)
Proteínas de la Membrana Bacteriana Externa/inmunología , Proteínas Bacterianas/inmunología , Vacunas Bacterianas/inmunología , Chlamydophila psittaci/inmunología , Epítopos/inmunología , Psitacosis/prevención & control , Traslado Adoptivo , Animales , Anticuerpos Antibacterianos/sangre , Carga Bacteriana , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas Bacterianas/genética , Vacunas Bacterianas/administración & dosificación , Vacunas Bacterianas/genética , Linfocitos T CD4-Positivos/inmunología , Modelos Animales de Enfermedad , Epítopos/genética , Inmunidad Celular , Inmunidad Humoral , Esquemas de Inmunización , Interferón gamma/metabolismo , Interleucina-6/metabolismo , Leucocitos Mononucleares/inmunología , Ratones Endogámicos BALB C , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Resultado del Tratamiento , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunología
15.
Int J Med Microbiol ; 308(7): 776-783, 2018 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-29891193

RESUMEN

Mycoplasma pneumoniae (M. pneumoniae), as an obligate parasite, has evolved a protective strategy for coping with oxidative challenges caused by M. pneumoniae itself as well as the host immune system. However, to date, few antioxidant enzymes have been identified in mycoplasmas. In this report, we identified a protein encoded by the mpn668 gene from M. pneumoniae with a putative function as an organic hydroperoxide reductase (Ohr). The results indicated that the recombinant 140 amino acid protein, designated rMPN668, displayed hydroperoxidase activity towards both organic (tert-butyl hydroperoxide) and inorganic (hydrogen peroxide) hydroperoxides in the presence of a reducing agent such as dithiothreitol. Moreover, the expression of mpn668 in M. pneumoniae is upregulated in response to oxidative stress. Additionally, homology modeling of MPN668 and a molecular dynamics simulation suggest that both Cys55 and Cys119 form part of the active site of the protein. Mutants in which Cys55 or Cys119 were replaced with a serine lack antioxidant activity, indicating that MPN668 is a Cys-based peroxidase, consistent with it representing a new member of the Ohr family.


Asunto(s)
Farmacorresistencia Bacteriana/genética , Peróxido de Hidrógeno/farmacología , Mycoplasma pneumoniae/genética , Peroxirredoxinas/genética , terc-Butilhidroperóxido/farmacología , Secuencia de Aminoácidos , Regulación Bacteriana de la Expresión Génica , Simulación de Dinámica Molecular , Mycoplasma pneumoniae/efectos de los fármacos , Mycoplasma pneumoniae/enzimología , Estrés Oxidativo/fisiología , Homología de Secuencia de Aminoácido
16.
Exp Cell Res ; 361(1): 46-55, 2017 12 01.
Artículo en Inglés | MEDLINE | ID: mdl-28982539

RESUMEN

Syphilis is a chronic disease caused by Treponema pallidum and the pathogenesis is still unclear. T. pallidum infection induced inflammatory responses are involved in the immunopathological damage in skin and other tissues. Flagellin, the monomeric subunit of bacterial flagella, is a classic pathogen associated molecular patterns (PAMPs) that interacts to TLR5 and induces inflammatory responses. Keratinocytes, as immune sentinels recognize the PAMPs via TLRs, play an important role in skin innate immune response. Matrix metalloproteinases (MMPs) expressed by keratinocytes are involved in skin inflammatory responses and promoting pathogens invasion. In this study, we demonstrate that FlaB1, FlaB2 and FlaB3, the flagellins of T. pallidum, induced MMP-9 and MMP-13 production in human immortalized keratinocytes cell line HaCaT. Silencing of TLR5, but not TLR2 and TLR4 attenuated MMP-9 and MMP-13 expressions induced by T. pallidum flagellins. MMP-9 and MMP-13 expressions were also be abrogated by transfection with a dominant negative (DN) plasmid of MyD88. We also found that treatment of HaCaT cells with FlaB1, FlaB2 and FlaB3 activate the MAPK and NF-κB signaling pathways. Inhibited of ERK, JNK, p38 and NF-κB suppressed MMP-9 expression induced by the FlaB1. MMP-13 expression was found to be suppressed by pretreatment with inhibitors of ERK, JNK and NF-κB, but not p38. These findings demonstrate that T. pallidum flagellins (FlaB1, FlaB2 or FlaB3) can stimulate MMP-9 and MMP-13 expression through TLR5 and MAPK/NF-κB signaling pathways in human epidermal keratinocytes, which could contribute to the pathogenesis of T. pallidum infection.


Asunto(s)
Flagelina/metabolismo , Queratinocitos/metabolismo , Metaloproteinasa 13 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , FN-kappa B/metabolismo , Receptor Toll-Like 5/metabolismo , Células Cultivadas , Humanos , Queratinocitos/citología , Transducción de Señal , Treponema pallidum/metabolismo
17.
BMC Microbiol ; 17(1): 153, 2017 Jul 11.
Artículo en Inglés | MEDLINE | ID: mdl-28693414

RESUMEN

BACKGROUND: Chlamydia pneumoniae (C. pneumoniae) is pathogenic to humans, by causing pulmonary inflammation or bronchitis in both adolescents and young adults. However, the molecular signals linking C. pneumoniae components to inflammation remain elusive. This study was to investigate the effect of Chlamydia-specific Cpn0423 of C. pneumoniae on C. pneumoniae-mediated inflammation. RESULTS: Cpn0423 was detected outside of C. pneumoniae inclusions, which induced production of several cytokines including macrophage inflammatory protein-2 (MIP-2) and interleukins (ILs). Production of the Cpn0423-induced cytokines was markedly reduced in cells pretreated with NOD2-siRNA, but not with negative control oligonucleotides. Mice treated with Cpn0423 through intranasal administration exhibited pulmonary inflammation as evidenced by infiltration of inflammatory cells, increased inflammatory scores in the lung histology, recruitment of neutrophils and increased cytokines levels in the BALF. CONCLUSION: Cpn0423 could be sensed by NOD2, which was identified as an essential element in a pathway contributing to the development of C. pneumoniae -mediated inflammation.


Asunto(s)
Proteínas Bacterianas/inmunología , Infecciones por Chlamydophila/inmunología , Chlamydophila pneumoniae/inmunología , Mediadores de Inflamación/inmunología , Proteína Adaptadora de Señalización NOD2/inmunología , Neumonía Bacteriana/microbiología , Animales , Proteínas Bacterianas/genética , Quimiocina CXCL2/genética , Quimiocina CXCL2/inmunología , Infecciones por Chlamydophila/genética , Infecciones por Chlamydophila/microbiología , Chlamydophila pneumoniae/genética , Humanos , Interleucinas/genética , Interleucinas/inmunología , Pulmón/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Proteína Adaptadora de Señalización NOD2/genética , Neumonía Bacteriana/genética , Neumonía Bacteriana/inmunología
18.
J Med Virol ; 88(10): 1804-13, 2016 10.
Artículo en Inglés | MEDLINE | ID: mdl-26991077

RESUMEN

Epstein-Barr virus (EBV) is a human oncogenic herpesvirus associated with lymphoma and nasopharyngeal carcinoma. Because the susceptible hosts of EB virus are limited to human and cotton-top tamarins (Saguinus oedipus), there have been no appropriate animal models until the lymphoma model induced by EBV in human peripheral blood lymphocyte (hu-PBL)/SCID chimeric mice was reported. However, it is still controversial whether the EBV-associated lymphoma induced in hu-PBL/SCID mice is a monoclonal tumor. In this study, we transplanted normal human peripheral blood lymphocytes (hu-PBL) from six donors infected with EBV into SCID mice to construct hu-PBL/SCID chimeric mice. The induced tumors were found in the mediastinum or abdominal cavity of SCID mice. Microscopic observation exhibited tumor cells that were large and had a plasmablastic, centroblastic or immunoblastic-like appearance. Immunophenotyping assays showed the induced tumors were LCA-positive, CD20/CD79a-positive (markers of B cells), and CD3/CD45RO-negative (markers of T cells). A human-specific Alu sequence could be amplified by Alu-PCR. This confirmed that induced tumors were B-cell lymphomas originating from the transplanted human lymphocytes rather than mouse cells. EBER in situ hybridization detected positive signals in the nuclei of the tumor cells. Expression of EBV-encoded LMP1, EBNA-1, and EBNA-2 in the tumors was significantly positive. PCR-based capillary electrophoresis analysis of IgH gene rearrangement revealed a monoclonal peak and single amplification product in all six cases of induced tumors. This indicated that EBV can induce monoclonal proliferation of human B lymphocytes and promotes the development of lymphoma. J. Med. Virol. 88:1804-1813, 2016. © 2016 Wiley Periodicals, Inc.


Asunto(s)
Infecciones por Virus de Epstein-Barr/complicaciones , Reordenamiento Génico , Cadenas Pesadas de Inmunoglobulina/genética , Inmunofenotipificación , Linfoma de Células B/virología , Elementos Alu , Animales , Modelos Animales de Enfermedad , Antígenos Nucleares del Virus de Epstein-Barr/genética , Herpesvirus Humano 4 , Humanos , Hibridación in Situ , Transfusión de Linfocitos , Ratones SCID , Proteínas de la Matriz Viral/genética , Proteínas Virales/genética
19.
Sex Transm Dis ; 43(5): 310-6, 2016 May.
Artículo en Inglés | MEDLINE | ID: mdl-27100768

RESUMEN

BACKGROUND: Over the past decade, the incidence of syphilis and widespread macrolide resistance in its etiological agent, Treponema pallidum subsp. pallidum, have become a major health concern across countries, including China. Regional trends in subtypes and antibiotic resistance can be monitored effectively by molecular surveillance programs. In this study, whole blood samples were used to assess circulating T. pallidum strains collected from various regions of Hunan, China, between 2013 and 2015. METHODS: Traditional polymerase chain reaction, targeting polA, tpp47, bmp, and tp0319 genes, was used as preliminary screening assay. About 455 polymerase chain reaction-positive specimens were obtained from 2253 whole blood samples of patients with secondary or latent syphilis. Molecular subtyping was performed using a Centers for Disease Control and Prevention-based typing method combined with an analysis of the variable region of tp0548 gene. Resistance to macrolides was analyzed by examining point mutations in 23S rRNA, and the presence of the G1058C point mutation within 16S rRNA associated with decreased susceptibility to doxycycline was assessed. RESULTS: Circulating T. pallidum strains were resolved into 32 subtypes, among which subtype 14d/f was predominant. A2059G mutation in 23S rRNA, and the G1058C mutation in 16S rRNA was absent, but the prevalence of A2058G mutation in 23S rRNA was 97.5%. CONCLUSIONS: We found that it is possible to use whole blood to evaluate molecular subtypes and monitor antibiotic resistance in circulating T. pallidum strains, especially when chancres are absent. High frequency of macrolide-resistant T. pallidum indicates that macrolide antibiotics, such as azithromycin, should be avoided as a treatment option for syphilis in Hunan, China.


Asunto(s)
Sífilis/epidemiología , Treponema pallidum/clasificación , Adulto , Anciano , Anciano de 80 o más Años , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Bacteriemia/microbiología , China/epidemiología , Farmacorresistencia Bacteriana/genética , Femenino , Genotipo , Humanos , Incidencia , Macrólidos/farmacología , Macrólidos/uso terapéutico , Persona de Mediana Edad , Tipificación Molecular , Mutación Puntual , Prevalencia , ARN Ribosómico 16S/genética , ARN Ribosómico 23S/genética , Sífilis/tratamiento farmacológico , Sífilis/microbiología , Sífilis Latente/tratamiento farmacológico , Sífilis Latente/epidemiología , Sífilis Latente/microbiología , Treponema pallidum/efectos de los fármacos , Treponema pallidum/genética , Treponema pallidum/aislamiento & purificación , Adulto Joven
20.
Appl Microbiol Biotechnol ; 100(14): 6385-6393, 2016 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-27052378

RESUMEN

Chlamydia psittaci is a zoonotic pathogen with a broad host range that can lead to severe respiratory and systemic disease in humans. Currently, an effective commercial vaccine against C. psittaci infection is not available. The chlamydial plasmid is an important virulence factor and encodes plasmid proteins that play important roles in chlamydial infection and the corresponding immune response. In this study, we assessed the efficacy of vaccination with plasmid proteins at preventing C. psittaci lung infection in a murine model. BALB/c mice were immunized intraperitoneally, three times at 2-week intervals, with purified recombinant CPSIT_p8 protein and then infected with C. psittaci. Immunization significantly decreased chlamydial load in the lungs of infected mice, resulted in a lower level of IFN-γ, and reduced the extent of inflammation. In vivo or in vitro neutralization of C. psittaci with sera collected from immunized mice did not reduce the amount of viable C. psittaci in the lungs of mice, indicating that CPSIT_p8-specific antibodies do not have neutralizing capacity. Furthermore, confocal fluorescence microscopy using a mouse anti-CPSIT_p8 antibody revealed that CPSIT_p8 was localized inside the inclusion of C. psittaci 6BC-infected cells. Our results demonstrate that CPSIT_p8 protein induces significant protective immunity against challenge with C. psittaci in mice and represents a promising new vaccine candidate for the prevention of C. psittaci infection.


Asunto(s)
Proteínas Bacterianas/inmunología , Vacunas Bacterianas/inmunología , Infecciones por Chlamydia/prevención & control , Chlamydophila psittaci/inmunología , Plásmidos/inmunología , Proteínas Recombinantes/inmunología , Animales , Anticuerpos Antibacterianos/sangre , Carga Bacteriana , Proteínas Bacterianas/genética , Infecciones por Chlamydia/inmunología , Clonación Molecular , Modelos Animales de Enfermedad , Femenino , Regulación de la Expresión Génica , Células HeLa , Humanos , Inmunización , Interferón gamma/inmunología , Ratones , Ratones Endogámicos BALB C , Proteínas Recombinantes/genética
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