Inmunogenicidad de proteínas recombinantes comparada con el empleo de herramientas bioinformáticas / Immunogenicity of recombinant proteins compared with the use of bioinformatics tools

Objetivo: para determinar la utilidad de herramientas inmunoinformáticas para detectar péptidos que puedan ser inmunodominantes, y evaluar las diferencias entre las respuestas inmunes de los modelos animales empleados en los estudios preclínicos y en los humanos. Métodos: se modeló la respuesta frente a dos proteínas exógenas: la estreptocinasa recombinante y el antígeno de superficie de la hepatitis B. A partir de sus secuencias primarias se emplearon algoritmos para identificar epítopes B y T frente a moléculas HLA de clase I y II (HLA-A*0201, HLA-DRB1*0301 y HLA-DRB1*0701) y los haplotipos murinos H2-Kd y H2-Kk. Se seleccionaron los péptidos de más alta puntuación. Resultados: el algoritmo ABCPred mostró una mejor capacidad de predicción de epítopes B, mientras fue mayor la coincidencia para los programas de modelación de la respuesta T. Los epítopes generados para el haplotipo H2-Kk tuvieron una similitud mayor con los presentados por las moléculas HLA seleccionadas. Conclusiones: se presenta una metodología aplicable al desarrollo de vacunas de subunidades y multiepitópicas, así como para otros fármacos biotecnológicos de naturaleza peptídica, que permite optimizar las etapas preclínicas y clínicas, a muy bajo costo, mínimos requerimientos tecnológicos, utilización óptima de medios, recursos y capital humano disponibles en cualquier institución del sistema nacional de salud

Objective: determine the usefulness of immunoinformatics tools to detect potentially immunodominant peptides, and evaluate the differences between the immune responses provided by the animal models used in preclinical and human studies. Methods: modeling was conducted of the response to two exogenous proteins: recombinant streptokinase and hepatitis B surface antigen. Based on their primary sequences, algorithms were used to identify B and T epitopes against HLA class I and II molecules (HLA-A*0201, HLA-DRB1*0301 and HLA-DRB1*0701), and murine haplotypes H2-Kd and H2-Kk. The highest scoring peptides were chosen. Results: ABCPred algorithm showed a better prediction capacity for B epitopes, whereas coincidence was greater in modeling programs for the T response. The epitopes generated for haplotype H2-Kk had greater similitude with those presented by the HLA molecules selected. Conclusions: a methodology is presented which is applicable to the development of subunit and multiepitope vaccines, as well as other peptidic biotechnological drugs. This methodology allows optimization of the preclinical and clinical phases at a very low cost, with minimal technological requirements, optimal use of media, and resources and human capital available at any institution of the national health system

Protective effects of recombinant Brucella abortus Omp28 against infection with a virulent strain of Brucella abortus 544 in mice

The outer membrane proteins (OMPs) of Brucella (B.) abortus have been extensively studied, but their immunogenicity and protective ability against B. abortus infection are still unclear. In the present study, B. abortus Omp28, a group 3 antigen, was amplified by PCR and cloned into a maltose fusion protein expression system. Recombinant Omp28 (rOmp28) was expressed in Escherichia coli and was then purified. Immunogenicity of rOmp28 was confirmed by Western blot analysis with Brucella-positive mouse serum. Furthermore, humoral- or cell-mediated immune responses measured by the production of IgG1 or IgG2a in rOmp28-immunized mice and the ability of rOmp28 immunization to protect against B. abortus infection were evaluated in a mouse model. In the immunogenicity analysis, the mean titers of IgG1 and IgG2a produced by rOmp28-immunized mice were 20-fold higher than those of PBS-treated mice throughout the entire experimental period. Furthermore, spleen proliferation and bacterial burden in the spleen of rOmp28-immunized mice were approximately 1.5-fold lower than those of PBS-treated mice when challenged with virulent B. abortus. These findings suggest that rOmp28 from B. abortus is a good candidate for manufacturing an effective subunit vaccine against B. abortus infection in animals.

Synergism/complementarity of recombinant adenoviral vectors and other vaccination platforms during induction of protective immunity against malaria

The lack of immunogenicity of most malaria antigens and the complex immune responses required for achieving protective immunity against this infectious disease have traditionally hampered the development of an efficient human malaria vaccine. The current boom in development of recombinant viral vectors and their use in prime-boost protocols that result in enhanced immune outcomes have increased the number of malaria vaccine candidates that access pre-clinical and clinical trials. In the frontline, adenoviruses and poxviruses seem to be giving the best immunization results in experimental animals and their mutual combination, or their combination with recombinant proteins (formulated in adjuvants and given in sequence or being given as protein/virus admixtures), has been shown to reach unprecedented levels of anti-malaria immunity that predictably will be somehow reproduced in the human setting. However, all this optimism was previously seen in the malaria vaccine development field without many real applicable results to date. We describe here the current state-of-the-art in the field of recombinant adenovirus research for malaria vaccine development, in particular referring to their use in combination with other immunogens in heterologous prime-boost protocols, while trying to simultaneously show our contributions and point of view on this subject.

Human immunodeficiency virus therapeutics and pharmacogenomics.

Pharmacogenomics and pharmacogenetics are promising in development of a personalized treatment approach They are of paramount importance for basic immunology, for peptide based vaccine design (vaccinomics) drug monitoring in clinical setting and molecular pathophysiology of multifactorial diseases like cancer, tuberculosis, cardiac disorders, diabetes, asthma, HIV, etc