Biomedical subjects

E C Segerson

Publications and source records attributed to E C Segerson.

At least 19 recordsLinked to original sources

PubMed2000

High-density ovine endometrial cells exhibit natural killer activity during early pregnancy.

Natural killer (NK)-like activity was assessed for peripheral blood lymphocytes (PBL) and unfractionated and fractionated endometrial cells recovered from ewes during the estrus cycle (Days 12 to 14) and early pregnancy (Days 16 to 18). The PBL and endometrial cells (each designated as effector cells) were cocultured with chromium-51 (51Cr) labeled NK-sensitive K-562 target cells in effector:target cell ratios ranging from 25:1 to 200:1, respectively. Lytic activity (i.e., release of 51Cr into the medium) was assessed at 22 h of culture. A high-density (> or = 1.088 g/mL) population of endometrial cells from the pregnant ewes exhibited NK-like activity, whereas endometrial cells from the cyclic ewes failed to exhibit activity. Lytic activity of these cells was greater (P < 0.05) for pregnant than for cyclic ewes (12.0 and 2.1%, respectively) at the effector:target cell ratio of 100:1, respectively. For both groups of ewes, PBL exhibited NK-like activity. These data indicate that the ovine endometrium contains NK-like cells with lytic activity between Days 16 and 18 of pregnancy.

Animals↗

PubMed2000

Immunosuppressive macromolecules of endometrial and conceptus origins in livestock species.

Since the mid- to late-1970s, intrauterine immunosuppressive macromolecules recovered from endometrial and conceptus secretions have been reported for livestock species. Using primarily in vitro assays, in conjunction with a limited number of techniques conducted in vivo, these macromolecules were shown to suppress various T-cell responses. Some macromolecules were also shown to suppress cytolytic activities of non T-cells. It remains unknown as to whether these macromolecules actually afford protection to the conceptus by suppressing cell-mediated immune responses directed toward conceptus tissues. Endometrial effector cells in the ewe respond to antigenic stimulation and preattachment trophoblastic cells of pigs and sheep can be lysed by effector cells. Consequently, these observations suggest a need for immunosuppression, either locally at placentation sites or within the entire uterus. This review describes the intrauterine macromolecules that have been shown to suppress lymphocyte responses. Additional information, although limited at this time, refers to their origin and possible mechanisms of action. As more reagents become available to complete the identification of the intrauterine immune cells in livestock animals, experiments (e.g. antibody-mediated depletion of cells) can be conducted to determine the precise functions of all these cells. Knowing their functions will help delineate whether or not immunosuppressive macromolecules have a role in the regulation and maintenance of conceptus tissues during pregnancy.

Animals↗

PubMed2000

Suppressor activity of bone marrow cells and localization of fluorescent-labeled bone marrow cells within ovine endometrial tissue.

Numbers of fluorescein isothiocyanate (FITC)-labeled bone marrow (BM) cells of donor lambs were quantified within endometrial cell suspensions following their administration to ovariectomized (OVX; control-and estradiol-17beta-treated) and intact (estrus, d-14 cyclic and pregnant) ewes. The numbers of fluorescent BM cells were greater (P < .05) for the estrous and d-14 cyclic ewes than for both groups of OVX ewes. Fractionation of the endometrial cells with Percoll revealed that the majority of fluorescent cells were low-density (1.002 to 1.056 g/mL) cells. In coculture experiments, low-density cells from lamb BM not only suppressed the incorporation of thymidine into phytohemagglutinin-treated peripheral blood lymphocytes, but the cells also released suppressor factor into the culture medium. Suppressor activity tended to be reversed (P < .1) by a pan-specific neutralization antibody to transforming growth factor-beta (TGF-beta); however, the activity was unaffected by a neutralization antibody to TGF-beta2. These findings suggest that ovine endometrial suppressor cells may represent a population of low-density BM-derived natural suppressor cells, and their trafficking and localization patterns may depend on an ovarian factor(s). Further, suppressor activity does not seem to be mediated by TGF-beta2.

Animals↗

PubMed2000

A porcine intrauterine 4-mda component with transforming growth factor-beta activity suppresses natural killer cell responses.

A 4-MDa component, recovered from uterine luminal secretions of gilts on d 15 of pregnancy, was assessed for suppression of the lytic responses from natural killer (NK) and lymphokine-activated killer (LAK) effector cells. Each cell type originated from preparations of peripheral blood lymphocytes (PBL), and the LAK cells were generated from the incubation of PBL with interleukin-2. The PBL and LAK cells were cultured for 5 d with and without the 4-MDa component. Following culture, the cells were incubated (22 h) with NK-sensitive K-562 target cells at varying effector:target cell ratios (25:1 to 200:1). Lytic activity was assessed with the chromium-51 release assay. Additional experiments were conducted in order to determine whether suppressor activity of the 4-MDa component was time-dependent and associated with transforming growth factor-beta2 (TGF-beta2). For effector:target cell ratios combined, the 4-MDa component suppressed the lytic activity of PBL but failed to affect the LAK cells. Suppression of NK-mediated lysis occurred by d 3 of the 5-d culture period. In addition, suppressor activity of the 4-MDa component was reversed by a neutralization antibody to TGF-beta2. In conclusion, the 4-MDa component with TGF-beta2 activity suppressed the lytic responses of porcine NK cells.

Animals↗

PubMed1999

Ovine endometrial cells fail to lyse K-562 target cells: a preliminary investigation.

The ability of unfractionated and fractionated endometrial cells to lyse K-562 target cells was investigated within ewes during the late luteal phase of the estrous cycle (Days 12 to 14) and pregnancy (Days 16 and 19). In separate experiments, lymphokine activated killer (LAK) cells and endometrial cells, both designated as effector cells, were co-cultured with chromium-51 (51Cr)-labeled K-562 target cells in varying effector: target cell ratios. At 22 h, lytic activity was assessed by the release of 51Cr into the culture medium. The LAK cells exhibited lytic activity in a ratio-dependent manner, whereas the unfractionated and fractionated endometrial cells failed to lyse the target cells. For ratios combined, the rate of cytotoxicity for unfractionated endometrial cells recovered from ewes in the cyclic and pregnant (Days 16 and 19 combined) groups was 13.9 and 5.4%, respectively. Although the findings are preliminary, they indicate that ovine endometrial cells recovered during the late luteal phase and early pregnancy failed to exhibit natural killer activity upon K-562 target cells.

Animals↗

PubMed1998

Endometrial suppressor cells in beef cattle.

Endometrial cells were recovered post mortem from cyclic and pregnant crossbred beef cattle (n = 5 each) on Days 16 to 18 after estrus, and were evaluated for their ability to suppress lymphocyte responses and release suppressor factor into the culture medium. The suppressor factor was assessed for transforming growth factor-beta (TGF-beta) activity. In addition, Percoll was used to fractionate endometrial cells from Angus cows (n = 4) on Days 16 to 18 of pregnancy to determine the density of the suppressor cells. Endometrial cells from cyclic and pregnant cows suppressed lymphocyte proliferative responses and released suppressor factor into the culture medium. The suppressor factor exhibited TGF-beta activity. Suppressor activities tended to be greatest for fractionated cells with densities of 1.01 and 1.095 g/mL. In conclusion, the bovine endometrium contains low- and high-density suppressor cells capable of releasing suppressor factor. The factor seems to be associated with TGF-beta.

Animals↗

PubMed1998

Partial characterization of ovine intrauterine suppressor cells.

Ovine uterine cells that represented Day 14 cyclic and pregnant endometrium were fractionated with Percoll and evaluated for suppression of cocultured phytohemagglutinin (PHA)-induced peripheral blood lymphocyte (PBL) proliferation and for the presence of T-lymphocyte markers. Uterine cells were then evaluated for suppressor activity following the depletion of conventional lymphocyte classes (i.e., T-, B-, and NK-like) with complement + antibody treatment. In addition, supernatant (derived from cultured uterine cells) was tested for transforming growth factor beta (TGFbeta) activity using neutralization antibodies to TGFbeta. Fractionated uterine cells (density range of 1.002-1.056 g/ml) from cyclic and pregnant ewes suppressed PHA-induced proliferation of PBL, and the majority (69.5%) of these cells were < or = 5.2 microm in diameter. Percentages of CD5+, CD4+, and CD8+ lymphocytes recovered from endometrial curettage were less for cells in this density range than for cells with greater densities. Uterine cells released suppressor factor(s) into the culture medium (supernatant); however, suppressor activity was unaffected by either anti-TGFbeta or complement + antibody treatment. In conclusion, low-density uterine cells from Day 14 cyclic and pregnant ewes suppressed the proliferation of cocultured PBL and released a suppressor factor(s) into the medium that did not exhibit TGFbeta activity. It is unlikely that the suppressor cells comprise conventional T-, B-, or NK-like lymphocyte lineages.

Animals↗

PubMed1997

Estradiol-17 beta and progesterone increase ovine uterine suppressor cell activity.

We evaluated the regulation of ovine uterine (UT) suppressor cell activity by progesterone (P4), estradiol-17 beta (E2), and P4 + E2 in ovariectomized (OVX) ewes. Following 14 d of steroid injections, endometrial cells (designated as UT cells) were recovered postmortem, and unfractionated and fractionated cells were assessed for suppression of autologous phytohemagglutinin (PHA)-treated peripheral blood lymphocytes (PBL). Supernatants from cultured UT cells were also assessed for suppressor activity. In other experiments, UT cells recovered from nontreated OVX ewes were cocultured with PHA-treated PBL and varying concentrations (1 x 10(-11) to 1 x 10(-5) M) of each steroid preparation. Supernatants from separate cultures that contained UT cells and steroids were evaluated for suppressor activity. Uterine cells from control and steroid-treated ewes suppressed proliferative responses of PHA-treated PBL; however, suppressor activity of UT cells was greater (P < .05) for E2-treated than for control and P4-treated ewes. Uterine suppressor cells from steroid-treated ewes sedimented in Percoll within a density range of 1.002 to 1.056 g/mL. Uterine cells from all ewes released suppressor factor(s) into the culture medium; however, the activity of the supernatant from the cultured cells was not increased for the steroid-treated ewes. For cocultures that contained steroids and cultures that contained supernatant, suppressor activity of the UT cells was increased by specific concentrations of each steroid preparation. These findings demonstrate that reproductive steroids augment ovine UT suppressor cell activity.

Animals↗

PubMed1997

Transforming growth factor-beta 2 activity is temporally associated with a porcine high-molecular-weight uterine component recovered during early pregnancy.

A high-molecular-weight component (> or = 4 MDa, eluted at void volume of a Sepharose CL-6B column) was recovered postmortem from uterine luminal secretions from crossbred gilts (4/d) on d 9, 12, 15, and 18 of pregnancy and d 15 of the cycle. It was tested for suppression of peripheral blood lymphocyte (PBL) proliferation and for transforming growth factor-beta (TGF-beta) activity. In Exp. 1, the > or = 4 MDa component was cultured with phytohemagglutinin (PHA)-treated PBL. Parallel cultures received a pan-specific neutralization antibody to TGF-beta. In Exp. 2, cultures contained PHA-treated PBL, the > or = 4 MDa component and neutralization antibodies to either TGF-beta 1 or -beta 2. In Exp. 1, the > or = 4 MDa component recovered from uterine secretions for d-12 to d-18 pregnant gilts and d-15 cyclic gilts suppressed (P < .001) the proliferation of PHA-treated PBL; however, suppressor responses were reversed (P < .001) by anti-TGF-beta only for the > or = 4 MDa component recovered from gilts on d 15 and 18 of pregnancy. In Exp. 2, anti-TGF-beta 2 reversed (P < .05) the suppressor activity of the > or = 4 MDa component recovered from gilts on d 15 of pregnancy. In conclusion, a temporal pattern of TGF-beta activity was associated with a > or = 4 MDa carrier recovered from porcine uterine luminal secretions during early pregnancy. For uterine secretions recovered on d 15, suppressor activity was at least partly attributed to TGF-beta 2.

Analysis of Variance↗

PubMed1995

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.

Animals↗

PubMed1994

In vitro and in vivo effects of lymphokine-activated killer cells upon preattachment ovine conceptuses.

An investigation assessed the cytolytic effect of autologous ovine lymphokine-activated killer (LAK) cells upon preattachment ovine conceptuses. For an in vitro experiment, conceptuses were recovered from six ewes on each of days 14, 16, and 19 (estrus = day 0). Dissociated conceptus cells (5 x 10(4), considered target cells) were incubated with autologous LAK cells at varying E:T ratios (5:1-200:1) and cytotoxicity was assessed by 51Cr release. In vivo, control (saline), cultured PBL (1 x 10(7)) and LAK cells (1- and 4- x 10(7)) were infused into the lumen of the uterine horn ipsilateral to the corpus luteum of mated ewes on day 14. Conceptuses were recovered on day 19 and placed within one of three morphologic categories (intact, large fragments, or highly fragmented). The numbers of mononucleated and binucleated giant cells and remaining trophoblastic cells were then quantitated within sections of conceptus tissue. Mitotic activity was also recorded. At E:T ratios of 100:1 and 200:1, respectively, percent 51Cr release for LAK cell cultures was greater (p < 0.05) for day-16 and 19- than day-14 conceptus cells. The frequencies of highly fragmented conceptuses recovered from saline + cultured PBL and LAK cell-infused ewes were 0/10 and 8/13, respectively. Within tissue sections, the numbers of morphologically normal mononucleated giant and mitotic cells were less (p < 0.05) for LAK cell (4 x 10(7)) than for saline-infused ewes. Data from additional in vitro and in vivo experiments revealed that supernatants from LAK cell (4 x 10(7)) cultures failed (p > 0.05) to affect % 51Cr release from K-562 target cells, conceptus morphology, and numbers of conceptus cells. In conclusion, autologous LAK cells exerted in vitro and in vivo lytic damage upon preattachment ovine conceptuses. A temporal pattern was observed for the in vitro lytic responses.

Animals↗

PubMed1993

Temporal patterns of secretion of porcine uterine suppressor and stimulatory macromolecules.

Temporal secretory patterns of porcine uterine suppressor (>/=230 kD) and stimulatory (29 kD) macromolecules were evaluated within uterine luminal protein (ULP) secretions recovered during early pregnancy. The ULP was recovered by uterine flushing from four Landrace gilts each on Days 9, 12, 15 and 18 of pregnancy. Unfractionated and fractionated ULP (using Sephacryl S-200) was tested for suppression or stimulation of phytohemagglutinin-induced peripheral blood T-lymphocyte proliferation. For all days of pregnancy, unfractionated ULP suppressed (P<0.002 to 0.0001) lymphocyte proliferative responses, with the greatest (P<0.05) activity observed for ULP collected on Day 9 of pregnancy. Suppressor activity resulted from the >/=230 kD component, in which the activity was greater (P<0.05) for ULP from gilts on Day 9 than Days 12, 15 and 18 of pregnancy. The 29 kD component failed (P>0.05) to stimulate lymphocyte proliferation, although there was a nonsignificant stimulatory trend for 2 of 4 gilts each at Days 12 and 15 of pregnancy. These findings demonstrate a temporal secretory pattern for the >/=230 kD lymphocyte suppressor component, which may be requisite for the immunological survival of the conceptus during early pregnancy. The inconsistent appearance of the lymphocyte stimulatory factor (29 kD component) tends to minimize its biological significance relative to the immunology of pregnancy.

Journal Article↗

PubMed1993

Interference with the cytolytic activity of interleukin-2-treated lymphocytes by bovine uterine luminal protein.

Bovine uterine luminal protein (ULP) components (> or = 4 x 10(6) and 21,000 M(r)) were obtained from uterine flushings on Day 17 of pregnancy. Experiments were conducted to determine the capability of these components to interfere with the cytolytic activity of interleukin-2 (IL-2)-treated peripheral blood lymphocytes (PBL), designated LAK (lymphokine-activated killer) cells, upon K-562 tumor cells. Proteins (10-100 micrograms) were added at the onset of a 5-day culture period to wells containing PBL (3 x 10(6) + bovine recombinant IL-2 (12,000 IU). After culture, percentage cytotoxicity (Cyt) was assessed by the 51Cr release assay at 4 and 22 h of incubation for ratios (5:1-200:1) of LAK:K-562 cells. Percentage Cyt was affected (p < 0.0001) by time, ratio, protein treatment, and all two-way interactions. Although trends were apparent, neither ULP component affected (p > 0.05) percentage Cyt at 4 h. By 22 h of incubation, mean percentage Cyt values for the > or = 4 x 10(6) and 21,000 M(r) components at effector:target cell ratios of 150:1 (p < 0.002) and 200:1 (p < 0.0001) were less than percentages for LAK cells alone. BSA and serum protein (control preparations) each failed (p > 0.05) to affect percentage Cyt. In conclusion, both ULP components interfered with the cytolytic activity of LAK cells, presumably by the suppression of LAK cell generation.

Animals↗

PubMed1992

Secretion of ovine lymphocyte suppressor factor from curetted uterine luminal cells.

The secretion of ovine lymphocyte suppressor factor from jugular vein (JV), uterine vein (UV), and curetted hemopoietic uterine luminal (UL) mononuclear cells was evaluated on d 14 of the cycle, following ovariectomy (OVX) and after 14 d of progesterone injections (OVX + P4, 1 mg/kg BW). Mononuclear cells (predominantly lymphocytes) were harvested by Ficoll-Paque and placed into culture (2.5 x 10(6).mL-1.well-1 in RPMI-1640). Cellular supernatants were obtained at 72 h and volumes (5 to 50 microL) were tested for suppression of phytohemagglutinin (PHA [.08 microgram l)-treated peripheral blood lymphocytes (1.0 x 10(5)). In a concurrent experiment, PHA-treated JV, UV, and UL cells (1.0 x 10(5)) were cultured singly and JV cells (1.0 x 10(5)) were also cocultured with each of 1.0 x 10(5), 5.0 x 10(4), and 2.5 x 10(4) UV and UL cells. The incorporation of thymidine into DNA was quantified at 60 h. For the cellular supernatant experiment, thymidine incorporation was affected by reproductive phase (P less than .036), lymphocyte source (P less than .0001), and phase x source (P less than .004). For UL cells, the degree of suppressor activity follows: d 14 greater than OVX greater than OVX+P4 (P less than .05). The UL supernatant from OVX+P4-treated ewes and supernatants of JV and UV cells, irrespective of reproductive phase, lacked suppressor activity. Sephacryl S-200 chromatography revealed that UL supernatant from d-14 ewes contained a greater than or equal to 248,000 molecular weight suppressor macromolecule. For the cellular coculture experiment, thymidine incorporation was affected by reproductive phase (P less than .05) and lymphocyte source (P less than .0001).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1991

Endometrial T-lymphocyte subset infiltration during the ovine estrous cycle and early pregnancy.

T-lymphocytes were quantitated within luminal, stromal and glandular areas of ovine endometrium. In experiment 1, ovariectomized (OVX), estrus (E) and day 13 (D13) ewes (six/group) received 500 micrograms of phytohemagglutinin (PHA) or vehicle in ligated right and left uterine horns, respectively. At 48 h, uteri were removed for the immunohistochemical evaluation of T-lymphocyte subsets. In experiment 2, T-lymphocytes were quantitated within non-pregnant and pregnant uterine horns on day 19. For experiment 1, mean numbers of T4 and T8 lymphocytes within luminal and stromal areas of PHA-treated horns were greatest (P less than 0.05) for D13 ewes and least (P less than 0.05) for E ewes. Numbers of T6 lymphocytes for these same areas were greatest (P less than 0.05) for PHA-treated horns of OVX ewes. Overall, the T4/T8 ratio (P less than 0.004) and mean number of T19 cells (P less than 0.009) were increased by PHA. Numbers of CD45R lymphocytes were not affected by PHA but were greater (P less than 0.05) in glandular and luminal than stromal areas. For experiment 2, mean numbers of endometrial T4, T6, T8 and T19 lymphocytes were similar (P greater than 0.05) between non-pregnant and pregnant horns; however, the number of CD45R lymphocytes was greater (P less than 0.05) in endometrial tissue of pregnant than non-pregnant horns. The data indicate that the in vivo response of specific ovine T-lymphocytes to PHA was generally dependent upon reproductive stage and the presence of conceptus tissue influenced the infiltration of CD45R lymphocytes.

Animals↗

PubMed1990

Interaction of bovine uterine luminal protein with interleukin-2 and the interleukin-2 receptor of T lymphocytes.

Bovine uterine luminal proteins (ULP) collected on Day 17 of pregnancy were tested for inhibition of binding of interleukin-2 (IL-2) to the IL-2 receptor (IL-2R) of bovine (CLC) and human (HLC) T lymphocytes and for binding to IL-2. Additional experiments assessed IL-2 binding to the p55 alpha chain (Tac protein) of the IL-2R of HLC. High- and low-molecular weight (Mr) ULP components (H-ULP greater than 248,000 Mr and L-ULP 21,000 Mr, respectively) inhibited (p less than 0.05 and 0.01, respectively) the binding of 125I-IL-2 to the IL-2R of CLC, whereas only H-ULP inhibited (p less than 0.05) binding to the IL-2R (presumably, the p75 beta chain) of HLC. H-ULP failed (p greater than 0.05) to bind to the p55 alpha chain of the IL-2R of HLC. For IL-2 binding, L-ULP failed (p greater than 0.05) to bind 125I-IL-2 in short (2 h)-term and long (45 h)-term experiments, whereas binding was evident (p less than 0.05) for H-ULP at 2 h of incubation. For H-ULP, mean (+/- SEM) percentages for bound and unbound 125I-IL-2 were 70.1 +/- 11.4 and 29.9 +/- 11.4, respectively. Further purification of H-ULP yielded a component (1.76 x 10(6) Mr) that bound 11.7% of 125I-IL-2 and inhibited (p less than 0.01) thymidine uptake and binding of 125I-IL-2 to the IL-2R of CLC. H-ULP-mediated suppression of lymphocyte proliferation may result from blocking IL-2R recognition of IL-2 as well as binding to IL-2, whereas suppression by L-ULP may predominantly result from blocking IL-2R.

Animals↗

PubMed1989

High molecular weight basic and acidic immunosuppressive protein components in uterine secretions of pregnant cows.

Immunosuppressive proteinaceous components were determined in bovine uterine milk (UTM) collected during late pregnancy. Crude UTM was separated by ion-exchange (carboxymethylcellulose-CMC) and gel filtration (Sephacryl S-200 and Sepharose CL-6B) chromatography. Basic (CMC+) and acidic (CMC-) protein molecular weight (Mr) components were tested for immunosuppressive activity in an in vitro mitogen (phytohemagglutinin-PHA)-treated lymphocyte blastogenesis assay. For most experiments, cultures containing 1 x 10(5) lymphocytes were incubated with 0.08 micrograms PHA and varying concentrations of test protein in RPMI-1640 with supplements. At 48 +/- 1 h, 0.1 microCi of [3H]thymidine was added to cultures and [3H]DNA was quantified at 60 +/- 1 h of culture. Results were expressed as percentage of control values. Crude UTM, CMC+, and CMC- components exhibited immunosuppressive activity. For immunosuppressive Sephacryl S-200 fractions, activity was greater (p less than 0.05-0.01) for CMC+, S-200 fraction I (greater than or equal to 250,000 Mr, void volume [Vo]) than for CMC-, S-200 fractions I (Vo) and III combined for protein concentrations of 20, 40, and 50 micrograms/ml. For the high Mr Sepharose CL-6B protein components, CMC+, CL-6B fraction I (greater than or equal to 4 x 10(6) Mr, Vo) exhibited greater (p less than 0.05-0.005) activity than CMC-, CL-6B fractions I (greater than or equal to 4 x 10(6) Mr, Vo) and II (2.8 x 10(6) Mr) combined at protein concentrations ranging from 20 to 80 micrograms/ml. In summary, bovine UTM contains basic and acidic immunosuppressive protein components, with the greatest activity being associated with a high Mr, basic component.

Animals↗

PubMed1988

Suppression of interleukin-2-mediated T-lymphocyte blastogenesis by ovine uterine luminal protein.

Ovine uterine luminal protein (ULP) obtained from ewes on Day 14 of pregnancy suppressed blastogenesis of interleukin-2 (IL-2)-dependent T-lymphocytes. Varying concentrations of ULP (4 to 96 micrograms/ml) followed by a 1:4 dilution of human IL-2 suppressed (p less than 0.001) IL-2 blastogenesis of IL-2-dependent T-lymphocytes with mean percentage of control values ranging from 55.3 to 34.5% (44.7 to 65.5% suppression, respectively). For two experiments, IL-2 was added at varying times (zero to 4 h) after the addition of ULP to cultures. Suppression was independent of IL-2 addition time. Mean (+/- SEM) percentage of control values for combined time periods for 40 and 120 micrograms ULP/ml were 43.3 +/- 1.0 and 27.8 +/- 1.9%, respectively. In another experiment, additional IL-2 (1:2 vs. 1:4 dilution) reduced (p less than 0.01) the immunosuppressive effect of ULP. Sephacryl S-200 chromatography of ULP and the phytohemagglutinin (PHA) blastogenesis assay revealed significant immunosuppressive activity for Fractions I (greater than or equal to 248,000 Mr), III (70,000 Mr), and V (14,000 Mr). These fractions also suppressed (p less than 0.001) IL-2-mediated blastogenesis of T-lymphocytes. Results indicate that immunosuppression of PHA-treated lymphocytes was associated with an alteration of the IL-2 system.

Animals↗