Research ArticleORIGINAL RESEARCH
Open Access

Systemic T Cell Subsets and Cytokines in Patients With Homozygous Sickle Cell Disease and Asymptomatic Urinary Tract Infections in Togo

Christèle Nguepou Tchopba, Gnatoulma Katawa, Essohana Padaro, Pélagie Edlom Tchadié, Simplice Damintoti Karou, Ahoefa Vovor and Yaovi Ameyapoh
Ochsner Journal June 2021, 21 (2) 163-172; DOI: https://doi.org/10.31486/toj.20.0061

Article Figures & Data

Figures

  • Figure 1.
    Figure 1.

    Gating strategy: (A) Lymphocyte gate; (B) CD4+ T lymphocyte population (LTCD4+) delineated among the lymphocytes; (C) (E) (G) are respectively CD4+Tbet+, CD4+GATA3+, and CD4+FoxP3+ dot plots; (D) CD4+Tbet+IFN-γ+ T cells are IFN-γ+ cells delineated among CD4+Tbet+ cells; (F) CD4+GATA3+IL-4+ T cells are IL-4+ cells delineated among CD4+GATA3+ cells; and (H) CD4+FoxP3+IL-10+ T cells are IL-10+ cells delineated among CD4+FoxP3+ cells.

  • Figure 2.
    Figure 2.

    Plasma cytokine profile measured by sandwich enzyme-linked immunosorbent assay method. The graphs show plasma levels of (A) interferon gamma (IFN-γ) and (B) interleukin-10 (IL-10) in HbSS subjects who had urinary tract infections (HbSS inf*, n=12), HbSS subjects who did not have urinary tract infections (HbSS, n=10), uninfected HbAS subjects (n=10), and uninfected HbAA volunteers (n=11). Bars indicate the concentration of cytokines in each group. Data are expressed as mean ± SEM. Asterisks show statistical differences (Mann-Whitney U test) between the groups (*P<0.05).

  • Figure 3.
    Figure 3.

    Plasma cytokine profile in females only measured by sandwich enzyme-linked immunosorbent assay method. The graphs show plasma levels of (left) interferon gamma (IFN-γ) and (right) interleukin-10 (IL-10) in HbSS subjects who had urinary tract infections (HbSS inf*, n=11), HbSS subjects who did not have urinary tract infections (HbSS, n=6), uninfected HbAS subjects (n=5), and uninfected HbAA volunteers (n=3). Bars indicate the concentration of cytokines in each group. Data are expressed as mean ± SEM. Asterisks show statistical differences (Mann-Whitney U test) between the groups (*P<0.05).

  • Figure 4.
    Figure 4.

    T helper type 1, T helper type 1, and natural regulatory T cells profile. Cells from HbSS subjects who had urinary tract infections (HbSS inf*, n=12), HbSS subjects who did not have a urinary tract infection (HbSS, n=10), uninfected HbAS subjects (n=10), and uninfected HbAA volunteers (n=11) were activated with phorbol 12-myristate 13-acetate and stained to determine the frequencies of (A) CD4+Tbet+IFN-γ+ T cells, (B) CD4+GATA3+IL-4+ T cells, and (C) CD4+FoxP3+IL-10+ T cells. Bars indicate the percentages of cells, expressed as mean ± SEM. Asterisks show statistical differences (Mann-Whitney U test) between the groups (*P<0.05).

  • Figure 5.
    Figure 5.

    Immunogenicity of bacterial strains isolated from the urine of infected HbSS subjects after 24 hours of culture. Peripheral blood mononuclear cells (PBMCs) from healthy volunteer donors (n=6) were stimulated with antigens (Ag) derived from bacterial strains (randomly named S1, S2, and S3). Bars indicate levels of type 1 cytokines: (A) tumor necrosis factor (TNF)-α and (B) interleukin (IL)-6. Medium indicates the well containing unstimulated cells. LPS indicates the well containing cells co-cultured with lipopolysaccharide. Asterisks show statistical differences (Mann-Whitney U test) between the different co-cultures or PBMC stimulations performed (*P<0.05).

  • Figure 6.
    Figure 6.

    Immunogenicity of bacterial strains isolated from the urine of infected HbSS subjects after 72 hours of culture. Peripheral blood mononuclear cells (PBMCs) from healthy volunteer donors (n=6) were stimulated with antigen (Ag) from bacterial strains (S1, S2, and S3). Bars indicate levels of type 2 cytokines: (A) interferon gamma (IFN-γ), (B) interleukin (IL)-5, and (C) IL-10. Medium indicates the well containing unstimulated cells. LPS indicates the well containing cells co-cultured with lipopolysaccharide. Asterisks show statistical differences (Mann-Whitney U test) between the different co-cultures or PBMC stimulations performed (*P<0.05).

Tables

  • Table.

    Demographic Characteristics of the Studied Populations

    Population
    VariableHbSS With Infectiona n=12HbSSb n=10HbASc n=10HbAAd n=11P Value
    Age, years, mean ± SD20.33 ± 3.5821.10 ± 2.3126.00 ± 2.8927.27 ± 1.700.106
    Age ratio: <18 years:≥18 years6:65:51:90:110.011
    Sex ratio: male:female1:114:65:58:30.018

    • ↵aHbSS with infection, subjects with homozygous sickle cell disease and with urinary tract infection.

    • ↵bHbSS, subjects with homozygous sickle cell disease and without urinary tract infection.

    • ↵cHbAS, heterozygous subjects without urinary tract infection.

    • ↵dHbAA, subjects without sickle cell disease or sickle cell trait and without urinary tract infection.

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