CD4 and CD8 T cell response to the rHSP60 from Klebsiella pneumoniae in peripheral blood mononuclear cells from patients with ankylosing spondylitis.
OBJECTIVE
To determine the processing pathways used by peripheral blood mononuclear cells (PBMC) and present the rHSP60Kp, and the T cell subpopulations involved in the response, in patients with ankylosing spondylitis (AS) METHODS: The lymphoproliferative response to the rHSP60Kp in PBMC from 14 HLA-B27+ AS patients and 15 B27- healthy controls was assessed by 3H-TdR incorporation. The processing pathways for the rHSP60Kp were analyzed by 3H-TdR incorporation in fresh PBMC from patients using homologous PBMC preincubated with the antigen and specific inhibitors: chloroquine, N-acetyl-L-leucil-L-leucil-L-nor-leucinal (LLnL) or brefeldin A (BFA), fixed with p-formaldehyde (fixed APC). The CD4+/CD8+ T cell subpopulation activated with the antigen was determined by three colours flow cytometry in PBMC from patients.


RESULTS
Eight out of fourteen patients showed positive lymphoproliferative responses to the rHSP60Kp while none of the healthy controls responded (p < 0.012). In five patients S.I. was above 4.0. In these patients lymphoproliferation was lower when chloroquine and LLnL was used and it became negative with BFA, indicating that both pathways are used. CD4+ and CD8+ T cells populations expressed CD69 when activated by the rHSP60Kp.


CONCLUSIONS
Our results suggest that CD4 and CD8 T cells participate in the response to the rHSP60Kp in B27+ AS patients.
Zambrano-Zaragoza
Francisco
F
Departamento de Inmunología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional.
García-Latorre
Ethel
E
Domínguez-López
María Lilia
ML
Cancino-Díaz
Mario Eugenio
ME
Burgos-Vargas
Rubén
R
Jiménez-Zamudio
Luis
L
D017951:Antigen Presentation / Q000187:drug effects; D000942:Antigens, Bacterial / Q000276:immunology*; D001327:Autoimmune Diseases / Q000097:blood / Q000235:genetics / Q000276:immunology*; D020126:Brefeldin A / Q000494:pharmacology; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D002738:Chloroquine / Q000494:pharmacology; D003600:Cytosol / Q000276:immunology; D004705:Endocytosis; D005434:Flow Cytometry; D015796:HLA-B27 Antigen / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007711:Klebsiella pneumoniae / Q000737:chemistry / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D007976:Leupeptins / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects; D013167:Spondylitis, Ankylosing / Q000097:blood / Q000235:genetics / Q000276:immunology*; D016176:T-Lymphocyte Subsets / Q000187:drug effects / Q000276:immunology*
D000942:Antigens, Bacterial; D015796:HLA-B27 Antigen; D007976:Leupeptins; C508888:rHSP60, Klebsiella pneumoniae; C063261:acetylleucyl-leucyl-norleucinal; D020126:Brefeldin A; D002738:Chloroquine