Recognition of B cells epitopes of the Klebsiella pneumoniae GroEL-like protein by HLA-B27 positive subjects.
The presence of antibodies against antigens of K. pneumoniae in HLA-B27 positive patients with ankylosing spondylitis (AS), has been well documented. We have previously reported that sera from HLA-B27 positive subjects react with the K. pneumoniae GroEL-like protein (HSP60Kp) and have higher titers than HLA-B27 negative individuals. We cloned the gene that codes for this protein, determined hydrophilic regions by computer analysis of the predicted amino acid sequence and found that residues 389-397, 360-368 and 282-290, were possible B cell epitopes. To test this prediction, and to determine if the HLA-B27 positive and negative AS patients recognize the same or different epitopes, we truncated the hsp60Kp gene, from the 3; terminal nucleotide, to obtain fragments having or not the predicted epitopes. Four polypeptides of 40, 37, 30 and 18 kDa were obtained and analysed, by ELISA and inhibition of ELISA, for their reactivity with IgG antibodies from three high responders HLA-B27 positive AS patients and three HLA-B27 negative subjects who recognized the rHSP60Kp. Sera from both HLA-B27 positive and negative subjects reacted equally well with rHSP60Kp or with the 40 and 37 kDa peptides, which do not have residues 389-397 and 360-368, respectively, but reactivity was lost with the 30 kDa peptide, which also lacks residues 282-290. Contrary to what we expected, antibodies from HLA-B27 negative and positive individuals recognized the same epitope of the HSP60Kp. Our results indicate that the important epitope for B cells could be the 282-290 region and that the contribution of the two other predicted regions is minimal. We also conclude that the differences in response to the HSP60Kp in HLA-B27 positive AS patients and HLA-B27 negative individuals is not qualitative, but only quantitative.
Cancino-Díaz
M
M
Departamento de Inmunología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Carpio y Plan de Ayala, México D.F., 11340, México. woischnik@bigfoot.com
Ayala-Narváez
H
H
Burgos-Vargas
R
R
Selene Reyes-López
A
A
Tovar-Castillo
L
L
Domínguez-López
L
L
Granados Arreola
J
J
Jiménez-Zamudio
L
L
García-Latorre
E
E
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D018834:Chaperonin 60 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D015796:HLA-B27 Antigen / Q000097:blood / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting; D007711:Klebsiella pneumoniae / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D013167:Spondylitis, Ankylosing / Q000276:immunology / Q000382:microbiology
D000907:Antibodies, Bacterial; D018834:Chaperonin 60; D018985:Epitopes, B-Lymphocyte; D015796:HLA-B27 Antigen; D011994:Recombinant Proteins