Effects of boar seminal immunosuppressive fraction on production of cytokines by Concanavalin A-stimulated spleen cells and on proliferation of B lymphoma cell lines.
OBJECTIVE
The immunosuppressive fraction (ISF) of boar seminal vesicle fluid has recently been demonstrated to inhibit mitogen-stimulated proliferation of lymphocytes and antibody response to corpuscular and soluble antigens. The effects of ISF on in vitro and in vivo production of cytokines as well as its possible inhibitory effect on proliferation of B lymphoma cells remain to be elucidated.


METHODS
The effect of ISF on proliferation of normal mouse spleen cells stimulated by Concanavalin A (Con A) and on mouse B lymphoma cells was measured by 3H-thymidine incorporation. Cytokines were determined in the supernatants of mouse spleen cells stimulated with Con A in the presence or absence of ISF by enzyme-linked immunosorbent assay (ELISA). In vivo cytokine production in the sera samples of mice treated with ISF and immunized with keyhole limpet hemocyanin (KLH) was followed by ELISA, too.


RESULTS
We confirmed the inhibitory effect of ISF on Con A-stimulated lymphocyte proliferation. ISF affected cytokine production in the Con A-stimulated spleen cells: production of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) was lowered, but production of IL-4, IL-6, and IL-10 was enhanced. Similarly, in the sera samples of mice immunized with keyhole limpet hemocyanin (KLH), IL-2 and IFN-gamma levels were decreased by ISF. ISF inhibited proliferation of Ag 8 and X 63-IL-2 B lymphoma cells as well.


CONCLUSIONS
ISF inhibited production of T helper1 (Th1) cytokines (IL-2 and IFN-gamma) and enhanced production of Th2 cytokines (IL-4, IL-6, and IL-10). ISF seems to shift the Th1/Th2 pattern in favor of Th2. ISF exhibited an antiproliferative activity on mouse B lymphoma cells.
Veselský
L
L
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic. veselsky@img.cas.cz
Holán
V
V
Zajícová
A
A
Dostál
J
J
Zelezná
B
B
D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D003208:Concanavalin A / Q000494:pharmacology; D016207:Cytokines / Q000096:biosynthesis* / Q000097:blood / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006133:Growth Substances / Q000276:immunology / Q000494:pharmacology*; D006433:Hemocyanins / Q000494:pharmacology; D008214:Lymphocytes / Q000187:drug effects / Q000378:metabolism*; D016393:Lymphoma, B-Cell / Q000378:metabolism; D051379:Mice; D017494:Proton Pumps; D006180:Proton-Translocating ATPases; D012661:Semen / Q000737:chemistry*; D034421:Sus scrofa
D000276:Adjuvants, Immunologic; C432920:Atp6v0a2 protein, mouse; D016207:Cytokines; D006133:Growth Substances; D017494:Proton Pumps; D003208:Concanavalin A; D006433:Hemocyanins; D006180:Proton-Translocating ATPases; C032808:keyhole-limpet hemocyanin