Botanical Core – Dr. Amar Chittiboyina, Core Leader
Before botanical dietary supplements can be meaningfully evaluated, the material must be properly authenticated, standardized, and safe to use. This can be adequately accomplished by implementing integrity profiles viz., NCCIH’s Natural Product Integrity Policy. While there has been an increased interest in science-based research of botanical dietary supplements, much of the research, unfortunately, has been plagued by studies being conducted on unauthenticated and uncharacterized products. Besides, there may be a misidentification of the collected plant, adulteration with other species, or contamination with extraneous ingredients. From the perspective of the scientific community, the lack of authentication of study materials has resulted in a considerable amount of published work that is misleading or not reproducible botanical research arena. Hence, the implementation of proper authentication tools such as the use of botanical reference materials, classical taxonomic techniques such as morphological evaluation, microscopic analysis, as well as chemical fingerprinting, and identification of quality chemical markers is necessary for botanical dietary supplement research.
Botanical raw materials exhibit inherent variability due to numerous factors, e.g., genetic/phenotypic variations, geographic/climatic variables, differences in agronomic conditions during growth, harvesting practices, etc. This variation in botanical raw materials can influence the level of active compounds and thereby result in a lack of consistent quality of the finished products. It is, therefore, possible that the inconsistent outcomes commonly reported for clinical trials associated with botanicals are due, in part, to variation in the level of active compounds within the products used in those studies. The use of properly standardized botanical test materials is therefore essential for quality research studies.
The overall purpose of the Botanical Core is to ensure product integrity and advance the chemistry research on the Spirulina-based product, Immulina™, by using a combination of bioassay- and chemical-based approaches.
The specific aims of the Botanical Core are as follows:
Aim 1.
Ensure the unambiguous identification and perform additional safety testing on Arthrospira (Spirulina) platensis raw material that will be used in the production of sufficient quantities of Immulina for the proposed research projects.
Aim 2.
Validate a selective in vitro bioassay for quantitation of the toll-like receptor 2-dependent activity exhibited by the Braun-type lipoproteins in Immulina extracts.
Aim 3.
Advance the chemistry research on Immulina by establishing chemical-based authentication and standardization approaches, characterization of immune-inhibitory substances, and detailed structural analysis of the active immune-enhancing Braun-type lipoproteins.
Aim 4.
Perform additional stability studies, provide sufficient quantities of well-characterized and safe Immulina for use in the proposed UM BDSRC research projects, and explore collaborative opportunities with other CARBON units.
Project 1 – Dr. Chalet Tan, Principal investigator
Built upon the significant knowledge gained by the scientists at the National Center for Natural Products Research over the past two decades as well as the exciting new findings from our studies, we propose a two-step mechanism of action for Immulina: 1) binding of Braun-type lipoproteins (bioactive molecules in Immulina) to TLR2 on the phagocytes, which are present abundantly in the Peyer’s patches of the small intestinal epithelium, induces coordinated expression of NFκB target genes in the phagocytes; 2) activated phagocytes leads to the stimulation of lymphoid cells in the circulation as well as in the disease site (such as influenza-infected lung and the draining lymph nodes), augmenting the proliferation and cytolytic activity of lymphocytes and resulting in enhanced immunity. To achieve better delivery of Immulina in vivo, we will also develop an optimized liquid formulation for Immulina.
The project will include 3 specific aims:
Aim 1.
Develop an optimized liquid formulation for Immulina.
Aim 2.
Evaluate the pharmacodynamics of Immulina and identify in vivo biomarker(s).
Aim 3.
Investigate the molecular mechanism on immune enhancement by Immulina.
Statement of Potential Impact.
The successful completion of this project will result in the development of an improved formulation of Immulina and the identification of in vivo biomarkers for Immulina treatment.
Project 2 – Dr. Gailen Marshall, Principal Investigator
Although oral ingestion of Immulina has been reported to exhibit a protective effect against influenza A (H1N1) viral infection using a mouse model (Pugh et al, 2015), it is unknown whether this protection is solely prophylactic, prodromal, therapeutic or a combination of these effect. Furthermore, very small human trials (Nielsen et al, 2010) indicate that the impact of Immulina on antiviral resilience could be through enhanced activity of natural killer cells. Building on these previous studies, there is a need to establish whether Immulina might be useful as a dietary supplement to modify human host antiviral immune mechanisms, in particular among individuals with immune compromised states. Further, it is important to determine whether such a supplement would need to be taken year-round or could be started at the first sign of viral infection.
Accordingly, this project will investigate the following hypothesis: Immulina given in its optimal oral form will alter the host antiviral immune response, manifested by increases in lymphocyte numbers and/or activity, anti-flu H and N antibody titers and CD8+ mediated cytotoxicity against flu-infected cells.
To investigate our hypothesis, we will achieve the following specific aims:
Mouse model (Years 1-2)
Aim 1.
Evaluate oral administration of Immulina in three non-lethal mouse models of resilience against influenza A virus infection (prophylaxis, prodrome and recovery) to determine the most effective utility of Immulina for enhancing host immunity to improve antiviral resilience.
Aim 2.
Confirm that activation of the TLR2 signaling pathway by Braun-type lipoproteins is a causal mechanism through which Immulina enhances host immunity against antiviral infection.
Human model (Years 3-5)
Aim 3.
Determine the optimal form and dosage of theImmulina-based supplement that will maximize effects on increasing lymphocyte cell numbers and/or activity, increased supporting cytokines, ADCC against influenza-infected target cell lines and influenza-specific antibody titers.
Aim 4.
Establish the timeline for optimal lymphocyte, cytokine and antibody responses in terms of both initial changes and maximal changes and duration of the change once the Immulina is discontinued in normal and immune compromised (elderly) human research participants.
Aim 5.
Examine the effects of routine influenzas vaccine given before, during, or after Immulina use to investigate influenza antigen-specific antibody responses in individuals receiving Immulina supplement vs placebo.
With this design, we anticipate developing the necessary pharmacokinetic parameters (dosage, duration needed for therapeutic-related biomarker alteration in prophylaxis vs prodromal vs. therapeutic models) to develop and perform future clinical trials that will study the potential for Immulina as an antiviral immune modulator for influenza and perhaps other viral infections.