Immunonutrition: Bioecological Control of Prebiotics, Probiotics and Symbolics to Beneficially Modify the Human Microbioma and Quality of Life

Introduction: Currently interest in bioecological and nutritional control arisen, for this reason our body had been in need of seeking help from microbes in our digestive system to process them, benefiting us with an energy source and essential vitamins. Objective: Verify through clinical evidence the capacity and usefulness of prebiotics, probiotics and symbiotics to modify the human microbiota and thus obtain benefit in human homeostasis. Method: Non-systematic bibliographic review was performed in PubMed, Elsevier, Academic Google, Scopus, Scielo databases, using the mentioned descriptors. Results: Review of 45 articles from the last 5 years, and 30 that were related to the topic and objective of this literature review study were included. Discussion: The relationship between the microorganism and the human being is found in the microbiome of the digestive tract, but this relationship can mean a beneficial contribution such as leading to diseases that are generated from genetic risk factors, the use of probiotics, prebiotics and Symbiotics beneficially minimize postoperative infectious risks and complications, in addition to enzyme activity, immune modulation. Conclusion: Immunonutrition was a beneficial tool that favors the growth of beneficial bacteria over harmful bacteria, in addition immunomodular functions usually interact with lymphocyte receptors and potentiate immune tolerance against intestinal pathogens, help in the treatment of various diseases and improve quality of life. Keywords: immunonutrition, prebiotics, probiotics, symbiotics, microbiota. RESUMEN Introducción: En la actualidad ha surgido interés por el control bioecológico y nutricional, por esta razón nuestro organismo se ha visto en la necesidad de buscar ayuda de microbios en nuestro aparato digestivo para procesarlos, beneficiándonos con una fuente de energía y vitaminas esenciales. Objetivo: Comprobar por medio de evidencia clínica la capacidad y utilidad que tienen los prebióticos, probióticos y simbióticos para modificar la microbiota humana y de esta forma obtener beneficio en la homeostasis humana. Método: Revisión bibliográfica no sistemática, se realizó en las bases de datos PubMed, Elsevier, Academic Google, Scopus, Scielo, usando los descriptores mencionados. Resultados: Revisión de 45 artículos de los últimos 5 años, y se incluyeron 30 que están en relación con el tema y objetivo de del presente estudio de revisión de la literatura. Discusión: La relación entre el microrganismo y el ser humano se encuentra en el microbioma del tracto digestivo, pero esta relación puede significar un aporte beneficioso tal como conllevar a enfermedades que se generan a partir de factores genéticos de riesgo, el uso de los probióticos, prebióticos y simbióticos minimizan beneficiosamente los riesgos y complicaciones infecciosas postoperatorias, además de la actividad enzimática, modulación inmune. Conclusiones: La inmunonutrición es una herramienta beneficiosa favorece el crecimiento de bacterias benéficas sobre las nocivas, además funciones inmunomodulares suelen interactuar con receptores linfocitarios y potencializan la tolerancia inmunológica frente e patógenos intestinales, ayudan en el tratamiento de diversas enfermedades y mejora la calidad de vida. Palabras clave: inmunonutrición, prebióticos, probióticos, simbióticos, microbiot

Did Tenascin-C Co-Evolve With the General Immune System of Vertebrates?

Tenascin-C plays important roles in immunity. Toll-like receptor 4, integrin α9β1 and chemokines have already been identified as key players in executing the immune regulatory functions of tenascin-C. Tenascin-C is also found in reticular fibers in lymphoid tissues, which are major sites involved in the regulation of adaptive immunity. Did the “tool box” for reading and interpreting the immune-regulating instructions imposed by tenascins and tenascin-C co-evolve? Though the extracellular matrix is ancient, tenascins evolved relatively recently. Tenascin-like genes are first encountered in cephalochordates and urochordates, which are widely accepted as the early branching chordate lineages. Vertebrates lacking jaws like the lamprey have tenascins, but a tenascin gene that clusters in the tenascin-C clade first appears in cartilaginous fish. Adaptive immunity apparently evolved independently in jawless and jawed vertebrates, with the former using variable lymphocyte receptors for antigen recognition, and the latter using immunoglobulins. Thus, while tenascins predate the appearance of adaptive immunity, the first tenascin-C appears to have evolved in the first organisms with immunoglobulin-based adaptive immunity. While a C-X-C chemokine is present in the lamprey, C-C chemokines also appear in the first organisms with immunoglobulin-based adaptive immunity, as does the major histocompatibility complex, T-cell receptors, Toll-like receptor 4 and integrin α9β1. Given the importance of tenascin-C in inflammatory events, the co-evolution of tenascin-C and key elements of adaptive and innate immunity is suggestive of a fundamental role for this extracellular matrix glycoprotein in the immune response of jawed vertebrates.

Immunoglobulins, MHC and T Cell receptors genes in Cetaceans

Cetaceans correspond to mammals that have returned to the marine environment. Adaptive changes are very significant with the conversion of the limbs into flippers. It is studied the changes that have occurred in immunoglobulins, MHC class I and II and T cell receptors genes. Constant regions of immunoglobulins are similar to those of the rest of mammals. An exception is the IgD gene, which is composed of three CH domains but CH1 similar to CH1 of immunoglobulin M. In the IGHV locus, it exist a decrease in the number of VH genes with the absence of genes within Clan I. The number of Vλ genes is greater than that of Vκ. In the genes for T lymphocyte receptors, it exists a decrease in the number of Vα genes with loss of significant clades and subclades. In Vβ and Vγ, there is also the loss of clades. These declines of Vα, Vβ and Vγ are not present Artiodactyla, and they are specific to Cetaceans. In MHC present tree evolutive lines of class I genes. These species have DQ, DR, DO and DM genes, but they are no present DP genes.

Generation of Lamprey Monoclonal Antibodies (Lampribodies) Using the Phage Display System

The variable lymphocyte receptors (VLRs) consist of leucine rich repeats (LRRs) and comprise the humoral antibodies produced by lampreys and hagfishes. The diversity of the molecules is generated by stepwise genomic rearrangements of LRR cassettes dispersed throughout the VLRB locus. Previously, target-specific monovalent VLRB antibodies were isolated from sea lamprey larvae after immunization with model antigens. Further, the cloned VLR cDNAs from activated lamprey leukocytes were transfected into human cell lines or yeast to select best binders. Here, we expand on the overall utility of the VLRB technology by introducing it into a filamentous phage display system. We first tested the efficacy of isolating phage into which known VLRB molecules were cloned after a series of dilutions. These experiments showed that targeted VLRB clones could easily be recovered even after extensive dilutions (1 to 109). We further utilized the system to isolate target-specific “lampribodies” from phage display libraries from immunized animals and observed an amplification of binders with relative high affinities by competitive binding. The lampribodies can be individually purified and ostensibly utilized for applications for which conventional monoclonal antibodies are employed.

SCIDOT-05. DEVELOPING VARIABLE LYMPHOCYTE RECEPTORS THAT TARGET PATHOLOGICALLY EXPOSED NEURAL ECM TO TREAT GLIOBLASTOMA

INTRODUCTION The median survival of gliobastoma (GBM) patients remains less than two years despite aggressive treatments. Current targeted GBM therapies demonstrate initial therapeutic benefit; however, patients relapse due to therapeutic resistance and failure to eliminate GBM cells at the invasive margin. Therefore, we propose a two-prong approach: first, target pathologic disruption of the blood brain barrier (BBB) via exposure of neural ECM rather than disease markers to overcome therapy-resistant GBM; and second, designing therapeutic payloads that extracellularly spread throughout the tumor volume. METHODS Variable Lymphocyte Receptors (VLRs, a lamprey-derived antigen recognition system) were identified with high specificity for neural ECM. Candidate VLRs underwent further refinement using ex vivo tissue staining. Utilizing pathologic disruption of BBB as an approach for targeting GBM was confirmed in vivo with intracranial murine glioblastoma models. Finally, an immunogenic peptide was attached via a cleavable linker to the neural ECM binding VLRs for conditional release extracellularly to spread throughout the tumor. RESULTS The lead neural ECM-binding VLR candidate, named P1C10, demonstrates diffuse binding to parenchymal neural ECM, without detectable binding to other tissues. P1C10 demonstrates nanomolar affinity for neural ECM, and preferentially accumulates in intracranial GL261 and U87 murine GBM models. Finally, P1C10-targeted doxorubicin-loaded liposomes significant increased survival of mice with intracranial GBM. In additional studies, treating murine GBM models with a P1C10 VLR linked to an immunogenic peptide reduced GBM proliferation and increased infiltration of cytotoxic T cells. CONCLUSIONS We present proof-of-concept demonstration for targeting intracranial GBM via neural ECM exposed at pathological BBB disrupted sites. Additionally, P1C10 neural ECM-targeting VLR delivers chemotherapy-loaded nanoparticles and immunogenic peptides designed to spread extracellularly throughout the tumor. Thus, this novel strategy links a physiological ECM targeting scheme with extracellular-released therapeutics to treat primary GBM, and has potential for delivering therapies to other CNS diseases with pathological BBB.

EXTH-67. IDENTIFICATION AND DEVELOPMENT OF NEURAL ECM-BINDING LAMPREY ANTIBODIES (VARIABLE LYMPHOCYTE RECEPTORS) THAT TARGET GLIOBLASTOMA

INTRODUCTION The median survival of glioblastoma (GBM) patients remains less than two years even with state-of-the-art treatment. Current targeted GBM therapies demonstrate initial therapeutic benefit; however, patients relapse due to therapeutic selection of treatment resistant GBM cellular populations. Therefore, we propose targeting pathologic disruption of the blood brain barrier (BBB) via exposure of neural ECM, rather than disease markers, to overcome therapy-resistant GBM. METHODS We identify Variable Lymphocyte Receptors (VLRs, the antigen recognition system used by lamprey) that demonstrate high specificity for neural ECM. Candidate VLRs underwent further refinement using in vitro binding assays and ex vivo tissue staining. Utilizing pathologic disruption of BBB as an approach for targeting GBM was confirmed in vivo using intracranial murine glioblastoma models. RESULTS The lead neural ECM-binding VLR candidate, named P1C10, demonstrates diffuse binding to parenchymal neural ECM, without detectable binding to other tissues. P1C10 demonstrates nanomolar affinity for in vitro derived neural ECM, and preferentially accumulates at intracranial GL261 and U87 lesions in murine GBM models. Finally, administration of P1C10-targeted doxorubicin-loaded liposomes significantly extends the survival of mice bearing intracranial U87 GBM. CONCLUSIONS We identified VLRs that bind neural ECM, and demonstrate their utility for delivering compounds and nanoparticles to sites of GBM induced blood brain barrier disruption. This novel strategy allows for targeting therapeutics via the underlying physiology of GBM rather than relying on cellular disease markers that are often lost in patients that relapse after targeting therapies.

Gouania willdenowi is a teleost fish without immunoglobulin genes

In the study of immunoglobulin V genes in fish genomes, we found that the species Gouania willdenowi does not possess any such regions, neither for the heavy chain nor for the light chains. Also, genes that code for the immunoglobulin constant regions were also not found. A detailed analysis of the chromosomal region of these genes revealed a deletion in the entire locus for regions of the heavy and light chains. These studies provide evidence that this species does not possess genes coding for immunoglobulins. Additionally, we found the genes that code for CD79a and CD79b protein molecules have also been deleted. Regions for the Tα/β lymphocyte receptors are present but the T γ/δ receptors were not found. In transcripts of two other Gobiesocidae species, Acystus sp. and Tomicodon sp., no antibody sequences could be detected, possibly indicating the absence of immunoglobulins in all species of this family.

Identification of variable lymphocyte receptors that can target therapeutics to pathologically exposed brain extracellular matrix

Diseases that lead to blood-brain barrier (BBB) disruption will pathologically expose normally inaccessible brain extracellular matrix (ECM) to circulating blood components. Therefore, we hypothesized that brain ECM-targeting moieties could specifically target the disrupted BBB and potentially deliver therapies. Variable lymphocyte receptors (VLRs) that preferentially associate with brain ECM were identified from an immune VLR library via yeast surface display biopanning coupled with a moderate throughput ECM screen. Brain ECM binding of VLR clones to murine and human brain tissue sections was confirmed. After systemic administration, P1C10, the lead brain ECM-targeting VLR candidate, specifically accumulated in brains with mannitol-disrupted BBB and at disrupted BBB regions in two different intracranial glioblastoma models. We also demonstrate P1C10’s ability to deliver doxorubicin-loaded liposomes, leading to significantly improved survival in glioblastoma-bearing mice. Thus, VLRs can be used to selectively target pathologically exposed brain ECM and deliver drug payloads.