Mayumi Nakagawa (United States of America)

University of Arkansas for Medical Sciences Pathology

Presenter of 1 Presentation

Conference Calendar

Basic Research / Papillomavirus Vaccines (i.e New Developments) ePoster

IDENTIFYING ACTIVE COMPONENTS OF CANDIDA SKIN TEST REAGENT USED AS AN ADJUVANT IN AN HPV THERAPEUTIC VACCINE, PEPCAN (ID 452)

Session Date

07/21/2020

Session Time

10:00 - 17:00

Room

ePoster

Session Type

Poster Viewing - 20-24 July

Session Name

Basic Research / Papillomavirus Vaccines (i.e New Developments)

Lecture Time

10:04 - 10:05

Presenter

Mayumi Nakagawa (United States of America)

Authors

Abstract

Abstract

Introduction

Candida skin test reagent has been used as an adjuvant for an HPV therapeutic vaccine due to its immunostimulatory effects. The goal of this project was to identify its active components and mechanisms.

Methods

Dialyzed Candida was fractionated in to 20 one minute parts using liquid chromatography tandem mass spectrometry. Each fraction was tested for induction of IL-12 transcription with quantative reverse transcriptase-polymerase chain reaction by monocyte-derived Langerhans cells from healthy donors (n=5) prescreened for IL-12 production with Candida. Five fractions induced IL-12 secretions in 4 of 5 donors. A total of 10 discrete small molecules (100-600 g/mole), disignated A-J, were identified. The fractions containing 3 of these molecules (A, F, and I) were consistently present in another lot of Candin, and were not present in PBS (dialysis fluid). Whole transcriptome profiling and multiplex cytokine analysis of supernatants [IL-1beta, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17, eotaxin, basic FGF, G-CSF, GM-CSF, IFN-gamma, IP-10, MCP-1 (MCAF), MIP-1alpha, MIP-1beta, PDGFBB, RANTES, TNF-alpha, VEGF] were performed in 10 screen-positive donors. RNA-seq data were demultiplexed, trimmed for adaptors, cleaned, mapped, annotated, and normalized. Principal component analysis (PCA), differential expression (DE) analysis, and over-representation analysis (ORA) were performed.

Results

The PCA did not reveal clustering; however, DE analyses revealed increased expression of 9 genes with the fraction containing A in comparison to PBS. Six these 9 genes significantly over-expressed were metallothineins. ORA revealed involvement of multiple pathways utilizing ions, such as copper, zinc, and cadmium, as well as those involved in cellular responses. Cytokine analysis revealed that the most number of significant increases (6 cytokines) occurred in the same fraction containing A.

Conclusions

The fraction containing A appears to be most biologically active. Future study would involve identification of this molecule, and a confirmation of its immunostimlulatory activities.

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Moderator of 1 Session

Conference Calendar

Public Health Oral Session

01050 - Oral Session 17: HEAD & NECK CANCERS. Epidemiology, natural history and prevention (ID 71)

Session Type

Public Health Oral Session

Session Date

07/24/2020

Session Time

14:15 - 15:15

Room

Hall D

Chair(s)

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Session Webcast

Session Webcast

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