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

Author

Romina Cecilia Russi, Diego del Balzo, Ivana Gabriela Reidel, Mariano Alonso Bivou, Noelia Flor, Agustín Lujan, Diego Sanchez, María Teresa Damiani, and Carolina Veaute

Subjects
PmpD, Chlamydia trachomatis, vaccine, adjuvants, CpG, gemini, Immunologic diseases. Allergy, RC581-607
Abstract

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.

Published
2023
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2. Heterologous prime-boost vaccination based on Polymorphic protein D protects against intravaginal Chlamydia trachomatis infection in mice

Author

Romina Cecilia Russi, Diego Del Balzo, Agustín Luján, Ivana Gabriela Reidel, María Inés García, Carolina Veaute, and María Teresa Damiani

Subjects
Medicine, Science
Abstract

Abstract The control of the worldwide spread of sexually transmitted Chlamydia trachomatis (Ct) infection urgently demands the development of a preventive vaccine. In this work, we designed a vaccine based on a fragment of polymorphic protein D (FPmpD) that proved to be immunogenic enough to generate a robust systemic and mucosal IgG humoral immune response in two strains of mice. We used a heterologous prime-boost strategy, including simultaneous systemic and mucosal administration routes. The high titers of anti-PmpD antibodies elicited by this immunization scheme did not affect murine fertility. We tested the vaccine in a mouse model of Ct intravaginal infection. Anti-PmpD antibodies displayed potent neutralizing activity in vitro and protective effects in uterine tissues in vivo. Notably, the humoral immune response of PmpD-vaccinated mice was faster and stronger than the primary immune response of non-vaccinated mice when exposed to Ct. FPmpD-based vaccine effectively reduced Ct shedding into cervicovaginal fluids, bacterial burden at the genitourinary tract, and overall infectivity. Hence, the FPmpD-based vaccine might constitute an efficient tool to protect against Ct intravaginal infection and decrease the infection spreading.

Published
2022
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3. In silico prediction of T- and B-cell epitopes in PmpD: First step towards to the design of a Chlamydia trachomatis vaccine

Author

Romina Cecilia Russi, Elian Bourdin, María Inés García, and Carolina Melania I. Veaute

Subjects
Chlamydia trachomatis, Vaccine, B-cell epitope, T-cell epitope, Molecular modeling, Epitope prediction, Medicine (General), R5-920, Biology (General), QH301-705.5
Abstract

Background: Chlamydia trachomatis is the most common sexually transmitted bacterial infection globally. Currently, there are no vaccines available despite the efforts made to develop a protective one. Polymorphic membrane protein D (PmpD) is an attractive immunogen candidate as it is conserved among strains and it is target of neutralizing antibodies. However, its high molecular weight and its complex structure make it difficult to handle by recombinant DNA techniques. Our aim is to predict B-cell and T-cell epitopes of PmpD. Method: A sequence (Genbank AAK69391.2) having 99–100% identity with various serovars of C. trachomatis was used for predictions. NetMHC and NetMHCII were used for T-cell epitope linked to MHC I or MHC II alleles prediction, respectively. BepiPred predicted linear B-cell epitopes. For three dimensional epitopes, PmpD was homology-modeled by Raptor X. Surface epitopes were predicted on its globular structure using DiscoTope. Results: NetMHC predicted 271 T-cell epitopes of 9-12aa with weak affinity, and 70 with strong affinity to MHC I molecules. NetMHCII predicted 2903 T-cell epitopes of 15aa with weak affinity, and 742 with strong affinity to MHC II molecules. Twenty four linear B-cell epitopes were predicted. Raptor X was able to model 91% of the three-dimensional structure whereas 57 residues of discontinuous epitopes were suggested by DiscoTope. Six regions containing B-cell and T-cell epitopes were identified by at least two predictors. Conclusions: PmpD has potential B-cell and T-cell epitopes distributed throughout the sequence. Thus, several fragments were identified as valuable candidates for subunit vaccines against C. trachomatis.

Published
2018
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4. Estudio de la respuesta inmune frente a infecciones genitales y su relación con infertilidad. Contribución al diseño de estrategias inmunoprofilácticas contra Chlamydia trachomatis

Author

Romina Cecilia Russi

Subjects
Gynecology, medicine.medical_specialty, business.industry, medicine, Prime boost, business, Chlamydia trachomatis, medicine.disease_cause
Abstract

Fil: Russi, Romina Cecilia. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas; Argentina.

Published
2021
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5. In silico prediction of T- and B-cell epitopes in PmpD: First step towards to the design of a Chlamydia trachomatis vaccine

Author

Elián D. Bourdin, Romina Cecilia Russi, Carolina Veaute, and María Inés García

Subjects
0301 basic medicine, CIENCIAS MÉDICAS Y DE LA SALUD, Immunogen, Protein Conformation, In silico, Inmunología, Molecular modeling, B-CELL EPITOPE, VACCINE, Epitopes, T-Lymphocyte, Chlamydia trachomatis, Biology, medicine.disease_cause, Epitope, law.invention, 03 medical and health sciences, Bacterial Proteins, law, T-CELL EPITOPE, MHC class I, medicine, Humans, MOLECULAR MODELING, CHLAMYDIA TRACHOMATIS, lcsh:QH301-705.5, Genetics, lcsh:R5-920, Membrane Proteins, purl.org/becyt/ford/3.1 [https], General Medicine, B-cell epitope, Medicina Básica, T-cell epitope, 030104 developmental biology, lcsh:Biology (General), Epitope prediction, GenBank, Bacterial Vaccines, Recombinant DNA, biology.protein, Epitopes, B-Lymphocyte, purl.org/becyt/ford/3 [https], Antibody, lcsh:Medicine (General), Vaccine, EPITOPE PREDICTION
Abstract

Background: Chlamydia trachomatis is the most common sexually transmitted bacterial infection globally. Currently, there are no vaccines available despite the efforts made to develop a protective one. Polymorphic membrane protein D (PmpD) is an attractive immunogen candidate as it is conserved among strains and it is target of neutralizing antibodies. However, its high molecular weight and its complex structure make it difficult to handle by recombinant DNA techniques. Our aim is to predict B-cell and T-cell epitopes of PmpD. Method: A sequence (Genbank AAK69391.2) having 99–100% identity with various serovars of C. trachomatis was used for predictions. NetMHC and NetMHCII were used for T-cell epitope linked to MHC I or MHC II alleles prediction, respectively. BepiPred predicted linear B-cell epitopes. For three dimensional epitopes, PmpD was homology-modeled by Raptor X. Surface epitopes were predicted on its globular structure using DiscoTope. Results: NetMHC predicted 271 T-cell epitopes of 9-12aa with weak affinity, and 70 with strong affinity to MHC I molecules. NetMHCII predicted 2903 T-cell epitopes of 15aa with weak affinity, and 742 with strong affinity to MHC II molecules. Twenty four linear B-cell epitopes were predicted. Raptor X was able to model 91% of the three-dimensional structure whereas 57 residues of discontinuous epitopes were suggested by DiscoTope. Six regions containing B-cell and T-cell epitopes were identified by at least two predictors. Conclusions: PmpD has potential B-cell and T-cell epitopes distributed throughout the sequence. Thus, several fragments were identified as valuable candidates for subunit vaccines against C. trachomatis. Fil: Russi, Romina Cecilia. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina Fil: Bourdin, Elian Daniel. Profesional independiente; Argentina Fil: Garcia, Maria Ines. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina Fil: Veaute, Carolina Melania Isabel. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina

Published
2018

6. Immune mediators associated to male infertility in a mouse model of DNA immunization with the sperm protease proacrosin

Author

Mónica H. Vazquez-Levin, Maria Florencia Veiga, Romina Cecilia Russi, Carolina Veaute, María Inés García, and Paulina Vignatti

Subjects
0301 basic medicine, Male, medicine.medical_treatment, Acrosome reaction, IMMUNOLOGY, Male infertility, Mice, 0302 clinical medicine, ACROSIN, Pregnancy, Vaccines, DNA, Immunology and Allergy, Birth Rate, Acrosin, Enzyme Precursors, 030219 obstetrics & reproductive medicine, biology, Obstetrics and Gynecology, Seminal Vesicles, Bioquímica y Biología Molecular, Cytokine, Cellular Microenvironment, GENE IMMUNIZATION, Cytokines, Female, Antibody, CIENCIAS NATURALES Y EXACTAS, Infertility, medicine.medical_specialty, Immunology, Immunization, Secondary, Mice, Inbred Strains, INFERTILITY, Ciencias Biológicas, Andrology, 03 medical and health sciences, Immune system, Antigen, Internal medicine, medicine, Animals, SEMINAL VESICLE, Infertility, Male, CYTOKINES, medicine.disease, Sperm, 030104 developmental biology, Endocrinology, Reproductive Medicine, Immunoglobulin G, biology.protein
Abstract

The immune response has relevant physiological functions both in the male and female reproductive system, and must be tightly controlled to achieve a successful pregnancy. Several immune factors have been related to infertility, among them humoral and cellular immune responses triggered by sperm antigens. The present study was aimed at evaluating the immune profile induced by DNA immunization against the sperm protease proacrosin in CF1 male mice and its effect upon fertility. Immunized animals exhibited higher anti-proacrosin antibodies levels than controls (indirect ELISA), both in serum (p

Published
2015

7. Detection of Staphylococcus aureus adhesion and biofilm-producing genes and their expression during internalization in bovine mammary epithelial cells

Author

Elizabet Amanda Lorena Pereyra, Florencia Picech, Cristina Diez, Celina Baravalle, Maria Sol Renna, Luis Fernando Calvinho, Carolina Soledad Andreotti, Bibiana Elisabet Dallard, and Romina Cecilia Russi

Subjects
0301 basic medicine, GENE EXPRESSION, Staphylococcus aureus, media_common.quotation_subject, ICAD, MAMMARY EPITHELIAL CELLS, Biology, medicine.disease_cause, Staphylococcal infections, Microbiology, Bacterial Adhesion, BOVINE MASTITIS, 03 medical and health sciences, Mammary Glands, Animal, Gene expression, medicine, Animals, Internalization, STAPHYLOCOCCUS AUREUS, Gene, Mastitis, Bovine, Phylogeny, media_common, General Veterinary, Ciencias Veterinarias, Biofilm, Epithelial Cells, General Medicine, Gene Expression Regulation, Bacterial, Staphylococcal Infections, medicine.disease, In vitro, Bacterial Typing Techniques, 030104 developmental biology, CIENCIAS AGRÍCOLAS, Genes, Bacterial, Biofilms, Cattle, Female
Abstract

Staphylococcus aureus is one of the most prevalent pathogens isolated from bovine mastitis,causing chronic intramammary infections (IMI) that limit profitable dairying. The course ofinfection is often associated with factors both related to the host and the bacterium. Aims of thisstudy were to select S. aureus isolates from bovine IMI with different genotypic profiles harboringgenes involved in adherence and biofilm production, to determine the behavior of these strains incontact with bovine mammary epithelial cells (MAC-T) and the expression of those genes duringbacterial-cell early interactions. The genetic diversity of 20 S. aureus strains that were isolated frommilk samples taken from cows with persistent-P and non-persistent-NP IMI was high, discriminatedinto 13 fingerprint groups. The occurrence of genes coding for S. aureus surface proteins (clfA, clfB,fnbA, fnbB, fib, cna) and biofilm formation (icaA, icaD, icaC, bap) and in vitro biofilm-formingability was not related to strain clinical origin (NP or P). Internalization of S. aureus into MAC-Tcells was strain-dependent and internalized bacteria overexpressed adherence and biofilm-forminggenes compared with those that remained in the supernatant of co-cultures; particularly those genesencoding FnBPs and IcaD. Strains yielding highest invasion percentages were those able tooverexpress fnBP, irrespectively of the presence of other evaluated genes. Strains from NP IMIshowed a greater multiplication capacity in vitro compared with strains from P IMI. These resultsprovide new insights about S. aureus differential gene expression of adhesion-internalization factorsduring early interaction with mammary epithelial cells. Fil: Pereyra, Elizabet Amanda Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Picech, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Ciencias de la Salud. Universidad Nacional de Córdoba. Instituto de Investigaciones en Ciencias de la Salud; Argentina Fil: Renna, Maria Sol. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Baravalle, Celina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Andreotti, Carolina Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Russi, Romina Cecilia. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Tecnología Inmunológica; Argentina Fil: Calvinho, Luis Fernando. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina Fil: Diez, Cristina Noemí. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Tecnología Inmunológica; Argentina Fil: Dallard, Bibiana Elisabet. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

Published
2015

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