Peripheral blood mononuclear cell proliferative responses to soluble and particulate heat shock protein 65 in health and inflammatory bowel disease.
OBJECTIVE
The aims of this study were to determine, in peripheral blood mononuclear cells (PBMC), whether particulate antigen triggers (i) an amplified cell proliferative response compared to soluble antigen and (ii) a dysfunctional response in cells derived from patients with chronic inflammation and specifically in those with inflammatory bowel disease (IBD).


METHODS
Healthy volunteers (n = 17), inflammatory controls (n = 8) and patients with IBD (n = 17) were recruited from St Thomas' and Guys' Hospital, London, UK.


METHODS
Following optimisation of experimental conditions (0.1-10.0 mug/ml antigen), PBMC were stimulated with (i) 10.0 mug/ml recombinant soluble heat shock protein 65 (hsp 65) and (ii) 1.0 and 10.0 mug/ml hsp 65 conjugated to microparticles (0.5 mum diameter). PBMC proliferative responses were measured by (3)H-Thymidine incorporation at day 5 and results compared between groups using unpaired t-test.


RESULTS
Conjugation to microparticles of low dose hsp 65 significantly increased overall proliferative responses by 2-11 fold compared to soluble antigen alone (p < 0.05). However, no specific PBMC proliferative dysregulation was noted in cells from subjects with IBD.


CONCLUSIONS
Low dose antigen, in microparticulate form, leads to amplified cell proliferation in primary human cells, as showed previously in cell lines and animal studies. However there is no abnormal proliferative response in cells from subjects with IBD.
Pele
L C
LC
MRC Human Nutrition Research, Elsie Widdowson Laboratory, Cambridge, CB1 9NL, UK.
Thompson
S J
SJ
Kirkham
B
B
Thompson
R P H
RP
Powell
J J
JJ
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000941:Antigens / Q000276:immunology; D016022:Case-Control Studies; D049109:Cell Proliferation; D005260:Female; D006360:Heat-Shock Proteins / Q000276:immunology*; D006801:Humans; D015212:Inflammatory Bowel Diseases / Q000276:immunology* / Q000473:pathology; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D010316:Particle Size; D013997:Time Factors; D018834:Chaperonin 60; D024101:Mitochondrial Proteins
D000941:Antigens; D006360:Heat-Shock Proteins; C060293:HSPD1 protein, human; D018834:Chaperonin 60; D024101:Mitochondrial Proteins