GRA8 DNA vaccine formulations protect against chronic toxoplasmosis

Toxoplasma gondii has a very wide host range and infects all warm-blooded animals including humans. The disease causes great economic losses both in animals and humans. Vaccination is the most effective approach to fight against toxoplasmosis however an effective vaccine has not been developed yet. In the present study, GRA8 protein of T. gondii that showed high immunogenicity in our previous microarray screening study was used to develop a DNA vaccine using pcDNA 3.3 vector for the first time. In order to increase the potency of the DNA vaccine, 10 times lower amount of GRA8 DNA vaccine was combined with molecular adjuvant CpG and formulated into a commercial liposome (pcDNA3.3-GRA8+CpG+Escort). Mice were vaccinated intramuscularly two times at three-week intervals and challenged orally with the T. gondii PRU strain tissue cysts. The humoral immune response was determined by Western Blot and ELISA. The cellular immune response was analyzed by flow cytometry, cytokine ELISA and MTT assay. Among the vaccine groups, pcDNA3.3-GRA8 and pcDNA3.3-GRA8+CpG+Escort induced strong IgG response compared to controls (P 0.001). The IgG1 and IgG2a responses showed a balanced Th1-Th2 polarization. The ratio of CD4(+) and CD8(+) T lymphocytes secreting IFN-gamma increased, and significantly higher extracellular IFN-gamma secretion was achieved compared to the controls (P 0.01). The amount of tissue cysts in the group of mice vaccinated with pcDNA3.3-GRA8 decreased significantly compared to control groups (P 0.0001). In the group vaccinated with pcDNA3.3-GRA8+CpG+Escort, the amount of tissue cysts also decreased significantly compared to PBS (P = 0.0086) and Empty plasmid+CpG+Escort (P = 0.0007) groups. This study showed for the first time that pcDNA 3.3. vector encoding GRA8 with or without CpG and Liposome can induce strong cellular and humoral immune responses and confer strong protection against mouse model of chronic toxoplasmosis.This study was supported by the Grant given by The Scientific and Technological Research Council of Turkey (119S367) to M.D. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Scientific and Technological Research Council of Turkey [119S367

Toxoplasma gondii destroys Her2/Neu-expressing mammary cancer cells in vitro using a continuous feed medium approach

Can, Huseyin/0000-0001-9633-9786WOS:000590262200021PubMed: 33175718Introduction: Toxoplasma gondii is an opportunistic protozoan and can be grown using several human cell lines. Breast cancer is the second leading cause of cancer death in women. Her2/Neu-expressing mammary cancer cell lines called TUBO can be grown in vitro. in recent years, protozoan parasites have become popular means of use in cancer therapy research. in this study, we analyzed whether T. gondii tachyzoites can destroy TUBO cells using a novel continuous feed medium approach. Methodology: Two sets of flasks (each containing four groups) containing TUBO cells were inoculated with T. gondii Ankara strain tachyzoites. First set containing 5x10(6) TUBO cells were inoculated with TUBO-tachyzoite ratios of 1:2, 1:1, 2:1, and 4:1 and second set containing 1x10(6) TUBO cells were inoculated with TUBO-tachyzoite ratios of 10:1, 100:1, 1000:1, and 2000:1. Thereafter, culture supernatants were harvested at various days until TUBO cells were destroyed and tachyzoites were counted. Results: in the first and second sets of flasks, TUBO cells were destroyed between days 8 to 12 and 12 to 25, respectively. in addition, the amount of tachyzoites increased 743 and 595 to 112500 times in the first and second set of flasks, respectively. Conclusions: These results show that T. gondii tachyzoites successfully destroy Her2/Neu-expressing mammary cancer cells using a continuous feed medium approach. Although this idea may be too premature for the moment, the approach defined herein may support future researchers investigating the relationship between cancer and parasites which can make important progress toward saving cancer patient lives.Scientific Research Projects Branch Directorate of Ege University, TurkeyEge University [2016TIP082]The authors would like to acknowledge Prof. Federica Cavallo and Irene Fiore Merighi (University of Torino, Torino, Italy) for providing TUBO cells and technical assistance for the maintenance of TUBO cells in our lab. This study was partly supported by the grant given by the Scientific Research Projects Branch Directorate of Ege University, Turkey (Grant No: 2016TIP082) to L.Y

Screening of immunocompromised patients at risk of strongyloidiasis in western Turkey using ELISA and real-time PCR

WOS: 000404381800027PubMed ID: 28618740Background/aim: Strongyloides stercoralis causes life-threatening hyperinfection or disseminated strongyloidiasis in immunocompromised patients such as HIV-positive, organ transplantation, and cancer patients. This study investigated the presence of strongyloidiasis in immunocompromised patients for the first time in Turkey. Materials and methods: Serum and stool samples were collected from 108 patients (25.9% of them were chronic renal failure and 74.1% were renal transplantation patients) who were admitted to Ege University Medical School in Izmir, located in western Turkey. Serum samples were analyzed by ELISA (DRG, Germany) and the presence of 18S rRNA gene of S. stercoralis was detected in stool samples by real-time PCR. Results: The analysis of serum samples showed that only one patient was anti-S. stercoralis IgG antibody and real-time PCR positive (0.92%). The patient was treated twice with albendazole (400 mg/day for 3 days) at 2-week intervals. Follow up real-time PCR was negative and the patient became seronegative 6 months after the initial diagnosis. Conclusion: This screening showed that the prevalence of strongyloidiasis in this small group of patients who were at risk of strongyloidiasis was 0.92%. Overall, the results showed that more systematic studies are required in Turkey to show the prevalence of strongyloidiasis.Ege University, TurkeyEge University [2013TIP022]This study received financial support through a grant given by Ege University, Turkey (Grant number: 2013TIP022) to Meltem Isikgoz Tasbakan

First time identification of Acanthamoeba genotypes in the cornea samples of wild birds; Is Acanthamoeba keratitis making the predatory birds a target?

WOS: 000418107300022Acanthamoeba is a free-living amoeba which can be isolated from environment and among others well known as an opportunist protozoan parasite causing infections in humans and animals. Eyes are extremely important for the wild birds and losing sight ability due to Acanthamoeba can be dangerous. The studies on Acanthamoeba infection in wild birds is very few in world and Turkey therefore we aimed to screen deceased wild birds found in Izmir and Manisa provinces located in western Turkey using PCR and non-nutrition agar (NNA) plate method. Cornea samples were obtained from 18 deceased wild birds. During the external examination, signs of keratitis were observed in two Eurasian sparrowhawks (Accipiter nisus). All of the corneal samples were analyzed by two PCR methods and NNA plate. According to results, the Acanthamoeba positivity in corneal samples was 16.6% and 5.5% by PCR and plate method, respectively. According to sequencing data, two of isolates belonged to genotype T5 and one was genotype T4. In conclusion, Acanthamoeba infection was detected in wild bird cornea samples with/without keratitis for the first time in the world. The result of this study also show that Acanthamoeba can be a cause of keratitis in wild birds of Turkey and thus these predator birds can be a target of other wild animals due to loss of sight ability. In terms of public health, these results show the importance of wild birds as a source of Acanthamoeba infection in nature. (C) 2017 Elsevier Inc. All rights reserved.Ege University, TurkeyEge University [2014-TIP-073]This study was partly supported by the grant given by the Scientific Research Projects Branch Directorate of Ege University, Turkey (Grant No: 2014-TIP-073) to Y.G. The authors state that they have no conflict of interest