Biomedical subjects

H Ueno

Publications and source records attributed to H Ueno.

At least 199 records · Page 11Linked to original sources

PubMed1998

Prognostic significance of extranodal microscopic foci discontinuous with primary lesion in rectal cancer.

PURPOSE: The most common recurrence after curative resection of rectal carcinoma originates from tiny, undetectable residual foci within the pelvic cavity. The significance and methods used to predict the presence of extramural and extranodal microscopic cancer foci discontinuous with the main lesion of rectal cancers were investigated. METHODS: Four hundred twenty-seven patients who underwent resection of rectal carcinoma were studied. All resected specimens were examined for histologic evidence of extramural cancer separate from the main lesion. RESULTS: Extramural cancers not in continuity with the main rectal lesion were classified as follows: 1) extranodal microscopic cancers; 2) large tumor nodules; 3) lymph node metastases. Each classification was found to influence long-term prognosis. Among them, microscopic cancer was thought to be especially relevant because, by virtue of its microscopic nature, it may be left in the pelvic cavity, causing local recurrence. The existence of large tumor nodules and metastatic lymph nodes correlated closely with the presence of microscopic cancer. Because large tumor nodules and lymph node metastases are possibly detectable during the operation by palpation and may be analyzed by microscopic frozen sections, they might be useful predictors of the presence of microscopic cancers. CONCLUSIONS: In cases with extensive local rectal cancer spread, the nerve-sparing rectal resection that omits lateral dissection may be insufficient for local control because of incomplete removal of occult microscopic cancer, resulting in local recurrence. Presence of microscopic cancer correlates closely with large tumor nodules and metastatic lymph nodes. Intraoperative frozen section investigations may, thus, help in deciding on extent of location resection.

Adenocarcinoma↗

PubMed1998

Intraperitoneal exfoliated cancer cells in patients with colorectal cancer.

PURPOSE: The aims of this study were to evaluate potential predictors of exfoliated free cancer cells in the peritoneal cavity and to assess intraoperative peritoneal lavage cytology as a prognostic indicator in patients with colorectal cancer. METHODS: From 1985 to 1987, intraoperative peritoneal lavage cytology was performed in 140 patients with colorectal cancer. Among them, 88 patients underwent curative resection and 52 patients had noncurative surgery. Cytology was examined twice, i.e., immediately after opening the peritoneal cavity (precytology) and just before closing the abdomen (postcytology). One hundred milliliters of saline was poured into the peritoneal cavity and it was retrieved by suction after irrigation. Cytologic examination was performed after staining with Papanicolaou, Giemsa, periodic acid-Schiff, and Alcian blue stains. RESULTS: Among the 140 patients examined, the incidence of positive cytology in the prelavage was 15 percent, and that in the postlavage was 9 percent, although it was 16 percent in either lavage. Among patients with curative resection, 10 percent had positive cytology. Seven characteristics were identified as features of tumors which are prone to exfoliate cells into the peritoneal cavity: 1) macroscopic peritoneal dissemination, 2) liver metastasis, 3) more than 20 ml of ascites, 4) ulcerated tumors without definite borders, 5) invasion of the serosal surface or beyond, 6) semiannular or annular shape, and 7) moderate or marked lymphatic invasion. In patients undergoing curative surgery, among these features, circumferential involvement was the only one correlated closely with positive cytology (P < 0.02). Positive cytology was associated with a worse outcome. In patients who were resected curatively, the postcytology had a stronger influence on local recurrence than the precytology; the local recurrence rate in patients with positive postcytology was higher than in those with negative postcytology, regardless of the precytology. All patients with cancer cells in the peritoneal cavity at the end of surgery had recurrence. CONCLUSIONS: Seven characteristics were identified as risk factors for exfoliation of cancer cells into the peritoneal cavity in patients with colorectal cancer. These findings may be helpful for the choice of laparoscopic surgery in this era of increasing port-site metastases after laparoscopic procedure. The results of peritoneal lavage cytology at the end of surgery were correlated with the long-term postoperative outcome of colorectal cancer. Thus, meticulous follow-up and possibly adjuvant chemotherapy may be beneficial for patients with free cancer cells in lavage fluid, even after curative surgery.

Adult↗

PubMed1998

Superiority of hepatic arterial infusion in preventing catabolism of 5-FU compared with portal vein infusion revealed by an in vivo 19F NMR study.

PURPOSE: The aim of this study was to identify the route of administration of 5-FU with the greatest pharmacological advantage in a rat model using non-invasive in vivo 19F nuclear magnetic resonance (NMR) spectroscopy. METHODS: 5-FU (50 mg/kg) was administered to anesthetized Wistar rats cannulated into the hepatic artery, portal vein or tail vein and 11 NMR spectra were acquired from the liver region to 60.5 min every 5.5 min. RESULTS: With systemic i.v. (tail vein) infusion, the 19F-NMR signal for 5-FU from the liver region peaked in the first spectrum (0-5.5 min), and then gradually decreased. The signal for the 5-FU catabolite alpha-fluoro-beta-alanine (FBAL) gradually increased to the sixth spectrum (0-33.0 min) and then plateaued. Following portal vein infusion the intensity of the first 5-FU spectrum was twice as high as that following i.v. infusion, but the intensity decreased and the FBAL signal increased gradually in the sixth spectrum as systemic i.v. infusion. In contrast, the intensity of the 5-FU signal following hepatic artery infusion was the same as that following portal vein infusion in the first spectrum, and maintained a strong intensity to the final spectrum (60.5 min). The FBAL signal was detected from the second spectrum following hepatic artery infusion, but its intensity was significantly weaker than that following i.v. or portal vein infusion. CONCLUSIONS: Hepatic arterial infusion resulted in the active form of 5-FU being present for a longer time and its degradation in the liver being suppressed compared with the results following portal vein infusion. This catabolic advantage of hepatic arterial infusion could lead to a more potent anti-tumor activity against liver metastases, but could also lead to significant host toxicity including biliary toxicity. We recommend that the dose/schedule of 5-FU administered via the hepatic artery should be adjusted carefully.

Animals↗

PubMed1998

Comparison of genetic susceptibility to experimental allergic/immune-mediated blepharoconjunctivitis between Lewis and Fischer rats.

BACKGROUND: Fischer rats were less susceptible to experimental autoimmune uveoretinitis (EAU) than Lewis rats, although both strains have the same MHC molecules. The purpose of this study was to compare the susceptibility of experimental allergic/immune-mediated blepharoconjunctivitis (EAC) between these two strains. METHODS: Male Lewis and Fischer rats were immunized with either ovalbumin (OVA) or OVA peptide (OVA323-339) in complete Freund's adjuvant (CFA). Three weeks later, they were challenged with OVA by eye drops. Twenty-four hours later, after clinical evaluation, they were killed and eyes, blood and lymph nodes were harvested for histology, antibody titer and proliferation assay, cytokine production or flow cytometric analysis, respectively. RESULTS: Fischer rats developed mild EAC compared with Lewis rats. Cellular proliferative responses, IFN-gamma production of culture supernatant and serum IgG specific for OVA were basically the same between the two strains. The same OVA peptides were selected as immunodominant. Flow cytometric analysis demonstrated the same cellular profile of lymph node cells in the two strains. CONCLUSION: EAC in Fischer rats was milder than that in Lewis rats, although no apparent differences in immunological parameters between these two strains were detected. These data suggest that factors unrelated to immunological parameters may depend on the susceptibility of EAC.

Allergens↗

PubMed1998

Type IIa early gastric cancer with proliferation of xanthoma cells.

We report a type IIa early gastric cancer associated with xanthoma cell proliferation in a 61-year-old man. The patient was admitted to our hospital because of a gastric polyp detected at a medical checkup. An irregular protruding lesion with xanthoma cell proliferation was detected endoscopically. Histological examination showed a well differentiated tubular adenocarcinoma in the mucosa associated with xanthoma cell proliferation. The distribution of the xanthoma cells in the stroma corresponded closely with that of the cancer cells. Neither atypism nor mitotic figures were recognized in the xanthoma cells. In an immunohistochemical study, almost all the xanthoma cells were stained positive for alpha 1-antitrypsin, while relatively few exhibited positive S-100 protein staining. Specific monocyte chemotactic and activating factor immunoreactivity was present only in the xanthoma cells, and not in the cancer cells. On the basis of these findings, it was speculated that the gastric cancer cells may have caused the xanthoma cell proliferation via an autocrine mechanism i.e., by a chemical mediator acting in a paracrine or juxtacrine manner.

Adenocarcinoma↗

PubMed1998

In vitro growth suppression of vascular smooth muscle cells using adenovirus-mediated gene transfer of a truncated form of fibroblast growth factor receptor.

Vascular smooth muscle cell (VSMC) proliferation associated with arterial injury causes restenosis, which remains to be resolved in cardiovascular and ischemic cerebrovascular disease, especially after balloon angioplasty. Fibroblast growth factor (FGF) is a potent mitogen and a trophic factor for a variety of cells, including VSMCs. We constructed a replication-deficient adenovirus vector, designated AxCA delta FR, coding a truncated form of fibroblast growth factor receptor-1 (FGFR-1) gene lacking the intracellular domain to interrupt receptor-mediated FGF signaling, and examined its effect on the proliferation of primary-cultured rat VSMCs. We transferred the truncated form of the FGFR-1 gene to the VSMCs and confirmed its expression and localization in infected cells by Western blotting and immunofluorescence study. The VSMCs infected with AxCA delta FR degenerated and the proliferation of these cells was suppressed markedly by the infection with this virus in vitro. Our results suggest that the receptor-mediated signal of FGFs has an important role in VSMC proliferation and gene transfer of a truncated form of FGFR using adenoviral vector may be useful for the treatment of the diseases caused by excessive proliferation of VSMCs like restenosis after percutaneous transluminal angioplasty or carotid endoarterectomy.

Adenoviridae↗

PubMed1998

A novel retinoic acid receptor (RAR)-selective antagonist inhibits differentiation and apoptosis of HL-60 cells: implications of RARalpha-mediated signals in myeloid leukemic cells.

Retinoic acid (RA) induces HL-60 cells to differentiate terminally into mature granulocytes, which subsequently die by apoptosis. The biological effects of RA are mediated by two distinct families of transcription factors: retinoic acid receptors (RARs) and retinoid X receptors (RXRs). RARs and RXRs form heterodimers and regulate retinoid-mediated gene expression. We have recently developed a novel RAR-selective antagonist (ER27191) which prevents RAR activation by retinoids. Using this RAR-selective antagonist, and RXR and RAR agonist, we demonstrate the RAR-mediated signaling pathway is important for differentiation and apoptosis of myeloid leukemic cells. Simple activation of RXRs is not sufficient to induce apoptosis of the cells. Interestingly, the combination of the RAR-selective antagonist and 9-cis RA resulted in partial differentiation and apoptosis of HL-60 and NB4 cells, whereas the RAR antagonist completely blocked all-trans RA-induced differentiation and apoptosis of the cells. Additional experiments showed that levels of BCL-2 protein decreased during differentiation of myeloid leukemic cells. Furthermore, HL-60 cells transduced with a bcl-2 expression vector showed the same differentiation response to retinoids as did parental HL-60 cells even though apoptosis was inhibited in these bcl-2-transduced cells, suggesting that differentiation and apoptosis are regulated independently in myeloid leukemic cells.

Alitretinoin↗

PubMed1998

IL-4 and prostaglandin E2 inhibit hypomethylation of the 5' regulatory region of IFN-gamma gene during differentiation of naive CD4+ T cells.

We have previously shown that prostaglandin E2 (PGE2) and IL-4 inhibit the priming of IFN-gamma-production during the differentiation of naive CD4+ T cells from human cord blood by different signal-transducing mechanisms. To compare and analyse the molecular mechanisms by which PGE2 and IL-4 inhibit the priming of IFN-gamma production, we investigated the effects of PGE2 and IL-4 on the methylation of the IFN-gamma gene during the in vitro differentiation of naive CD4+ T cells. In human naive CD4+ T cells, which produce primarily IL-2 and a little amount of IFN-gamma, the IFN-gamma gene was methylated. After stimulation via TCR, CD4+ T cells produced IFN-gamma and the CpG dinucleotide contained within the TATA proximal regulatory element of the IFN-gamma gene was partially hypomethylated. Both IL-4 and PGE2 inhibited the hypomethylation of this site and the acquisition of IFN-gamma-producing ability. In contrast to the SnaBI site in the TATA proximal regulatory element, the HpalI site in the first intron of the IFN-gamma gene of the CD4+ T cells from cord blood was completely methylated even after stimulation via TCR. 5-azacytidine restored the IFN-gamma-producing ability of these cells treated with IL-4 and PGE2. These findings suggest that, although the signal transduction that inhibits the priming of IFN-gamma-production is different for each reagent, the protection from hypomethylation of the regulatory region of the IFN-gamma gene is involved in the molecular mechanisms by which these reagents inhibit the priming of IFN-gamma-production during the differentiation of human naive CD4+ T cells.

Azacitidine↗

PubMed1998

Impingement after total knee arthroplasty caused by cement extrusion and proximal tibiofibular instability.

A 57-year-old patient with rheumatoid arthritis showed posterolateral impingement after total knee arthroplasty. The radiographs showed bone cement extrusion posterolateral to the tibial tray. Arthrotomy through a posterolateral approach revealed that the impingement was caused not only by cement extrusion against the fibular head but also by proximal tibiofibular joint instability. It was speculated that rheumatoid arthritis had caused proximal tibiofibular instability, active knee motion had caused fibular head shift by tension of biceps femoris and the fibular head had been impinged on the extruded cement. In cementing the tibial tray, especially in a rheumatoid patient, it is of paramount importance to take caution against posterolateral cement extrusion in order to minimize the risk of fibular head impingement during total knee arthroplasty.

Arthritis, Rheumatoid↗

PubMed1998

Structure-activity relationship and conformational analysis of monoglycosylceramides on the syngeneic mixed leukocyte reaction.

We examined effects of alpha-, beta-galactosylceramides (CalCers) and alpha-, beta-glucosylceramides (GlcCers) on the syngeneic mixed leukocyte reaction (MLR) using spleen cells (responder cells) and dendritic cells (DC, stimulator cells). The DC pretreated with these alpha-monoglycosylceramides markedly stimulated the proliferation of spleen cells, in contrast to the little stimulatory effects produced by the DC pretreated with the corresponding beta-anomers. In addition, when we compared the effects of alpha-GalCer derivatives on the syngeneic MLR, it appeared that the 2"- and 3-hydroxyl groups in alpha-GalCers play a critical role in their stimulation of the MLR response. Based on these results, we performed a computer-aided molecular modeling study, and found that the orientations of the 2"-, 4"- and 3-hydroxyl groups common to alpha-GalCer and alpha-GlcCer are not accessible to those of inactive monoglycosyleeramides such as beta-GalCer. These results suggest that there might be a receptor-like site for alpha-monoglycosylceramides on the cells which are involved in the MLR response.

Animals↗

PubMed1998

The levels of integrin alpha v beta 5 may predict the susceptibility to adenovirus-mediated gene transfer in human lung cancer cells.

Adenovirus mediated transfer of growth-inhibiting molecules, such as p53 shows promise as an effective method of suppressing the growth of cancer cells. As the basis for in vivo studies, we examined transfection efficiency using 15 human lung cancer cell lines that differ in their endogenous p53 status. When infected with an adenovirus expressing bacterial beta-galactosidase, the different cell lines showed different levels of beta-galactosidase activity. We found a correlation between the level of integrin alpha v beta 5, which is thought to be an adherence receptor for adenoviruses, and the expression level of the transferred gene, suggesting that gene expression is largely dependent on the infection efficiency. Growth inhibition was induced in all cell lines tested following infection with an adenovirus containing p53, regardless of the genetic status of their endogenous p53 provided a sufficient amount of p53 protein was expressed. Our results (1) confirm that the examination of the susceptibility of target cancer cells to an adenovirus is important when considering performing adenovirus-mediated gene transfer and for evaluating its therapeutic effects; and (2) suggest that the quantification of integrin alpha v beta 5 may be a good way of predicting the susceptibility of cells to adenoviral vectors.

Adenoviridae↗

PubMed1998

A vitrectomy improves the transfection efficiency of adenoviral vector-mediated gene transfer to Müller cells.

The neural retina is a logical target of gene therapy for various ocular diseases. We developed a new gene delivery method to the neural retina using an adenoviral vector with a high degree of gene transfection efficiency and less functional damage. An adenoviral vector bearing the lacZ gene (AdCALacZ) was injected into the eyes of adult Wistar rats after an SF6 compression gas vitrectomy and left for 30 min followed by washing with balanced salt solution (BSS) (method A). Three other methods, comprising a simple intravitreal injection (method B), an intravitreal injection after an SF6 compression gas vitrectomy (method C) or a subretinal injection (method D), were also studied. The gene expression was examined 6 days after the AdCALacZ injection. An immunohistochemical study for antivimentin, antiglial fibrillary acidic protein and anti S100 protein antibodies showed the neural retinal cells (Müller cells) to be primarily transfected by methods A, B and C, while only a few cells were transfected by method D. The expression of beta-galactosidase was visualized by X-gal staining and the positive areas on each hemiflat mount specimen were measured by an image analyzer and then were adopted as a value of gene transfer efficiency. The highest degree of gene expression was obtained by methods A (23.2% of total retinal area) and C (19.8%), while the lowest degree was obtained by method B (8.9%). The inflammation was observed in all eyes and the value of inflammation was quantified as the average inflammatory cell number in each microscopic field (cells per fields). A moderate degree of inflammation was induced by methods B (28.3 cells per field) and C (27.5 cells per field) and a minimal degree of inflammation was induced by method A (11.2 cells per field). We evaluated the retinal function by measuring an electroretinogram (ERG). The amplitudes of the ERG were depressed in all eyes treated with AdCALacZ. This depression was manifested most by methods B and C, and least by method A. The deterioration in the ERG findings seemed to correlate with the intensity of inflammation. Our study showed that an intravitreal injection with an adenoviral vector can transfer the genes to the neural retinal cells and therefore a vitrectomy and the subsequent removal of the adenoviral vector, can thus significantly improve the transfection efficiency and also reduce the degree of functional damage.

Adenoviridae↗

PubMed1998

Bovine spleen cathepsin A: characterization and comparison with the protective protein.

Cathepsin A was purified approximately 550-fold with an overall yield of 4% from bovine spleen crude extracts by successive chromatographies on DEAE-Sephadex A-50, phenyl-Toyopearl 650C, and Con A-agarose. PAGE of the purified enzyme without 2-mercaptoethanol revealed an apparent molecular size of 110 kDa, and SDS-PAGE with 2-mercaptoethanol gave two polypeptide bands corresponding to 32 and 25 kDa and without 2-mercaptoethanol a single polypeptide 52 kDa band. These results indicate that the enzyme has an (alpha beta)2 tetrameric structure in which the alpha (32 kDa) and beta (25 kDa) subunits are linked by disulfide bond(s). The enzyme exhibited peptidase activities, hydrolyzing various Z-dipeptides with optimum pHs between 5.0 and 5.8. The hydrolytic rate for Z-Phe-Ala was 15 times higher than that for Z-Glu-Tyr, the traditional cathepsin A substrate. The enzyme also catalyzed the hydrolysis of the C-terminal amino acids of RCM-RNase A and showed esterase activity toward BTEE at pH around 7.5. DFP and TPCK completely inhibited both peptidase and esterase activities, and [1,3-3H]DFP was bound to the alpha subunit. All these results support the fact that the enzyme is a serine carboxypeptidase. The N-terminal amino acid sequences of the alpha and beta subunits are highly homologous to those of the human protective protein in galactosialidosis, strongly supporting the identity between cathepsin A and the protective protein.

Amino Acid Sequence↗

PubMed1998

Role of Phe120 in the activity and structure of bovine pancreatic ribonuclease A.

Phenylalanine120 is a candidate residue juxtaposing catalytic His12 and His119 in ribonuclease A (RNase A). To clarify its role in construction of the catalytic center, Phe120 was replaced by alanine, tryptophan, leucine, or glutamic acid by site-directed mutagenesis. The transphosphorylation and hydrolysis activities of the mutant RNase As, respectively, toward cytidinyl 3',5' adenosine (CpA) and cytidine 2',3' cyclic monophosphate (C>p) were compared with those of the wild type enzyme. The Km values of the two reactions increased markedly with slight changes in the Kcat values. The pKe values of His12 and His119 in the wild type and mutant enzymes, estimated from the pH dependence of the kcat/Km values, showed little change. The rate of carboxymethylation was reduced markedly by the mutations. The Ki values of the phosphate anion as to hydrolysis activity increased only slightly when Phe120 was replaced by leucine, tryptophan, or alanine. These findings suggest that Phe120 participates in the binding of the substrate, juxtaposing His12 and His119, and in stabilizing the transition state intermediate in the hydrolysis reaction. Furthermore, the decreases in the thermal denaturation temperatures of all the mutants, particularly F120E, indicate that Phe120 also helps maintain the conformational stability of RNase A.

Animals↗

PubMed1998

Proton-relay system of carboxypeptidase Y as a sole catalytic site: studies on mutagenic replacement of his 397.

His397 was replaced with alanine by site-directed mutagenesis of the cloned PRC1 gene in order to confirm the role of this residue in the proton-relay system of carboxypeptidase Y (CPY). The expressed and purified H397A showed a CD spectrum almost identical to that of the wild type enzyme, but its heat stability and conformation on heating differed somewhat. Kinetic analysis showed that the kcat values of the purified H397A toward the peptide substrates, Z-Phe-Leu and Z-Gly-Phe, were reduced to approximately 4 x 10(-5)-fold, whereas the Km values remained almost unchanged. The activity of the H397A preparation with the ester substrate, Ac-Phe-OEt, was negligible. The low activity of our H397A was lost on treatment with DFP and Z-Phe-CH2Cl, site-specific inhibitors, respectively, for Ser146 and His397, and with the HgCl2 and PCMB, SH-reagents for Cys341. After treatment with these inhibitors, the kcat value for the H397A preparation toward Z-Phe-Leu decreased 1 x 10(3)-fold or more. The value was approximately 10(-8) for the wild type enzyme. This level of activity is 10(3)-fold lower than the reported value for the same mutant of CPY [Carlsberg Res. Commun. 54, 165-171 (1989)], and more than 10-fold lower than the values for the corresponding His-to-Ala mutants of trypsin [J. Am. Chem. Soc. 114, 1784-1790 (1992)] and subtilisin [Nature 332, 564-568 (1988)]. These findings, together with the pH profiles and chromatographic behavior, are evidence that the low activity of the H397A preparation is due to contamination by wild type CPY. The decreased kcat value of our H397A mutant is the lowest reported among the corresponding histidine mutants of serine proteases. We conclude that the proton-relay system composed of Ser146 and His397 is the sole catalytic center of CPY, and that its destruction leads to complete inactivation.

Binding Sites↗

PubMed1998

Cellular injury score for multiple organ failure severity scoring system.

BACKGROUND: Cellular Injury Score (CIS) is an index of cellular injury, being calculated from three parameters of intracellular metabolism: arterial ketone body ratio, osmolality gap, and blood lactate. METHODS: The usefulness of CIS as a severity scoring system for patients with multiple organ failure was prospectively evaluated in 157 consecutive patients with MOF (58 survivors, 99 nonsurvivors). RESULTS: CISs in nonsurvivors were significantly higher compared with those in survivors throughout the clinical courses. CIS was significantly correlated with the number of failing organs and mortality rate. The optimal cutoff point of CIS from receiver operating characteristics curve analysis was 4 for the maximal value during the clinical course. The changes in CIS well reflected the severity of injury in survivors and nonsurvivors who died within 2 weeks. CONCLUSION: CIS could be a useful index for mortality risk prediction and is potentially applicable as a severity scoring system for individual patients with MOF.

3-Hydroxybutyric Acid↗

PubMed1998

Detection of Coxiella burnetii from dust in a barn housing dairy cattle.

We attempted to detect Coxiella burnetii in dust samples collected from a barn housing dairy cattle by the polymerase chain reaction (PCR) method. Ten dust samples (five from ventilation fans and five from crossbeams) were collected from two areas in a barn on a farm near Sapporo, Hokkaido. C. burnetii was detected in 5 of the 10 dust samples. It was believed that aerial contamination by C. burnetii occurred in the barn.

Air Microbiology↗

PubMed1998

Analysis of effects of stimulation in vitro of ovalbumin primed lymph node cells on adoptive transfer of experimental immune mediated blepharoconjunctivitis in Lewis rats.

AIM: To analyse the role of stimulation in vitro of lymphocytes on the augmentation of experimental immune mediated blepharoconjunctivitis (EC, formerly EAC) in Lewis rats induced by adoptive transfer. METHODS: Two weeks after immunisation with ovalbumin (OVA), rat draining lymph nodes were collected and 50 x 10(6) cells were injected into naive syngeneic recipients either directly or after culture in vitro with OVA, concanavalin A (Con A), or purified protein derivative (PPD) for 3 days. Four days after injection the rats were topically challenged with OVA. 24 hours later, they were sacrificed and eyes and spleens were harvested for histology and proliferation assay. In some experiments, naive recipient rats were irradiated with 7 Gy gamma ray before transfer. The expression of adhesion molecules and cytokine profile of OVA primed lymph node cells were also investigated. RESULTS: Both infiltrated cell number and splenocyte proliferation in the recipients of stimulated cells were higher than those of unstimulated cells. In vitro stimulation with OVA or Con A induced a severe cellular infiltration, while stimulation with PPD did not. Irradiation markedly diminished cellular infiltration. Stimulation in vitro upregulated the CD4/CD8 ratio by four times and augmented expression of CD25, I-A, ICAM-1 molecules on OVA primed lymph node cells by about five times. IFN-gamma was detected in OVA primed cells by stimulation in vitro, while IL-4 mRNA was extinguished by stimulation in vitro. CONCLUSIONS: Augmentation of EC by stimulation in vitro of transferred lymphocytes might depend on the upregulation of expression of cell surface molecules and cytokine shift as well as augmented antigen specificity.

Adoptive Transfer↗