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Mycobacterial ESAT-6 and katG are recognized by sarcoidosis CD4+ T cells when presented by the American sarcoidosis susceptibility allele, DRB1*1101.

Oswald-Richter K, Sato H, Hajizadeh R, Shepherd BE, Sidney J, Sette A, Newman LS, Drake WP - J. Clin. Immunol. (2009)

Bottom Line: We previously reported immune recognition of Mycobacterium peptides from peripheral cells of 26 sarcoidosis subjects, 24 PPD- healthy volunteers, and eight with latent tuberculosis infection.Recognition was inhibited by monoclonal antibody against HLA-DR and HLA-DQ, but not HLA-DP.Immune recognition of ESAT-6 peptide NNALQNLARTISEAG was associated with possession of DRB1*1101.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Division of Infectious Diseases, Vanderbilt University School of Medicine, Nashville, TN, USA.

ABSTRACT

Introduction: Genetic associations of American sarcoidosis susceptibility implicate MHC class II allele, DRB1*1101. We previously reported immune recognition of Mycobacterium peptides from peripheral cells of 26 sarcoidosis subjects, 24 PPD- healthy volunteers, and eight with latent tuberculosis infection.

Materials and methods: In order to further link these genetic and immunologic pillars of sarcoidosis pathogenesis, we performed flow cytometry on these same subjects to identify the cells responsible for immune responses to ESAT-6 and katG peptides, followed by HLA typing to determine allelic associations with recognition.

Discussion and conclusion: Sarcoidosis CD4+ T cells were primarily responsible for the systemic responses. Recognition was inhibited by monoclonal antibody against HLA-DR and HLA-DQ, but not HLA-DP. Immune recognition of ESAT-6 peptide NNALQNLARTISEAG was associated with possession of DRB1*1101. ESAT-6 and katG presented by antigen-presenting cells expressing DRB1*1101-induced Th-1 responses from sarcoidosis T cells, thus providing a mechanistic insight for the association of HLA DRB1*1101 with sarcoidosis, and sarcoidosis T cell interaction with microbial antigens.

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HLA-DRB1*1101 presents ESAT-6 to sarcoidosis CD4+ T cells. Intracellular cytokine staining for IFN-γ was performed on expanded ESAT-6 and katG cell lines derived from sarcoidosis subject 6, after stimulation with either ESAT-6 peptide 14, katG peptide 13, or SEB. No recognition was observed in the expanded cell lines alone, demonstrating the absence of baseline IFN-γ production. Expanded cells stimulated with ESAT-6 or katG peptide in the absence of Sweig cells revealed minimal responses, confirming the loss of antigen-presenting cells during expansion. Significant responses to ESAT-6 and katG were observed in the respective cell line when antigen was presented using the DRB1*1101 expressing Sweig cell lines. Shown are representative flow cytometry dot plots indicating percentage of sarcoidosis CD4+ T cell responses to the different stimulation conditions
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Fig4: HLA-DRB1*1101 presents ESAT-6 to sarcoidosis CD4+ T cells. Intracellular cytokine staining for IFN-γ was performed on expanded ESAT-6 and katG cell lines derived from sarcoidosis subject 6, after stimulation with either ESAT-6 peptide 14, katG peptide 13, or SEB. No recognition was observed in the expanded cell lines alone, demonstrating the absence of baseline IFN-γ production. Expanded cells stimulated with ESAT-6 or katG peptide in the absence of Sweig cells revealed minimal responses, confirming the loss of antigen-presenting cells during expansion. Significant responses to ESAT-6 and katG were observed in the respective cell line when antigen was presented using the DRB1*1101 expressing Sweig cell lines. Shown are representative flow cytometry dot plots indicating percentage of sarcoidosis CD4+ T cell responses to the different stimulation conditions

Mentions: Sarcoidosis T Cells Recognize ESAT-6 and katG Antigens Presented by DRB1*1101-Expressing Antigen-Presenting Cells The observation that monoclonal antibody against HLA-DR partially inhibited immune responses to ESAT-6, along with the detection of immune responses to ESAT-6 and katG in a subject homozygous for DRB1*1101 (PPD+ 2) suggested that the peptides could be presented by this allele. Using the Sweig cell line which presents using the DRB1*1101 allele, we assessed for immune responses to ESAT-6 and katG peptides from an expanded cell line of Sarcoidosis 6. This subject had the MHC class II alleles DRB1*1101/1501. The expanded cell line was generated from PBMC stimulated with ESAT-6 peptide 14 or katG peptide 13. The expanded ESAT-6 line demonstrated no response to ESAT-6 peptide 14 without the presence of Sweig cells, confirming the loss of the patient’s own antigen-presenting cells during expansion. The addition of Sweig cells and ESAT-6 peptide to the ESAT-6 expanded cell line displayed a strong CD4+ IFN-γ response (Fig. 4). The ESAT-6 expanded cell line stimulated with katG resulted in a minimal response. The katG-expanded cell line resulted in minimal response, after stimulation with ESAT-6. It was noteworthy also that PBMC expanded with katG peptide 13 also demonstrated strong CD4+ responses to katG peptides if presented by the Sweig cells. This data confirms that mycobacterial virulence factors ESAT-6 and katG can be presented by the sarcoidosis susceptibility allele, DRB1*1101.Fig. 4


Mycobacterial ESAT-6 and katG are recognized by sarcoidosis CD4+ T cells when presented by the American sarcoidosis susceptibility allele, DRB1*1101.

Oswald-Richter K, Sato H, Hajizadeh R, Shepherd BE, Sidney J, Sette A, Newman LS, Drake WP - J. Clin. Immunol. (2009)

HLA-DRB1*1101 presents ESAT-6 to sarcoidosis CD4+ T cells. Intracellular cytokine staining for IFN-γ was performed on expanded ESAT-6 and katG cell lines derived from sarcoidosis subject 6, after stimulation with either ESAT-6 peptide 14, katG peptide 13, or SEB. No recognition was observed in the expanded cell lines alone, demonstrating the absence of baseline IFN-γ production. Expanded cells stimulated with ESAT-6 or katG peptide in the absence of Sweig cells revealed minimal responses, confirming the loss of antigen-presenting cells during expansion. Significant responses to ESAT-6 and katG were observed in the respective cell line when antigen was presented using the DRB1*1101 expressing Sweig cell lines. Shown are representative flow cytometry dot plots indicating percentage of sarcoidosis CD4+ T cell responses to the different stimulation conditions
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2821522&req=5

Fig4: HLA-DRB1*1101 presents ESAT-6 to sarcoidosis CD4+ T cells. Intracellular cytokine staining for IFN-γ was performed on expanded ESAT-6 and katG cell lines derived from sarcoidosis subject 6, after stimulation with either ESAT-6 peptide 14, katG peptide 13, or SEB. No recognition was observed in the expanded cell lines alone, demonstrating the absence of baseline IFN-γ production. Expanded cells stimulated with ESAT-6 or katG peptide in the absence of Sweig cells revealed minimal responses, confirming the loss of antigen-presenting cells during expansion. Significant responses to ESAT-6 and katG were observed in the respective cell line when antigen was presented using the DRB1*1101 expressing Sweig cell lines. Shown are representative flow cytometry dot plots indicating percentage of sarcoidosis CD4+ T cell responses to the different stimulation conditions
Mentions: Sarcoidosis T Cells Recognize ESAT-6 and katG Antigens Presented by DRB1*1101-Expressing Antigen-Presenting Cells The observation that monoclonal antibody against HLA-DR partially inhibited immune responses to ESAT-6, along with the detection of immune responses to ESAT-6 and katG in a subject homozygous for DRB1*1101 (PPD+ 2) suggested that the peptides could be presented by this allele. Using the Sweig cell line which presents using the DRB1*1101 allele, we assessed for immune responses to ESAT-6 and katG peptides from an expanded cell line of Sarcoidosis 6. This subject had the MHC class II alleles DRB1*1101/1501. The expanded cell line was generated from PBMC stimulated with ESAT-6 peptide 14 or katG peptide 13. The expanded ESAT-6 line demonstrated no response to ESAT-6 peptide 14 without the presence of Sweig cells, confirming the loss of the patient’s own antigen-presenting cells during expansion. The addition of Sweig cells and ESAT-6 peptide to the ESAT-6 expanded cell line displayed a strong CD4+ IFN-γ response (Fig. 4). The ESAT-6 expanded cell line stimulated with katG resulted in a minimal response. The katG-expanded cell line resulted in minimal response, after stimulation with ESAT-6. It was noteworthy also that PBMC expanded with katG peptide 13 also demonstrated strong CD4+ responses to katG peptides if presented by the Sweig cells. This data confirms that mycobacterial virulence factors ESAT-6 and katG can be presented by the sarcoidosis susceptibility allele, DRB1*1101.Fig. 4

Bottom Line: We previously reported immune recognition of Mycobacterium peptides from peripheral cells of 26 sarcoidosis subjects, 24 PPD- healthy volunteers, and eight with latent tuberculosis infection.Recognition was inhibited by monoclonal antibody against HLA-DR and HLA-DQ, but not HLA-DP.Immune recognition of ESAT-6 peptide NNALQNLARTISEAG was associated with possession of DRB1*1101.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Division of Infectious Diseases, Vanderbilt University School of Medicine, Nashville, TN, USA.

ABSTRACT

Introduction: Genetic associations of American sarcoidosis susceptibility implicate MHC class II allele, DRB1*1101. We previously reported immune recognition of Mycobacterium peptides from peripheral cells of 26 sarcoidosis subjects, 24 PPD- healthy volunteers, and eight with latent tuberculosis infection.

Materials and methods: In order to further link these genetic and immunologic pillars of sarcoidosis pathogenesis, we performed flow cytometry on these same subjects to identify the cells responsible for immune responses to ESAT-6 and katG peptides, followed by HLA typing to determine allelic associations with recognition.

Discussion and conclusion: Sarcoidosis CD4+ T cells were primarily responsible for the systemic responses. Recognition was inhibited by monoclonal antibody against HLA-DR and HLA-DQ, but not HLA-DP. Immune recognition of ESAT-6 peptide NNALQNLARTISEAG was associated with possession of DRB1*1101. ESAT-6 and katG presented by antigen-presenting cells expressing DRB1*1101-induced Th-1 responses from sarcoidosis T cells, thus providing a mechanistic insight for the association of HLA DRB1*1101 with sarcoidosis, and sarcoidosis T cell interaction with microbial antigens.

Show MeSH
Related in: MedlinePlus