Proliferative Response Of Spleen Cells Research Articles

Immune potentiation after fractionated exposure to very low doses of lonizing radiation and/or caloric restriction in autoimmune-prone and normal [formula omitted] mice

Very low doses of ionizing radiation can enhance immune responsiveness and extend life span in normal mice. Total lymphoid irradiation at relatively high doses of radiation can retard autoimmune disease in genetically susceptible mice, but may impair immune function. In order to determine whether fractionated low dose exposure would enhance immune response and retard lymphadenopathy in autoimmune-prone mice, groups of C57B1 6 lpr lpr mice were sham irradiated, exposed 5 days/week for 4 weeks to 0.04 Gy/day (0.8 Gy cumulative dose), or to 0.1 Gy/day (2.0 Gy cumulative dose). After the radiation protocol, the mice were evaluated for splenic T cell proliferative capacity, T cell subset distribution, and total spleen cell numbers. The independent and additive effect of caloric restriction was additionally assessed since this intervention has been shown to increase immune responsiveness and retard disease progression in autoimmune-prone mice. The congenic C57B1 6 +/+ immunologically normal strain was evaluated in parallel as congenic control. The results indicated that mitogen-stimulated proliferation was up-regulated in both strains of mice after exposure to 0.04 Gy/day. The proliferative capacity was additively enhanced when radiation at this dose level was combined with caloric restriction. Exposure to 0.1 Gy/day resulted in further augmentation of proliferative response in the lpr lpr mice, but was depressive in the +/+ mice. Although the proportions of the various T cell subpopulations were altered in both strains after exposure to LDR, the specific subset alterations were different within each strain. Additional experiments were subsequently performed to assess whether the thymus is required for LDR-induced immune potentiation. Thymectomy completely abrogated the LDR effect in the +/+ mice, suggesting that thymic processing and/or trafficking is adaptively altered with LDR in this strain. In contrast, augmentation in proliferative activity after LDR in the lpr lpr mice was maintained, although attenuated, in thymectomized mice. Taken together, these results indicate that fractionated exposure to LDR augments the proliferative response of spleen cells in both autoimmune-prone and immunologically normal mice; however, within each strain, the mechanisms appear to be different.

Suppressive macrophages occurring in murine Trypanosoma brucei infection inhibit T‐cell responses in vivo and in vitro

Intraperitoneal injection of Trypanosoma brucei AnTat 1.1 into mice of the C3H.He, BALB/c or C57BL/6 strains resulted in impaired immune responses from day 3 onwards, as measured by the reduction in DNA synthesis in spleen cell populations stimulated with concanavalin A (Con-A) in vitro. Adherent cells from the peritoneum (PC) or from the spleen of infected mice, consisting predominantly of macrophages, caused a 60-80% reduction of the Con-A response in spleen cells from syngeneic recipients 3-4 days after transfer in vivo. Adherent PC from irradiated or athymic mice were equally suppressive. Spleen cells from infected mice reduced the proliferative response of spleen cells from uninfected mice upon co-cultivation in vitro. This dominant suppressive effect was abolished after the selective removal of macrophages from the spleen cell population by treatment with L-leucine methylester. Moreover, the macrophage-depleted spleen cells from infected mice responded normally to Con-A provided they were supplemented with splenic adherent cells from naive mice as a source of accessory cells. Both the cell transfer and co-cultivation experiments suggest that infection with African trypanosomes changes the properties of macrophages to a state which allows them actively to suppress immune responses.

Immune Deficiency of Cataract Shionogi (Cts) Mouse. H. Impaired in Vivo T Cell-Mediated Immune Response

Our previous study demonstrated that cataract Shionogi (CTS) mice, an inbred strain related to non-obese diabetic (NOD) mice, are T lymphocytopenic and that their T cell-mediated in vitro reactions, such as proliferative responses of spleen cells to T cell mitogens and alloantigens or production of IL 2 and IL 2 receptors after stimulation of spleen cells with Con A, are greatly reduced. To confirm these in vitro characteristics, in vivo immune responses of CTS mice to T-dependent and T-independent antigens were compared with those of some reference strains including NOD mice. Antibody responses of CTS mice after one injection of a high dose (10(8)) or one or two injections of a low dose (10(5)) of sheep red blood cells (SRBC) were markedly lower than those of the reference strains. The decrease was particularly striking in the IgM antibody production at primary response to both high and low doses, and the IgG antibody production at the secondary response to low dose. Similar lower antibody production was observed in CTS mice against bovine serum albumin (BSA). Little production of IgE antibody was observed from 1 through 3 weeks after an injection of BSA plus Bordetella pertussis. IgG1 response was observed at high incidence but lower in titer than those in the reference strains. Unexpectedly, in spite of the poor antibody production to BSA, potent systemic sensitization for anaphylactic shock was easily established; incidence of lethal shock being comparable with those in the reference strains. This suggests that CTS mice are highly susceptible to the effector phase of active anaphylactic shock. Cell-mediated immunity was also impaired. Delayed type of hypersensitivity to SRBC was low, and the rejection of the skin graft from NOD mouse did not occur. In contrast to the reduced T cell-mediated responses, no difference was found between CTS and reference strains with regard to the antibody production to LPS, a T-independent antigen. These in vivo findings are consistent with the previous in vitro study.

Effect of luteotrophic hormone (LH) and thymosin on mitogen-induced blast transformation of Balb/c mouse lymphoid cells

The present study investigates the effect of a pituitary hormone (LH) and a thymic factor (Thymosin Fraction 5 = TF5) on in vitro and in vivo spleen cell proliferative response of 2, 6 and 8-month-old Balb/c mice. In vitro experiments showed that the addition of LH at various concentrations (0.5, 5, 50 or 500 ng/ml) to cultures increased significantly the proliferative response to some known mitogens (ConA, PHA, PWM). The LH stimulation was further enhanced by subsequent addition of TF5 to these cultures. However, the addition of LH to LPS cultures resulted in a blockage of the cell growth which persisted after the addition of TF5. In vivo experiments showed that injection of LH (5 or 50 ng/ml) to 2, 6 and 8-month-old Balb/c mice had a significant increase on blast transformation of lymphoid cells following their incubation with ConA, PHA and PWM, and a significant decrease when they were incubated with LPS. The physiological significance of these findings which point out an intimate connection between immune and endocrine functions is discussed.

Treatment of mice with human recombinant interleukin-2 augments resistance to the facultative intracellular pathogen Listeria monocytogenes.

The effects of exogenously administered human recombinant IL-2 (hrIL-2) on resistance to Listeria monocytogenes infection were examined. Intravenous injection of hrIL-2 significantly enhanced antibacterial resistance in both BDF1 and C3H/HeJ mice. The beneficial effect of hrIL-2 was observed with as little as 0.6 micrograms per mouse, whereas optimum protection occurred at 6 micrograms per mouse, hrIL-2 was equally protective when administered concomitant with the listeriae or up to 24 h prior to infection; it had little effect if given after the bacterial challenge. Kinetic experiments indicated that both the peak bacterial burden and the time lag before L. monocytogenes began to be cleared from the spleen and liver were reduced in hrIL-2-treated mice as compared with control mice. Histopathological examination of spleens and livers confirmed that hrIL-2-treated Listeria-infected mice experienced considerably less damage to these organs than did control mice. Spleen cells from Listeria-infected mice exhibited depressed levels of mitogen-induced proliferation coincident with the peak bacterial burden in the spleen and liver and during the subsequent recovery from the infection. Administration of hrIL-2 to uninfected mice had no effect on spleen cell proliferation in response to mitogens in vitro, nor did hrIL-2 treatment restore normal levels of splenocyte proliferative responses to Listeria-infected mice. In addition, hrIL-2 treatment resulted in attenuated levels of serum colony-stimulating activity in infected mice as compared with control infected mice. Coadministration of both hrIL-2 and human recombinant interleukin-1 alpha at various dose and time combinations had no detectable additive or synergistic effect. Although these data do not suggest an obvious mechanism of action, they clearly demonstrate that hrIL-2 can augment host defense against the facultative intracellular pathogen L. monocytogenes.

Neonatal injection of CD3 antibody into nonobese diabetic mice reduces the incidence of insulitis and diabetes.

Over 80% of nonobese diabetic (NOD) mice develop lymphocytic infiltrates of their pancreatic islets (insulitis) by 6 wk of age and 50% of the females are diabetic by 6 mo of age. The incidence of insulitis in NOD mice injected once as neonates with 250 micrograms of the CD3 antibody, 145.2C11, was 8% at 10 wk of age, increasing to 25% at 32 wk of age. Fewer than 10% of these animals developed diabetes by 8 mo of age. Neonatal administration of 145.2C11 reduced the proliferative responses of spleen cells to mitogen stimulation 2 and 4 wk postinjection and expression of TCR was reduced 1 to 5 wk postinjection. The percentage of CD4 and CD8 cells in the spleen was transiently reduced after injection and the frequency of Pgp-1+-high cells (putative memory cells) was increased 2 to 4 wk postinjection, suggesting that in vivo administration of the antibody caused some T cells to divide as well as transiently reducing T cell numbers. IL-2R expression was not detected on spleen cells in the 4 wk after antibody injection. The phenotypic and functional changes after neonatal CD3 antibody injection resolved by 8 wk of age. The control and injected mice grew normally and made equivalent IgG antibody responses to injected human IgG. Neonatally injected 145.2C11 antibody was cleared from the circulation with a terminal half-life approximating to 21 days but greater than 90% of antigen binding activity was lost 6 days after injection. Protection from diabetes did not follow neonatal elimination of T cells with CD4 and CD8 antibodies, nor the injection of a TCR subset antibody, F23.1. Our data suggest that the neonatal T cell repertoire is open to modulation by a single injection of a CD3 antibody and they offer a new experimental approach to immunotherapy in an animal model of type 1 diabetes.

Mechanisms of resistance/susceptibility to murine encephalitozoonosis

Mechanisms of resistance/susceptibility to the obligate intracellular protozoan Encephalitozoon cuniculi were studied in resistant BALB/c and susceptible C57BL/6 mice. Three immunological functions were examined: the production of lymphokine(s) (LK) by T-lymphocytes, the proliferative response of spleen cells to parasite spore fragments, and the ability of splenic and thioglycollate-induced peritoneal macrophages to act as accessory cells in antigen-induced T-cell proliferation. The two strains showed differences in the time required for LK production in vitro but not in their ability to generate LK. Spore fragment-induced lymphoblastogenesis was found in spleen cells of infected BALB/c and C57BL/6 mice. There was no difference between the two strains in dose response and time of maximal response, but the magnitude of maximal response was significantly less in C57BL/6 mice. Indomethacin was found to augment the lymphoproliferative response of C57BL/6 but not BALB/c mice, suggesting that prostaglandin production may be involved in immunosuppression in C57BL/6 mice. C57BL/6 mice required more splenic adherent cells to achieve the same proliferative response as found in BALB/c mice. The ability of thioglycollate-induced peritoneal macrophages to act as accessory cells in antigen-induced T-cell proliferation was less in C57BL/6 mice than in BALB/c mice. Thus, it appeared that the relative susceptibility of C57BL/6 mice to encephalitozoonosis may be due to defective accessory cell function of splenic and peritoneal macrophages, depressed lymphoproliferation against spore fragments (possibly due to prostaglandin-mediated suppression) and a delay in LK production. There was no significant difference between the survival times of BALB/c-nu and C57BL/6-nu mice, suggesting that non-immune mediated resistance did not play a role in determining resistance/susceptibility.

In vivo effects in mice of an anti-T cell immunotoxin.

We have evaluated both the proliferative response as well as the Thy-1 Ag expression of lymphocytes from mice treated in vivo with an anti-Thy-1 immunotoxin (IT). The IT was a rat IgG2c mAb recognizing the Thy-1 Ag, disulfide-linked to a ribosome-inactivating protein isolated from the seeds of the plant Saponaria officinalis (soapwort). Toxicity studies showed that a single i.v. injection of doses up to 20 micrograms IT/mouse was well tolerated and allowed indefinite survival. The Con A-induced proliferative response of spleen cells from mice killed 1 day after treatment with sublethal doses of IT was inhibited in a dose-dependent manner, with complete inhibition observed at doses of greater than or equal to 5 micrograms IT/mouse. Control experiments showed that the inhibition was due to the IT and not to its single components. Moreover, the IT effect was abolished by a large (100-fold) excess of anti-Thy-1 mAb alone given concurrently, but not by an unrelated, isotype-matched rat mAb. At all IT doses, the proliferative response to a B cell mitogen (LPS) was normal. Kinetic studies showed a time- and dose-dependent reconstitution of Con A responsiveness. In limiting dilution cultures of spleen cells from mice treated with 5 micrograms IT 1 or 4 days before death, a 97% depletion of T lymphocytes capable of proliferation was observed. Limiting dilution cultures showed that also the thymus of IT-treated mice was depleted by more than 90% of growth-competent T lymphocytes. Cytofluorographic studies of Thy-1+ cells from the spleens of IT-treated mice gave results which did not correlate with those obtained in functional assays. Thus, a dose-dependent reduction, followed by a time-dependent reconstitution of Thy-1+ cells was observed in this case too, but the depletion occurred at later time points and was less complete than that observed in functional assays. Moreover, the mean fluorescence intensity of the residual Thy-1+ cells decreased below normal levels.

MEDIATION OF THE INDUCTION OF IMMUNOLOGIC UNRESPONSIVENESS FOLLOWING ANTIGEN PRETREATMENT BY A CD4 (W3/25+) T CELL APPEARING TRANSIENTLY IN THE SPLENIC COMPARTMENT AND SUBSEQUENTLY IN THE TDL

We have previously demonstrated in a rat renal allograft model, using adoptive transfer studies, that a single blood transfusion results in the generation of blood donor-specific W3/25+ (CD4) T suppressor cells detectable in the thoracic duct lymph, but not in the spleen, 7 days after transfusion. In this study, again using the LEW-to-DA strain combination, we have investigated the time of appearance of suppressor activity in different lymphoid compartments in vivo and in vitro following a single blood transfusion. Cells (5 x 10(7) unfractionated TDL or 1 x 10(8) unfractionated spleen cells) were harvested from DA rats on days 1-6 after a single LEW blood transfusion and were adoptively transferred into syngeneic (DA), lightly irradiated (200 rads) hosts. The following day these animals received a kidney allograft (LEW or PVG). Specific suppressor activity, as demonstrated by the prolongation of survival of LEW, but not PVG, renal allografts was noted in the spleen 4 days after transfusion (MST greater than 100 days). Suppressor cells were not found in the TDL at this stage--but 6 days after transfusion suppressor activity could be detected in the TDL (median survival time [MST]greater than 100 days), while at this time no such activity was demonstrable in the spleen. The suppressor activity in the spleen on day 4 after transfusion was shown to be mediated by a W3/25+ (CD4) T cell. The proliferative response in a mixed lymphocyte culture of DA spleen cells, prepared 4 days after transfusion, or TDL cells prepared 6 days after transfusion, to LEW stimulator cells was significantly suppressed when compared with the response of cells harvested from untreated controls. In contrast, the proliferative response of spleen cells examined 6 days after transfusion, when no suppressor activity was detected in vivo, was significantly augmented in comparison with untreated controls. Thus we have demonstrated that a DST results in the generation of W3/25-positive (CD4) T cells capable of suppressing renal allograft rejection that appear transiently in the splenic compartment (4 days after transfusion) and subsequently in the TDL (six days after transfusion).

Modulation of mitogen-induced spleen cell proliferation and the antibody-forming cell response by beta-endorphin in vivo

Experiments were conducted which compared the in vivo effects of beta-endorphin (BEP), gamma-endorphin (γEP), methionine-enkephalin (Met-ENK), and acetylated BEP(1–27) on the in vitro proliferative response of rat spleen cells to concanavalin A (ConA). In addition, the influence of BEP administration on the primary and secondary antibody-forming cell (AFC) response to the soluble antigen keyhole-limpet hemocyanin (KLH) was examined. Intravenous administration of BEP enhanced the spleen cell proliferative response to ConA assessed 3 hr after a single bolus infusion. Conversely, infusion with AcBEP(1–27) suppressed the proliferative response, whereas no effects of intravenous γEP or Met-ENK treatment were observed. The enhancing effect of BEP administration was not detectable 24 hr after a single infusion, but could be maintained over a 44 hr period by multiple infusions. The primary AFC response to KLH was suppressed by a dose of 1 nmole BEP only. On the other hand, the secondary IgG AFC response to KLH was enhanced by 10 pmoles BEP, while the IgM and IgA AFC responses remained unaltered by BEP treatment. The anamnestic in vitro proliferative response of spleen cells cultured with KLH was not altered if BEP was administered at the time of secondary KLH immunization. These results extend previous observations of BEP-induced modulation of in vitro immune function by demonstrating that opioid and nonopioid forms of BEP administered in vivo alter the capacity of spleen cells to proliferate and develop antibody responses to antigen.

Differential effect of cyclosporine in vivo on the distribution of T cell subsets in the thymus, spleen, and lymph nodes.

Cyclosporine treatment of BALB/c mice (at a dose of 20 mg/kg every other day for 3 weeks) caused a remarkable reduction in the PNA-, L3T4+Lyt-2- subset of thymocytes. A significant reduction of the L3T4+Lyt-2-subset was also observed in both the lymph node and spleen cells of CsA-treated mice, though the degree of the reduction was lower than that in thymocytes. Both lymph node and spleen cells from CsA-treated mice showed a significant increase in the percentage of Thy-1.2 negative, L3T4-Lyt-2- cells (perhaps B cells). Thymocytes from CsA-treated mice showed a reduction in the in vitro proliferative responses to Con A and PHA. On the other hand, there was a slight, but not significant, decrease in the responses of lymph node cells to Con A, PHA and LPS. Spleen cells from CsA-treated mice showed a significant reduction in the responses to Con A and PHA, though the degree of the reduction was lower than that of thymocytes. There was a significant decrease in the proliferative response of spleen cells to LPS. These results suggest that CsA affects both thymus and spleen cells in vivo, preferentially impairing the L3T4+Lyt-2- subset (helper T cells or their precursors) within the thymus. The lymph node cells seem to be relatively spared from the in vivo effect of CsA compared with cells in the thymus and spleen.

Regulation of parasite antigen-induced T cell growth factor activity and proliferative responsiveness in Brugia pahangi-infected jirds.

Previous studies have demonstrated that the induction of immunoregulatory mechanisms in the spleens of Brugia pahangi-infected jirds is correlated with the onset of microfilaremia. This study investigated the relationship between production of a factor with IL-2-like activity and the regulation of T cell-mediated responses in jirds experimentally infected with B. pahangi. A factor present in culture supernatants of mitogen-stimulated jird lymphocytes supported the proliferation of murine CTLL cells and provided the basis for an IL-2 assay. Mitogen induced proliferative responses and IL-2 production of spleen cells but not lymph node cells from pre-patent and microfilaremic jirds were suppressed. Both B. pahangi Ag-induced proliferative responsiveness and IL-2 production of spleen cells from microfilaremic jirds were also suppressed relative to lymph node cells from the same animals or spleen cells from B. pahangi immunized or prepatent jirds. Depletion of histamine receptor-bearing cells restored the ability of spleen cells from microfilaremic jirds to produce significant levels of IL-2. In addition, in add-mixture experiments, spleen cells from microfilaremic jirds suppressed Ag-induced IL-2 production by cells from either B. pahangi- or KHL-immunized jirds. Exogenous IL-2 failed to reconstitute the suppressed Ag-induced proliferative response of spleen cells from microfilaremic jirds. This study demonstrates that the down-regulation of immune responses in B. pahangi infection is a cell-mediated event and is associated with an inability to produce IL-2.

Immunosuppressive effects of dietary wortmannin on rats and mice.

In order to assess the effects of the fungal toxin wortmannin on the immune system, rats and mice were fed wortmannin-containing cultures of Fusarium oxysporum for 1 or 2 weeks. Wortmannin caused significant decreases in thymic weight, thymic lymphocyte numbers, serum IgG and IgM levels, the primary humoral response to T-dependent and T-independent antigens and the proliferative response of spleen cells to pokeweed mitogen. In vitro administration of wortmannin did not produce evidence of cytotoxicity to spleen or thymus cells. The data indicate that wortmannin inhibits immune function in rats and mice and suggest that metabolic modification of the toxin is necessary for toxicity.

Immunomodulation by orally administered β-glucan in mice

Orally administered SSG, a β-1,3-glucan obtained from the culture filtrate of the fungus Sclerotinia sclerotiorum IFO 9395, was examined for effects on immune responses in mice. The proliferative responses of spleen cells from SSG-administered mice (40 or 80 mg/kg, daily for 5 or 10 consecutive days) to a T-cell mitogen, concanavalin A (Con A), or a B-cell mitogen, lipopolysaccharide (LPS), were higher than those from normal mice. Oral administration of SSG (80 mg/kg) to mice also enhanced the activities of both natural killer (NK) cells in spleen and the lysosomal enzyme of peritoneal macrophages. Furthermore, significant inhibition of tumor growth was observed in syngeneic tumor systems when SSG was administered directly after tumor implantation. The inhibiting effect required high doses of SSG (over 80 mg/kg). These results demonstrate that SSG can potentiate the immune response of mice following oral administration.

Immunomodulatory effects of neurotropin through the recovery of interleukin-2 production in autoimmune-prone (NZB/NZW) F 1 mice

The immunomodulatory effects of Neurotropin, a substance extracted from inflammatory skin of rabbits inoculated with vaccinia virus, were assessed in autoimmune-prone (NZB/NZW) F 1 (B/W F 1) mice. The concanavalin A (Con A)-induced proliferative response of spleen cells was markedly decreased in aged B/W F 1 mice as compared with young B/W F 1 mice. Neurotropin, when administered i.p. to aged B/W F 1 mice, significantly increased the Con A-induced proliferative response. In aged B/W F 1 mice, interleukin-2 (IL-2) production by Con A-stimulated spleen cells was severely impaired and IL-2 responsiveness of Con A-activated spleen cells was partially decreased in comparison with young B/W F 1 mice. Neurotropin, administered to the aged B/W F 1 mice, restored IL-2 production by Con A-stimulated spleen cells to the level of young B/W F 1 mice. Furthermore, Neurotropin completely restored the IL-2 responsiveness of Con A-activated spleen cells from aged B/W F 1 mice. To test whether Neurotropin exerts its immunoregulatory activities in B/W F 1 mice by restoring IL-2 production, we directly examined the effect of recombinant IL-2 on the immune functions of spleen cells in vitro. Recombinant IL-2 markedly enhanced Con A-induced proliferative response of aged B/W F 1 mice. Furthermore, the suppressive activity of spleen cells which had been activated by Con A in the presence of rIL-2 was significantly increased. These results indicate that some immunoregulatory activity in B/W F 1 mice can be corrected by IL-2 and suggest that Neurotropin restores immunoregulatory activity in B/W F 1 mice by the recovery of IL-2 production.

脂溶性プロドラッグパルミチン酸デキサメタゾンの免疫抑制作用

Corticosteroids which are incorporated in lipid emulsion are easily taken up by reticulo-endothelial system (RES) much more than free corticosteroids. In this study, the immunosuppressive effect of dexamethasone palmitate incorporated in lipid emulsion was compared with that of the free dexamethasone disodium phosphate. It showed that the antibody response in spleen to SRBC and DNP-Ficoll, the proliferative response of spleen cells to mitogens, DTH response to SRBC, ADCC activity of spleen cells and the effect on RES function in mice were strongly suppressed by dexamethasone palmitate incorporated in lipid emulsion when compared with the free corticosteroid. In addition, the effect in delaying mortality by preventing the spontaneous autoimmune disease of MRL/l mice was observed by the treatment with dexamethasone palmitate incorporated in lipid emulsion.From these results, dexamethasone palmitate incorporated in lipid emulsion is a potent long-acting immunosuppressant and is thought to be of value in the treatment of autoimmune disease such as idiopathic thrombocytopenic purpura (ITP) .

Modulation of cell-mediated immunity in mice with chronic unisexual or bisexualSchistosoma mansonicercarial infection

Three groups of C57BL/6J female mice were infected with female Schistosoma mansoni cercariae alone, male cercariae alone, or both sexes of cercariae. In a longitudinal study, the spleen cell proliferative responses to the mitogens phytohaemagglutinin (PHA) and Escherichia coli lipopolysaccharide (LPS) were monitored. Significant immune suppression was found in the three infected groups when compared with uninfected controls. Within the infected groups, mice inoculated with both sexes of cercariae were significantly more suppressed than those with single-sex cercarial infection. Thus, in addition to schistosome eggs, either sex of S. mansoni worms is capable, although to a lesser extent, of inducing hyporesponsiveness of cell-mediated immunity (CMI) in chronic schistosomiasis mansoni.

Thymosin, interleukins, isoprinosine and imuthiol do not reconstitute T-cells in athymic nude mice

Previous reports suggest that immunotherapy can induce T-cell development in athymic nude mice. Eight-week-old BALB/c nu/nu (athymic nude) mice were treated for 3, 6 and 12 weeks with thymosin fraction 5 (TF-5), buffy coat interleukins (BC-IL), recombinant IL-2 (rIL-2), a combination of TF-5 and BC-IL, isoprinosine or imuthiol. Control animals were treated with sterile saline or were given a syngeneic thymus graft. Spleen weight, cell yields and Thy 1.2 + T-cell markers were monitored and spleen cell proliferative responses to rIL-2, concanavalin A (Con A), Con A + rIL-2, phytohemagglutinin (PHA) and endotoxin (LPS) were assessed. Only mice transplanted with a syngeneic thymus showed progressive increases in both T-cell number and function. Minor changes in proliferative responses were noted at 3 weeks with all treatments; however, no biologically significant reconstitution of T-cell number or function was observed.

A soluble factor produced by bone marrow natural suppressor cells blocks interleukin 2 production and activity

We previously reported that a population of Fcγ-receptor + (FcγR +) suppressor cells present in normal unstimulated rabbit bone marrow inhibited the growth of autologous rapidly proliferating bone marrow cells devoid of FcγR. It is now reported that the FcγR + bone marrow cells produced a soluble, nondialyzable suppressor factor(s) (SF) which blocked the proliferation of FcγR − bone marrow cells. In addition, the FcγR + cells and SF significantly inhibited spleen cell proliferation in response to concanavalin A (Con A), phytohemagglutinin, and pokeweed mitogen. The bone marrow SF exhibited a dose-dependent suppression of the growth of IL-2-dependent T lymphocytes in the presence of IL-2. SF also completely blocked the production or release of IL-2 by Con A-stimulated T cells. Thus, these bone marrow natural suppressor cells produced a soluble factor, which regulated the growth of rapidly proliferating bone marrow cells and also regulated T cell reactivity by modulating IL-2 production and activity.

Specific suppression of allograft rejection after treatment of recipient mice with ultraviolet radiation and allogeneic spleen cells.

Sensitization of UV-irradiated mice with alloantigen results in suppression of the immune response against that antigen. The induction of delayed hypersensitivity and the proliferative response of spleen cells isolated from the UV-irradiated mice are significantly suppressed. We demonstrate here that the rejection of tissue allografts is also suppressed after treatment of the recipient animals with UVR. Two signals, UVR and antigenic sensitization are required to suppress allograft rejection, exposure to UVR alone is not sufficient. The resulting immunosuppression is specific for the antigen used to sensitize the UV-irradiated animal. The induction of graft-versus-host disease was also significantly suppressed when spleen cells from UV-irradiated alloantigen sensitized mice were used to reconstitute lethally irradiated allogeneic mice. Our interpretation of these data is that the antigen-specific suppressor T cells present in the spleens of the UV-irradiated alloantigen sensitized mice are suppressing the immune response against the alloantigen. Thus, the induction of suppression by UVR serves as a novel method of suppressing graft rejection.