Polymorphic Membrane Protein Research Articles

Clustering of Polymorphic Membrane Protein E Clade in Chlamydia trachomatis Lineages from Men Who Have Sex with Men.

Several Chlamydia trachomatis lineages identified through outer membrane protein A genotyping or multilocus sequence typing have been circulating worldwide among men who have sex with men. In a study in Tokyo, Japan, we demonstrate that such lineages commonly belong to a specific polymorphic membrane protein E clade across genotypes.

Sequence, structure prediction, and epitope analysis of the polymorphic membrane protein family in Chlamydia trachomatis.

The polymorphic membrane proteins (Pmps) are a family of autotransporters that play an important role in infection, adhesion and immunity in Chlamydia trachomatis. Here we show that the characteristic GGA(I,L,V) and FxxN tetrapeptide repeats fit into a larger repeat sequence, which correspond to the coils of a large beta-helical domain in high quality structure predictions. Analysis of the protein using structure prediction algorithms provided novel insight to the chlamydial Pmp family of proteins. While the tetrapeptide motifs themselves are predicted to play a structural role in folding and close stacking of the beta-helical backbone of the passenger domain, we found many of the interesting features of Pmps are localized to the side loops jutting out from the beta helix including protease cleavage, host cell adhesion, and B-cell epitopes; while T-cell epitopes are predominantly found in the beta-helix itself. This analysis more accurately defines the Pmp family of Chlamydia and may better inform rational vaccine design and functional studies.

The greenbeard gene tgrB1 regulates altruism and cheating in Dictyostelium discoideum

Greenbeard genetic elements encode rare perceptible signals, signal recognition ability, and altruism towards others that display the same signal. Putative greenbeards have been described in various organisms but direct evidence for all the properties in one system is scarce. The tgrB1-tgrC1 allorecognition system of Dictyostelium discoideum encodes two polymorphic membrane proteins which protect cells from chimerism-associated perils. During development, TgrC1 functions as a ligand-signal and TgrB1 as its receptor, but evidence for altruism has been indirect. Here, we show that mixing wild-type and activated tgrB1 cells increases wild-type spore production and relegates the mutants to the altruistic stalk, whereas mixing wild-type and tgrB1-null cells increases mutant spore production and wild-type stalk production. The tgrB1-null cells cheat only on partners that carry the same tgrC1-allotype. Therefore, TgrB1 activation confers altruism whereas TgrB1 inactivation causes allotype-specific cheating, supporting the greenbeard concept and providing insight into the relationship between allorecognition, altruism, and exploitation.

In Silico Annotation and Immunoinformatics Guided Epitope Mapping of Potential Antigenic Proteins of Trichomonas foetus

Bovine trichomonosis is one of the neglected tropical diseases of cattle that is resulting in severe reproductive failure. With present knowledge, disease diagnosis and maintaining the infected animals in the quarantine are the only available strategies. Several spillover incidences of Trichomonas foetus had also resulted in zoonotic transmission to humans. In spite of above circumstances, till date there are no point of care diagnostics developed for screening bovine trichomoniasis in cattle. In the light of above circumstances, there exists a demand for cost-effective diagnostic kits to be provided to farming community. This current study highlights evaluation of few surface proteins of Trichomonas foetus for the suitability as sero-diagnostic markers. Few target Proteins such as Adhesin, Immuno-dominant variable surface antigen-like protein, Polymorphic membrane protein - like protein, GP-63-like (Clan MA, family M8) protein and Hypothetical protein (OHS95735.1) were evaluated for suitable pH, Signal Peptide, protein glycosylation pattern using freely available Bioinformatics tools. Mapping of potential epitopes of all the target proteins was done using immunoinformatics tools. Among the above proteins, GP63-like protein, immuno-dominant variable antigenic domain-like protein, and polymorphic membrane protein-like proteins are most suitable as diagnostic targets, owing to their higher levels of glycosylation, large epitope domains, and showing structural similarities with the domains of known toxic proteins. On the other hand, adhesin protein has the potential to be exploited as a vaccine candidate. The above proteins are suitable to be expressed in suitable host system and validated the immunogenic potential by animal inoculation and by testing with the real samples.

Interferon-γ Responses to Chlamydia trachomatis Vaccine Candidate Proteins in Women With Different Chlamydia Outcomes.

Chlamydia trachomatis testing and treatment strategies have not decreased infection rates, justifying need for a chlamydia vaccine. A murine study showed that a vaccine consisting of major outer membrane protein (MOMP) and polymorphic membrane proteins (Pmps) E, F, G, and H elicited protective immunity; studies on human cellular immune responses to Pmps are sparse. Interferon gamma (IFN-γ) responses to these 5 proteins were measured by ELISPOT in peripheral blood mononuclear cells from women returning for treatment of a positive chlamydia test. Responses were compared in those with spontaneous chlamydia clearance versus persisting infection at baseline and no reinfection versus reinfection at a 3-month follow-up visit. IFN-γ response to 1 or more proteins was detected in 39% at baseline and 51.5% at follow-up, most often to PmpE and MOMP. IFN-γ responses to MOMP were detected less often at follow-up versus baseline in women with reinfection, but were maintained in those without reinfection. Women with spontaneous clearance had a higher magnitude of IFN-γ response to PmpE and MOMP. IFN-γ responses to these 5 C. trachomatis vaccine candidate proteins were heterogenous and primarily directed against MOMP and PmpE. Spontaneous chlamydia clearance and absence of reinfection may be clinical correlates of protection.

Draft genomes of novel avian Chlamydia abortus strains from Australian Torresian crows (Corvus orru) shed light on possible reservoir hosts and evolutionary pathways.

Chlamydia abortus, an obligate intracellular bacterium, is a major causative agent of reproductive loss in ruminants, with zoonotic potential. Though this pathogen is primarily known to infect livestock, recent studies have detected and isolated genetically distinct avian strains of C. abortus from wild birds globally. Before this study, only five avian C. abortus genomes were publicly available. Therefore, we performed culture-independent probe-based whole-genome sequencing on clinical swabs positive for avian C. abortus obtained from Australian Torresian crows (Corvus orru) in 2019 and 2020. We successfully obtained draft genomes for three avian C. abortus strains (C1, C2 and C3), each comprising draft chromosomes with lengths of 1 115 667, 1 120 231 and 1 082 115 bp, and associated 7 553 bp plasmids, with a genome completeness exceeding 92 %. Molecular characterization revealed that these three strains comprise a novel sequence type (ST333), whilst phylogenetic analyses placed all three strains in a cluster with other avian C. abortus genomes. Interestingly, these three strains share a distant genomic relation (2693 single nucleotide variants) with the reference strain 15-58d/44 (ST152), isolated from a Eurasian magpie (Pica pica) in Poland, highlighting the need for more publicly available genomes. Broad comparative analyses with other avian C. abortus genomes revealed that the three draft genomes contain conserved Chlamydia genomic features, including genes coding for type III secretion system and polymorphic membrane proteins, and potential virulence factors such as the large chlamydial cytotoxin, warranting further studies. This research provides the first avian C. abortus draft genomes from Australian birds, highlighting Torresian crows as novel reservoir hosts for these potential pathogens, and demonstrates a practical methodology for sequencing novel Chlamydia genomes without relying on traditional cell culture.

Evaluation of three formulations based on Polymorphic membrane protein D in mice infected with Chlamydia trachomatis.

The significant impact of Chlamydia trachomatis(Ct) infections worldwide highlights the need to develop a prophylactic vaccine that elicits effective immunity and protects the host from the immunopathological effects of Ct infection. The aim of this study was to evaluate a vaccine based on a fragment of the Polymorphic membrane protein D (FPmpD) of C. trachomatis as an immunogen using a heterologous DNA prime-protein boost strategy in female mice Three different formulations were evaluated as protein boost: free recombinant FPmpD (rFPmpD) or rFPmpD formulated with a liposomal adjuvant alternatively supplemented with CpG or a cationic gemini lipopeptide as immunostimulants. The three candidates induced an increase in the cervicovaginal and systemic titers of anti-rFPmpD antibodies in two strains of mice (BALB/c and C57BL/6), with no evidence of fertility alterations. The three formulations induced a rapid and robust humoral immune response upon the Ct challenge. However, the booster with free rFPmpD more efficiently reduced the shedding of infective Ct and prevented the development of immunopathology. The formulations containing adjuvant induced a strong inflammatory reaction in the uterine tissue. Hence, the prime-boost strategy with the adjuvant-free FPmpD vaccine formulation might constitute a promissory candidate to prevent C. trachomatis intravaginal infection.

A structural foundation for studying chlamydial polymorphic membrane proteins.

Polymorphic membrane proteins (Pmps) are chlamydial cell surface proteins with signature repeating FxxN and GGA (I, V, L) tetrapeptide motifs. These autotransporter proteins are key immune targets and mediators of host-microbe interactions but by unknown mechanisms. AlphaFold predictions reveal that Pmp passenger domains have two distinct β-helical parts: a C-terminal globular "Pmp middle region" that forms six β-helical rungs, and an N-terminal highly-regular "Pmp repeat region" that is from 9 to 25 rungs long. In the repeat region, paired FxxN and GGA (I, V, L) motifs - or close variants - are fully buried and packed together to form novel β-helical rungs from which variable-length loops extend out from one face. Similar motifs in proteins from diverse organisms are predicted to form analogous structures. In Pmps, the structures imply that the tetrapeptide motifs are not directly involved in host-cell adhesion, but position protein loops on a common face where they can interact with ligands important for infection. These structural descriptions provide a guide for mutagenesis studies to identify those parts of Pmps involved in infection.IMPORTANCEInfections by bacteria in the genus Chlamydia cause a range of widespread and potentially debilitating conditions in humans and other animals. We analyzed predicted structures of a family of proteins that are potential vaccine targets found in all Chlamydia spp. Our findings deepen the understanding of protein structure, provide a descriptive framework for discussion of the protein structure, and outline regions of the proteins that may be key targets in host-microbe interactions and anti-chlamydial immunity.

LA PREDICCIÓN IN SILICO DETERMINA LA PRESENCIA DE EPÍTOPOS INMUNOGÉNICOS ALTAMENTE ESPECÍFICOS EN FRAGMENTOS DE LA PROTEÍNA DE LA MEMBRANA POLIMÓRFICA (PMP17G) DE Chlamydia abortus

<p><strong>Background.</strong> Ovine Enzootic Abortion is a contagious infectious disease caused by a Gram negative and obligate intracellular bacterium, <em>Chlamydia abortus</em>. For field diagnosis, commercial serological tests are used; however, some of these tests show low sensitivity and specificity rates, due to the cross-reactions that the antigens used have against other pathogens. For the most accurate diagnosis, it is necessary to develop tests with more specific antigens such as polymorphic membrane proteins (Pmp's), that allow to determine the presence of specific epitopes using new technologies. <strong>Objective.</strong> To determine <em>in silico</em> the presence of epitopes with specific immunogenic potential against <em>Chlamydia abortus</em> of two fragments of the PMP17G protein. <strong>Methodology.</strong> The cloning and sequencing of the fragments was carried out from a field isolate of <em>Chlamydia abortus</em>, and from the analysis of these sequences, with the help of two bioinformatics software’s. <strong>Results.</strong> Several epitopes from <em>Chlamydia abortus</em> were found, rPOMP90-3 (eight epitopes) and rPOMP90-4 (one epitope). <strong>Implications.</strong> Bioinformatics analysis indicated that both fragments of the protein have the capacity to activate the immune system, which would be useful for the development of diagnostic kits and immunogens. <strong>Conclusions.</strong> The <em>in silico</em> analysis allowed to efficiently predict and identify specific epitopes against <em>Chlamydia abortus</em> in both fragments of the protein. </p>

Expression comparisons of defense-related genes in resistant and susceptible chickpea cultivars in response to Ascochyta rabiei

Ascochyta blight caused by Ascochyta rabiei is a destructive disease affecting chickpea crops worldwide. Numerous studies have sought to identify resistance genes involved in the defense response against Ascochyta blight, but the function of potential resistant genes across different chickpea genetic backgrounds remains poorly understood. This study evaluated the potential role of 16 genes involved in defense responses in partially resistant (ILC482) and susceptible (Sarı98) chickpea cultivars. The leaf samples were collected from 2-week-old seedlings at 6, 12, 24, 48, and 72 h post-inoculation (hpi) with A. rabiei and analyzed for differential expression of defense genes using qRT-PCR. Twelve out of 16 genes were differentially regulated between chickpea cultivars. The qRT-PCR analysis indicated that the expression of defense genes was significantly higher in the partially resistant cultivar than that in the susceptible cultivar. The time point of the highest upregulation ratio for defense genes was variable between the cultivars. The expression of glutathione S-transferase, serine/threonine-protein kinase, WRKY gene (CaWRKY16), and wall associated-receptor kinase genes reached maximum levels in the resistant cultivar at 6 hpi, while antimicrobial peptide precursor (SNAKIN2), polymorphic antigen membrane protein, ethylene receptor-like sequences (CaETR1), and two nucleotide-binding site-leucine-rich repeat genes showed the highest activity at 24 hpi. In particular, CaWRKY16, CaWRKY50, glutathione S-transferase, CaETR1, and wall-associated-receptor kinase genes provided valuable information for assessing chickpea cultivars with different resistance levels. The results indicated that the expression changes of these genes involved in defense pathways play a crucial role in enhanced resistance of chickpea plants against Ascochyta blight and, therefore could be potential candidate genes for future breeding studies.

Whole Genome Sequencing and Comparative Genomic Analysis of Chlamydia gallinacea Field Strains Isolated from Poultry in Poland.

Chlamydia gallinacea is an intracellular bacterium belonging to the Chlamydiaceae family. Poultry is considered to be the major reservoir of this agent, which has worldwide distribution and a particularly consistent worldwide occurrence in chicken flocks. The bacterium has been linked to respiratory disease in humans but without definitive confirmation; nevertheless, while it has not been proved to be the cause of human respiratory disease, a recent report from Italy verified its bird-to-human transmission. This aspect being significant for public health, more research is needed to gain insight into the infection biology of C. gallinacea. In this study, the genomes of eleven novel C. gallinacea field strains from different regions of Poland were analyzed comparatively. It was confirmed that C. gallinacea strains are closely related, with at least 99.46% sequence identity. They possess a conservative genome structure involving the plasticity zone with a complete cytotoxin, the type three secretion system, inclusion membrane proteins, polymorphic membrane proteins, hctA and hctB histone-like proteins, and the chlamydial protease-like activating factor exoenzyme, as well as plasmids. Genetic diversity seems to be restricted. However, some genetic loci, such as ompA and multi-locus sequence typing target genes, are diverse enough to enable high-resolution genotyping and epidemiological tracing.

Genomic analysis of 61 Chlamydia psittaci strains reveals extensive divergence associated with host preference

BackgroundChlamydia (C.) psittaci, the causative agent of avian chlamydiosis and human psittacosis, is a genetically heterogeneous species. Its broad host range includes parrots and many other birds, but occasionally also humans (via zoonotic transmission), ruminants, horses, swine and rodents. To assess whether there are genetic markers associated with host tropism we comparatively analyzed whole-genome sequences of 61 C. psittaci strains, 47 of which carrying a 7.6-kbp plasmid.ResultsFollowing clean-up, reassembly and polishing of poorly assembled genomes from public databases, phylogenetic analyses using C. psittaci whole-genome sequence alignment revealed four major clades within this species. Clade 1 represents the most recent lineage comprising 40/61 strains and contains 9/10 of the psittacine strains, including type strain 6BC, and 10/13 of human isolates. Strains from different non-psittacine hosts clustered in Clades 2– 4. We found that clade membership correlates with typing schemes based on SNP types, ompA genotypes, multilocus sequence types as well as plasticity zone (PZ) structure and host preference. Genome analysis also revealed that i) sequence variation in the major outer membrane porin MOMP can result in 3D structural changes of immunogenic domains, ii) past host change of Clade 3 and 4 strains could be associated with loss of MAC/perforin in the PZ, rather than the large cytotoxin, iii) the distinct phylogeny of atypical strains (Clades 3 and 4) is also reflected in their repertoire of inclusion proteins (Inc family) and polymorphic membrane proteins (Pmps).ConclusionsOur study identified a number of genomic features that can be correlated with the phylogeny and host preference of C. psittaci strains. Our data show that intra-species genomic divergence is associated with past host change and includes deletions in the plasticity zone, structural variations in immunogenic domains and distinct repertoires of virulence factors.

The Polymorphic Membrane Protein G Has a Neutral Effect and the Plasmid Glycoprotein 3 an Antagonistic Effect on the Ability of the Major Outer Membrane Protein to Elicit Protective Immune Responses against a Chlamydia muridarum Respiratory Challenge.

Chlamydia trachomatis is the most common bacterial sexually transmitted pathogen. The number of chlamydial infections continuous to increase and there is an urgent need for a safe and efficacious vaccine. To assess the ability of the Chlamydia muridarum polymorphic membrane protein G (PmpG) and the plasmid glycoprotein 3 (Pgp3) as single antigens, and in combination with the major outer-membrane protein (MOMP) to induce protection, BALB/c mice were immunized utilizing CpG-1826 and Montanide ISA 720 VG as adjuvants. Following vaccination with MOMP, significant humoral and cell-mediated immune responses were observed, while immunization with PmpG, or Pgp3, elicited weaker immune responses. Weaker immune responses were induced with MOMP+Pgp3 compared with MOMP alone. Following the intranasal challenge with C. muridarum, mice vaccinated with MOMP showed robust protection against body-weight loss, inflammatory responses in the lungs and number of Chlamydia recovered from the lungs. PmpG and Pgp3 elicited weaker protective responses. Mice immunized with MOMP+PmpG, were no better protected than animals vaccinated with MOMP only, while Pgp3 antagonized the protection elicited by MOMP. In conclusion, PmpG and Pgp3 elicited limited protective immune responses in mice against a respiratory challenge with C. muridarum and failed to enhance the protection induced by MOMP alone. The virulence of Pgp3 may result from its antagonistic effect on the immune protection induced by MOMP.

Comparative analysis of two genomes of Chlamydia pecorum isolates from an Alpine chamois and a water buffalo

BackgroundTo date, whole genome sequencing has been performed mainly for isolates of Chlamydia trachomatis, C. pneumoniae, C. psittaci and C. abortus, but only a few isolates of C. pecorum have been entirely sequenced and this makes it difficult to understand its diversity and population structure. In this study the genome of two C. pecorum strains isolated from the lung of an Alpine chamois affected with pneumonia (isolate PV7855) and the brain of a water buffalo affected with meningoencephalomyelitis (isolate PV6959), were completely sequenced with MiSeq system (Illumina) and analyzed in their most polymorphic regions.ResultsThe genome length and GC content of the two isolates were found to be consistent with other C. pecorum isolates and the gene content of polymorphic membrane proteins and plasticity zone was found to be very similar. Some differences were observed in the phospholipase genes for both isolates and in the number of genes in the plasticity zone, such as the presence of some hypothetical proteins in PV6959, not present in any other genomes analyzed in this study. Interestingly, PV6959 possesses an extra pmp and has an incomplete tryptophan biosynthesis operon. Plasmids were detected in both isolates.ConclusionsGenome sequencing of the two C. pecorum strains did not reveal differences in length and GC content despite the origin from different animal species with different clinical disease. In the plasticity zone, the differences in the genes pattern might be related to the onset of specific symptoms or infection of specific hosts. The absence of a tryptophan biosynthesis pathway in PV6959 may suggest a strict relationship between C. pecorum and its host.

ICAM-1-binding Plasmodium falciparum erythrocyte membrane protein 1 variants elicits opsonic-phagocytosis IgG responses in Beninese children

Members of the highly polymorphic Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) family expressed on the surface of infected erythrocytes (IEs) are important virulence factors, which mediate vascular adhesion of IEs via endothelial host receptors and are targets of naturally acquired immunity. The PfEMP1 family can be divided into clinically relevant subgroups, of which some bind intercellular adhesion molecule 1 (ICAM-1). While the acquisition of IgG specific for ICAM-1-binding DBLβ domains is known to differ between PfEMP1 groups, its ability to induce antibody-dependent cellular phagocytosis (ADCP) is unclear. We therefore measured plasma levels of DBLβ-specific IgG, the ability of such IgG to inhibit PfEMP1-binding to ICAM-1, and its ability to opsonize IEs for ADCP, using plasma from Beninese children with severe (SM) or uncomplicated malaria (UM). IgG specific for DBLβ from group A and B ICAM-1-binding PfEMP1 were dominated by IgG1 and IgG3, and were similar in SM and UM. However, levels of plasma IgG inhibiting ICAM-1-binding of group A DBLβ of PFD1235w was significantly higher in children with UM than SM, and acute UM plasma induced a higher ADCP response than acute SM plasma.

Evaluation of the protective efficacy of a novel rVCG-based Chlamydia abortus vaccine in a pregnant mouse model

Abstract Chlamydia abortus (Cab) is the cause of ovine enzootic abortion (OEA) in small ruminants and poses a zoonotic risk to pregnant women, which may result in spontaneous abortion. Although antibiotics are effective against Cab infection, most infections are asymptomatic, suggesting alternative strategies may be required for controlling these infections. We tested the ability of a recombinant Vibrio cholerae ghost (rVCG)-based Cab subunit vaccine expressing the truncated Cab polymorphic membrane protein 18D (rVCG-Pmp18.3) to protect against Cab-induced abortion in a pregnant mouse model. Thus, groups of female C57bl/6J mice were immunized intranasally (IN) with rVCG-Pmp18.3 and mated with males 3 weeks post immunization. Immunized and control mice were infected IN or transcervically (TC) at day 10 of pregnancy and the number of pups per mouse was enumerated 7 days postpartum. Mice were sacrificed on day 8 and Cab burden in the lungs and spleens was determined. The results showed IN immunization protected mice against abortion and neonatal death while IN infected control mice lost 20% of their litter and 60% of TC infected controls lost their entire litter 7 days post-parturition. Immunized mice completely cleared the infection in the lungs and showed a significant reduction (p ≤0.0001) in bacterial burden in the spleen compared to non-immunized, infected controls. Furthermore, immunized mice were protected from uterine dilation compared to 60% of non-immunized controls that developed tubal pathology. These results demonstrate the ability of rVCG-Pmp18.3 to prevent respiratory/systemic infection and protect against Cab-induced abortion and tubal pathologies in mice, suggesting its potential development as a vaccine for ruminants. Supported by PHS grant R01AI41231 and NIH R01AI26897

A Novel Intranasal Vaccine With PmpGs + MOMP Induces Robust Protections Both in Respiratory Tract and Genital System Post Chlamydia psittaci Infection

Chlamydia psittaci (C. psittaci) is a crucial zoonotic pathogen that causes severe respiratory and reproductive system disease in humans and animals. In our pioneer study, polymorphic membrane protein G (PmpG) mediated attachment to host cells as the adhesions and induced immunity against C. psittaci infection. We hypothesize that multiple PmpG antigens adjuvanted with Vibrio cholerae ghost (VCG) and chitosan gel might trigger full protection via the intranasal route (i.n). In the present study, 40 SPF chickens were randomly divided into four groups, including the PmpGs + MOMP group (i.n), major outer membrane protein (MOMP) group (i.n), PmpGs (Pmp17G + Pmp20G + Pmp21G) group (i.n), and control groups (VCG + chitosan gel) (i.n). Post twice immunizations, the PmpGs + MOMP group yielded highly level-specific IgG, IgA antibodies, and lymphocyte proliferation. As for cytokines, IFN-γ expression was upregulated significantly, while IL-10 concentration was downregulated in the PmpGs + MOMP group compared with other groups. Post challenge, exudate inflammations in air sacs, bacterial loads in lungs, and bacterial shedding in throat swabs were reduced significantly in the PmpGs + MOMP group. In the second experiment, 100 breeder ducks were divided into the PmpGs + MOMP group (i.n), the commercial MOMP group (via intramuscular injection, i.m), the inactivated EBs group (i.n), and the control group (i.n), 25 ducks per group. Post challenge, the reduced egg production recovered soon in the inactivated EBs group and the PmpGs + MOMP group. Moreover, the aforementioned two groups induced higher robust IgG antibodies, lymphocyte proliferation, and IFN-γ secretions than the commercial MOMP vaccine did. Postmortem, lower bacterial loads of spleens were determined in the PmpGs + MOMP group and the inactivated EBs group. However, bacterial clearance of follicular membranes and shedding from the vaginal tract were not significant differences among the three tested groups. Furthermore, the PmpGs + MOMP group induced lower inflammations in the follicles and oviducts. Based on the above evidence, the combination of PmpGs and MOMP adjuvanted with chitosan gel and VCG via intranasal route could induce full protection both in the respiratory system and genital tract post C. psittaci infection. More importantly, the combination antigens are superior to the inactivated EBs antigen due to no contamination to the environment and less genital inflammation. The combination of PmpGs + MOMP adjuvanted with VCG and chitosan gel might be a promising novel vaccine by blocking C. psittaci infection from animals to human beings.

Expression Level of the mip, pmp18D, and ompA Genes in Chlamydia abortus Isolated from Aborted Ewes

In this manuscript, we report the proteins macrophage infectivity potentiator (mip, CAB080), major outer membrane protein (momp, CAB048), and polymorphic outer membrane protein (pmp18D, CAB776) that are expressed in different times of pregnancy in mice infected with Chlamydia abortus. Enzootic abortion of ewes (EAE) by C. abortus, an obligate intracellular pathogen, is a critical zoonotic disease-causing significant economic loss to livestock farming globally. This study was carried out for the detection and characterization of macrophage infectivity potentiator (mip, CAB080), major outer membrane protein (momp, CAB048), and polymorphic outer membrane protein (pmp18D, CAB776) using RT-qPCR. These proteins are believed to be expressed as virulence factors in C. abortus isolated from aborted ewes. BALB/c mice (pregnant and nonpregnant) were used as an animal model to be injected intraperitoneally with C. abortus culture in Vero cells since the endometrial lymphoid tissues of these animals resembles that of ewes. Also, the short duration of pregnancy in mice makes them a suitable animal model for obstetric studies. Tissue samples were taken from the mice after 10, 15, and 20 days of pregnancy to compare the expression of the genes mip, pmp18D, and ompA. Transcription level was quantified using RT-qPCR, the GAPDH transcription quantification, as a normalization signal. Abortion occurred in pregnant mice, and apparent differences between the transcriptional levels of the mip, pmp18D, and ompA genes in the samples taken during different time intervals of pregnancy were not observed (p > 0.05). The result indicated that the three bacterial genes, mip, pmp18D, and ompA, play a role as virulence factors in abortion and are differentially expressed in pregnant and nonpregnant animals. Inactivation of the genes is suggested to confirm the hypothesis.

Serum antibodies to surface proteins of Chlamydia trachomatis as candidate biomarkers of disease: results from the Baltimore Chlamydia Adolescent/Young Adult Reproductive Management (CHARM) cohort.

We previously observed that the nine-member family of autotransported polymorphic membrane proteins (Pmps) of Chlamydia trachomatis is variably expressed in cell culture. Additionally, C. trachomatis-infected patients display variable Pmp-specific serum antibody profiles indirectly suggesting expression of unique Pmp profiles is an adaptive response to host-specific stimuli during infection. Here, we propose that the host response to Pmps and other outer surface proteins may correlate with disease severity. This study tests this hypothesis using an ELISA that measures serum IgG antibodies specific for the nine C. trachomatis Pmp subtypes and four immunodominant antigens (MOMP, OmcB, Hsp60, ClpP) in 265 participants of the Chlamydia Adolescent/Young Adult Reproductive Management (CHARM) cohort. More C. trachomatis-infected females displayed high Pmp-specific antibody levels (cut-off Indexes) than males (35.9%-40.7% of females vs. 24.2%-30.0% of males), with statistical significance for PmpC, F and H (P < 0.05). Differences in Pmp-specific antibody profiles were not observed between C. trachomatis-infected females with a clinical diagnosis of pelvic inflammatory disease (PID) and those without. However, a statistically significant association between high levels of OmcB-specific antibody and a PID diagnosis (P< 0.05) was observed. Using antibody levels as an indirect measure of antigen expression, our results suggest that gender- and/or site-specific (cervix in females vs. urethra in males) stimuli may control pmp expression in infected patients. They also support the possible existence of immune biomarkers of chlamydial infection associated with disease and underline the need for high resolution screening in human serum.

Polymorphic Membrane Protein 17G of Chlamydia psittaci Mediated the Binding and Invasion of Bacteria to Host Cells by Interacting and Activating EGFR of the Host.

Chlamydia psittaci (C. psittaci) is an obligate intracellular, gram-negative bacterium, and mainly causes systemic disease in psittacine birds, domestic poultry, and wild fowl. The pathogen is threating to human beings due to closely contacted to employees in poultry industry. The polymorphic membrane proteins (Pmps) enriched in C. psittaci includes six subtypes (A, B/C, D, E/F, G/I and H). Compared to that of the 1 pmpG gene in Chlamydia trachomatis (C. trachomatis), the diverse pmpG gene-coding proteins of C. psittaci remain elusive. In the present study, polymorphic membrane protein 17G (Pmp17G) of C. psittaci mediated adhesion to different host cells. More importantly, expression of Pmp17G in C. trachomatis upregulated infections to host cells. Afterwards, crosstalk between Pmp17G and EGFR was screened and identified by MALDI-MS and Co-IP. Subsequently, EGFR overexpression in CHO-K1 cells and EGFR knockout in HeLa 229 cells were assessed to determine whether Pmp17G directly correlated with EGFR during Chlamydial adhesion. Finally, the EGFR phosphorylation was recognized by Grb2, triggering chlamydial invasion. Based on above evidence, Pmp17G possesses adhesive property that serves as an adhesin and activate intracellular bacterial internalization by recognizing EGFR during C. psittaci infection