SBIR-STTR Award

Endotoxin neutralization in human plasma is an excellent indicator of chronic immune activation, which is the most accurate predictor of mortality in HIV. Recently we developed an assay using endotoxin neutralization as an indicator of bacterial translocation. This assay is accurate in discriminating control patients from those with inflammatory bowel disease in addition to indicating disease severity. We propose to adapt our endotoxin neutralization assay for the evaluation of HIV disease severity and progression. In the 6 month Phase I part of the SBIR project, we will (1) collect plasma from 20 19-50 year old HIV-positive males and an equivalent number of demographically similar HIV-negative male subjects and test for viral load and CD4+ T cell count, (2) measure the levels of specific plasma markers indicative of bacterial translocation, immune system activation and/or inflammatory response, and (3) measure immune activation using our endotoxin neutralization biomarker monitor system. Once a neutralization profile of each sample is established, Pearson correlation will be used to determine the relationship between endotoxin neutralization and all factors established in the first two aims. Student's T-test will be used to determine statistical significance between HIV- positive and control groups. The results from Phase I will show whether our endotoxin neutralization system is a convenient, reproducible and inexpensive method to assess the severity of HIV disease. In Phase II of the project, we will use our system to monitor patients over a 1-2 year period to correlate endotoxin neutralization with treatment.

Thesaurus Terms:
Acquired Immunodeficiency Syndrome;Address;Affect;Age;Bacterial Translocation;Base;Biological Assay;Biological Markers;Biological Monitoring;Blood Circulation;Cd14 Gene;Cd4 Positive T Lymphocytes;Cell Count;Chronic;Clinic;Clinical;Collection;Control Groups;Cost;Cytokine;Demographics;Detection;Disease;Disease Progression;Elements;Endotoxins;Evaluation;Funding;Gastrointestinal Tract Structure;Health;Hiv;Hiv Infections;Hiv Seropositivity;Human;Immune Activation;Immune System;Immunoglobulins;Inflammatory;Inflammatory Bowel Diseases;Inflammatory Response;Interleukin-1;Intestines;Lipopolysaccharide-Binding Protein;Male;Measures;Medical;Meetings;Methods;Monitor;Mortality Determinants;Mortality Vital Statistics;Novel;Pathology;Patient Monitoring;Patient Population;Patients;Permeability;Phase;Plasma;Preparation;Public Health Relevance;Publishing;Recording Of Previous Events;Research Personnel;Response;Sampling;Severities;Severity Of Illness;Small Business Innovation Research Grant;Source;Specimen;Students;System;Technology;Testing;Tool;Tumor Necrosis Factor-Alpha;United States;User-Friendly;Viral;Viral Load Result;

Endotoxin neutralization in human plasma is an excellent indicator of chronic immune activation, which is an accurate predictor of mortality in HIV-1 infection. Recently we developed an assay using endotoxin neutralization as an indicator of immune activation due to bacterial translocation. This assay is accurate in discriminating healthy patients from those with inflammatory bowel disease in addition to indicating disease severity. Phase I of this project expanded the assay to patients with HIV-1 infection. Those experiments showed statistically significant differences between healthy controls and HIV-1-infected patients as well as between anti-retroviral therapy (ART)-naïve patients and those actively receiving treatment. Additionally, endotoxin neutralization correlated as expected with markers of viral load, CD4+ T cell number, inflammatory response, macrophage activation, endotoxin-specific immunoglobulin sequestration and endotoxin-specific protein production. In Phase II of this SBIR project we will follow a group of HIV-1-infected, ART-naïve patients as they initiate ART and continue treatment over a 6 month time period. At specific time points we will collect blood plasma to measure the level of endotoxin neutralization (Specific Aim #2), a panel of traditional biomarkers indicating specific sequelae of the disease (Specific Aim #3) and both a quantitative and qualitative analysis of circulating 16S rDNA (Specific Aim #3). With this data, we will use Pearson correlation and Student’s t-test to determine (1) differences in endotoxin neutralization between HIV-1-infected patients and a demographically similar healthy control group, (2) the effect of ART on endotoxin neutralization, (3) the correlation of endotoxin neutralization with viral load, CD4+ T cell number, intestinal permeability, bacterial translocation, immune activation, inflammatory response and coagulopathy, and (4) the relationship of endotoxin neutralization with the species composition of the microbiome. Additionally, this project will include experiments to optimize the assay and prepare it for commercialization (Specific Aim #1).

Public Health Relevance Statement:
NARRATIVE This project addresses the major medical problem of HIV infection which affects over 1 million people in the United States at an annual cost of $40 billion. The proposed assay depends on the novel observation that the level of endotoxin neutralization in human plasma correlates with immune system activation due to intestinal permeability, a significant determinant of mortality in HIV infection. This technology will provide a fast, low-cost biomonitor that could be implemented in research and clinical applications.

Project Terms:
Address; Affect; Anti-Retroviral Agents; Anticoagulants; antiretroviral therapy; Bacterial Translocation; base; Biological Assay; Biological Markers; Blood Circulation; Blood Coagulation Disorders; Buffers; CD14 gene; CD4 Positive T Lymphocytes; CD80 gene; Cell Count; Chronic; Clinical; clinical application; clinical predictors; Collection; commercialization; Control Groups; cost; Data; Detection; Digestion; Disease; Disease Progression; Divalent Cations; Elements; Endotoxins; Evaluation; experimental study; Fibrin fragment D; Gastrointestinal tract structure; Health; Heating; HIV; HIV Infections; HIV-1; Human; IL2RA gene; immune activation; Immune Cell Activation; Immune response; Immune system; Immunoglobulins; Immunologic Markers; Infection; Inflammatory Bowel Diseases; Inflammatory Response; inhibitor/antagonist; Intestines; Kinetics; Macrophage Activation; Measures; Medical; Methods; microbial; microbiome; monocyte; mortality; Mortality Determinants; novel; Patients; Permeability; Phase; Plasma; Preparation; Production; Proteins; Protocols documentation; Recombinant DNA; Recombinants; Reproducibility; Research; response; RNA; Sampling; Severities; Severity of illness; Small Business Innovation Research Grant; Source; Students; T cell response; Technology; Temperature; Testing; Time; TNF gene; United States; user-friendly; Viral Load result; Viral Markers; zonulin