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Epitopes of microbial and human heat shock protein 60 and their recognition in myalgic encephalomyelitis.

Elfaitouri A, Herrmann B, Bölin-Wiener A, Wang Y, Gottfries CG, Zachrisson O, Pipkorn R, Rönnblom L, Blomberg J - PLoS ONE (2013)

Bottom Line: A HSP60-based panel of 25 antigens was selected.When evaluated with 61 other ME and 399 non-ME samples (331 BD, 20 Multiple Sclerosis and 48 Systemic Lupus Erythematosus patients), a peptide from Chlamydia pneumoniae HSP60 detected IgM in 15 of 61 (24%) of ME, and in 1 of 399 non-ME at a high cutoff (p<0.0001).IgM to specific cross-reactive epitopes of human and microbial HSP60 occurs in a subset of ME, compatible with infection-induced autoimmunity.

View Article: PubMed Central - PubMed

Affiliation: Section of Clinical Microbiology, Department of Medical Sciences, Uppsala University, Uppsala, Sweden.

ABSTRACT
Myalgic encephalomyelitis (ME, also called Chronic Fatigue Syndrome), a common disease with chronic fatigability, cognitive dysfunction and myalgia of unknown etiology, often starts with an infection. The chaperonin human heat shock protein 60 (HSP60) occurs in mitochondria and in bacteria, is highly conserved, antigenic and a major autoantigen. The anti-HSP60 humoral (IgG and IgM) immune response was studied in 69 ME patients and 76 blood donors (BD) (the Training set) with recombinant human and E coli HSP60, and 136 30-mer overlapping and targeted peptides from HSP60 of humans, Chlamydia, Mycoplasma and 26 other species in a multiplex suspension array. Peptides from HSP60 helix I had a chaperonin-like activity, but these and other HSP60 peptides also bound IgG and IgM with an ME preference, theoretically indicating a competition between HSP60 function and antibody binding. A HSP60-based panel of 25 antigens was selected. When evaluated with 61 other ME and 399 non-ME samples (331 BD, 20 Multiple Sclerosis and 48 Systemic Lupus Erythematosus patients), a peptide from Chlamydia pneumoniae HSP60 detected IgM in 15 of 61 (24%) of ME, and in 1 of 399 non-ME at a high cutoff (p<0.0001). IgM to specific cross-reactive epitopes of human and microbial HSP60 occurs in a subset of ME, compatible with infection-induced autoimmunity.

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Phyloscan with G20c homologs of 25 bacterial and protozoal HSP60s for IgG (A) and IgM (B) binding.The average binding of patient (pt-av) and BD (bd-av) samples is shown. The relatively high NTC values (C) were partially subtracted as described in Materials and Methods. A significance test (D) using the Fisher exact test is also shown. The red line depicts a p value of 0.001. The peptides G20 (lane 1), G20c (2), recombinant human HSP60 (3) and E. coli GroEL (4) were also included. The names of the G20c homolog peptides are explained in File S1. Lanes 24-29 refer to eukaryotes. The rest are from prokaryotes. The order is the same as in the alignment of HSP60 sequences (Figure SF4 in file S1). Ordinates of sections A-C show MFI. E-F: Phylogenetic epitope definition. For each peptide pair, the correlation coefficient, the degree of sequence similarity (calculated with the BLOSUM62 scoring matrix), and the two peptide sequences, is shown. Identical peptide sequences get around 150 points with this scoring system. Three discordant sequence pairs with high IgG (E) and IgM (F) serological correlation are shown.
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pone-0081155-g002: Phyloscan with G20c homologs of 25 bacterial and protozoal HSP60s for IgG (A) and IgM (B) binding.The average binding of patient (pt-av) and BD (bd-av) samples is shown. The relatively high NTC values (C) were partially subtracted as described in Materials and Methods. A significance test (D) using the Fisher exact test is also shown. The red line depicts a p value of 0.001. The peptides G20 (lane 1), G20c (2), recombinant human HSP60 (3) and E. coli GroEL (4) were also included. The names of the G20c homolog peptides are explained in File S1. Lanes 24-29 refer to eukaryotes. The rest are from prokaryotes. The order is the same as in the alignment of HSP60 sequences (Figure SF4 in file S1). Ordinates of sections A-C show MFI. E-F: Phylogenetic epitope definition. For each peptide pair, the correlation coefficient, the degree of sequence similarity (calculated with the BLOSUM62 scoring matrix), and the two peptide sequences, is shown. Identical peptide sequences get around 150 points with this scoring system. Three discordant sequence pairs with high IgG (E) and IgM (F) serological correlation are shown.

Mentions: When the G20c peptide had been identified we were led to perform phylogenetic scanning of HSP60 G20c homologs for ME and BD antibody binding. Amino acid sequences of HSP60 from bacteria, protozoa, fungi and humans were aligned (Figure SF4 in file S1). A set of 25 G20c homolog 30-mer peptides (Table ST3 in file S1) were synthesized and tested in the IgG and IgM tests. The highest IgG values were with the Schistosoma mansoni, Plasmodium falciparum, Cryptosporidium, Staphylococcus, Borrelia, Leishmania, Chlamydia and Leptospira homologs. In the IgM test, especially strong reactions were seen with Leishmania, Treponema, Entamoeba and Chlamydia homologs (Figure 2). The five G20c homolog peptides with the highest positive (100%) and negative predictive values (62-68%) for ME in the IgM test were from Staphylococcus, Plasmodium, Listeria, Burkholderia and Chlamydia (Figure SF5 in file S1)


Epitopes of microbial and human heat shock protein 60 and their recognition in myalgic encephalomyelitis.

Elfaitouri A, Herrmann B, Bölin-Wiener A, Wang Y, Gottfries CG, Zachrisson O, Pipkorn R, Rönnblom L, Blomberg J - PLoS ONE (2013)

Phyloscan with G20c homologs of 25 bacterial and protozoal HSP60s for IgG (A) and IgM (B) binding.The average binding of patient (pt-av) and BD (bd-av) samples is shown. The relatively high NTC values (C) were partially subtracted as described in Materials and Methods. A significance test (D) using the Fisher exact test is also shown. The red line depicts a p value of 0.001. The peptides G20 (lane 1), G20c (2), recombinant human HSP60 (3) and E. coli GroEL (4) were also included. The names of the G20c homolog peptides are explained in File S1. Lanes 24-29 refer to eukaryotes. The rest are from prokaryotes. The order is the same as in the alignment of HSP60 sequences (Figure SF4 in file S1). Ordinates of sections A-C show MFI. E-F: Phylogenetic epitope definition. For each peptide pair, the correlation coefficient, the degree of sequence similarity (calculated with the BLOSUM62 scoring matrix), and the two peptide sequences, is shown. Identical peptide sequences get around 150 points with this scoring system. Three discordant sequence pairs with high IgG (E) and IgM (F) serological correlation are shown.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3842916&req=5

pone-0081155-g002: Phyloscan with G20c homologs of 25 bacterial and protozoal HSP60s for IgG (A) and IgM (B) binding.The average binding of patient (pt-av) and BD (bd-av) samples is shown. The relatively high NTC values (C) were partially subtracted as described in Materials and Methods. A significance test (D) using the Fisher exact test is also shown. The red line depicts a p value of 0.001. The peptides G20 (lane 1), G20c (2), recombinant human HSP60 (3) and E. coli GroEL (4) were also included. The names of the G20c homolog peptides are explained in File S1. Lanes 24-29 refer to eukaryotes. The rest are from prokaryotes. The order is the same as in the alignment of HSP60 sequences (Figure SF4 in file S1). Ordinates of sections A-C show MFI. E-F: Phylogenetic epitope definition. For each peptide pair, the correlation coefficient, the degree of sequence similarity (calculated with the BLOSUM62 scoring matrix), and the two peptide sequences, is shown. Identical peptide sequences get around 150 points with this scoring system. Three discordant sequence pairs with high IgG (E) and IgM (F) serological correlation are shown.
Mentions: When the G20c peptide had been identified we were led to perform phylogenetic scanning of HSP60 G20c homologs for ME and BD antibody binding. Amino acid sequences of HSP60 from bacteria, protozoa, fungi and humans were aligned (Figure SF4 in file S1). A set of 25 G20c homolog 30-mer peptides (Table ST3 in file S1) were synthesized and tested in the IgG and IgM tests. The highest IgG values were with the Schistosoma mansoni, Plasmodium falciparum, Cryptosporidium, Staphylococcus, Borrelia, Leishmania, Chlamydia and Leptospira homologs. In the IgM test, especially strong reactions were seen with Leishmania, Treponema, Entamoeba and Chlamydia homologs (Figure 2). The five G20c homolog peptides with the highest positive (100%) and negative predictive values (62-68%) for ME in the IgM test were from Staphylococcus, Plasmodium, Listeria, Burkholderia and Chlamydia (Figure SF5 in file S1)

Bottom Line: A HSP60-based panel of 25 antigens was selected.When evaluated with 61 other ME and 399 non-ME samples (331 BD, 20 Multiple Sclerosis and 48 Systemic Lupus Erythematosus patients), a peptide from Chlamydia pneumoniae HSP60 detected IgM in 15 of 61 (24%) of ME, and in 1 of 399 non-ME at a high cutoff (p<0.0001).IgM to specific cross-reactive epitopes of human and microbial HSP60 occurs in a subset of ME, compatible with infection-induced autoimmunity.

View Article: PubMed Central - PubMed

Affiliation: Section of Clinical Microbiology, Department of Medical Sciences, Uppsala University, Uppsala, Sweden.

ABSTRACT
Myalgic encephalomyelitis (ME, also called Chronic Fatigue Syndrome), a common disease with chronic fatigability, cognitive dysfunction and myalgia of unknown etiology, often starts with an infection. The chaperonin human heat shock protein 60 (HSP60) occurs in mitochondria and in bacteria, is highly conserved, antigenic and a major autoantigen. The anti-HSP60 humoral (IgG and IgM) immune response was studied in 69 ME patients and 76 blood donors (BD) (the Training set) with recombinant human and E coli HSP60, and 136 30-mer overlapping and targeted peptides from HSP60 of humans, Chlamydia, Mycoplasma and 26 other species in a multiplex suspension array. Peptides from HSP60 helix I had a chaperonin-like activity, but these and other HSP60 peptides also bound IgG and IgM with an ME preference, theoretically indicating a competition between HSP60 function and antibody binding. A HSP60-based panel of 25 antigens was selected. When evaluated with 61 other ME and 399 non-ME samples (331 BD, 20 Multiple Sclerosis and 48 Systemic Lupus Erythematosus patients), a peptide from Chlamydia pneumoniae HSP60 detected IgM in 15 of 61 (24%) of ME, and in 1 of 399 non-ME at a high cutoff (p<0.0001). IgM to specific cross-reactive epitopes of human and microbial HSP60 occurs in a subset of ME, compatible with infection-induced autoimmunity.

Show MeSH
Related in: MedlinePlus