Data from: NK-CD11c+ cell crosstalk in diabetes enhances IL-6-mediated inflammation during mycobacterium tuberculosis infection

Cheekatla SS, Tripathi D, Venkatasubramanian S, Nathella PK, Paidipally P, Ishibashi M, Welch E, Tvinnereim AR, Ikebe M, Valluri VL, Babu S, Kornfeld H, Vankayalapati R

Date Published: November 4, 2016

DOI: http://dx.doi.org/10.5061/dryad.qn42t

 

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Title S1
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Description Figure S1. Mtb infection enhances the dissemination of bacteria in T2DM. Control and T2DM mice were infected with 50-100 CFU of aerosolized Mtb H37Rv A & B. Bacterial burden in the spleen and liver six months post-infection. C & D. Random blood glucose levels and body weight were measured at monthly intervals for up to 6 months.
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Title S2
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Description Figure S2. Type 2 diabetes enhances pro- and anti-inflammatory responses during Mtb infection. Control and T2DM mice were infected with 50-100 CFU of aerosolized Mtb H37Rv. One and six months p.i., lung homogenates were collected from uninfected control and diabetic mice and from Mtb-infected control and diabetic mice and subjected to multiplex ELISA to determine the various cytokines and chemokines. Mean values, p-values and SEs are shown. *P < 0.05, **P < 0.01, ***P < 0.001.
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Title S3
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Description Figure S3. Type 2 diabetes enhances pro- and anti-inflammatory responses during Mtb infection. Control and T2DM mice were infected with 50-100 CFU of aerosolized Mtb H37Rv. A to M. Six months p.i., lung cytokine mRNA expression was determined by real-time PCR. Mean values, p-values and SEs are shown. *P < 0.05, **P < 0.01, ***P < 0.001.
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Title S4
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Description Figure S4. CD11c+ dendritic cells are the major source of IL-6 in Mtb-infected type 2 diabetic mice. Control and T2DM mice were infected with 50-100 CFU of aerosolized Mtb H37Rv. A. Six months p.i., pooled CD4+, CD8+, DX5+ and CD11c+ cells from the lung, spleen and lymph nodes were isolated by magnetic selection. IL-6 mRNA expression was determined by real-time PCR. The results of three independent experiments are shown. B & C. Splenic DX5+ and CD11C+ cells were isolated by magnetic selection and cultured (1 NK cell and 4 dendritic cells) with or without g-irradiated Mtb (10 µg/ml). Some of the g-irradiated Mtb H37Rv cultured cells were cultured in the presence of neutralizing antibodies (10 µg/ml) against DNAM-1 or rat IgG2a, κ (the isotype control antibody for the anti-DNAM-1 antibody) or against NKG2D or rat IgG1, κ (the isotype control antibody for the anti-NKG2D antibody). After 18 hours, cell-free culture supernatants were collected and IL-6 levels were measured by ELISA. Mean values, p-values and SEs are shown. *P ≤ 0.05, **P ≤0.01, ***P ≤ 0.001.
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Title S5
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Description Figure S5 Natural killer and dendritic cell interaction enhanced IL-6 production in Mtb-infected type 2 diabetic mice. Control and T2DM mice were infected with 50-100 CFU of aerosolized Mtb H37Rv. Six months p.i., lungs from Mtb-infected control and T2DM mice were isolated and formalin-fixed. Paraffin-embedded tissue sections were prepared and confocal microscopy analysis was performed to determine NK (pink), IL-6+ (green) and dendritic (red) cell co-localization. Scale bar: 20 µm (yellow bar) and 5 µm (white bar). Representative images of staining pattern of three independent experiments, each with three per group were shown.
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Title S6
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Description Figure S6. IL-6 is responsible for the increased pro-inflammatory cytokine production in type 2 diabetic patients with pulmonary tuberculosis. Blood from 20 pulmonary tuberculosis patients with T2DM and 20 pulmonary tuberculosis patients without diabetes was obtained. Whole blood was cultured with 10 µg/ml of purified protein derivative (PPD) as indicated in the methods section. A representative flow cytometric contour plot showing the frequency of cells expressing IFN-, TNF-, IL-17 and IL-2 is shown.
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When using this data, please cite the original publication:

Cheekatla SS, Tripathi D, Venkatasubramanian S, Nathella PK, Paidipally P, Ishibashi M, Welch E, Tvinnereim AR, Ikebe M, Valluri VL, Babu S, Kornfeld H, Vankayalapati R (2016) NK-CD11c+ Cell Crosstalk in Diabetes Enhances IL-6-Mediated Inflammation during Mycobacterium tuberculosis Infection. PLOS Pathogens 12(10): e1005972. http://dx.doi.org/10.1371/journal.ppat.1005972

Additionally, please cite the Dryad data package:

Cheekatla SS, Tripathi D, Venkatasubramanian S, Nathella PK, Paidipally P, Ishibashi M, Welch E, Tvinnereim AR, Ikebe M, Valluri VL, Babu S, Kornfeld H, Vankayalapati R (2016) Data from: NK-CD11c+ cell crosstalk in diabetes enhances IL-6-mediated inflammation during mycobacterium tuberculosis infection. Dryad Digital Repository. http://dx.doi.org/10.5061/dryad.qn42t
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