The ΦCPG1 chlamydiaphage can infect Chlamydia trachomatis and significantly reduce its infectivity.

Recent years have seen a significant increase in rates of persistent, antibiotic-resistant infection of Chlamydia trachomatis (CT) infections, representing an increasingly serious public health threat. At present there are no effective vaccines or antibodies available to treat CT, prompting the need for novel treatment strategies. One potential solution to this issue is the use of ΦCPG1, a chlamydia-specific lytic phage which has over 90% nucleotide sequence identity with other chlamydiaphages. Previous work has shown the Vp1 capsid protein of ΦCPG1 to exhibit broad inhibitory activity against all CT serotypes, inhibiting CT-mediated host cell toxicity. Patients with CT infections exhibit circulating antibodies against this Vp1 protein, suggesting that this or similar phages may be present in vivo in the context of CT infections, even though no phages have been specifically detected to date. Given these previous findings, we hypothesized that the ΦCPG1 chlamydiaphage may be able to infect CT, thereby inhibiting its growth and proliferation. To test this, we generated a recombinant pGFP-ΦCPG1 phage which we used to explore its effects on CT and chlamydia conjunctivitis of guinea pigs (GPIC). We found that pGFP insertion did not alter the packaging or infectivity of ΦCPG1, and that this recombinant phage was readily able to infect CT and GPIC and inhibit CT and GPIC in a dose-dependent fashion. This inhibition was most pronounced during the mid and late stages of the CT infection, disrupting the reticular body (RB) to EB transition, leading to the formation of enlarged RBs. These results indicate that ΦCPG1 is able to infect CT, highlighting this phage as a novel potential therapeutic agent for treating chlamydia infections. In addition, by engineering pGFP to express ΦCPG1, we have produced an valuable experimental tool useful for future studies of drug resistance, pathogenicity, and vaccine research aimed at improving CT treatment.

Identification of proteins differentially expressed by Chlamydia trachomatis treated with chlamydiaphage capsid protein VP1 during intracellular growth.

Chlamydia trachomatis infection is one of the most prevalent sexually transmitted diseases. Our research pertains to the inhibitory effect and molecular mechanism of the chlamydiaphage capsid protein VP1 on the growth of Chlamydia trachomatis. In this research, the capsid protein VP1 of the guinea-pig conjunctivitis chlamydiaphage phiCPG1 was expressed, purified and identified, and then, it was applied to the cultivation of different serovars of Chlamydia trachomatis and Chlamydia psittaci. The inhibitory effect was observed in each serovar of Chlamydia trachomatis (D, E, F, G, H, I, K, and L2) and Chlamydia psittaci inoculated with VP1 protein. The inhibition affection of VP1 on the growth of Chlamydia trachomatis was caused by the changes of expressions of some related proteins including 36 proteins up-regulated and 81 proteins down-regulated in the development cycle of Ct through the label-free test, and the transcription levels of these proteins, including Hc1, pmpD, and MOMP, were confirmed by RT-PCR. It provides information that is essential for understanding the mechanism of chlamydiaphage capsid protein VP1 on chlamydia and a new direction for further clinical treatment of chlamydial infection.

Biological effects of chlamydiaphage phiCPG1 capsid protein Vp1 on chlamydia trachomatis in vitro and in vivo.

The researches on chlamydia in recent years show that chlamydia bacteriophage may be a potential and effective means to solve the clinical infection of chlamydia trachomatis (Ct). We investigated the biological effect of chlamydiaphage phiCPG1 capsid protein Vp1 on Ct both in McCoy cells and genital tract of mice. Different concentrations of Vp1 were co-incubated with Ct E serotype strain in McCoy cells. Female BALB/c mice were used to establish Ct E strain-induced urogenital infection model. They were randomly divided into five groups and given different treatments on the fifth day after Ct inoculation. Animals in groups 1 and 2 were given 30 µL different concentrations of Vp1 in the genital tract respectively, those in group 3 were intramuscularly injected with 30 µL Vp1, those in the infected group did not receive any intervention, and those in the control group received 30 µL PBS in the genital tract. The vaginal discharge was collected to identify the live chlamydia by cell culture and gene fragment by real time PCR different days after infection. Inhibition rate of 100 µg/mL and 50 µg/mL Vp1 proteins against Ct E strain in the McCoy cell cultures was 91% and 79% respectively. The number of intracellular Ct inclusion in the McCoy cells co-cultured with vaginal discharge of group 1 and group 2 was less than in the infected group, and that in group 1 was less than in group 2, on the 7th day after Ct inoculation. Real-time PCR showed that chlamydia concentration of the vaginal discharge in group 2 was lower than in the infected group, and that in group 1 was lower than in group 2 on the 10th day. It was suggested that Vp1 capsid proteins had inhibitory effect on the proliferation of Ct serovar E strain in cell culture and mouse genital tract.

The presence of Chlamydia phage PhiCPG1 capsid protein VP1 genes and antibodies in patients infected with Chlamydia trachomatis.

Chlamydia phage PhiCPG1 has been found in Chlamydia caviae in a guinea pig model for inclusion conjunctivitis, raising the possibility that Chlamydia phage is also present in patients infected with C. trachomatis (Ct). In the present study, we assayed for presence of Chlamydia phage capsid protein VP1 genes and antibodies in 84 non-Ct controls and 206 Ct patients using an enzyme-linked immunoassay (ELISA), followed by verification with Western blot. None of the subjects were exposed to an antibiotic treatment or had a C. pneumoniae infection. The VP1 antibody test was positive in both, the ELISA and Western blot assay, in 4 Ct patients. PCR amplification experiments revealed presence of the VP1 gene in 5 Ct patients. The results suggest that Chlamydia phage capsid protein VP1 may exist in some Ct patients.

Chlamydiaphage φCPG1 Capsid Protein Vp1 Inhibits Chlamydia trachomatis Growth via the Mitogen-Activated Protein Kinase Pathway.

Chlamydia trachomatis is the most common cause of curable bacterial sexually transmitted infections worldwide. Although the pathogen is well established, the pathogenic mechanisms remain unclear. Given the current challenges of antibiotic resistance and blocked processes of vaccine development, the use of a specific chlamydiaphage may be a new treatment solution. φCPG1 is a lytic phage specific for Chlamydia caviae, and shows over 90% nucleotide sequence identity with other chlamydiaphages. Vp1 is the major capsid protein of φCPG1. Purified Vp1 was previously confirmed to inhibit Chlamydia trachomatis growth. We here report the first attempt at exploring the relationship between Vp1-treated C. trachomatis and the protein and gene levels of the mitogen-activated/extracellular regulated protein kinase (MAPK/ERK) pathway by Western blotting and real-time PCR, respectively. Moreover, we evaluated the levels of pro-inflammatory cytokines interleukin (IL)-8 and IL-1 by enzyme-linked immunosorbent assay after Vp1 treatment. After 48 h of incubation, the p-ERK level of the Vp1-treated group decreased compared with that of the Chlamydia infection group. Accordingly, ERK1 and ERK2 mRNA expression levels of the Vp1-treated group also decreased compared with the Chlamydia infection group. IL-8 and IL-1 levels were also decreased after Vp1 treatment compared with the untreated group. Our results demonstrate that the inhibition effect of the chlamydiaphage φCPG1 capsid protein Vp1 on C. trachomatis is associated with the MAPK pathway, and inhibits production of the pro-inflammatory cytokines IL-8 and IL-1. The bacteriophages may provide insight into a new signaling transduction mechanism to influence their hosts, in addition to bacteriolysis.

Reactivation of chromosomally integrated human herpesvirus-6 by telomeric circle formation.

More than 95% of the human population is infected with human herpesvirus-6 (HHV-6) during early childhood and maintains latent HHV-6 genomes either in an extra-chromosomal form or as a chromosomally integrated HHV-6 (ciHHV-6). In addition, approximately 1% of humans are born with an inheritable form of ciHHV-6 integrated into the telomeres of chromosomes. Immunosuppression and stress conditions can reactivate latent HHV-6 replication, which is associated with clinical complications and even death. We have previously shown that Chlamydia trachomatis infection reactivates ciHHV-6 and induces the formation of extra-chromosomal viral DNA in ciHHV-6 cells. Here, we propose a model and provide experimental evidence for the mechanism of ciHHV-6 reactivation. Infection with Chlamydia induced a transient shortening of telomeric ends, which subsequently led to increased telomeric circle (t-circle) formation and incomplete reconstitution of circular viral genomes containing single viral direct repeat (DR). Correspondingly, short t-circles containing parts of the HHV-6 DR were detected in cells from individuals with genetically inherited ciHHV-6. Furthermore, telomere shortening induced in the absence of Chlamydia infection also caused circularization of ciHHV-6, supporting a t-circle based mechanism for ciHHV-6 reactivation.

Infección por Chlamydia trachomatis / Chlamydia trachomatis infection

Determinar la prevalencia de la infección por Chlamydia trachomatis en una población de parejas infértiles. Validar la eficacia del diagnóstico de anticuerpos anti Chlamydia para tratar, prevenir y controlar la infección por C. trachomatis. Justificar la necesidad de implementar el monitoreo de rutina para administrar el tratamiento oportuno de la infección por C. trachomatis. Se determinó la prevalencia de la infección por Chlamydia trachomatis en 4 619 pacientes, 2607 mujeres y 2012 hombres en edades reproductivas, entre 1999 y 2008 por problemas de infertilidad. Se detectaron anticuerpos anti-Chlamydia trachomatis (IgG, IgA e IgM) por SeroELISA (Savyon Diagnostics Ltd.) de 1999 a 2005; desde 2006 hasta 2008 se utilizó el kit InmunoComb II (Orgenics). Centro de fertilidad UNIFERTES, Caracas, Venezuela. Se encontró una prevalencia en mujeres de 25,40 ± 6,26 por ciento; y en los hombres de 31,12 ± 2,88 por ciento. La prevalencia de la infección por C. trachomatis en parejas infértiles es alta y no ha disminuido en los últimos 10 años. Se recomienda implementar en Venezuela un monitoreo de rutina para el diagnóstico de C. trachomatis, incluyendo su determinación en el control ginecológico anual y en las evaluaciones urológicas a hombres jóvenes, a fin de prevenir que la infección pase a ser crónica

To determine the prevalence of Chlamydia trachomatis infection in infertile couples. To evaluate the efficiency of the Chlamydia trachomatis screening programs. To establish the need of implementing the routine early diagnosis and opportune treatment of the infection. Prevalence of Chlamydia trachomatis infection was determined in 4619 patients, 2607 women and 2012 men in reproductive agesbetween 1999 and 2008. Anti-chlamydia and C.trachomatis antobodies (IgG, IgA and IgM) were detected by SeroELISA (Savyon Diagnostics Ltd.) from 1999 to 2005; and by the ImmunoComb II kit (Orgenics) from 2006 to 2008. Fertility clinic UNIFERTES in Caracas, Venezuela. A prevalence of 25,40 ± 6,26 percent was found in women and a prevalence of 31,12 ± 2,88 percent was found in men. Prevalence of the C. trachomatis infection in infertile couples is high and has not decreased over the last 10 years. Implementation of routine screening programs for C. trachomatis detection is recommended, including its assessment in annual gynecological controls, as well as in urologic evaluations in young men, in order to prevent the infection from being chronic

Frecuencia de anticuerpos IgA e IgM anti Chlamydia trachomatis: en mujeres embarazadas procedentes de una consulta prenatal Cumaná, Estado Sucre, Venezuela, marzo-junio de 2006 / Frequency of IgA and IgM anti Chlamydia trachomatis antibodies: in pregnant women coming from a prenatal consult in Cumana, State of Sucre, Venezuela, march-june of 2006

Con el propósito de establecer la frecuencia de anticuerpos IgA e IgM anti-C-trachomatis en mujeres embarazadas se realizó un estudio en 84 mujeres con esa condición, en edades comprendidas entre 14 y 43 años, que acudieron a la consulta prenatal, del Servicio Autónomo Hospital Universitario “Antonio Patricio de Alcalá”, en Cumaná, estado Sucre, Venezuela, durante el período marzo-junio de 2006. Para ello se obtuvieron 84 muestras de suero para la determinación de anticuerpos IgA e IgM anti C-trachomatis a través del método de inmunoabsorción ligado a enzimas ELISA (Diagnostic Automation INC). Del total de muestras analizadas 16 (19,05 por ciento) y 55 (65,48 por ciento) resultaron positivas para la determinación de anticuerpos IgA e IgM anti C-trachomatis respectivamente. No se encontró asociación entre la presencia de estos anticuerpos con la edad de las pacientes, aunque el mayor número de pacientes positivas se ubicó en el intervalo de edades comprendidas entre 14 a 23 años. Asimismo al asociarse las manifestaciones clínicas genitales con la presencia de anticuerpos IgA e IgM anti C- trachomatis no se encontraron valores estadísticamente significativos. Por lo anteriormente expuesto se concluye que la infección genital por Chlamydia trachomatis en mujeres embarazadas es extremadamente frecuente, de manera especial en las edades comprendidas entre 24 a 33 años, y ocurre habitualmente en forma asintomática con las graves repercusiones que esto acarrea a la paciente, al feto y a su pareja.

In order to establish the frequency of IgA and IgM anti-C. Trachomatis antibodies in expectant women, a study was made of 84 women between the ages of 14 and 43, who attended prenatal consults in the Autonomous Service at the University Hospital “Antonio Patricio of Alcalá,” Cumaná, State of Sucre, during the March-June period, 2006. 84 serum samples were obtained to determine IgA and IgM anti-C. trachomatis antibodies using the immunoabsorption method connected to ELISA enzymes (Diagnostic Automation INC). Of the total samples studied, 16 (19.05 percent) and 55 (65.48 percent) resulted positive for the IgA and IgM anti-C. trachomatis antibodies, respectively. No association was found between the presence of these antibodies and the age of the patients, although the greater number of positive patients was in the 14 to 23 year age interval. Likewise, no statistically significant values were found between the association of clinical genital manifestations and the presence of IgA and IgM anti C- trachomatis antibodies; therefore, it was shown that Chlamydia trachomatis is presented asymptomatically in most cases. Conclusions were that genital infection by Chlamydia trachomatis in pregnant women is extremely frequent, especially for ages between 24 and 33 years, and it occurs habitually in an asymptomatic form with the serious repercussions that this produces on the patient, the fetus and the partner.

Infecção genital por Chlamydia trachomatis em casais atendidos em laboratório de esterelidade conjugal / Genital infection by Chlamydia trachomatis in couples attending in conjugal sterelity ambulatory

Introdução: as infecções sexualmente transmissíveis são relativamente freqüentes e constituem sério problema de saúde pública, em quase todo o mundo. Dentre essas, a causada por Chlamydia trachomatis (CT) é uma das mais prevalentes e de alto índice de complicação para a saúde reprodutiva. Objetivo: investigar a taxa de infecção genital por CT em casais que procuram tratamento para infertilidade e possíveis fatores associados. Métodos: estudo transversal englobou 100 casais atendidos no ambulatório de esterelidade da Clínica Ginecológica do Hospital Agamenon Magalhães - Recife-PE, submetidos a um questionário contendo informações sócio-econômicas e demográficas, sinais e sintomas relacionados com possível infecção genitourinária. A propedêutica rotineira foi empregada, incluindo espermograma e histerossalpingografia. A técnica laboratorial foi a reação em cadeia de polimerase (PCR), com amostra de 1º jato urinário dos homens e endocervicais das mulheres. Resultados: observou-se infecção genital por CT em 17 casais (9% dos homens e 10% das mulheres). Houve coincidência de infecção entre parceiros em dois casais. Não houve nenhuma associação com características sócio-demográficas e econômicas, sinais e sintomas genitourinários e as causas básicas de infertilidade. Conclusão: a infecção por CT em casais inférteis mostrou freqüência elevada na amostra. Portanto, sugere-se seu rastreio rotineiro em clínicas de infertilidade.

Utilizing Chlamydia trachomatis IgG serology with HSG to diagnose tuboperitoneal-factor infertility.

To evaluate if Chlamydia trachomatis IgG serology combined with hysterosalpingography can make it easier to detect tuboperitoneal factor infertility, we conducted a chart review of 76 consecutive patients at an infertility practice at West Virginia University from 1999-2001. We checked the charts for results of Chlamydia trachomatis IgG serology, Hysterosalpingography (HSG) and laparoscopy. Results of these tests were reviewed along with age, parity, previous reproductive tract disease surgery and duration of infertility. Complete data was found on 32 of the 76 patients. Chlamydia serology in conjunction with the HSG had a sensitivity of 80% for tuboperitoneal factor (tubal obstruction or pelvic adhesions), and a specificity of 82.3%. The positive predictive value was 80% and the negative predictive value was 82%. Since Chlamdia trachomatis IgG serologic testing is non-invasive and relatively inexpensive, we recommend combining it with hysterosalpingography as an infertility work-up. More invasive testing such as laparoscopy may be postponed or completely eliminated.

Joint effect of HPV16 with Chlamydia trachomatis and smoking on risk of cervical cancer: antagonism or misclassification (Nordic countries).

OBJECTIVES: To estimate the joint effects of infections with human papillomavirus type 16 (HPV16) and Chlamydia trachomatis and smoking on the risk of cervical cancer. To study whether the joint effects can be accounted for by misclassification in the HPV serology. METHODS: A nested case-control study with incidence density sampling was conducted in three cohorts of 530,000 women, who donated serum samples to three Nordic serum banks in 1973-1994. The main outcome measure is the odds ratio (OR) of incidence rates of invasive cervical squamous cell carcinoma (SCC) among those seropositive for HPV16 and/or C. trachomatis and/or with increased levels of cotinine in serum compared to those negative for all the three exposures. RESULTS: Two hundred eight women with SCC and 624 matched controls were identified during a mean follow-up of 5 years through linkage to the national cancer registries. Exposure to past infections and smoking was defined by presence of specific IgG antibodies to HPV16 and C. trachomatis and increased levels of serum cotinine. Observed ORs were compared to OR = 20 for HPV16 and accounting the differences for by misclassification bias. OR = 20 was elected as a gold standard on the basis of other studies with PCR-based analyses and a follow-up design. Each of the three exposures was associated with an increased risk of SCC (OR = 5.4 for HPV16, 3.4 for C. trachomatis and 1.8 for cotinine). The interaction was antagonistic (observed OR = 2.5 among those positive for all three exposures as compared to OR = 33 expected on the basis of multiplicative single effects (p = 0.047)). The antagonism could not totally be accounted for by any credible combination of sensitivity and specificity of HPV16 serology. CONCLUSION: HPV16, C. trachomatis, and smoking are likely to be risk factors of SCC with strong antagonistic joint effect. Non-differential misclassification in serology for HPV16 could be ruled out (but only some types of differential) as an alternative explanation for the observed antagonism.