B cells and tertiary lymphoid structures are associated with survival in papillary thyroid cancer.

PURPOSE: The function of B cells in papillary thyroid cancer (PTC) is controversial. The role of B-cell-related tertiary lymphoid structures (TLSs) is still unclear. Whether B cells exert their anti-tumor effect through forming TLS in PTC needs further investigation. METHODS: We detected the percentage of B cells in PTC tissues by multi-parameter flow cytometry. Paraffin-embedded tumor tissues of 125 PTC patients were collected and stained with Haematoxylin-Eosin (H&E) for inflammatory infiltration analysis in combination with clinical features. Multiplexed immunohistochemistry (mIHC) was performed to verify the TLSs in above inflammatory infiltration. Correlation of B cells and TLSs with prognosis was analyzed using the TCGA database. RESULTS: We observed that PTC patients with higher expression of B lineage cell genes had improved survival and the percentage of B cells in the PTC tumor tissues was variable. Moreover, PTC tumor tissues with more B cells were surrounded by immune cell aggregates of varying sizes. We furtherly confirmed the immune cell aggregates as TLSs with different maturation stages. By analyzing PTC data from TCGA database, we found the maturation stages of TLSs were associated with genders and clinical stages among PTC patients. Moreover, patients with high TLSs survived longer and had a better prognosis. CONCLUSION: B cells are associated with the existence of TLSs which have different maturation stages in PTC. Both B cells and TLSs are associated with the survival rate of PTC. These observations indicate that the anti-tumor effects of B cells in PTC are associated with TLSs formation.

Reversible phospho-Smad3 signalling between tumour suppression and fibrocarcinogenesis in chronic hepatitis B infection.

Transforming growth factor (TGF)-ß, type I receptor (TßRI) and c-Jun N-terminal kinases (JNK) phosphorylate Smad3 differentially to create 2 isoforms phosphorylated (p) at the COOH-terminus (C) or at the linker region (L) and regulate hepatocytic fibrocarcinogenesis. This study aimed to compare the differences between how hepatitis B virus (HBV) infection affected hepatocytic Smad3 phosphorylated isoforms before and after anti-viral therapy. To clarify the relationship between Smad3 phosphorylation and liver disease progression, we studied 10 random patients in each stage of HBV-related fibrotic liver disease (F1-4) and also 10 patients with HBV-associated HCC. To examine changes in phosphorylated Smad3 signalling before and after anti-HBV therapies, we chose 27 patients with chronic hepatitis B who underwent baseline and follow-up biopsies at 52 weeks from the start of nucleoside analogue treatments (Lamivudine 100 mg daily or Telbivudine 600 mg daily). Fibrosis stage, inflammatory activity and phosphorylated Smad3 positivity in the paired biopsy samples were compared. Hepatocytic pSmad3C signalling shifted to fibrocarcinogenic pSmad3L signalling as the livers progressed from chronic hepatitis B infection to HCC. After nucleoside analogue treatment, serum alanine aminotransferase (ALT) and HBV-DNA levels in 27 patients with HBV-related chronic liver diseases were decreased dramatically. Decrease in HBV-DNA restored pSmad3C signalling in hepatocytes, while eliminating prior fibrocarcinogenic pSmad3L signalling. Oral nucleoside analogue therapies can suppress fibrosis and reduce HCC incidence by successfully reversing phosphorylated Smad3 signalling; even liver disease progressed to cirrhosis in chronic hepatitis B patients.

Successful immunotherapy of autoimmune cholangitis by adoptive transfer of forkhead box protein 3(+) regulatory T cells.

Treatment of primary biliary cirrhosis (PBC) has lagged behind that of other autoimmune diseases. In this study we have addressed the potential utility of immunotherapy using regulatory T cells (Treg ) to treat murine autoimmune cholangitis. In particular, we have taken advantage of our ability to produce portal inflammation and bile duct cell loss by transfer of CD8(+) T cells from the dominant negative form of transforming growth factor beta receptor type II (dnTGF-ßRII) mice to recombination-activating gene (Rag)1(-/-) recipients. We then used this robust established adoptive transfer system and co-transferred CD8(+) T cells from dnTGF-ßRII mice with either C57BL/6 or dnTGF-ßRII forkhead box protein 3 (FoxP3(+) ) T cells. Recipient mice were monitored for histology, including portal inflammation and intralobular biliary cell damage, and also included a study of the phenotypical changes in recipient lymphoid populations and local and systemic cytokine production. Importantly, we report herein that adoptive transfer of Treg from C57BL/6 but not dnTGF-ßRII mice significantly reduced the pathology of autoimmune cholangitis, including decreased portal inflammation and bile duct damage as well as down-regulation of the secondary inflammatory response. Further, to define the mechanism of action that explains the differential ability of C57BL/6 Treg versus dnTGF-ßRII Treg on the ability to down-regulate autoimmune cholangitis, we noted significant differential expression of glycoprotein A repetitions predominant (GARP), CD73, CD101 and CD103 and a functionally significant increase in interleukin (IL)-10 in Treg from C57BL/6 compared to dnTGF-ßRII mice. Our data reflect the therapeutic potential of wild-type CD4(+) FoxP3(+) Treg in reducing the excessive T cell responses of autoimmune cholangitis, which has significance for the potential immunotherapy of PBC.

Serum secretory phospholipase A2-IIa (sPLA2-IIA) levels in patients surviving acute myocardial infarction.

BACKGROUND: The increase of secretory phospholipase A2-IIa (sPLA2-IIa) in culprit coronary lesions is associated with myocardial infarction, and the increase of sPLA2-IIa in peripheral plasma levels has a significant risk and prognostic value in patients with coronary artery disease. Little is known about the prognostic significance of elevated serum sPLA2-IIa in post-acute myocardial infarction (post-AMI) patients. OBJECTIVES: The present study is designed to investigate the potential association between serum sPLA2-IIa and prognosis in post-acute myocardial infarction (post-AMI) patients. PATIENTS AND METHODS: From Oct 1998 to Sep 2008, a total of 964 post-AMI patients with serum samples collected in the convalescent stage were studied. Serum levels of sPLA2-IIa were measured by ELISA. According to the optimal cut-off value for sPLA2-IIa concentration, patients were then divided into 2 groups. Categorical variables were compared between the 2 groups using the χ2 test. All continuous variables are described as mean ± SD and were compared using Student's t-test. Statistical associations between clinicopathological observations and sPLA2-IIa levels were determined using the Mann-Whitney U test. The clinical value of sPLA2-IIa level as a prognostic parameter was evaluated using the Cox's proportional hazards model. RESULTS: During a median follow-up period of 1,462 days, 123 patients (12.7%) had adverse events (all-cause mortality, n=52; non-fatal MI, n=31; readmission for heart failure [HF], n=40). Patients were divided into 2 groups according to a serum sPLA2-IIa level of 360 ng/dl, which was determined to be the optimal cut-off for discriminating all-cause mortality based on the maximum value of the area under the receiver operating characteristic curve. Patients with elevated sPLA2-IIa (> 360 ng/dl, n=164) had a significantly higher prevalence of death (18.3% [30/164] vs. 2.75% [22/800] p < 0.001) and readmission for HF (14% [23/164/ vs. 2.1% [17/800], p < 0.0001), but not of non-fatal MI (4.88% [8/164]vs. 2.87% [23/800], p = 0.096), compared to those with sPLA2-IIa < 360 ng/dl. Multivariate Cox regression analysis indicated that elevated serum sPLA2-IIa was associated with an increased risk of mortality and readmission for HF. CONCLUSIONS: Elevated serum sPLA2-IIa during the convalescent stage of AMI predicted long-term mortality and readmission for HF after survival discharge in the post-AMI patients.

Monocytes-macrophages phagocytosis as a potential marker for disease resistance in generation 1 of dwarf chickens.

Monocytes-macrophages play an indispensable role in the immune system. The current study investigated the effect of selection for monocytes-macrophages phagocytosis on disease resistance in generation 1 (G1) of dwarf chickens. Five hundred dwarf chickens of generation 0 (G0) were divided into high and low phagocytic index (PI) groups (HPIG and LPIG, respectively) based on their PI of monocytes-macrophages at 290 d of age. Then, 2 x 2 mating combinations were conducted. Sixty G0 chickens from another dwarf chicken group were used to measure the levels of monocytes-macrophages phagocytosis at different developmental stages. Among a total of 2,500 randomly selected G1 chickens, 2,100 individuals were used for a surviving and growing test under adverse feeding circumstances, and the other 400 individuals were tested for Salmonella Pullorum challenge. The results showed that progenies of HPIG hens (female symbol) were more resistant to Salmonella Pullorum. After challenge, the death rate of progeny from HPIG female symbol (28.9%) was only 58% that of progeny from LPIG female symbol (49.4%, P < 0.001). In addition, the natural infection rate of Salmonella Pullorum before 207 d for offspring from HPIG female symbol (35.0%) was significantly lower than that for offspring from LPIG female symbol (48.3%, P < 0.001). The natural mortality before 56 d in progeny of HPIG female symbol (22.6%) was significantly lower than that in progeny of LPIG female symbol (29.1%) with a P-value of 0.001. The G1 chickens of HPIG G0 female symbol weighed more than those born to LPIG G0 female symbol at 28 and 42 d of age, whereas the difference was not statistically significant at 56 d of age. The heritability of monocytes-macrophages phagocytosis was 0.40, which was moderate. The PI values were at a low level before 126 d and increased dramatically until they declined significantly after 294 d. It could be concluded that phagocytosis of monocytes-macrophages is a marker for breeding excellent progeny with strong disease resistance.

Beta-glucosylceramide ameliorates liver inflammation in murine autoimmune cholangitis.

We have demonstrated spontaneous development of autoimmune cholangitis, similar to human primary biliary cirrhosis, in mice expressing a dominant negative form of the transforming growth factor-beta receptor (dnTGF-betaRII) restricted to T cells. The autoimmune cholangitis appears to be mediated by autoreactive CD8(+) T lymphocytes that home to the portal tracts and biliary system. Because the liver pathology is primarily secondary to CD8(+) T cells, we have determined herein whether administration of beta-glucosylceramide (GC), a naturally occurring plant glycosphingolipid, alters the natural history of disease in this model. We chose GC because previous work has demonstrated its ability to alter CD8(+) T cell responses and to down-regulate tissue inflammation. Accordingly, dnTGF-betaRII mice were treated with either GC or control for a period of 18 weeks beginning at 6 weeks of age. Importantly, in mice that received GC, there was a significant decrease in the frequency and absolute number of autoreactive liver-infiltrating CD8(+) T cells, accompanied by a significant decrease in activated CD44(high) CD8(+) T cell populations. Further, there was a significant reduction in portal inflammation in GC-treated mice. Interestingly, there were no changes in anti-mitochondrial antibodies, CD4(+) T cells, CD19(+) B cells or natural killer (NK) T cell populations, indicating further that the beneficial effects of GC on liver inflammation were targeted specifically to liver-infiltrating CD8(+) T cells. These data suggest that further work on GC in models of CD8(+) T-mediated inflammation are needed and point to a new therapeutic venue for potentially treating and/or modulating autoimmune disease.

Induction of autoimmune cholangitis in non-obese diabetic (NOD).1101 mice following a chemical xenobiotic immunization.

Our laboratory has suggested that loss of tolerance to pyruvate dehydrogenase (PDC-E2) leads to an anti-mitochondrial antibody response and autoimmune cholangitis, similar to human primary biliary cirrhosis (PBC). We have suggested that this loss of tolerance can be induced either via chemical xenobiotic immunization or exposure to select bacteria. Our work has also highlighted the importance of genetic susceptibility. Using the non-obese diabetic (NOD) congenic strain 1101 (hereafter referred to as NOD.1101 mice), which has chromosome 3 regions from B6 introgressed onto a NOD background, we exposed animals to 2-octynoic acid (2OA) coupled to bovine serum albumin (BSA). 2OA has been demonstrated previously by a quantitative structural activity relationship to react as well as or better than lipoic acid to anti-mitochondrial antibodies. We demonstrate herein that NOD.1101 mice immunized with 2OA-BSA, but not with BSA alone, develop high titre anti-mitochondrial antibodies and histological features, including portal infiltrates enriched in CD8(+) cells and liver granulomas, similar to human PBC. We believe this model will allow the rigorous dissection of early immunogenetic cause of biliary damage.

Effects of methyltestosterone on immunity against Salmonella Pullorum in dwarf chicks.

This study was conducted to determine effects of methyltestosterone on innate immunity and adaptive immunity against Salmonella Pullorum in dwarf chicks. In vivo experiment, comparisons of pathological sections, viable counts of bacteria, specific antibody levels, and subsets of T lymphocytes were set forth between chicks with or without 10(-7) M methyltestosterone treatment (2 d of age through 21 d of age) and challenged with 5 x 10(8) virulent Salmonella Pullorum (7 d of age), and in vitro experiment, phagocytic and killing abilities, reactive oxygen intermediate production, and reactive nitrogen intermediate production of monocytes-macrophages treated with high (10(-8) M/10(6) cell) or physiological (10(-14) M/10(6) cell) concentration of methyltestosterone were examined after Salmonella Pullorum infection. The results showed that (1) in vivo, administration of methyltestosterone enhanced susceptibility to Salmonella Pullorum infection and depressed cellular immunity against Salmonella Pullorum, whereas it had no effect on humoral immunity in dwarf chicks; (2) in vitro, at high concentration, methyltestosterone reduced (P < 0.05) monocytes-macrophages mediated reactive oxygen intermediate-dependent killing of Salmonella Pullorum, whereas low concentration of methyltestosterone enhanced (P < 0.05) reactive oxygen intermediate-dependent killing of Salmonella Pullorum in male dwarf chicks but not in females; and (3) although challenged with Salmonella Pullorum, phagocytic ability and monocytes-macrophages mediated reactive nitrogen intermediate-dependent killing were not affected by methyltestosterone in vitro. The results indicated that methyltestosterone affected the immune response to Salmonella Pullorum in dwarf chicks by changing monocytes-macrophages mediated reactive oxygen intermediate-dependent killing and cellular immunity, and the effects were dose-dependent; furthermore, the former 2 pathways played important roles in preventing Salmonella Pullorum infection in dwarf chicks, although the mechanism needs further study.

Effect of selection for phagocytosis in dwarf chickens on immune and reproductive characters.

In current study, phagocytosis product (PP) of peripheral blood monocytes was detected among 920 dwarf chickens (460 per sex) at 20 wk of age, and based on discrepancies of PP, the flock was grouped (the highest group, the medium group, and the lowest group). Then serum hemagglutination inhibition antibody titers and subpopulations of T-lymphocytes of each group were examined after inoculations of avian influenza virus H5N2 inactivated vaccine (20 wk of age), avian influenza virus H9 inactivated vaccine (24 wk of age), and Newcastle disease virus-egg drop syndrome virus bigeminal inactivated vaccine (28 wk of age), respectively, to study the relationship between PP and immune response. To gain insight into effects of selection for PP on number of eggs, mean egg weight, fertilization rate, hatchability, and rate of healthy chicks, 9 (3 x 3) mating combinations were conducted. The results showed that (1) selection for higher PP in both sexes benefited to humoral immunity but not CD8(+) T-lymphocyte mediated immunity in dwarf chickens; (2) there were effects of selection for higher PP in hens on fertilization rate (P < 0.05), hatchability (P < 0.05), rate of healthy chicks (P < 0.05), and level of IgY antibody (P < 0.0001); however, hens' PP had no effects on number of eggs (P > or = 0.05) or egg weight (P > or = 0.05) and cocks' PP had no effect (P > or = 0.05) on any trait mentioned above. The results indicated that phagocytosis of peripheral blood monocytes might be an indicator of humoral immunity in dwarf chickens; furthermore, selection of hens with higher PP was not only beneficial to fertilization rate, but also benefited to hatchability and rate of healthy chicks in that the hens had stronger humoral immunity, which might contribute to maternal antibody in eggs.

Stronger in vitro phagocytosis by monocytes-macrophages is indicative of greater pathogen clearance and antibody levels in vivo.

Monocytes-macrophages are crucial players in specific and nonspecific immune responses to protect organisms from invasion of bacteria or viruses. In this study, monocytes in circulation from 2 lines of Silky and Starbro chickens with different disease resistance were separated and cultured in vitro. After identification with acridine orange (AO), Giemsa staining, and CD14 immunostaining, monocytes-macrophages were used for adherence and phagocytosis test. The overall percentages of adherence of Silky monocytes was 1.5 times greater than that of Starbro (P < 0.01), which were 26.85% +/- 8.24% and 18.34% +/- 8.15%, respectively (mean +/- SD). The monocytes-macrophages phagocytic index, phagocytic product, and percentage of phagocytosis in Silkies were greater than in Star-bros, respectively. The difference of phagocytic index was significant (P < 0.05), that is, 3.70 +/- 1.75 and 1.97 +/- 0.31, respectively (mean +/- SD). Then, 20 Silkies were divided into 2 groups according to phagocytic index: high phagocytic index (HPI) group and low phagocytic index (LPI) group, to study the relationship between phagocytic activity in vitro and pathogen clearance. After being challenged against Salmonella Pullorum C79-13, the Silky birds with HPI produced a 3-fold greater level of specific antibodies compared with those with LPI (P < 0.01), 50.21 +/- 6.67 and 16.85 +/- 4.52, respectively (mean +/- SD). In contrast to LPI birds, HPI birds shed less Salmonella Pullorum bacteria (P < 0.05), that is, 168.98 x 10(8) +/- 294.74 x 10(8) compared to 385.40 x 10(8) +/- 399.94 x 10(8) (mean +/- SD), and the shedding peak of Salmonella Pullorum in the test span appeared 4 d earlier. These results indicated that phagocytosis of monocytes-macrophages had strong effects on antibody titer and bacteria shedding postchallenge, which could be used to predict the disease resistance in animals.

Generation of functionally distinct B lymphocytes from common myeloid progenitors.

Current models of adult haematopoiesis propose that haematopoietic stem cells (HSCs) differentiate into common lymphoid (CLP) and common myeloid (CMP) progenitors and establish an early separation between myeloid and lymphoid lineages. Nevertheless, the developmental potential of CMP-associated B cells suggests the existence of alternate pathways for B lymphopoesis. The aim of this study was to compare the developmental and functional properties of CMP- and CLP-derived B cells. While both populations matured through pro-B cell and transitional B cell intermediates in the bone marrow and spleen, respectively, following transfer into irradiated mice, mature CMP- and CLP-derived B cells exhibit distinct functional responses. Specifically, CMP-derived B cells did not respond to mitogenic stimulation to the same degree as their CLP-derived counterparts and secrete lower levels of IgM and the inflammatory cytokines such as interleukin (IL)-6 and IL-10. Together, these data suggest the existence of multiple pathways for generating functionally distinct B cells from bone marrow precursors.

RAPID COMMUNICATION: Generation of FGF5 knockout sheep via the CRISPR/Cas9 system.

Sheep are an important source of fiber production. Fibroblast growth factor 5 (FGF5) is a dominant inhibitor of length of the anagen phase of the hair cycle. Knockout or silencing of the gene results in a wooly coat in mice, donkeys, dogs, and rabbits. In sheep breeding, wool length is one of the most important wool quality traits. However, traditional breeding cannot accurately and efficiently mediate an advanced genotype into the sheep genome. In this study, we generated 3 knockout sheep via the 1-step clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system. Sequencing analysis confirmed that mutations in the gene existed in all germ lines of 3 founders: besides the intact sequence, 3 kinds of deletions in the gene (including 5, 13, and 33 bp) were detected. The changes in the primary and senior structure of the FGF5 protein due to the 3 deletions in founders suggested that the FGF5 protein was dysfunctional. In addition, the expression level of intact mRNA in heterozygous individuals decreased compared with the wild types ( < 0.01). Functionally, we discovered that wool length in founders was significantly longer than in wild types ( < 0.05). Collectively, the knockout sheep with the longer wool length phenotype will provide an efficient way for fast genetic improvement of sheep breeding and promote the development of wool industry.

[The genetic analysis of amplitude of progesterone secretion in serial blood collection from tail in min sows].

28 adult Min sows (3-7 parities), represented 9 sire families were selected to conduct the continual blood collection from the tail for two hours at the interval of ten minutes between 9:00-11:00 in the morning on the sixth day after the last mating. The content of progesterone was assayed by radio-immunological assay. The amplitudes of the secretion of progesterone were generated by the procedure of HORM fft. exe complied by the author. It is difficult to get a reasonable conclusion from the content analysis, because the content varied widely between different points of blood collection in single sow. After Fourier conversion, the heritability of amplitude for the fifth partial wave was high (h2 = 0.932); middle for the basal, 2nd and 3rd partial waves; lower for others. This supported the idea that prolific sows had more active secretion of progesterone in luteal cells. The coexist of low and high heritability components in amplitude of progesterone secretion showed that the activities of progesterone secretion were influenced by both the genetics and environments. The genetic correlation of basal, 1st, 5th partial waves were negative, the others were positive, with the litter size and litter size alive. It showed that the reproductive performance of Min pigs were not expressed perfectly, as the balance of secretion of progesterone did not reach the best. The genetic correlation of all partial waves with duration of estrus was negative which supported that the progesterone suppresses the estrus. The genetic correlation of 3rd, 5th partial waves were stronger than others, but selection for 5th partial wave would make high responses than the 3rd wave, because the genetic correlation of 3rd partial wave with litter size and litter size alive was positive, therefore, the more appropriats method of selection for progesterone was adopted, the better results would be achieved in the improvement of indirect selection response for litter size.

[Production of transgenic lamb integrated with modified human anti-trypsin gene].

This experiment is to produce the human mAAT(modified anti-trypsin) which cures the emphysema specifically through mammalian galactophore of transgenic goat. 56 goats were selected as donor for superovulation by FSF + LH microinjection in this experiment. The pronucleic embryos were injected with human mAAT gene after fertilization in vivo, and transferred to the donors or receptors directly. The superovulation was better in March and May than in December with the number of ovulation of 19.50, 21.70 and 16.06, and number of fertilized embryos of 4.31, 6.48 and 3.57 per-animal respectively. The pregnant rates were 18.18% and 25% respectively after transferred to donors and receptors with natural estrus. The donors also can be used as the embryo receptor with no remarkable decrease of pregnant rate. 29 lamb were labored. 4 positive transgenic lamb were checked by PCR, PCR-Southern and Southern analysis. The integrated efficiency of foreign DNA was 13.79% with microinjection of high copy number of foreign DNA fragment.

Comparative immunobiology of thymic DC mRNA in autoimmune-prone mice.

New Zealand Black (NZB) mice have multiple defects in both innate and acquired immunity. A fundamental defect, described more than 25 years ago, is premature thymic involution. Subsequent studies have disclosed multiple defects in thymic epithelial cells, and it has been proposed that thymic dendritic cells (DCs) play an important role not only in thymic involution but also in the appearance of immunopathology. However, the number of available thymic DCs makes this population extremely difficult to study. We have taken advantage of our ability to isolate pure populations of thymic DCs and have examined several key mRNA levels of enzymes involved in signal transduction. Our data on NZB mice was compared to that of NZB x NZW F1 (B/WF1), BXSB-Yaa, MRL/lpr, NOD and control mice. Importantly, we demonstrate herein that a common feature in autoimmune-prone mice is an increase of thymic DC c-met mRNA. Indeed, the increase in c-met mRNA levels appeared specific to the thymus and was not noted in the spleen. Additionally, we demonstrate that E-cadherin, a downstream molecule of c-met, is also reduced. Finally, we note that the levels of HGF mRNA are normal in the autoimmune strains examined herein, confirming that the abnormality of c-met mRNA is not due to primary defects in thymic stromal cells. We submit that these results highlight the possibility of a selective defect in thymic DCs which will be a pivotal step in loss of tolerance, and suggest that future studies focus on adoptive cell transfer involving this population.

The role of CD11c(+) hepatic dendritic cells in the induction of innate immune responses.

The role of the liver in the initiation and maintenance of tolerance is a critical immune function that involves multiple lineages of immune cells. Included within these populations are liver dendritic cells (DCs). Although there has been significant work on the phenotypic and functional roles of splenic and bone marrow dendritic cells, as well as their subsets, comparable studies in liver have often been difficult. To address this issue we have isolated, from C57BL/6 mice, relatively pure populations of DCs and compared phenotype and function to the data from spleen using flow cytometry, cell sorter assisted purification and culture, morphology by cytospin and May-Giemsa staining, cell cycle progression, antigen uptake, cytokine production and allo-activation potential. natural killer (NK)1.1(-)CD11c(+) liver DC subsets (conventional DCs, T cell receptor (TcR)beta(-)NK1.1(-)CD11c(+)B220(-) and plasmacytoid DCs, TcRbeta(-)NK1.1(-)CD11c(+)B220(+)) efficiently endocytose dextran and produce significant levels of tumour necrosis factor (TNF)-alpha, interleukin (IL)-6 and IL-12 p40 in response to Toll-like receptor (TLR) ligands, with responses higher than splenic DCs. There is also a differential capability of hepatic DCs to respond to innate signals. Indeed, CD11c(+) hepatic DCs have a greater capacity to respond to innate stimulation but are less capable of inducing CpG activated-allogeneic T cells. These data suggest that hepatic dendritic cells function as a critical bridge between innate and adaptive immunity and are capable of inducing stronger innate responses with a lower capacity for allo-stimulation than splenic dendritic cells. These properties of liver dendritic cells contribute to their unique role in the induction of tolerance.

Successful allogeneic bone marrow transplantation (BMT) by injection of bone marrow cells via portal vein: stromal cells as BMT-facilitating cells.

We examined the importance of the coadministration of bone marrow (BM) stromal cells with BM cells via the portal vein. A significant increase in the number of day-14 colony-forming unit-spleen (CFU-S) was observed in the recipient mice injected with hemopoietic stem cells (HSCs) along with donor BM stromal cells obtained after three to four weeks of culture. Histological examination revealed that hematopoietic colonies composed of both donor hemopoietic cells and stromal cells coexist in the liver of these mice. However, when donor HSCs plus BM stromal cells were administered i.v., neither the stimulatory effects on CFU-S formation nor the hemopoietic colonies in the recipient liver were observed. These findings suggest that the interaction of HSCs with stromal cells in the liver is the first crucial step for successful engraftment of allogeneic HSCs. It is likely that donor stromal cells and HSCs trapped in the liver migrate into the recipient BM and spleen, where they form CFU-BM and CFU-S, respectively.