Antibody response to Klebsiella pneumoniae 60 kDa protein in familial and sporadic ankylosing spondylitis: role of HLA-B27 and characterization as a GroEL-like protein.
OBJECTIVE
To study the antibody response of HLA-B27+ patients with ankylosing spondylitis (AS) and their first degree relatives to the 60 kDa protein of Klebsiella pneumoniae and to characterize this protein.


METHODS
Sera from 84 individuals were analyzed by ELISA to determine the titer of antibodies against the 60 kDa protein of K. pneumoniae. Subjects were divided into 3 categories: Group 1: 44 HLA-B27+ AS related individuals (35 patients, 9 healthy controls); Group 2: 28 healthy B27- AS related individuals; and Group 3: 12 healthy B27- non-AS related subjects. The 60 kDa protein of K. pneumoniae was induced at 45 degrees C and purified by electroelution from sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was characterized as a GroEL-like heat shock protein (HSP). The recognition of GroEL-like protein was confirmed by immunoblot of 2 dimension electrophoresis. The response to GroEL-like protein from other bacteria and the response to lipopolysaccharide (LPS) was also analyzed by immunoblot.


RESULTS
HLA-B27+ individuals (Group 1), independent of their disease status, showed a significant higher response to the 60 kDa protein of K. pneumoniae than HLA-B27- subjects from Groups 2 and 3 (p < 0.0001). This protein was characterized as a HSP of the GroEL family and designated HSP60Kp. The GroEL of other enterobacteria as well as that of Mycobacterium leprae were recognized by HLA-B27+ individuals by immunoblot, whereas HLA-B27- individuals did not. LPS was not recognized by HLA-B27 positive or negative subjects.


CONCLUSIONS
These findings suggest a relationship between HLA-B27 and the response to a GroEL-like protein that could have implications in AS.
Cancino-Díaz
M E
ME
Departamento de Inmunología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Hospital General de México SSA, DF, México.
Pérez-Salazar
J E
JE
Domínguez-López
L
L
Escobar-Gutiérrez
A
A
Granados-Arreola
J
J
Jiménez-Zamudio
L
L
Burgos-Vargas
R
R
García-Latorre
E
E
D001426:Bacterial Proteins; D001886:Bordetella pertussis / Q000276:immunology; D018834:Chaperonin 60 / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D018833:Chaperonins / Q000096:biosynthesis / Q000276:immunology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D015796:HLA-B27 Antigen / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007711:Klebsiella pneumoniae / Q000276:immunology*; D009166:Mycobacterium leprae / Q000276:immunology; D013167:Spondylitis, Ankylosing / Q000097:blood / Q000235:genetics / Q000276:immunology*
D001426:Bacterial Proteins; D018834:Chaperonin 60; D015796:HLA-B27 Antigen; D007074:Immunoglobulin G; C074119:heat-shock protein 65, Mycobacterium; D018833:Chaperonins