Immunosuppressive activity of a porcine high-molecular weight uterine macromolecule is associated with transforming growth factor-beta.
A > or = 230 kDa lymphocyte suppressor component recovered from uterine luminal protein (ULP) secretions of Landrace and Yorkshire gilts on day 15 of pregnancy was further purified by anion-exchange (DEAE Sepharose CL-6B) and gel filtration (Sepharose CL-6B) chromatography. Macromolecules within each peak fraction were tested for suppression of thymidine (TdR) incorporation into phytohemagglutinin (PHA)-treated peripheral blood lymphocytes (PBL). A neutralizing antibody to transforming growth factor-beta (TGF-beta) was added to additional cultures in order to determine whether suppressor activity of high-molecular weight macromolecules was associated with TGF-beta. The > or = 230 kDa component suppressed (P < 0.0005) TdR incorporation into PHA-treated PBL and anti-TGF-beta reversed (P < 0.034) the suppressor response. Following anion-exchange chromatography of this component, suppressor activity was greatest (P < 0.05) for an acidic component (fraction III), which comprised 45.9% of eluted macromolecules. Sepharose CL-6B chromatography of fraction III resulted in a major (58.7% of eluted macromolecules) component (> or = 4 x 10(6) Da (> or = 4 MDa), eluted at the void volume) which suppressed (P < 0.05) TdR incorporation into PHA-treated PBL. Anti-TGF-beta reduced the suppressor activity of this macromolecule by 34.2%. Suppressor responses were not associated with viabilities of PBL. These data demonstrate that ULP secretions from gilts on day 15 of pregnancy contain a > or = 4 MDa macromolecule which suppresses proliferative responses of PBL and likely serves as a carrier for TGF-beta.
Department of Animal Science, North Carolina Agricultural and Technical State University, Greensboro 27411, USA.
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Segerson
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005260:Female; D006023:Glycoproteins / Q000302:isolation & purification / Q000494:pharmacology*; D007108:Immune Tolerance; D007166:Immunosuppressive Agents / Q000302:isolation & purification / Q000494:pharmacology*; D008213:Lymphocyte Activation / Q000187:drug effects; D008970:Molecular Weight; D011247:Pregnancy; D011270:Pregnancy, Animal / Q000276:immunology*; D015843:Serpins; D013552:Swine; D016212:Transforming Growth Factor beta / Q000276:immunology / Q000502:physiology*; D014599:Uterus / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D007166:Immunosuppressive Agents; D015843:Serpins; D016212:Transforming Growth Factor beta; C029708:uterine luminal fluid proteins