PubMed Health⌕ Search

Biomedical subjects

C S Chen

Publications and source records attributed to C S Chen.

At least 73 records · Page 4Linked to original sources
PubMed1999

A novel acidotropic pH indicator and its potential application in labeling acidic organelles of live cells.

BACKGROUND: Ratio imaging has received intensive attention in the past few decades. The growing potential of ratio imaging is significantly limited, however, by the lack of appropriate fluorescent probes, for acidic organelles in particular. The classic fluorescent dyes (such as fluoresceins, rhodamines and coumarins) are not suitable for studying acidic organelles (such as lysosomes) because their fluorescence is significantly decreased under neutral or acidic conditions. This has motivated us to develop probes that can be used in ratio imaging that are strongly fluorescent even in acidic media. RESULTS: The compound 2-(4-pyridyl)-5-((4-(2-dimethylaminoethyl-aminocarbamoyl) methoxy)phenyl)oxazole (PDMPO) was prepared and characterized as a new acidotropic dual-excitation and dual-emission pH indicator. It emits intense yellow fluorescence at lower pH and gives intense blue fluorescence at higher pH. This unique pH-dependent fluorescence property was readily explored to selectively stain lysosomes and to determine the pH of the organelle in an emission-ratio-imaging mode. PDMPO is selectively localized to lysosomes and exhibits a pH-dependent dual excitation and emission. CONCLUSIONS: PDMPO selectively labels acidic organelles (such as lysosomes) of live cells and the two distinct emission peaks can be used to monitor the pH fluctuations of live cells in ratio measurements. Additionally, the very large Stokes shift and excellent photostability of PDMPO make the compound an ideal fluorescent acidotropic probe. The unique fluorescence properties of PDMPO might give researchers a new tool with which to study acidic organelles of live cells.

Acids↗
PubMed1999

Tensegrity and mechanoregulation: from skeleton to cytoskeleton.

OBJECTIVE: To elucidate how mechanical stresses that are applied to the whole organism are transmitted to individual cells and transduced into a biochemical response. DESIGN: In this article, we describe fundamental design principles that are used to stabilize the musculoskeletal system at many different size scales and show that these design features are embodied in one particular form of architecture that is known as tensegrity. RESULTS: Tensegrity structures are characterized by use of continuous tension and local compression; architecture, prestress (internal stress prior to application of external force), and triangulation play the most critical roles in terms of determining their mechanical stability. In living organisms, use of a hierarchy of tensegrity networks both optimizes structural efficiency and provides a mechanism to mechanically couple the parts with the whole: mechanical stresses applied at the macroscale result in structural rearrangements at the cell and molecular level. CONCLUSION: Due to use of tensegrity architecture, mechanical stress is concentrated and focused on signal transducing molecules that physically associate with cell surface molecules that anchor cells to extracellular matrix, such as integrins, and with load-bearing elements within the internal cytoskeleton and nucleus. Mechanochemical transduction may then proceed through local stress-dependent changes in molecular mechanics, thermodynamics, and kinetics within the cell. In this manner, the entire cellular response to stress may be orchestrated and tuned by altering the prestress in the cell, just as changing muscular tone can alter mechanical stability and structural coordination throughout the whole musculoskeletal system.

Biomechanical Phenomena↗
PubMed1999

Activation of protein kinase B and induction of adipogenesis by insulin in 3T3-L1 preadipocytes: contribution of phosphoinositide-3,4,5-trisphosphate versus phosphoinositide-3,4-bisphosphate.

Ectopic expression of activated protein kinase B (PKB) induces the differentiation of confluent 3T3-L1 preadipocytes into adipocytes. PKB is regulated by the lipid products of phosphoinositide 3-kinase (PI 3-kinase), phosphatidylinositol-3,4-bisphosphate [PI(3,4)P2], and phosphatidylinositol-3,4,5-trisphosphate [PI(3,4,5)P3]. However, the relative contribution of each 3-phosphorylated phosphoinositide species in activating PKB remains unclear. Treatment of intact 3T3-L1 preadipocytes with synthetic 3-phosphorylated phosphoinositides revealed that only PI(3,4)P2 stimulated PKB activity. PKB was also activated by insulin, in a dose- and time-dependent manner. This activation was associated with an isolated rise in PI(3,4,5)P3, without any detectable change in PI(3,4)P2, demonstrating that this lipid was sufficient to activate PKB. Wortmannin and LY294002, inhibitors of PI 3-kinase, reduced insulin-dependent activation of PKB, whereas rapamycin, an inhibitor of p70 S6 kinase, had no effect. Platelet-derived growth factor (PDGF), which is not adipogenic, stimulated the production of both 3-phosphorylated phosphoinositide species, and this was associated with a greater activation of PKB than that observed with insulin. A low dose of PDGF (1 ng/ml), which increased the production of only PI(3,4,5)P3 and mirrored the insulin effect, was unable to induce adipocyte differentiation. In summary, insulin and PDGF differ with respect to the accumulation of 3-phosphorylated phosphoinositides and to PKB activation in 3T3-L1 preadipocytes, but these responses do not themselves explain why insulin, but not PDGF, is adipogenic.

3T3 Cells↗
PubMed1999

Prognostic factors and strategy of treatment in Fournier's gangrene: a 12-year retrospective study.

BACKGROUND: Fournier's gangrene (FG) is a fulminant and fatal infection of the genitalia. However, the clinical course is unpredictable. This study retrospectively analyzed the possible prognostic factors of FG. METHODS: Data obtained from 57 patients treated for FG from January 1985 through December 1996 were retrospectively analyzed. Possible prognostic factors including age, diagnostic delay, hospital stay, underlying diseases, clinical symptoms, origins, extents, bacteriologic findings, diverting colostomy and mortality rate were all considered in the analysis. RESULTS: Patients with extensive or localized FG had mortality rates of 31.3% and 16.0%, respectively (p = 0.227). The mortality rates of patients with FG of anorectal, urogenital and non-specific origin were 30.3%, 0% and 40.0%, respectively (p = 0.712). The mortality rates of patients with FG of anorectal origin who received primary or secondary diverting colostomy were 16.7% and 40.0%, respectively. However, the mortality rate of patients with FG of anorectal origin who did not undergo diversion was 29.4%. The mortality rate of patients with FG presenting with septic shock at emergency was 53.8% as compared with 0% in those without septic shock (p < 0.001). CONCLUSION: Fournier's gangrene is a rapidly progressive and life threatening infection of the genitalia. Age, underlying diseases, origin, extent and fecal diversion can not be regarded as prognostic factors of FG. Early primary diverting colostomy may reduce the mortality rate in those with severe infection of anorectal origin. Presence of septic shock in those with FG is the most important and the only factor related to death.

Adult↗
PubMed1999

Application of the holmium yttrium-aluminum-garnet laser for complicated impacted ureteral stones: a preliminary report.

BACKGROUND: Minimally invasive surgery is a current trend in all kinds of surgical fields. Endoscopic stone manipulation with electrohydraulic lithotripsy (EHL) is the preferred method for treating lower ureteral stones or calculi which cannot be resolved with extracorporeal shock wave lithotripsy (SWL). We evaluated the efficiency and safety of holmium yttrium-aluminum-garnet (YAG) laser for treatment of large impacted ureteral stones. METHODS: In a clinical trial period including April and May 1998, 13 patients with ureteral stones underwent holmium YAG lasertripsy. Six patients had large impacted ureteral stones (1.3 to 3.8 cm). We conducted lasertripsy with a 6.5-Fr. rigid tapered ureteroscope and a 550-nm SlimLine laser fiber under the guidance of a 3-Fr. ureteral catheter. The irrigation pump was set at 300 mmHg to increase the surgeon's visual clearance and to help keep the operative field ston-free. RESULTS: The postoperative stone-free rate was 100%. There were no intraoperative complications, even in the treatment for a 3.8-cm steinstrasse in the upper ureter. All preoperative hydronephrosis improved. In general, the operative time, not including anesthesia, was less than 30 minutes. There was no intraoperative or postoperative flank pain or fever when the procedure was combined with pressure irrigation for visual clearance and keeping the area stone-free. CONCLUSION: Holmium YAG lasertripsy is an excellent treatment modality, especially for a large impacted ureteral stone or steinstrasse. The treatment is efficient and safe. Furthermore, there is no learning curve necessary for an experienced endo-urologist. It helps the urologist to carry out a minimally invasive lithotripsy instead of an open surgery for large impacted ureteral stones.

Adult↗
PubMed1999

Enterovesical fistula: experiences with 41 cases in 12 years.

BACKGROUND: A retrospective analysis of enterovesical fistula treated at Chang Gung Memorial Hospital was conducted to determine the optimal diagnosis and management of this disease. METHODS: The records of 41 patients who presented from 1984 to 1996 and had a final diagnosis of enterovesical fistula were retrospectively reviewed. The etiology, symptoms on presentation, diagnostic tools, and modality of treatment were analyzed. RESULTS: The majority of these cases were associated with malignancy (38, 92.7%), and the others with diverticulitis (2, 4.9%) and iatrogenic causes (1, 2.4%). In those with malignancy, 15 patients (39.5%) were found to have tumor recurrence. The most frequent symptom in enterovesical fistula was fecaluria (58.5%), followed by abdominal pain (22%) and dysuria (14.6%). Diagnostic tools included the barium enema, cystography, and cystoscopy; these methods could identify the fistula in 63.2%, 60%, and 53.8% of the patients, respectively. Methods of management included diversion only (39%), one-stage fistula repair (36.6%), and watchful surveillance (24.4%). CONCLUSION: Enterovesical fistula should be considered if fecaluria, pneumaturia, or persistent non-specific urinary tract infection present as the initial complaint. A thorough surgery for a possible underlying malignancy is mandatory when confronted with enterovesical fistula, since the incidence of inflammatory bowel disease is low in this area. An abdominal computer tomography (CT) scan, barium enema, and cystogram can be useful diagnostic tools. Treatment of this entity should be individualized according to each patients clinical status.

Adult↗
PubMed1998

Synthesis and potent antifolate activity and cytotoxicity of B-ring deaza analogues of the nonpolyglutamatable dihydrofolate reductase inhibitor Nalpha-(4-amino-4-deoxypteroyl)-Ndelta-hemiphthaloyl- L-ornithine (PT523).

Six new B-ring analogues of the nonpolyglutamatable antifolate Nalpha-(4-amino-4-deoxypteroyl)-Ndelta-hemiphthaloy l-L-ornithine (PT523, 3) were synthesized with a view to determining the effect of modifications at the 5- and/or 8-position on dihydrofolate reductase (DHFR) binding and tumor cell growth inhibition. The 5- and 8-deaza analogues were prepared from methyl 2-L-amino-5-phthalimidopentanoate and 4-amino-4-deoxy-N10-formyl-5-deaza- and -8-deazapteroic acid, respectively. The 5,8-dideaza analogues were prepared from methyl 2-L-[(4-aminobenzoyl)amino]-5-phthalimidopentanoate and 2, 4-diaminoquinazoline-6-carbonitriles. The Ki for inhibition of human DHFR by the 5-deaza and 5-methyl-5-deaza analogues was about the same as that of 3 (0.35 pM), 11-fold lower than that of aminopterin (AMT, 1), and 15-fold lower than that of methotrexate (MTX, 2). However the Ki of the 8-deaza analogue was 27-fold lower than that of 1, and that of the 5,8-dideaza, 5-methyl-5,8-dideaza, and 5-chloro-5,8-dideaza analogues was approximately 50-fold lower. This trend was consistent with the published literature on the corresponding DHFR inhibitors with a glutamate side chain. In colony formation assays against the human head and neck squamous carcinoma cell line SCC25 after 72 h of treatment, the 5- and 8-deaza analogues were approximately as potent as 3, whereas the 5,8-dideaza analogue was 3 times more potent. 5-Methyl and 5-chloro substitution was also favorable, with the 5-methyl-5-deaza analogue being 2. 5-fold more potent than the 5-deaza analogue. However the effect of 5-methyl substitution was less pronounced in the 5,8-dideaza analogues than in the 5-deaza analogues. The 5-chloro-5,8-dideaza analogue of 3 was the most active member of the series, with an IC50 = 0.33 nM versus 1.8 nM for 3 and 15 nM for MTX. The 5-methyl-5-deaza analogue of 3 was also tested at the National Cancer Institute against a panel of 50 human tumor cell lines in culture and was consistently more potent than 3, with IC50 values in the low-nanomolar to subnanomolar range against most of the tumors. Leukemia and colorectal carcinoma cell lines were generally most sensitive, though good activity was also observed against CNS tumors and carcinomas of the breast and prostate. The results of this study demonstrate that B-ring analogues of 3 inhibit DHFR activity and tumor cell colony formation as well as, or better than, the parent compound. In view of the fact that 3 and its B-ring analogues cannot form polyglutamates, their high cytotoxicity relative to the corresponding B-ring analogues of AMT is noteworthy.

Antineoplastic Agents↗
PubMed1998

Regulation of protein kinase C zeta by PI 3-kinase and PDK-1.

BACKGROUND: Protein kinase C zeta (PKC zeta) is a member of the PKC family of enzymes and is involved in a wide range of physiological processes including mitogenesis, protein synthesis, cell survival and transcriptional regulation. PKC zeta has received considerable attention recently as a target of phosphoinositide 3-kinase (PI 3-kinase), although the mechanism of PKC zeta activation is, as yet, unknown. Recent reports have also shown that the phosphoinositide-dependent protein kinase-1 (PDK-1), which binds with high affinity to the PI 3-kinase lipid product phosphatidylinositol-3,4,5-trisphosphate (Ptdins-3,4,5-P3), phosphorylates and potently activates two other PI 3-kinase targets, the protein kinases Akt/PKB and p70S6K. We therefore investigated whether PDK-1 is the kinase that activates PKC zeta. RESULTS: In vivo, PI 3-kinase is both necessary and sufficient to activate PKC zeta. PDK-1 phosphorylates and activates PKC zeta in vivo, and we have shown that this is due to phosphorylation of threonine 410 in the PKC zeta activation loop. In vitro, PDK-1 phosphorylates and activates PKC zeta in a Ptdins-3,4,5-P3-enhanced manner. PKC zeta and PDK-1 are associated in vivo, and membrane targeting of PKC zeta renders it constitutively active in cells. CONCLUSIONS: Our results have identified PDK-1 as the kinase that phosphorylates and activates PKC zeta in the PI 3-kinase signaling pathway. This phosphorylation and activation of PKC zeta by PDK-1 is enhanced in the presence of Ptdins-3,4-5-P3. Consistent with the notion that PKCs are enzymes that are regulated at the plasma membrane, a membrane-targeted PKC zeta is constitutively active in the absence of agonist stimulation. The association between PKC zeta and PDK-1 reveals extensive cross-talk between enzymes in the PI 3-kinase signaling pathway.

3-Phosphoinositide-Dependent Protein Kinases↗
PubMed1998

Phosphatidylinositol 3,4,5-trisphosphate triggers platelet aggregation by activating Ca2+ influx.

Exogenous phosphatidylinositol 3,4,5-trisphosphate [PtdIns(3,4,5)P3] stimulates the aggregation of washed rabbit platelets in a Ca2+- and dose-dependent manner. This aggregation is reversible at low PtdIns(3,4,5)P3 levels, but becomes irreversible when the concentration exceeds a threshold of about 20 microM. Other D-3 and D-4 phosphoinositides examined, including phosphatidylinositol 3, 4-bisphosphate [PtdIns(3,4)P2], phosphatidylinositol 4, 5-bisphosphate [PtdIns(4,5)P2], and phosphatidylinositol 3-monophosphate [PtdIns(3)P], fail to exert appreciable platelet activation at comparable concentrations. In addition, PtdIns(3,4, 5)P3 can reverse the inhibitory effect of wortmannin on thrombin-induced platelet aggregation. Taken together with the observation that PtdIns(3,4,5)P3 is readily incorporated into cell membranes, these findings reaffirm the second messenger role of PtdIns(3,4,5)P3 in thrombin receptor activation. The existence of a PtdIns(3,4,5)P3-dependent Ca2+ entry system on platelet membranes is supported by the partial inhibition of thrombin-induced Ca2+ influx by wortmannin. Evidence suggests that this system differs from receptor-operated nonselective Ca2+ channels. However, the mechanism by which PtdIns(3,4,5)P3 facilitates Ca2+ entry remains unclear. Although PtdIns(3,4,5)P3 has been known to stimulate phospholipase C-gamma (PLC-gamma), internal Ca2+ mobilization does not play a significant role in the cytosolic Ca2+ increase in response to PtdIns(3,4,5)P3 stimulation. Collectively, these data provide a putative link between PtdIns(3,4,5)P3 and Ca2+ signaling, which may, in part, account for the regulatory function of PtdIns(3,4,5)P3 during platelet aggregation. Moreover, this study bears out the notion that individual PI 3-kinase lipid products play distinct roles in the regulation of cellular functions.

Animals↗
PubMed1998

Use of BODIPY-labeled sphingolipids to study membrane traffic along the endocytic pathway.

In this chapter we discuss the use of BODIPY-labeled sphingolipids to study lipid transport along the endocytic pathway of cultured mammalian cells. The unique spectral properties of the BODIPY fluorophore allow the investigator to distinguish various populations of labeled endosome and lysosomes within the living cell by fluorescence microscopy, and in conjunction with quantitative fluorescence microscopy, to estimate the concentration of these lipids in different intracellular compartments. This methodology is particularly useful for visualizing the accumulation of lipids in the lysosomes of storage disease fibroblasts and may provide a useful method for screening various agents that abrogate this accumulation.

Animals↗
PubMed1998

Abnormal transport along the lysosomal pathway in mucolipidosis, type IV disease.

Mucolipidosis, type IV (ML-IV) is an autosomal recessive storage disease that is characterized by lysosomal accumulation of sphingolipids, phospholipids, and acid mucopolysaccharides. Unlike most other storage diseases, the lysosomal hydrolases participating in the catabolism of the stored molecules appear to be normal. In the present study, we examined the hypothesis that the ML-IV phenotype might arise from abnormal transport along the lysosomal pathway. By using various markers for endocytosis, we found that plasma membrane internalization and recycling were nearly identical in ML-IV and normal fibroblasts. A fluorescent analog of lactosylceramide (LacCer) was used to study plasma membrane lipid internalization and subsequent transport. Lipid internalization at 19 degreesC was similar in both cell types; however, 40-60 min after raising the temperature to 37 degreesC, the fluorescent lipid accumulated in the lysosomes of ML-IV cells but was mainly concentrated at the Golgi complex of normal fibroblasts. Biochemical studies demonstrated that at these time points, hydrolysis of the lipid analog was minimal ( approximately 7%) in both cell types. A fluorescence ratio imaging assay was developed to monitor accumulation of fluorescent LacCer in the lysosomes and showed that the apparent concentration of the lipid increased more rapidly and to a greater extent in ML-IV cells than in normal fibroblasts. By 60 min, LacCer apparently decreased in the lysosomes of normal fibroblasts but not in ML-IV cells, suggesting that lipid efflux from the lysosomes was also impaired. These results demonstrate that there is a defect in ML-IV fibroblasts that affects membrane sorting and/or late steps of endocytosis.

Antigens, CD↗
PubMed1998

A dynamic local bump system for producing synchrotron radiation with an alternating elliptical polarization.

To facilitate high-sensitivity soft X-ray magnetic circular dichroism experiments, a dynamic local bump system has been developed at the SRRC storage ring. This system was devised to vary dynamically the vertical slope of the electron beam in a bending magnet, producing, in the electron orbit plane, soft X-rays with an alternating elliptical polarization. The local bump was created by using two pairs of vertical correctors located on each side of the bending magnet. The bump strength coefficient was obtained both from calculated estimation and from measured beam-response matrices. Control electronics for proper bump strength settings were designed to incorporate the existing orbit-corrector function. A corresponding graphic user interface was implemented so that the bump amplitude could be easily adjusted. The performance of this system is presented. Disturbance on the stored electron beam orbit was observed while flipping the corrector polarity during EPBM (elliptical polarization from bending magnets) operation. A local feedback loop, developed to eliminate such disturbance on other beamlines, is also described.

Journal Article↗
PubMed1998

Phosphoinositide 3-kinase in rat liver nuclei.

Biochemical and immunochemical data from the present investigation reveal the existence of a p85/p110 phosphoinositide 3-kinase (PI 3-kinase) in rat liver nuclei. 32P-Labeling of membrane phosphoinositides by incubating intact nuclei with [gamma-32P]ATP results in the formation of [32P]phosphatidyl-inositol 3,4, 5-trisphosphate [PtdIns(3,4,5)P3], accompanied by small quantities of [32P]phosphatidylinositol 3-phosphate [PtdIns(3)P]. Studies with subnuclear fractions indicate that the PI 3-kinase is not confined to nuclear membranes. The nuclear soluble fraction also contains PI 3-kinase and an array of inositide-metabolizing enzymes, including phospholipase C (PLC), phosphoinositide phosphatase, and diacylglycerol (DAG) kinase. As a result, exposure of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] to the nuclear extract in the presence of [gamma-32P]ATP generates a series of 32P-labeled D-3 phosphoinositides and phosphatidic acid (PA) in an interdependent manner. On the basis of the immunological reactivity and kinetic behavior, the nuclear PI 3-kinase is analogous, if not identical, to PI 3-kinase alpha, and constitutes about 5% of the total PI 3-kinase in the cell. Moreover, we test the premise that nuclear PI 3-kinase may, in part, be regulated through the control of substrate availability by PtdIns(4,5)P2-binding proteins. Effect of CapG, a nuclear actin-regulatory protein, on PI 3-kinase activity is examined in view of its unique Ca2+-dependent PtdIns(4, 5)P2-binding capability. In vitro data show that the CapG-mediated inhibition of nuclear PI 3-kinase is prompted by PKC phosphorylation of CapG and elevated [Ca2+]. This CapG-dependent regulation provides a plausible link between nuclear PLC and PI 3-kinase pathways for cross-communications. Taken together, these findings provide definite data concerning the presence of an autonomous PI 3-kinase cycle in rat liver nuclei. The nuclear location of PI 3-kinase may lead to a better understanding regarding its functional role in transducing signals from the plasma membrane to the nucleus in response to diverse physiological stimuli.

Animals↗
PubMed1998

Regulation of nuclear calcium uptake by inositol phosphates and external calcium.

Factors affecting Ins(1,3,4,5)P4-mediated nuclear Ca2+ uptake are investigated, which include Ins(1,3,4,5)P4 receptor ligand specificity and free external Ca2+ concentrations. Among various inositol phosphates examined, Ins(1,3,4,5)P4, Ins(3,4,5,6)P4, and Ins(1,3,4,5,6)P5 can also stimulate 45Ca2+ influx into isolated rat liver nuclei by activating the Ins(1,3,4,5)P4 receptor-mediated Ca2+ uptake into the nucleus. The EC50 values of these polyphosphates range between 200 and 300 nM, which are 3-4 folds higher than that of Ins(1,3,4,5)P4. It is plausible that these polyphosphates in conjunction with Ins(1,3,4,5)P4 take part in the regulation of nuclear Ca2+ uptake in view of their intracellular levels during cell activation. Moreover, the inositol phosphate-induced Ca2+ uptake is facilitated by increasing Ca2+ levels in the uptake milieu, suggesting a possible link between cytosolic and nuclear Ca2+ signals through the Ins(1,3,4,5)P4 receptor.

Animals↗
PubMed1998

Activation of phospholipase C-gamma by phosphatidylinositol 3,4,5-trisphosphate.

Signal transduction across cell membranes often involves the activation of both phosphatidylinositol (PI)-specific phospholipase C (PLC) and phosphoinositide 3-kinase (PI 3-kinase). Phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2), a substrate for both enzymes, is converted to phosphatidylinositol 3,4, 5-trisphosphate (PI(3,4,5)P3) by the action of PI 3-kinase. Here, we show that PI(3,4,5)P3 activates purified PLC-gamma isozymes by interacting with their Src homology 2 domains. Furthermore, the expression of an activated catalytic subunit of PI 3-kinase in COS-7 cells resulted in an increase in inositol phosphate formation, whereas platelet-derived growth factor-induced PLC activation in NIH 3T3 cells was markedly inhibited by the specific PI 3-kinase inhibitor LY294002. These results suggest that receptors coupled to PI 3-kinase may activate PLC-gamma isozymes indirectly, in the absence of PLC-gamma tyrosine phosphorylation, through the generation of PI(3,4,5)P3.

Animals↗
PubMed1998

CD72-mediated B cell activation involves recruitment of CD19 and activation of phosphatidylinositol 3-kinase.

Occupancy of the B cell glycoprotein, CD72 results in syk-independent activation of phospholipase-C gamma and calcium mobilization. The cytoplasmic tail of CD72 does not contain an immunoreceptor tyrosine-based activation motif to directly transduce signals into the B lymphocyte. Hence, we investigated whether other coreceptors such as CD19 and its associated phosphatidylinositol 3-kinase (PI 3-K) were involved in CD72 signaling. Two specific inhibitors of PI 3-K inhibited CD72-stimulated B cell proliferation in a dose-dependent manner. Activation of B lymphocytes via CD72 resulted in recruitment and activation of PI 3-K, which was mediated by CD19. Accordingly, CD72 ligation induced CD19 tyrosine phosphorylation. Thus, lipid products generated as a result of PI 3-K activation may have an important function in CD72-mediated B lymphocyte activation. The kinetics of CD19 tyrosine phosphorylation induced by CD72 ligation were strikingly different from those seen following B cell antigen receptor (BCR) stimulation. A transient increase in the tyrosine phosphorylation of the complement receptors, CD21 and CD35 was observed in BCR- but not CD72-stimulated cells. Co-cross-linking of CD72 and CD19 failed to induce syk tyrosine phosphorylation suggesting that even under these conditions, CD72 signaling was independent of syk activation. A transient and stimulation-dependent physical association between CD19 and CD72 was observed in CD72-ligated cells. These observations suggest a mechanism by which CD72 can recruit CD19 and influence activation of CD19-associated PI 3-K, which appears to be critical for CD72-mediated B cell activation.

Androstadienes↗