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1.
The proinflammatory cytokine, interleukin-1, plays a prominent role in the inflammatory reactions that characterize numerous diseases. In this study, we examined the gene expression for bovine IL-1 ligands and receptors by bovine peripheral blood mononuclear cells (MNCs) and neutrophils (PMNs) in response to E. coli lipopolysaccharide (LPS) in vitro. Gene expression of mRNA for IL-1alpha, IL-1beta, IL-1 receptor antagonist (IL-1ra), type 1 IL-1 receptor, type 2 IL-1 receptor, and IL-1 beta converting enzyme (ICE), were measured by a semi-quantitative RT-PCR technique. LPS had little effect on type 1 IL-1R expression in MNC, whereas, it strongly up-regulated type 1 IL-1R expression in PMNs. Co-incubation of PMNs with LPS and bovine recombinant IL-1beta had little additional effect on type 1 IL-1R expression. Incubation of MNCs with LPS resulted in up-regulation of IL-1beta, IL-1ra, and type 2 IL-1R, no change in IL-1alpha, and a decrease in ICE gene expression. Incubation of PMNs with LPS up-regulated IL-1beta gene expression, whereas, IL-1alpha, IL-1ra, type 2 IL-1R and ICE were unchanged. This study provides evidence for differential regulation of gene products of the bovine IL-1 family by peripheral blood mononuclear cells (MNC) and neutrophils (PMNs) in response to E. coli LPS.  相似文献   

2.
ACTH is the primary regulator of adrenal function during acute stress. However, during chronic inflammatory stress additional factors play a major role in the regulation of adrenal secretion. Many cytokines circulate in the blood and are synthesized and released from adrenal tissue. Furthermore, these peptides modify adrenal function. Recently, interleukin-4 (IL-4) was demonstrated to be released from a human adrenal tumor cell line. Therefore, we hypothesized that normal bovine adrenocortical cells could express IL-4 and that this cytokine may modify adrenal function. We determined that IL-4 and IL-4 receptors (IL-4R) are expressed in the bovine adrenal cortex whereas the expression of IL-4 and IL-4R in the adrenal medulla was not apparent. Exposure of dispersed bovine adrenocortical cells isolated from the zona fasciculate to IL-4 did not modify basal release of cortisol. However, the ACTH-stimulated release of cortisol from the bovine adrenal cells was augmented by IL-4. IL-4 exposure had no affect on adrenal androgen release from bovine zona reticularis cells, but IL-4 inhibited the ACTH-stimulated release of adrenal androgens from these cells. The effects of IL-4 on ACTH-stimulated cortisol and adrenal androgen release were dependent upon the IL-4 incubation interval and the IL-4 concentration. Because communication between the immune and endocrine systems is important in inflammatory conditions, IL-4 may play a role in coordinating the adrenal response to inflammatory stress.  相似文献   

3.
Respiratory infection of cattle with bovine herpesvirus type 1 (BHV-1) predisposes cattle to secondary pneumonia with Mannheimia haemolytica as part of the bovine respiratory disease complex (BRD). One cell type that has received limited investigation for its role in the inflammation that accompanies BRD is the respiratory epithelial cell. In the present study we investigated mechanisms by which BHV-1 infection of respiratory epithelial cells contributes to the recruitment and activation of bovine polymorphonuclear neutrophils (PMNs) in vitro. Primary cultures of bovine bronchial epithelial (BBE) cells were infected with BHV-1 and assessed for cytokine expression by real-time PCR. We found that BHV-1 infection elicits a rapid IL-1, IL-8 and TNF-α mRNA response by BBE cells. Bovine PMNs exhibited greater adherence to BHV-1 infected BBE cells than uninfected cells. The increased adherence was significantly reduced by the addition of an anti-IL-1β antibody or human soluble TNF-α receptor (sTNF-αR). Pre-incubation of bovine PMNs with conditioned media from BHV-1 infected BBE cells increased PMN migration, which was inhibited by addition of an anti-IL-1β antibody, sTNF-αR, or an IL-8 peptide inhibitor. Conditioned media from BHV-1 infected BBE cells activated bovine PMNs in vitro as demonstrated by PMN shape change, production of reactive oxygen species and degranulation. PMNs also exhibited increased LFA-1 expression and susceptibility to M. haemolytica LKT following incubation with BHV-1 infected BBE cell conditioned media. Our results suggest that BHV-1 infection of BBE cells triggers cytokine expression that contributes to the recruitment and activation of neutrophils, and amplifies the detrimental effects of M. haemolytica LKT.  相似文献   

4.
cDNA generated from lipopolysaccharide-stimulated equine peripheral blood mononuclear cells was used to amplify and clone type I and type II equine interleukin-1 receptors (IL-1RI and IL-1RII) using primers derived from semi-conserved regions between human and mouse IL-1RI and IL-1RII sequences, respectively. 5' and 3' terminal sequences of equine IL-1RI and IL-1RII were amplified by 5' and 3' rapid amplification of cDNA ends. The deduced amino acid sequence of equine IL-1RI demonstrated 77, 64 and 63% similarity with human, mouse and rat sequences, respectively. The predicted amino acid sequence of equine IL-1RII demonstrated 70, 60 and 58% similarity with human, mouse and rat sequences, respectively. Recombinant equine soluble IL-1RI and IL-1RII produced in insect cells bound recombinant equine IL-1alpha and IL-1beta. Furthermore, both receptors suppressed the growth inhibitory activities of equine IL-1alpha and IL-1beta toward A375 cells in a dose-dependent manner, indicating that the present equine IL-1RI and IL-1RII cDNA encodes biologically active proteins.  相似文献   

5.
Interleukin (IL)-8 and its receptors, CXCR1 and CXCR2, are key regulators of inflammation. However, knowledge of these receptors at the genomic level is limiting or absent in cattle. Therefore, our objective was to identify bovine orthologs of human CXCR1 and CXCR2. Alignment of bovine CXCR2 reference mRNA to the bovine genome revealed two regions of similarity on BTA2 approximately 20 kb apart and on opposite strands. Comparison with the human genome suggested the more centromeric region to be CXCR2 and the more telomeric region to be CXCR1 which contradicts the current annotation of the bovine CXCR2 reference mRNA. This observation was verified by sequencing RT-PCR products of specific regions within each predicted IL-8 receptor and comparing with human sequences using ClustalW. Further examination of coding and non-coding regions within the IL-8 receptor genome complex revealed that both bovine and canine CXCR1 and CXCR2 genes had more conserved sequences in common with the human genes than either mouse or rat, and may offer more suitable animal models for certain applications. This molecular information provides a stepping stone for greater understanding of the role each IL-8 receptor plays in inflammation and will enhance our ability to develop strategies against inflammatory based diseases.  相似文献   

6.
Bovine interleukin 2: regulatory mechanisms   总被引：1，自引：0，他引：1  
A cDNA clone of the bovine interleukin-2 (IL-2) gene has been isolated and demonstrated to produce a functional bovine IL-2 protein when transfected into either CV-1 or COS-1 monkey cells. Homology comparisons of both the nucleotide and predicted amino acid sequences of bovine IL-2 with those of the human and mouse show extensive regions of sequence conservation between the species. The amino acid sequence of the mature bovine IL-2 protein shares about 60-63% homology with those of the human and mouse, but the 3' untranslated regions of the human and mouse gene share as much, if not greater, sequence homology with the 3' untranslated regions of the human and mouse genes. In particular, a tandemly repeated sequence (TATT), n, found in the 3' untranslated tail of the bovine IL-2 clone is also found in the 3' untranslated region of a large group of cytokine genes and other inducible genes of the lymphoid and immune response systems. This sequence may serve a specific regulatory function in the immune system.  相似文献   

7.
The three members of the T1R class of taste-specific G protein-coupled receptors have been proven to function in combination with heterodimeric sweet and umami taste receptors in many mammals that affect food intake. This may in turn affect growth traits of livestock. We performed a comprehensive evaluation of single-nucleotide polymorphisms (SNPs) in the bovine TAS1R gene family, which encodes receptors for umami and sweet tastes. Complete DNA sequences of TAS1R1-, TAS1R2-, and TAS1R3-coding regions, obtained from 436 unrelated female cattle, representing three breeds (Qinchuan, Jiaxian Red, Luxi), revealed substantial coding and noncoding diversity. A total of nine SNPs in the TAS1R1 gene were identified, among which seven SNPs were in the coding region, and two SNPs were in the introns. All five SNPs in the TAS1R2 gene and all three SNPs in the TAS1R3 gene were identified in the coding region. Four SNPs (TAS1R1 g.5081C>T, TAS1R1 g.5110C>A, TAS1R2 g.288A>G, TAS1R2 g.2552T>C) were significantly associated with body height of Qinchuan cattle (P<0.05). The heterozygous genotypes of the four SNPs showed a molecular heterosis on cattle heights at hip cross and sacra. The individuals with different genotypic combinations of the four SNPs had significant association with heights at hip cross and sacra (P<0.05).  相似文献   

8.
Bovine viral diarrhea (BVD) virus inhibited phytohemagglutinin (PHA)-stimulated bovine peripheral blood mononuclear cell (PBMC) proliferation and bovine interleukin-2 (IL-2) production. In the controls, the heat-inactivated BVD virus was not capable of suppressing the PHA-stimulated PBMC proliferation. Presence of exogenous cytokines, such as purified human IL-2, recombinant bovine interleukin-1 (rbovIL-1), recombinant bovine IL-2, and recombinant human IL-6 failed to reverse the BVD virus-induced immunosuppression. Also, we found that the BVD virus inhibited PHA and IL-2 induced proliferation of bovine PBMC in the early and late stages of activation. In summary, our data suggest that BVD virus induced immunosuppression was not due to destruction of the PBMC but may be inhibiting one or more of the important intracellular enzymes that may regulate PBMC proliferation.  相似文献   

9.
The interaction of bovine cells with lipopolysaccharide (LPS) was explored using human embryo kidney (HEK) 293 cell line stably transduced with bovine toll-like receptor-4 (TLR4) alone or in combination with bovine MD-2. These lines and mock-transduced HEK293 cells were tested by flow cytometry for LPS-fluorescein isothiocyanate (LPS-FITC) binding, nuclear factor kappa B (NFkappaB) activation, interleukin-8 (IL-8) production and interferon-beta mRNA expression/interferon (IFN) type I production. Whereas bovine TLR4 was sufficient to promote binding of high concentrations of LPS-FITC, both bovine TLR4 and MD-2 were required for activation by LPS, as assessed by NFkappaB activation and IL-8 production. Induction of IFN bioactivity was not observed in doubly transduced HEK293 cells, and no evidence for IFN-beta mRNA induction in response to LPS was obtained, although cells responded by IFN-beta mRNA expression to stimulation by Sendai virus and poly-inosinic acid-poly-cytidylic acid (poly(I:C)). Cells stably transduced with both bovine TLR4 and bovine MD-2 responded to LPS by IL-8 production, in decreasing order, in the presence of fetal bovine serum (FCS), of human serum, and of human serum albumin (HSA). The reduced activity in the presence of HSA could be restored by the addition of soluble CD14 (sCD14) but not of LPS binding protein (LBP). This is in contrast to macrophages which show a superior response to LPS in the presence of HSA when compared with macrophages stimulated by LPS in the presence of FCS. This suggests that macrophages but not HEK293 cells express factors rendering LPS stimulation serum-independent. Stably double-transduced cells reacted, in decreasing order, to LPS from Rhodobacter sphaeroides, to LPS from Escherichia coli, to synthetic lipd-IVa (compound 406), to diphosphoryl-lipid-A (S. minnesota) and to monophosphoryl-lipid-A (S. minnesota). They failed to react to the murine MD-2/TLR4 ligand taxol. This resembles the reactivity of bovine macrophages with regard to sensitivity (ED(50)) and order of potency but is distinct from the reactivity pattern of other species. This formally establishes that in order to react to LPS, cattle cells require serum factors (e.g. sCD14) and cell-expressed factors such as MD-2 and TLR4. The cell lines described are the first of a series expressing defined pattern recognition receptors (PRR) of bovine origin. They will be useful in the study of the interaction of the bovine TLR4-MD-2 complex and Gram-negative bovine pathogens, e.g. the agents causing Gram-negative bovine mastitis.  相似文献   

10.
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11.
IL-4 and IL-13 share a wide range of activities on monocytes, epithelial cells and B cells and thus play an important role in host defense. Many of these activities are not conserved among species as human, but not murine, B cells are thought to be responsive to IL-13. We previously demonstrated that human IL-13 is highly conserved at the nucleic acid level with a candidate bovine IL-13 cDNA homologue. Moreover, recombinant human IL-13 stimulates Ig secretion by appropriately activated bovine B cells. These studies have been extended to examining Ig class switching at both the protein and mRNA levels in addition to examining other markers of cellular activation. Our results suggest that IL-13 influences B cell differentiation by enhancing IgM, IgG1, and IgE production. IL-13 stimulation alone increases MHC class II expression and progression through cell cycle, although at lower levels in comparison to rboIL-4. The biology of the receptors for IL-4 and IL-13 is complex and raises several key questions with regard to IL-4-dependent and -independent mechanisms of host immunomodulation. Recent studies suggest that at least four chains are involved. These include the p140 IL-4 binding chain (IL-4Ralpha), the common gamma chain (gammac chain), IL-13 receptor alpha- chain (IL-13Ralpha-1) and the IL-13 receptor alpha-2 chain (IL-13Ralpha-2). We have recently cloned cDNAs for the bovine homologues of the IL-13Ralpha-1 and IL-4Ralpha chains and evaluated mRNA expression for a variety of cell types following stimulation. The expression patterns and their implications for receptor chain utilization in signaling via these key TH2 signature cytokines will be discussed.  相似文献   

12.
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13.
1-(5-Isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride (H-7), a potent and selective inhibitor of protein kinase C (PKC), inhibited PHA-stimulated bovine peripheral blood mononuclear cell (PBMC) proliferation, interleukin-2 (IL-2) production, and cytosolic PKC activity without affecting the cell viabilities. Presence of exogenous cytokines, such as purified human IL-2 or recombinant bovine IL-2 (rbovIL-2), reversed the H-7 inhibitory effects on PHA-stimulated PBMC proliferation. We conclude that the PKC enzyme plays an important role as a second messenger in bovine PBMC proliferation in the early stages of cell activation.  相似文献   

14.
The bovine haemal nodes are lymphatic organs located in the haemal circulation. Their parenchyma is represented by plasma cells, macrophages and B and T lymphocytes. The T helper type 2 (Th2) CD4 lymphocyte can be found within the T lymphocytes. The activated Th2 CD4 lymphocyte produces interleukin-4 (IL-4), a peptidic hormone involved in the acute-phase immune response. This interleukin can promote either B-lymphocyte differentiation and T-lymphocyte proliferation or it can promote the type of immunoglobulin that can be liberated. Our results have shown, by immunostaining with anti-IL-4, not only the presence and localization of these lymphocytes in bovine haemal nodes but also the participation of polymorphonuclear cells (neutrophils) in the storage of IL-4. These results give value to the humoral and cellular immunological importance of haemal nodes in bovines and they can serve as a contribution to determine the cross-reactivity of bovine IL-4 with the human anti-serum used in this work.  相似文献   

15.
The objectives of this study were to investigate if insulin receptors (IR) and insulin-like growth factor-1 receptors (IGF-1R) could be detected on bovine leukocytes using fluorescence antibodies and flowcytometry, and use this method to investigate whether the amount of receptors differed among heifers at different ages. Twenty Danish Holstein heifers in the following three age groups were included in the investigation: (1) heifers aged 25-45 days (n = 8), (2) heifers aged 185-205 days (n = 7), and (3) heifers aged 780-900 days (approximately one month prepartum; n = 5). Antibodies against human IR and IGF-1R were used for indirect immunofluorescence staining of cells, and were found to cross-react with the bovine receptors. IR were found on lymphocytes, monocytes, and neutrophils in all animals, while IGF-1R were found only on monocytes and neutrophils. The percentage of IR+ lymphocytes was almost doubled from stage 1 to 3 (34% versus 57%) and IR expression on lymphocytes and monocytes increased significantly (P<0.01) from 6.80 and 13.60 to 8.24 and 17.50 in heifers aged 185-205 days and heifers aged 780-900 days, respectively. No differences were observed for neutrophil IR or IGF-1R expression. In conclusion, surface IR and IGF-1R can be detected on bovine leukocytes by flowcytometry. Interestingly, the highest numbers of IR were found in heifers one month prepartum. The regulation of IR and IGF-1R expression on bovine leukocytes, and their role in host immunity, needs further investigation.  相似文献   

16.
Interleukin-2 and interleukin-12 have been used independently to successfully treat the induced and the spontaneous tumours in animals. This trial was done to determine if a combination of IL-2 and IL-12 in the treatment of spontaneous bovine ocular squamous cell carcinomas (BOSCC) would be more successful than IL-2 or IL-12 therapy by themselves. For this trial, we selected 25 BOSCC tumours seen on Holstein Fresian cows in Beatrice, Zimbabwe. The cows were randomly assigned to a treatment group of 5 days of IL-2 (200,000 U/day), 5 days of IL-12 (0.5 microg/day) or 5 days of IL-2 (200,000 U/day) and IL-12 (0.5 microg/day). At 20 months after treatment, the IL-2 therapy group had 63% complete regressions; the combination group had 38% complete regressions, which were significantly higher than the IL-12 group, which had 0% complete regressions at 20 months, despite having 29% complete regressions at 6 months. These results show that IL-2 therapy by itself and in combination with IL-12 is more successful than IL-12 by itself. However, combination therapy does not improve the outcome in comparison to IL-2 as a single therapy. It also proves that IL-2 is consistently successful in the therapy of BOSCC with over 60% complete regression, which corresponds to a number of other studies we have done on IL-2 therapy of BOSCC [Rutten, V.P.M.G., Klein, W.R., De Jong, W.A., Misdorp, W., Den Otter, W., Steerenberg, P.A., De Jong, W.H., Ruitenberg, E.J., 1989. Local interleukin-2 therapy in bovine ocular squamous cell carcinoma. A pilot study. Cancer Immunol. Immunother. 30, 165--169; Stewart, R.J.E., Hill, F.W.G., Masztalerz, A., Jacobs, J.J.L., Koten, J.W., Den Otter, W., 2003. Local low dose interleukin-2 therapy of bovine ocular squamous cell carcinomas in cattle in Zimbabwe, submitted for publication; Den Otter, W., Hill, F.W.G., Klein, W.R., Koten, J.W., Steerenberg, P.A., De Mulder, P.H.M., Rutten, V.P.M.G., Ruitenberg, E.J., 1993. Low doses of interleukin-2 can cure large bovine ocular squamous cell carcinoma. Anticancer Res. 13, 2453-2455; Den Otter, W., Hill, F.W.G., Klein, W.R., Koten, J.W., Steerenberg, P.A., De Mulder, P.H., Rhode, C., Stewart, R., Faber, J.A., Ruitenberg, E.J., 1995. Therapy of bovine ocular squamous cell carcinoma with local doses of interleukin-2: 67% complete regressions after 20 months of follow-up. Cancer Immunol. Immunother. 41, 10-14].  相似文献   

17.
This study describes a method for detecting canine interleukin-2 receptors (IL-2R) by flow cytometry, using human recombinant IL-2 labeled with phycoerythrin (IL-2-PE). Peripheral blood mononuclear cells from four normal dogs were washed, incubated with IL-2-PE, and then washed to remove any unbound IL-2-PE. Flow cytometric analysis of the cells was performed with a 488 nm argon laser while gating on lymphocytes. Cells expressing the IL-2R were identified by their fluorescence as compared to cells stained with an anti-mouse immunoglobulin-G conjugated to phycoerythrin. The average percentage of resting cells expressing the IL-2R was found to be 21%. The addition of unlabeled human recombinant IL-2 to Day 3 phytohemagglutinin (PHA)-stimulated cells reduced the fluorescence intensity four-fold, thereby demonstrating the specificity of IL-2-PE for canine IL-2R. Following stimulation with optimal concentrations of PHA, the percentage of cells expressing the IL-2R increased daily and reached a maximum on Day 3 (76.4%). IL-2R density, as measured by mean fluorescence intensity, also increased and reached maximal levels on Days 2-3 (twenty-fold greater than resting cells). The binding, inhibition, and kinetic experiments provide evidence that human recombinant IL-2-PE is a useful tool for studying canine IL-2R expression. Thus, a one-step direct method for the flow cytometric detection and quantification of the canine IL-2R is now available.  相似文献   

18.
Cloning,expression, and tissue distribution of bovine interleukin-21   总被引：3，自引：0，他引：3  
Bovine interleukin-21 (IL-21) cDNA was cloned and sequenced from bovine peripheral blood lymphocytes (PBLs) stimulated with 10 microg/ml concanavalin A (ConA), 10 microg/ml phytohemagglutinin (PHA), and 50 ng/ml phorbol 12-myristate 13-acetate (PMA) for 48 h. The open reading frame of the bovine IL-21 cDNA is 459 bp in length and encodes 152 amino acids. The predicted amino acid sequence is 78.2 and 58.5% homologous to the human and murine IL-21 amino acid sequences, respectively. Recombinant bovine IL-21 was expressed by a baculovirus expression system. The bovine IL-21 was processed to the mature form in insect cells and secreted to the supernatant confirmed by N-terminal amino acid sequencing. The recombinant bovine mature IL-21 induced the proliferation of human IL-2-dependent cells, ILT-MAT. The mRNA expression for bovine IL-21 was observed in the spleen, but not in the brain, heart, lung, liver, and kidney. The bovine IL-21 identified in this study may provide new methods for the enhancement of innate immunity in cows.  相似文献   

19.
Mannheimia haemolytica, one of the agents associated with bovine respiratory disease complex, can cause severe lung pathology including the leakage of vascular products into the airways and alveoli. Previous work by this laboratory has demonstrated that bovine lung endothelial and epithelial cells undergo dramatic permeability increases when exposed to adenosine-5'-triphosphate (ATP). Therefore, we wanted to determine if ATP levels were elevated in bronchoalveolar lavage (BAL) samples from calves experimentally infected with M. haemolytica. In addition, cultured bovine pulmonary epithelial (BPE) cells were stimulated with heat-killed and live M. haemolytica bacteria, lipopolysaccharide (LPS), lipoteichoic acid (LTA), interleukin-1 (IL-1), and zymosan activated plasma (ZAP) to determine whether they might release extracellular ATP during in vitro infection. Calves experimentally exposed to M. haemolytica had an approximately 2-fold higher level of ATP in their BAL samples compared to control. BPE cells exposed to increasing numbers of heat-killed or live M. haemolytica had significantly increased levels of ATP release as compared to time-matched controls. Finally, BPE cells treated with several concentrations of LPS and IL-1 had increases in ATP release as compared to time-matched controls. This increase appeared to be a result of active ATP secretion by the cells, as cell viability was similar between treated and non-treated cells. Neither ZAP nor LTA induced any ATP release by the cells. In conclusion, ATP levels are elevated in lung secretions from calves infected with M. haemolytica. In addition, lung epithelial cells can actively release ATP when exposed to heat-killed or live M. haemolytica, LPS or IL-1.  相似文献   

20.
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