# Activation of semi-invariant and diverse NKT cells with an adjuvant combination

> **NIH NIH R01** · UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR · 2021 · $367,120

## Abstract

ABSTRACT
There are several safe candidate adjuvants that potentiate beneficial immunity following vaccination. However,
single adjuvant formulations have performed quite modestly in terms of stimulating the desired blend of
humoral versus cellular immunity and/or Th1 versus Th2 immunity. Combination adjuvants offer a possible way
forward, but the mechanisms of action of such combinations remain poorly defined. Therefore, we will combine
the FDA-approved adjuvant Alum (alhydrogel) with the CD1d-binding glycolipid adjuvant α-galactosylceramide
(α-GC) and delineate the mechanisms of action with a particular emphasis on CD1d-restricted Natural Killer T
(NKT) cells and humoral immunity. Alum is safe and stimulates excellent Th2 but poor Th1 immunity. We
discovered that the Th2 response to Alum depends in large part upon diverse TCR-expressing Type II NKT
cells (dNKT) which recognize CD1d/glycolipid complexes but do not recognize the α-GC adjuvant. The α-GC
adjuvant stimulates semi-invariant TCR-expressing Type I NKT cells (iNKT) and results in a mixed Th1/Th2
response. It is already known from Phase I clinical trials in patients with cancer that α-GC is well-tolerated and
safe. Furthermore, modification of the α-GC structure can readily be employed to skew the Type I NKT-
dependent Th1/Th2 balance. Our objective for this proposal is to test the central hypothesis that the
combination of Alum and α-GC leads to CD1d/glycolipid presentation and a coordinated dNKT and iNKT-
driven mixed Th1/Th2 response against vaccine antigens.
In Specific Aim 1, we will perform a series of in vitro studies in primary murine and human cells to determine
how the adjuvant combination affects CD1d Ag presentation and activation and functional differentiation of
NKT cells into different effector subsets. We will also examine the effect of adjuvant combination on class I and
class II presentation of co-administered protein antigens.
In Specific Aim 2, we will undertake a series of in vivo experiments in mice to examine the functional
consequences of adjuvant combination for Ab-mediated protection against bacterial toxins. Experiments to
analyze cellular as well as humoral immunity will be included as the project develops and in vivo toxicity (or
lack thereof) will be determined by measurement of pro-inflammatory cytokines as well as markers of
autoimmunity and organ damage.
We feel this project is innovative because it will provide the vaccine community with mechanistic information
when CD1d-binding adjuvants are combined with Alum adjuvant and give careful consideration to NKT cell-
driven components of the immune response. We feel the project is significant because it will illuminate
potential avenues for inclusion of CD1d-binding glycolipids in mixed adjuvant platforms.

## Key facts

- **NIH application ID:** 10053313
- **Project number:** 5R01AI134719-04
- **Recipient organization:** UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
- **Principal Investigator:** Mark L Lang
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $367,120
- **Award type:** 5
- **Project period:** 2017-11-06 → 2022-10-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10053313

## Citation

> US National Institutes of Health, RePORTER application 10053313, Activation of semi-invariant and diverse NKT cells with an adjuvant combination (5R01AI134719-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10053313. Licensed CC0.

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