This proposal intends to characterize Tfh cells, a population that is critical for disease development and maintenance in SLE and test the novel hypothesis that SLE-Tfh are uniquely sensitive to metabolic inhibition, a feature that can be exploited to therapeutic intervention. The scientific premise of this proposal is based on three lines of evidence: 1. The necessary role of Tfh cells for the production of pathogenic autoAbs in lupus has been well-established in SLE patients and mouse models; 2. Tfh cells induced by immunization are metabolically quiescent, a fact that we have confirmed showing that they are not affected by glucose inhibitors in lupus-prone and control mice; and 3. We have strong preliminary data showing that SLE-Tfh cells from four different mouse models of lupus are sensitive to glucose. We propose the hypothesis that spontaneous Tfh cells supporting the production of autoAbs (SLE-Tfh) have a different metabolism than Tfh cells providing protective humoral immunity against pathogens (TD-Tfh) in either lupus or normal mice. Based on similarities of CD4+ T cell functions and metabolism in lupus-prone mice and SLE patients, we also hypothesize that the expanded Tfh cells in SLE patients also have a different metabolism than that of health controls (HCs). Consequently, we predict that targeting Tfh cellular metabolism provides an effective approach to treat lupus without compromising the patients? protection against TD- pathogens. We propose to test these hypotheses using cellular and molecular immunology approaches in mouse models as well as with human PBLs with the three following specific aims 1. To define the molecular and metabolic signatures of SLE-Tfh as compared to TD-Tfh cells elicited by TD-dependent Ags (PR8 flu virus and NP-OVA) in lupus mice and B6 controls. The immunophenotypes, anatomical location, gene expression, and metabolic profile of spontaneous Tfh cells in TC and B6.lpr mice will be compared to that of flu-specific I-A(b) NP-tetramer positive Tfh cells in TC, B6.lpr and B6 mice infected with PR8 virus. In addition to these polyclonal T cell models, we will use an adoptive transfer model of OVA-specific OT-II T cells carrying the Sle1 lupus susceptibility allele that favors the expansion of Tfh cells (9) into NP-OVA immunized mice (10).
This aim will define the functional differences between TD-Tfh and SLE-Tfh cells in two models of lupus and with two different TD-immunizations. 2. To define the response of SLE-Tfh and TD-Tfh cells to glucose inhibition in the mouse. Using the same experimental systems as in SA1, we will compare the responses of SLE-Tfh cells and TD-Tfh cells to glucose inhibition in mice treated with 2DG, a glucose analog that blocks the first reaction of glycolysis. 3. To compare the molecular and metabolic signatures of circulating cTfh cells in SLE patients and HCs, as well as their response to metformin. We hypothesize that the expansion of cTfh cells in SLE patients is largely driven by autoAgs, and that SLE-cTfh cells share functional signatures with murine SLE-Tfh cells. On the other hand, cTfh cells from HCs have been largely induced by TD-Ags, and HC-cTfh cells should overlap with murine TD-Tfh cells. We will compare the immunophenotypes and gene expression of these two types of Tfh cells, as well as their response to the metabolic inhibitor metformin in vitro. Our first goal is to advance our understanding of SLE-Tfh cells, and ultimately to advance the treatment of lupus, based on discrete, achievable goals focusing on one cell population. This project will advance our understanding of disease mechanisms, and yield results with a high translational potential.

Public Health Relevance

The production of autoantibodies that are responsible for tissue damage in lupus requires the contribution of a specific type of T cells called follicular helper T cells (Tfh cells). The expansion of the number of Tfh cells correlates with disease severity in lupus patients, and there is ample evidence from mouse models that reducing the number of Tfh cells eliminates disease. We have shown that glucose inhibition normalizes the frequency of Tfh cells that are responsible for the production of autoantibodies in lupus-prone mice, while Tfh cells that are responsible for the production of protective antibodies following immunization are largely unaffected. We propose the hypothesis that the elimination of lupus Tfh cells through glucose inhibition represents a safe therapeutic approach. We propose to test this hypothesis by characterizing lupus Tfh cells relative to Tfh cells induced by immunization in mouse models and cells obtained from lupus patients.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI128901-02
Application #
9391652
Study Section
Hypersensitivity, Autoimmune, and Immune-mediated Diseases Study Section (HAI)
Program Officer
Johnson, David R
Project Start
2016-12-01
Project End
2021-11-30
Budget Start
2017-12-01
Budget End
2018-11-30
Support Year
2
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of Florida
Department
Pathology
Type
Schools of Medicine
DUNS #
969663814
City
Gainesville
State
FL
Country
United States
Zip Code
32611