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1.
Purification of GP-83, a glycoprotein secreted by the human epididymis and conjugated to mature spermatozoa 总被引:4,自引:0,他引:4
Epididymal secretions are critical for mammalian spermatozoa to acquire both forward motility and an ability to recognize and penetrate oocytes. Previous studies identified two glycoproteins, GP-83 and GP-39, which were secreted by the human epididymis and may be related to maturation of sperm function. In this study, GP-83 was purified from human seminal fluid by DEAE-ion exchange, gel filtration chromatography and preparative gel elution. The isoelectric point (pI) of purified GP-83 was 6.57. Monospecific antiserum to GP-83 was induced in male New Zealand rabbits and confirmed on immunoblots. GP-83 was found in fluid, tissue and sperm extracts of corpus and cauda epididymis, but not in the caput. Immunohistochemical localization identified GP-83 in the luminal contents and in the supranuclear region and cell membrane of principal cells of the corpus and cauda epididymis. GP-83 was found on the anterior acrosome in ejaculated spermatozoa, and shifted to the equatorial region after capacitation and the acrosome reaction. 相似文献
2.
Mouse and guinea pig epididymal tissues have been investigated by light and electron microscopic autoradiography after long intervals ranging from 24 h to 5 days postinjection (p.i.) of the glycoprotein precursors, L-fucose-6-3H or D-glucosamine-1-3H. Using modified fixations to enhance glycoprotein preservation in situ, we found intense labelling of luminal contents in at least some of the epididymal segments after all the intervals investigated. At 24 h p.i., the label in guinea pig was associated with spermatozoa during remodelling of the acrosome in segment II, and at 3 days p.i., radioactivity was trapped within sperm head associations ("rouleaux") in segment IV of the epididymis. At this time, similar rouleau labelling extended from segment IV to segment VIII. In mouse, the luminal contents of the cauda epididymis were still intensely labelled at 5 days p.i.; analysis of the electron microscopic autoradiograms showed that relative grain concentration over the spermatozoa was twice that of the epididymal plasma. This concentration was especially elevated in the region of the sperm head. These findings taken together were interpreted as the binding of secreted epididymal glycoproteins to spermatozoa during sperm transit through the epididymis. In contrast to luminal contents, the labelling of the epididymal epithelium was generally lower, except on the clear cells which showed more pronounced labelling than the neighboring principal cells in mouse cauda epididymis at 5 days p.i. This label probably originated from the resorption of luminal glycoproteins. 相似文献
3.
Changes in lipids and membrane anisotropy in human spermatozoa during epididymal maturation 总被引:3,自引:3,他引:3
Previously it was demonstrated that immature and immotile human spermatozoa
from the caput epididymides developed a good progressive motility after
in-vitro stimulation with phosphatidylcholine (PC). In order to define the
role of PC and membrane anisotropy in epididymal maturation and to
determine the exact lipid composition of human spermatozoa during
epididymal maturation, spermatozoa from seven epididymides from patients
who underwent orchiectomy because of prostatic cancer were investigated.
Lipids were determined by high- performance thin-layer chromatography and
gas chromatography. Membrane anisotropy was measured by fluorescence
polarization. The ratio between PC and phosphatidylserine (PS) plus
phosphatidyl ethanolamine (PE) plus sphingomyelin (SM) was significantly
higher in spermatozoa from the cauda compared to those from the caput and
corpus. This was due to an increase of PC and a decrease of the
concentration of PS plus PE plus SM. With regard to fatty acids, those with
saturated chains predominated in caput spermatozoa while the highest
concentration of unsaturated long-chain fatty acids was in cauda
spermatozoa. A lower membrane anisotropy of cauda spermatozoa compared with
caput or corpus spermatozoa was found. In conclusion, during epididymal
maturation human spermatozoa integrate lipids, particularly PC, which is
strongly associated with the induction of progressive motility. A change in
the pattern of fatty acids and a decrease in the cholesterol/phospholipid
molar ratio cause a decrease in membrane anisotropy in cauda spermatozoa.
相似文献
4.
5.
T Inage H Shimokawa Y Teranishi T Iwase Y Toda I Moro 《Archives of histology and cytology》1989,52(3):213-229
Rabbit polyclonal antibodies against bovine amelogenins and enamelins which did not show any cross-reaction were raised, and ultrathin sections of rat incisors were examined by the protein A-gold and ABC methods. The immunoreactivity of amelogenins was found in dense granules in the intercellular spaces between preameloblasts, and later over the fine- and coarse-textured material. The immunoreactivity was present over the cell organelles associated with the secretory pathway, as well as pale and dark lysosomes of the presecretory and secretory ameloblasts. Here the enamel was immunolabeled in the intercrystal spaces. The immunoreactivity in multivesicular bodies was stronger in preameloblasts than in secretory ameloblasts. In the region of second ruffle-ended ameloblasts at the maturation stage, the immunolabeling was intense in the ruffled-border, but in the rough endoplasmic reticulum and Golgi apparatus, the immunolabeling was much weaker than at the secretory stage. The immunolabeling for enamelins showed essentially the same intracellular topographical pattern as that for amelogenins by the secretory stage, but was weaker. The immunoreactivity was found mainly attached to the enamel crystals. Double immunostaining of amelogenins and enamelins revealed that both immunoreactivities were present over the same cell organelles associated with secretion and lysosomal systems. It is suggested that the presecretory and secretory ameloblasts are actively involved in the secretion, degradation and resorption of enamel proteins, and that multivesicular bodies and lysosomes in the cells take part in these processes. Ameloblasts are considered to be related to the synthesis of enamelins. 相似文献
6.
Testicular and epididymal spermatozoa are routinely used with in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) to achieve pregnancies. In addition, excess cryopreserved spermatozoa can be thawed and used for ICSI. However, information on the recovery of epididymal and testicular spermatozoa after freeze-thaw is lacking. This is important to determine the feasibility of using previously cryopreserved aspirated spermatozoa for ICSI. We prospectively compared the viability of fresh and frozen-thawed spermatozoa from the vas deferens, epididymis and testicle by several measures. Testis spermatozoa were obtained from men with non-obstructive azoospermia (n = 5), epididymal spermatozoa from men with obstructive azoospermia (n = 8), and vasal spermatozoa from fertile men by vasal irrigation at vasectomy (n = 5). The viability of fresh spermatozoa was assessed by motility, two vital stains (carboxyfluorescein, 0.08 mg/ml and propidium iodide, 20 mg/ml) and the hypo-osmotic swelling assay (HOS; 100 mmol/l citrate and fructose). After cryopreservation, spermatozoa were thawed and all viability measures repeated. Although fresh vasal spermatozoa were the most motile, testicular spermatozoa exhibited similar, high viability (91 and 86% respectively) by vital stain. Spermatozoa from testis, epididymis and vas deferens survived cryopreservation equally well by vital stain, but not by motility. As a selection measure, the HOS assay identified significantly more viable epididymal and testicular spermatozoa than did motility in both fresh and frozen-thawed populations. It appears feasible to use frozen-thawed extracted spermatozoa for ICSI when motility and a selection measure such as the HOS assay are used. With fresh testis spermatozoa, selection methods may not be necessary prior to ICSI, as cell viability is high. 相似文献
7.
The human cationic antimicrobial protein (hCAP-18) is expressed in the epithelium of human epididymis, is present in seminal plasma at high concentrations, and is attached to spermatozoa 总被引:18,自引:0,他引:18
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Malm J Sørensen O Persson T Frohm-Nilsson M Johansson B Bjartell A Lilja H Ståhle-Bäckdahl M Borregaard N Egesten A 《Infection and immunity》2000,68(7):4297-4302
Innate immunity is important for the integrity of the host against potentially invasive pathogenic microorganisms in the environment. Antibiotic peptides with broad antimicrobial activity are part of the innate immune system. We investigated the presence of the cathelicidin, human cationic antimicrobial protein (hCAP-18), in the male reproductive system. We found strong expression of the hCAP-18 gene by in situ hybridization and hCAP-18 protein, as detected by immunohistochemistry, in the epithelium of the epididymis, but not in the testis. The highest expression in the epididymis was in the caudal part. Western blotting showed a doublet band, the upper part corresponding to the size of hCAP-18 in plasma and neutrophils. Using a specific enzyme-linked immunosorbent assay (ELISA), levels of 86.5 +/- 37.8 microg/ml (mean +/- standard deviation; range, 41.8 to 142.8 microg/ml; n = 10) were detected in seminal plasma from healthy donors, which is 70-fold higher than the level in blood plasma. Flow cytometry and immunocytochemistry revealed the presence of hCAP-18 on spermatozoa. ELISA measurement showed levels of 196 ng/10(6) spermatozoa, corresponding to 6.6 x 10(6) molecules of hCAP-18 per spermatozoon. Our results suggest a key role for hCAP-18 in the antibacterial integrity of the male reproductive system. The attachment of hCAP-18 to spermatozoa may implicate a role for hCAP-18 in conception. 相似文献
8.
S. S. Raitsina N. A. Chuich Z. S. Khlystova A. I. Davydova 《Bulletin of experimental biology and medicine》1977,83(4):548-551
Cells forming rosettes with homologous or heterologous spermatozoa were found in the thymus, spleen, and bone marrow of sexually mature guinea pigs and of 14–30-week-old human fetuses, and also in the peripheral blood of men suffering from sterility. On the development of autoimmune orchitis after measured trauma to the testis in rats or after immunization of guinea pigs with testicular tissue homogenate mixed with Freund's complete adjuvant, cells forming rosettes with spermatozoa were found to appear in the spleen and thymus of the rats, and their number in the lymphoid organs of the guinea pigs increased. These procedures had no effect on the number of cells forming rosettes with sheep red blood cells in the lymphoid organs of rats and guinea pigs. The possible use of this newly discovered ability of human and animal lymphocytes to form spontaneous and immune rosettes with spermatozoa as a means of assessing the degree of differentiation of lymphocytes and of their sensitization to spermatozoal antigens in cases of disturbance of spermatogenesis of autoimmune nature is discussed.Laboratory of Human Embryonic Histogenesis, Institute of Human Morphology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 4, pp. 464–467, April 1977. 相似文献
9.
Andrology: Prevention of osmotic injury to human spermatozoa during addition and removal of glycerol
Gao D.Y.; Liu J.; Liu C.; McGann L.E.; Watson P.F.; Kleinhans F.W.; Mazur P.; Critser E.S.; Critser J.K. 《Human reproduction (Oxford, England)》1995,10(5):1109-1122
Use of a cryoprotective agent is indispensable to prevent injuryto human spermatozoa during the cryopreservation process. However,addition of cryoprotective agents to spermatozoa before coolingand their removal after warming may create severe osmotic stressfor the cells, resulting in injury. The objective of this studywas to test the hypothesis that the degree (or magnitude) ofhuman sperm volume excursion can be used as an independent indicatorto evaluate and predict possible osmotic injury to spermatozoaduring the addition and removal of cryoprotective agents. Glycerolwas used as a model cryoprotective agent in the present study.To test this hypothesis, first the tolerance limits of spermatozoato swelling in hypoosmotic solutions (iso-osmotic medium dilutedwith water) and to shrinkage in hyperosmotic solutions (iso-osmoticmedium with sucrose) were determined. Sperm plasma membraneintegrity was measured by fluorescent staining, and sperm motilitywas assessed by computer-assisted semen analysis before, duringand after the anisosmotic exposure. The results indicate firstlythat motility was much more sensitive to anisosmotic conditionsthan membrane integrity, and secondly that motility was substantiallymore sensitive to hypotonic than to hypertonic conditions. Basedon the experimental data, osmotic injury as a function of spermvolume excursion (swelling or shrinking) was determined. Thesecond step, using these sperm volume excursion limits and previouslymeasured glycerol and water permeability coefficients of humanspermatozoa, was to predict, by computer simulation, the cellosmotic injury caused by different procedures for the additionand removal of glycerol. The predicted sperm injury was confirmedby experiment. Based on this study, an analytical methodologyhas been developed for predicting optimal protocols to reduceosmotic injury associated with the addition and removal of hypertonicconcentrations of glycerol in human spermatozoa. 相似文献
10.
Prolyl oligopeptidase (POP) is an endopeptidase which cleaves short proline-containing neuropeptides, and it is involved in memory and learning. POP also has an intercellular function mediated through the inositol pathway, and has been involved in cell death. POP has been early considered as a housekeeping enzyme, but the recent research indicates that POP expression is regulated across tissues and intracellularly. In the brain, POP is exclusively expressed in neurons and most abundantly in pyramidal neurons of cerebral cortex, in the CA1 field neurons of hippocampus and in cerebellar Purkinje's cells. Intracellularly, POP is mainly present in the cytoplasm and some in intracellular membranes, like rough endoplasmic reticulum and Golgi apparatus. In this paper, we systematically studied the levels of expression of POP along the life of cerebellar granule cells (CGC) in culture and the distribution of POP within different intracellular compartments. We used the tight-binding inhibitor JTP-4819 covalently coupled with fluorescein (FJTP) as a tool to study the changes on expression and localization of POP protein. Our results indicate that POP activity levels are regulated during the life of the neurons. POP was found mainly in cytoplasm and neuronal projections, but at an early developmental phase significant amounts were found also in nuclei. Along the life of the neurons, POP activity fluctuated in 7-day cycles. In young neurons, the cytosolic POP activity was low but increased by maturation so that the activity peak coincided with full differentiation. Over aging, cytoplasmic POP was concentrated around nucleus, but the activity decreased with time. POP was also present in vesicles across the neuron. No major changes were seen in the nuclear or membrane bound POP over aging until activity disappeared upon neuronal death. This is the first time when POP was found in the nuclei of human neuronal cells. 相似文献
11.
Intracellular calcium increase and acrosome reaction in response to progesterone in human spermatozoa are correlated with in-vitro fertilization 总被引:2,自引:1,他引:1
Krausz Csilla; Bonaccorsi Lorella; Luconi Michaela; Fuzzi Beatrice; Criscuoli Luciana; Pellegrini Sandra; Forti Gianni; Baldi Elisabetta 《Human reproduction (Oxford, England)》1995,10(1):120-124
In this study we have investigated responsiveness to progesteronein spermatozoa from a group of unselected male partners of couplesundergoing in-vitro fertilization (IVF). We evaluated progesterone-stimulatedintracellular Ca2+ ([Ca2+]i) and percentage increase in acrosomereaction in the same sperm sample used for oocyte inseminations.[Ca2+]i was measured with a fluorimetric method, while the acrosomereaction was assessed using a fluorescent probe (fluoresceinisothiocyanate-labelled peanut lectin). The average percentage[Ca2+]i as well as the rate of increase in the frequency ofacrosome reaction following progesterone challenge were significantlylower (P < 0.005) in the group of patients with a fertilizationrate <50%. In addition, significant correlations betweenthe fertilization rate and the progesterone-stimulated [Ca2+]iand acrosome reaction increases (r = 0.78 and r = 0.79 respectively)were observed. Furthermore, in cases of fertilization failure,no increase of [Ca2+]i or acrosome reaction was observed inresponse to progesterone with the exception of one case. Ourresults indicate that [Ca2+]i and acrosome reaction increasesin response to progesterone can be of value in the predictionof sperm fertilizing ability. As the two parameters were significantlycorrelated to each other (r = 0.86), the two assays have similarIVF predictive value and might be used interchangeably as adiagnostic tool in the assignment of male patients to the differentkinds of assisted fertilization techniques. 相似文献
12.
Lundell AC Andersson K Josefsson E Steinkasserer A Rudin A 《Infection and immunity》2007,75(8):4097-4104
CD14 is expressed on the cell surface of various antigen-presenting cells, and CD83 is a maturation marker for dendritic cells (DC). CD14 and CD83 are also present as soluble proteins, and both have immunoregulatory functions. We examined whether neonatal cord blood monocytes or DC released soluble CD14 (sCD14) or sCD83 when exposed to the commensal intestinal bacteria Clostridium perfringens, Staphylococcus aureus, Lactobacillus rhamnosus, Escherichia coli, and Bacteroides fragilis. We found that the gram-positive bacteria C. perfringens and S. aureus, but not gram-negative bacteria, induced the release of sCD14 from monocytes. DC, on the other hand, released sCD14 in response to both gram-positive and gram-negative bacteria. Moreover, the expression of the virulence factor staphylococcal protein A seemed to be important for S. aureus-induced sCD14 production from both monocytes and DC. Soluble CD83 was released from DC, but not from monocytes, when exposed to both gram-positive and gram-negative bacteria. Finally, to investigate whether sCD14 or sCD83 could modulate neonatal allergen-induced T-cell differentiation, DC were exposed to birch allergen alone or in the presence of sCD14 or sCD83 and then cocultured with autologous T cells. We demonstrate that sCD14 and sCD83 inhibited the birch allergen-induced Th2 differentiation by suppressing interleukin 13 production. Together, these results suggest that the commensal intestinal flora may be an important stimulus for the developing immune system by inducing the immunoregulatory proteins sCD14 and sCD83, which may be involved in preventing T-cell sensitization to allergens in infants. 相似文献
13.
Intracellular Ca(2+) oscillations at fertilization are responsible for triggering egg activation. The aim of this study was to examine the effect of the age of the oocyte donor and in-vitro maturation on the generation of Ca(2+) transients at fertilization. The results show that <10% of in-vivo and in-vitro matured oocytes from 19-day old mice develop to the blastocyst stage in vitro. In contrast, 43% of in-vivo and 25% of in-vitro matured oocytes from 24-day old mice developed to the blastocyst stage. In parallel experiments, intracellular Ca(2+) was monitored at fertilization. Oocytes from 19-day old mice generate significantly fewer transients than oocytes from 24-day old mice. In-vitro maturation significantly decreased the ability of oocytes from 19-day old mice but not 24-day old mice to generate Ca(2+) transients in response to spermatozoa. Furthermore, we investigated the effect of oocyte maturation on Ca(2+) signalling. Immature oocytes generated fewer Ca(2+) oscillations and ceased oscillating earlier than mature oocytes. These studies suggest that the ability to generate Ca(2+) transients in response to spermatozoa increases in the final stages of oocyte development and during oocyte maturation. This may contribute to the acquisition of developmental competence in the final stages of oogenesis. 相似文献
14.
Wróblewski S Berenson M Kopecková P Kopecek J 《Journal of biomedical materials research》2000,51(3):329-342
Lectins are proteins that bind glycoproteins; binding patterns are altered with changes in glycoprotein expression accompanying maturation or disease. Binding of two lectins, wheat germ agglutinin (WGA) and peanut agglutinin (PNA), in human and rodent colon were previously examined. Normal tissue showed intense WGA binding; PNA binding was minimal. Diseased tissues showed increased PNA binding. We hypothesized that N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer-lectin-drug conjugates could deliver therapeutic agents to diseased tissues by targeting colonic glycoproteins. We examined biorecognition of free and HPMA copolymer-conjugated WGA and PNA and anti-Thomsen-Friedenreich (TF) antigen antibody binding in normal neonatal, adult, and diseased rodent tissues, human specimens of inflammation, and Barrett's esophagus. Neonatal WGA binding was comparable to the adult, with additional luminal columnar cell binding. PNA binding was more prevalent; luminal columnar cell binding existed during the first 2.5 weeks of life. WGA binding was strong in both normal and diseased adult tissues; a slight decrease was noted in disease. PNA binding was minimal in normal tissues; increases were seen in disease. Anti-TF antigen antibody studies showed that PNA did not bind to the antigen. The results suggest that HPMA copolymer-lectin-drug conjugates may provide site-specific treatment of conditions such as colitis and Barrett's esophagus. 相似文献
15.
Several techniques have been used for selecting motile spermatozoa including Percoll and albumin gradients, swim-up, and glass wool filtration. A high yield of motile spermatozoa as well as an enhancement of motility are the most desirable features of a practical method. An equally important consideration is whether or not these techniques select functionally normal spermatozoa. In this study we have compared two methods for separation of motile cells, swim-up and Percoll gradient. Normal semen samples from 12 different men were used in this study. Each sample was simultaneously processed by swim-up and Percoll gradient using modified Tyrode's medium. After the sperm concentration was adjusted to 1 x 10(7) spermatozoa/ml, the suspensions were incubated at 37 degrees C, 5% CO2 in air. In each suspension the percentage of sperm recovery, percentage of motile spermatozoa, percentage of acrosome reacted spermatozoa (either spontaneously or stimulated with human follicular fluid), percentage of zona-free hamster oocytes penetrated, and number of spermatozoa bound to the human zona pellucida were determined. The results obtained indicated that the percentage of sperm recovery was higher with the Percoll gradient than with the swim-up procedure (P less than 0.001). However, no significant differences were found between these two sperm populations in the percentage of motile cells, in the percentage of acrosome reacted spermatozoa, and in the percentage of zona-free hamster oocytes penetrated. In addition, the number of spermatozoa bound per zona pellucida was similar for spermatozoa selected by Percoll or swim-up. We conclude that there were no functional differences between the spermatozoa selected by either method. 相似文献
16.
Cyril Le Nouën Stéphanie Losq Thomas McCarty Kevin L. Holmes Ronald L. Rabin Ursula J. Buchholz 《Virology》2009,385(1):169-182
Human respiratory syncytial virus (HRSV), human metapneumovirus (HMPV), and human parainfluenza virus type 3 (HPIV3) are common, important respiratory pathogens, but HRSV has a substantially greater impact with regard to acute disease, long-term effects on airway function, and frequency of re-infection. It has been reported to strongly interfere with the functioning of dendritic cells (DC). We compared HRSV to HMPV and HPIV3 with regard to their effects on human monocyte-derived immature DC (IDC). Side-by-side analysis distinguished between common effects versus those specific to individual viruses. The use of GFP-expressing viruses yielded clear identification of robustly infected cells and provided the means to distinguish between direct effects of robust viral gene expression versus bystander effects. All three viruses infected inefficiently based on GFP expression, with considerable donor-to donor-variability. The GFP-negative cells exhibited low, abortive levels of viral RNA synthesis. The three viruses induced low-to-moderate levels of DC maturation and cytokine/chemokine responses, increasing slightly in the order HRSV, HMPV, and HPIV3. Infection at the individual cell level was relatively benign, such that in general GFP-positive cells were neither more nor less able to mature compared to GFP-negative bystanders, and cells were responsive to a secondary treatment with lipopolysaccharide, indicating that the ability to mature was not impaired. However, there was a single exception, namely that HPIV3 down-regulated CD38 expression at the RNA level. Maturation by these viruses was anti-apoptotic. Inefficient infection of IDC and sub-optimal maturation might result in reduced immune responses, but these effects would be common to all three viruses rather than specific to HRSV. 相似文献
17.
Summary Human antibody responses to human cytomegalovirus (HCMV) envelope glycoproteins were analyzed using immunoaffinity purified glycoproteins and Western blotting. Two families of disulfide linked glycoprotein complexes, designated gC-I and gC-II, were isolated. These complexes were reduced and their individual glycoproteins separated by polyacrylamide gel electrophoresis prior to electroblotting. The reactivity of adult convalescent sera with individual glycoproteins was compared to that of sera from congenitally infected infants. All sera tested reacted with a 52,000 molecular weight glycoprotein from these complexes, but only 75% reacted with a 93,000 to 130,000 molecular weight glycoprotein from gC-I complexes. Most adult convalescent sera reacted with glycoproteins from gC-II complexes. However, 14 of 16 infant sera did not have high enough levels of gC-II antibodies to give a positive reaction with Western blotting. A longitudinal study was done with several infants and their mothers. These studies indicated a failure by the infants and their mothers to develop detectable levels of gC-II antibodies months to years after the initial infection or after repeated stimulation with HCMV due to persistent infection. The inability of these infants to develop significant levels of gC-II antibodies was not due to an inability to respond to viral glycoproteins since they had antibodies to gC-I glycoproteins. We also determined that the strains of HCMV infecting some of these infants expressed gC-II glycoproteins. Thus, the lack of response by these infants was not due to lack of expression of gC-II glycoproteins by their infecting strain. 相似文献
18.
Matthijs van de Rijn Peter G. Lerch Ben R. Bronstein Robert W. Knowles Atul K. Bhan Cox Terhorst 《Human immunology》1984,9(4):201-210
Monoclonal antibodies anti-T6 and anti-M241 define unique cell populations within different lineages: cortical thymocytes and dendritic cells in the skin. T6 positive cutaneous dendritic cells are located predominantly in the epidermis and belong to the Langerhans/indeterminate lineage, whereas, most of the M241 positive cells are located in the perivascular regions of the dermis. Biochemical analysis of thymocytes and cutaneous dendritic cells was performed in order to determine weather the reactivity of these antibodies with these cell types is due to sharing of antigenic determinants by two unrelated proteins, or whether similar proteins are present on cells of different lineages. Our results indicate that T6 antigens are borne by the same glycoprotein (49K) on cortical thymocytes and Langerhans/indeterminate cells. Similarly, M241 antigens isolated from thymocytes and cutaneous dendritic cells are found on the same glycoprotein (43K). 相似文献
19.
Maturation of dendritic cells (DCs) by TLR ligands induces expression of IFN-beta and autocrine activation of IFN-inducible Stat1-dependent genes important for DC function. In this study, we analyzed the regulation of STAT signaling during maturation of human DCs by TNF-alpha and PGE2, which induced maturation of human DCs comparably with LPS but did not induce detectable IFN-beta production or Stat1 tyrosine phosphorylation. Consistent with these results, TNF-alpha and PGE2 did not induce Stat1 DNA binding to a standard Stat1-binding oligonucleotide. Instead, TNF-alpha and PGE2 increased Stat1 serine phosphorylation and Stat4 tyrosine phosphorylation and activated expression of the NF-kappaB and Stat1 target gene IFN regulatory factor 1 (IRF1), which contributes to IFN responses. TNF-alpha and PGE2 induced a complex that bound an oligonucleotide derived from the IRF1 promoter that contains a STAT-binding sequence embedded in a larger palindromic sequence, and this complex was recognized by Stat1 antibodies. These results suggest that TNF-alpha and PGE2 activate STAT-mediated components of human DC maturation by alternative pathways to the IFN-beta-mediated autocrine loop used by TLRs. 相似文献
20.
Molluscum contagiosum virus (MCV) is a human poxvirus that produces small, benign skin tumors primarily in young children and encodes proteins that modulate the host immune response. The MC51L, MC53L, and MC54L open reading frames of MCV have significant amino acid sequence similarities and related proteins are encoded by other poxviruses. These three MCV genes were individually expressed in mammalian cells as glycosylated and secreted proteins. A database search detected partial sequences of homologous human and mouse cDNAs; determination of the complete sequences confirmed the homology and indicated potential signal peptides and N-glycosylation sites as well as a pattern of cysteines that are conserved in the viral proteins. The human gene, which was mapped to chromosome 11q13, was highly expressed in spleen and lymph nodes, suggesting an immune modulatory role. The latter finding is consistent with a recent report (Novick et al., 1999, Immunity 10, 127-136) that the human protein binds IL-18, suggesting an anti-inflammatory role for the poxvirus homologs. 相似文献