L-10 has been recognized as an irnmune suppressive cytokine which inhibits Ag-specific activation and proliferation of T cells. It also inhibits Ag presenting capacity of monocyte/macrophage and down-regulates monokine production. However it has also shown that IL-10 has stimulatory effect on immune effector cells in recent studies. This report shows that IL-10 has direct stimulatory effect on antitumor cytolytic activity suppressed by TGF-B. To assess the effect of IL-10 on cytolytic activity against tumor, spleen cells prepared from tumor-bearing mice were cultured with mitomycin C-treated MOPC-315 cells in the presence of IL-10. Unexpectedly, IL-10 was able to reverse the cytolytic activity suppressed with TGF-B. The stimulatory effect of IL-10 was dependent on the addition time of IL-10. At day 0, 4, those effects were shown higher than those of the other days. Also, the stimulatory effect of IL-10 showed specificity against MOPC-315 tumor cells. To elucidate the role of endogenous IL-10, TGF-B in MLTC cultures, anti-IL-10 and anti-TGF-B mAb were used. The inhibition of IL-10 release in MLTC cultures by using anti-IL-10 mAb resulted in the suppression of cytolytic activity against MOPC-315 tumor cells. Taken together, although IL- 10 has been recognized as a strong immunosuppressive cytokine derived of tumor cells, IL-10 showed the direct stimulatory effect on the antitumor cytolytic activity of spleen cells.
Animals ; Interleukin-10* ; Mice ; Mitomycin ; Sensitivity and Specificity ; Spleen* ; T-Lymphocytes ; Transforming Growth Factor beta

BACKGROUND: Transforming growth factor-beta1 (TGF-beta1) directs class switch recombination (CSR) to IgA isotype, which is a predominant antibody in mucosal surfaces. Although IgA is preferentially committed in mucosal lymphoid tissues, it is not definitely established whether hallmarks of IgA CSR such as IgA germ-line transcripts (GLTalpha), post-switch transcripts (PSTalpha) and circle transcripts (CTalpha) are readily expressed in such tissues. Therefore, we compared the expression of these transcripts among mouse Peyer's patches (PP), mesenteric lymph nodes (MLN), and spleen. METHODS: Levels of GLTs, PSTs and CTs were measured by RT-PCR in isolated PPs, MLNs and spleen cells. RESULTS: GLTalpha and PSTalpha were well expressed in PP and MLN cells but in spleen cells. Similar patterns were observed in the expression of GLgamma2b and PSTgamma2b. On the other hand, these transcripts were only inducible in spleen cells upon stimulated with LPS and TGF-beta1. In addition, CTalpha and CTgamma2b were detected in PP cells. CONCLUSION: PP B cells readily express IgA GLT, PST, and CT. Overall expression patterns of these transcripts were similar in MLN cells. Thus, these results suggest that microenvironment of PP and MLN influences spontaneous IgA CSR, which lacks in systemic lymphoid tissues such as spleen.
Animals ; B-Lymphocytes ; Hand ; Immunoglobulin A* ; Lymph Nodes ; Lymphoid Tissue* ; Mice* ; Peyer's Patches ; Recombination, Genetic ; Spleen ; Transforming Growth Factor beta1

Eimeria tenella, an intracellular protozoan parasite infecting the epithelial cells of the ceca of chickens, causes severe diarrhea and bleeding that can lead its host to death. It is of interest that E. tenella first penetrate into the mucosal intraepithelial lymphocytes (IEL) before they parasitize crypt or villous epithelial cells. This in vitro study was undertaken to know whether the penetration of E. tenella into such a lymphoid cell is a beneficial step for the parasite survival and development. Three sequential experiments were performed. First, the in vitro established bovine kidney cell line, MDBK cells, were evaluated for use as host cells for E. tenella, through morphological observation. Second, the degree of parasite development and multiplication in MDBK cells was quantitatively assayed using radioisotope-labelled uracil (3H-uracil). Third, the E. tenella sporozoites viability was assayed after preincubation of them with chicken spleen cells. E. tenella oocysts obtained from the ceca of the infected chickens were used for the source of the sporozoites. Spleen cells (E) obtained from normal chickens (FP strain) were preincubated with the sporozoites (T) at the E:T ratio of 100:1, 50:1 or 25:1 for 4 or 12 hours, and then the mixture was inoculated into the MDBK cell monolayer. Morphologically the infected MDBK cells revealed active schizogonic cycle of E. tenella in 3-4 days, which was characterized by the appearance of trophozoites, and immature and mature schizonts containing merozoites. The 3H-uracil uptake by E. tenella increased gradually in the MDBK cells, which made a plateau after 48-60 hours, and decreased thereafter. The uptake amount of 3H-uracil depended not only upon the inoculum size of the sporozoites but also on the degree of time delay (preincubation; sporozoites only) from excystation to inoculation into MDBK cells. The 3H-uracil uptake became lower as the preincubation time was prolonged. In comparison, after preincubation of sporozoites with spleen cells for 4 or 12 hours, the 3H-uracil uptake was significantly increased compared with that of control group. From the results, it was inferred that, although the penetration of E. tenella sporozoites into the lymphoid cells such as IEL is not an essential step, it should be at least a beneficial one for the survival and development of sporozoites in the chicken intestine.
Cattle- ; Cell-Line ; Cells,-Cultured ; Chickens- ; English-Abstract ; *Eimeria-growth-and-development ; *Kidney-parasitology ; *Lymphocytes-parasitology ; *Spleen-cytology

It is well established that TGF-beta1 and retinoic acid (RA) cause IgA isotype switching in mice. We recently found that lactoferrin (LF) also has an activity of IgA isotype switching in spleen B cells. The present study explored the effect of LF on the Ig production by mouse peritoneal B cells. LF, like TGF-beta1, substantially increased IgA production in peritoneal B1 cells but little in peritoneal B2 cells. In contrast, LF increased IgG2b production in peritoneal B2 cells much more strongly than in peritoneal B1 cells. LF in combination with RA further enhanced the IgA production and, interestingly, this enhancement was restricted to IgA isotype and B1 cells. Similarly, the combination of the two molecules also led to expression of gut homing molecules alpha4beta7 and CCR9 on peritoneal B1 cells, but not on peritoneal B2 cells. Thus, these results indicate that LF and RA can contribute to gut IgA response through stimulating IgA isotype switching and expression of gut-homing molecules in peritoneal B1 cells.
Animals ; B-Lymphocytes ; Immunoglobulin A* ; Immunoglobulin Class Switching ; Immunoglobulin G ; Lactoferrin* ; Mice ; Spleen ; Transforming Growth Factor beta1 ; Tretinoin*

OBJECTIVETo study on the effect of been pollen on development of immune organ of animal.

METHODA total of 144 one day-old broilers were randomly divided into 2 groups, in which each group included 72 chickens. The control group was fed on the basal diet for 42 days, and that of experiment group supplemented 1.5% bee pollen. Six chickens in each group were selected and slaughtered at 7, 14, 21, 28, 35, 42 days respectively, and the thymuses, cloacal bursa and spleens were obtained, weighted, fixed in Bouin liquid and made into paraffin section.

RESULTCompared with control group, the weight and the relative weight of thymuses, cloacal bursa and spleens of experiment group increased significantly (P < 0.05) or extremely significantly (P < 0.01). In experiment group, the cortex of thymic lobule, bursa nodule and Periarterial Lymphatic Sheaths thicken obviously; the volume of bursa nodule, splenic nodule and ellipsoid augmented, and the germinal center of splenic nodule were obvious; the thymic corpuscle increased; the plica of cloacal bursa developed well and the degenerating of it retarded.

CONCLUSIONThe diet supplemented bee pollen could boost the early development of thymus and cloacal bursa, retard the degenerating of cloacal bursa and promote the immune response of spleen.

Animal Nutritional Physiological Phenomena ; Animals ; Bees ; Bursa of Fabricius ; anatomy & histology ; growth & development ; Chickens ; growth & development ; immunology ; Female ; Male ; Organ Size ; Pollen ; Random Allocation ; Spleen ; anatomy & histology ; growth & development ; Thymus Gland ; anatomy & histology ; growth & development

OBJECTIVE: To investigate the dosage of bovine type II collagen (BnCII) for the induction of oral tolerance in CIA animals, and to verify the changes of immune response and TGF-beta production of mesenteric lymph node cells in tolerized CIA animals. METHODS: Oral tolerance was induced by feeding of variable doses (5 microgram, 10 microgram, 20 microgram and 40 microgram) of BnCII to DBA/1 mice 4 times per week during 2 weeks, and control mice were given ovalbumin (1000 microgram), before immunization. We examed clinical assessment ; incidence of arthritis, severity of arthritis, arthritic limb by visual analysis. IgG antibodies to BnCII were measured by ELISA, T cell responses to BnCII and PHA were quantified by antigen (CII)-induced 3H-thymidine incorporation into lymphocytes of mesenteric lymph node, draining lymph node, and spleen. TGF-beta in supernatants obtained from lymph node culture medium was measured by ELISA. RESULTS: Arthritis limbs were observed in 100% of control at 5 weeks after subcutaneous BnCII injection. The incidences of CIA in all tolerized group were significantly lower than that in control 5 weeks after immunization (control 100 % vs. 5 microgram feeding group: 50%, 10 microgram feeding group: 50%, 20 microgram feeding group: 50%, 40 microgram feeding group: 55.5%, P<0.01). In comparison to control, mean articular indices were lower in all tolerized groups (control 5.13 : 5 microgram feeding group 3.50, 10 microgram feeding group 2.75, 20 microgram feeding group 2.87, 40 microgram feeding group 2.63, P<0.05). Arthritic limbs were also significantly lower in tolerized groups (control 58.3 : 5 microgram feeding group 20.8, 10 microgram feeding group 16.7, 20 microgram feeding group 20.8, 40 microgram feeding group 20.8, P<0.05). The titers of IgG antibody to CII were lower in tolerized group than that in control [tolerized group ; median 10 (min. 0, max. 48), control ; median 33 (min. 8.6, max. 101), P<0.05]. The proliferative responses to BnCII were significantly suppressed in tolerized (control 8010+/-2319cpm, tolerized group 4500+/-2060cpm, P<0.01). High TGF-beta production was noted in tolerized group (control ; 28pg/ml, BnCII feeding group ; 73pg/ml). CONCLUSION: Oral tolerance in DBA/1 mice was successfully induced from low doses of BnCII (5 microgram) and suppressed T and B cell function in conjunction with increased TGF-beta production may play an important role for the induction of CII induced oral tolerance in DBA/1 mice.
Animals* ; Antibodies ; Arthritis ; Arthritis, Experimental ; Collagen Type II* ; Enzyme-Linked Immunosorbent Assay ; Extremities ; Immunization ; Immunoglobulin G ; Incidence ; Lymph Nodes ; Lymphocytes ; Mice ; Ovalbumin ; Spleen ; Transforming Growth Factor beta

OBJECTIVE: To investigate the dosage of bovine type II collagen (BnCII) for the induction of oral tolerance in CIA animals, and to verify the changes of immune response and TGF-beta production of mesenteric lymph node cells in tolerized CIA animals. METHODS: Oral tolerance was induced by feeding of variable doses (5 microgram, 10 microgram, 20 microgram and 40 microgram) of BnCII to DBA/1 mice 4 times per week during 2 weeks, and control mice were given ovalbumin (1000 microgram), before immunization. We examed clinical assessment ; incidence of arthritis, severity of arthritis, arthritic limb by visual analysis. IgG antibodies to BnCII were measured by ELISA, T cell responses to BnCII and PHA were quantified by antigen (CII)-induced 3H-thymidine incorporation into lymphocytes of mesenteric lymph node, draining lymph node, and spleen. TGF-beta in supernatants obtained from lymph node culture medium was measured by ELISA. RESULTS: Arthritis limbs were observed in 100% of control at 5 weeks after subcutaneous BnCII injection. The incidences of CIA in all tolerized group were significantly lower than that in control 5 weeks after immunization (control 100 % vs. 5 microgram feeding group: 50%, 10 microgram feeding group: 50%, 20 microgram feeding group: 50%, 40 microgram feeding group: 55.5%, P<0.01). In comparison to control, mean articular indices were lower in all tolerized groups (control 5.13 : 5 microgram feeding group 3.50, 10 microgram feeding group 2.75, 20 microgram feeding group 2.87, 40 microgram feeding group 2.63, P<0.05). Arthritic limbs were also significantly lower in tolerized groups (control 58.3 : 5 microgram feeding group 20.8, 10 microgram feeding group 16.7, 20 microgram feeding group 20.8, 40 microgram feeding group 20.8, P<0.05). The titers of IgG antibody to CII were lower in tolerized group than that in control [tolerized group ; median 10 (min. 0, max. 48), control ; median 33 (min. 8.6, max. 101), P<0.05]. The proliferative responses to BnCII were significantly suppressed in tolerized (control 8010+/-2319cpm, tolerized group 4500+/-2060cpm, P<0.01). High TGF-beta production was noted in tolerized group (control ; 28pg/ml, BnCII feeding group ; 73pg/ml). CONCLUSION: Oral tolerance in DBA/1 mice was successfully induced from low doses of BnCII (5 microgram) and suppressed T and B cell function in conjunction with increased TGF-beta production may play an important role for the induction of CII induced oral tolerance in DBA/1 mice.
Animals* ; Antibodies ; Arthritis ; Arthritis, Experimental ; Collagen Type II* ; Enzyme-Linked Immunosorbent Assay ; Extremities ; Immunization ; Immunoglobulin G ; Incidence ; Lymph Nodes ; Lymphocytes ; Mice ; Ovalbumin ; Spleen ; Transforming Growth Factor beta

This study was aimed to investigate the effect of exogenous VEGF on hematopoietic stem cell mobilization and immune system. The C57BL/6J mice were randomly divided into the normal control group, VEGF short-term group (5 d) and VEGF long-term group (27 d). Mice in the experimental group were injected ip with VEGF (100 ng/d); mice in control group were injected ip with PBS. The white blood cell (WBC) count and the ratio of lymphocyte in the peripheral blood at different time point were assayed by hemacytometer. The percentage of hematopoietic stem cell (HSC), lymphocyte subgroup, regulatory T cell (Treg), myeloid-derived suppressor cells (MDSC) in the peripheral blood and spleen of different groups were detected by flow cytometry. The morphological changes of spleen and spleen index of mice in the control and long-term group were observed by microscopy. The results showed that the absolute number of WBC in the peripheral blood of mice significantly increased after injection of VEGF, and the peak value was at day 3. The percentage of Lin(-)Sca-1(+)CD117(+) cells in the peripheral blood and spleen of the long-term group were significantly higher than that in the normal control group (P < 0.05). The spleen of the mice in VEGF long-term group was larger than that of the control group, the spleen index also increased (P < 0.05), and remarkable extramedullary hematopoietic signs were found in the HE stained sections. There was no significant change in the total ratio of lymphocytes in the peripheral blood after injection, but the percentage of CD3(+) cells and the CD3(+)/B220(+) ratio in the long-term group deceased; the percentages of Treg and Gr-1(+)CD11b(+) MDSC in the experimental groups increased (P < 0.05), which more significantly increased in the long-term group than that in the short-term group (P < 0.05). It is concluded that the exogenous VEGF promotes hematopoietic stem cell mobilization, and at same time up-regulates the many kinds of suppressive immune cell levels which leads to changes of immuno-function.
Animals ; Hematopoietic Stem Cell Mobilization ; methods ; Male ; Mice ; Mice, Inbred C57BL ; Spleen ; cytology ; T-Lymphocytes, Regulatory ; Vascular Endothelial Growth Factor A ; pharmacology

recombinant human IGF-I in energy-restriction model. Experimental design; Sprague-Dawley rats(n=20) weighing 90-100g were used. Rats were fed a control diet two times a day(AM 8-11, PM 5-8) for four days after arrival and then assigned to one of three groups: control, energy-restricted, energy-restricted IGF-I treatment group. Energy restricted group was given with a decrese of 25% in the energy without changes in the protein by feeding 88% by weight to energy-restricted diet. During the 10days of energy restriction, the growth rate was reduced by 35%(2.70+-0.18g/day in energy restricted group vs. 4.13+-0.75g/day in the control group). At sacrifice, the tail lengh and weight of organs were not significantly decreased except the spleen and thymus(-17%: P<0.05). Serum IGF-I was reduced by 19% at the end of 10days of energy restriction. The glycemia, measured each day by glucometer from blood collected at the tail, was not reduced by energy restriction(105.4+-7.7 in control group vs. 101.3+-4.1mg/dl). The abundance of serum IGF-BPs was unchanged by this restriction.Despite the 1.5 fold increase of IGF-I concentration in energy restricted IGF-I injection group at sacrifice(1994+-172ng/ml vs. 1221+-110 ng/ml energy restricted group), IGF-I treatment(300 ug/day in twice sc injection for 6day) did not significantly accelerate the growth rate(body weight)(2.87+-0.20 vs. 2.70+-0.18g/day in energy restricted group).The glycemia was slightly reduced by IGF-I treatment(91.7+-5.0 mg/dl vs. 101.3+-4.5 mg/dl in energy restricted group), but it was not significant. However, the spleen and thymus weight, decreased by energy restriction, was completely normalized by IGF-I treatment.In summary, lack of a significant anabolic response to injection of IGF-I during energy restriction in this study may be associated with the compensatory growth response(alterations in dietary protein utilization) which followed initial period of energy restriction.
Animals ; Diet ; Dietary Proteins ; Humans ; Insulin ; Insulin-Like Growth Factor I ; Rats ; Rats, Sprague-Dawley ; Research Design ; Spleen ; Tail ; Thymus Gland

Leishmania (L.) tropica is a causative agent of cutaneous leishmaniasis, and occasionally of visceral or viscerotropic leishmaniasis in humans. Murine models of Leishmania infection have been proven to be useful for elucidation of mechanisms for pathogenesis and immunity in leishmaniasis. The aim of this study was to establish a murine model for human viscerotropic leishmaniasis, and the growth pattern of L. tropica was studied in different tissues of BALB/c mice in order to find out whether the parasite visceralizes in this murine model. L. major was used as a control as this species is known to cause a progressive infection in BALB/c mice. L. tropica or L. major was injected into the footpad of mice, and thickness of footpad, parasite loads in different tissues, and the weight of the spleen and lymph node were determined at different intervals. Results showed that L. tropica visceralizes to the spleen and grows there while its growth is controlled in footpad tissues. Dissemination of L. tropica to visceral organs in BALB/c mice was similar to the growth patterns of this parasite in human viscerotropic leishmaniasis. The BALB/c model of L. tropica infection may be considered as a good experimental model for human diseases.
Animals ; *Disease Models, Animal ; Female ; Foot/parasitology ; Humans ; Leishmania major/growth & development ; Leishmania tropica/*growth & development ; Leishmaniasis/*parasitology ; Lymph Nodes/parasitology/pathology ; Mice ; Mice, Inbred BALB C ; Organ Size ; Spleen/parasitology/pathology