Antibody responses of Macaca fascicularis against a new inactivated polio vaccine derived from Sabin strains (sIPV) in DTaP-sIPV vaccine.

Antibody responses of Macaca fascicularis against a new tetravalent vaccine composed of diphtheria toxoid, tetanus toxoid, acellular pertussis antigens, and inactivated poliovirus derived from Sabin strains (sIPV) was investigated to predict an optimal dose of sIPV in a new tetravalent vaccine (DTaP-sIPV) prior to conducting a dose-defined clinical study. Monkeys were inoculated with DTaP-sIPVs containing three different antigen units of sIPVs: Vaccine A (types 1:2:3 = 3:100:100 DU), Vaccine B (types 1:2:3 = 1.5:50:50 DU), and Vaccine C (types 1:2:3 = 0.75:25:25 DU). There was no difference in the average titers of neutralizing antibody against the attenuated or virulent polioviruses between Vaccines A and B. The average neutralizing antibody titers of Vaccine C tended to be lower than those of Vaccines A and B. The sIPV antigens did not affect the anti-diphtheria or anti-tetanus antibody titers of DTaP-sIPV. Furthermore, the average neutralizing antibody titers of Vaccine A against the attenuated and virulent polioviruses were comparable between M. fascicularis and humans. These results suggest that M. fascicularis may be a useful animal model for predicting the antibody responses to sIPVs in humans, and that it may be likely to reduce the amount of sIPVs contained in DTaP-sIPVs, even for humans.

Inhibition of cell growth and induction of apoptotic cell death by the human tumor-associated antigen RCAS1.

Tumor-associated antigens that can be recognized by the immune system include the MAGE-family, p53, MUC-1, HER2/neu and p21ras. Despite their expression of these distinct antigens, tumor elimination by the immune system is often inefficient. Postulated mechanisms include insufficient expression of co-stimulatory or adhesion molecules by tumor cells, or defective processing and presentation of antigens on their cell surfaces. Tumor cells may also evade immune attack by expressing CD95 (APO-1/Fas) ligand or other molecules that induce apoptosis in activated T cells. Here we describe RCAS1 (receptor-binding cancer antigen expressed on SiSo cells), a membrane molecule expressed on human cancer cells. RCAS1 acts as a ligand for a putative receptor present on various human cell lines and normal peripheral lymphocytes such as T, B and NK cells. The receptor expression was enhanced by activation of the lymphocytes. RCAS1 inhibited the in vitro growth of receptor-expressing cells and induced apoptotic cell death. Given these results, tumor cells may evade immune surveillance by expression of RCAS1, which would suppress clonal expansion and induce apoptosis in RCAS1 receptor-positive immune cells.

CD1-reactive natural killer T cells are required for development of systemic tolerance through an immune-privileged site.

Systemic tolerance can be elicited by introducing antigen into an immune-privileged site, such as the eye, or directly into the blood. Both routes of immunization result in a selective deficiency of systemic delayed type hypersensitivity. Although the experimental animal model of anterior chamber-associated immune deviation (ACAID) occurs in most mouse strains, ACAID cannot be induced in several mutant mouse strains that are coincidentally deficient in natural killer T (NKT) cells. Therefore, this model for immune-privileged site-mediated tolerance provided us with an excellent format for studying the role of NKT cells in the development of tolerance. The following data show that CD1-reactive NKT cells are required for the development of systemic tolerance induced via the eye as follows: (a) CD1 knockout mice were unable to develop ACAID unless they were reconstituted with NKT cells together with CD1(+) antigen-presenting cells; (b) specific antibody depletion of NKT cells in vivo abrogated the development of ACAID; and (c) anti-CD1 monoclonal antibody treatment of wild-type mice prevented ACAID development. Significantly, CD1-reactive NKT cells were not required for intravenously induced systemic tolerance, thereby establishing that different mechanisms mediate development of tolerance to antigens inoculated by these routes. A critical role for NKT cells in the development of systemic tolerance associated with an immune-privileged site suggests a mechanism involving NKT cells in self-tolerance and their defects in autoimmunity.

Factors influencing progressive collapse of the transposed necrotic lesion after transtrochanteric anterior rotational osteotomy for osteonecrosis of the femoral head.

BACKGROUND: Transtrochanteric anterior rotational osteotomy (ARO) for osteonecrosis of the femoral head (ONFH) can preserve for a long-time collapsed femoral head. Progressive collapse of anteriorly-transposed necrotic lesion leads to secondary arthritic changes and clinical failure. Critical factors influencing collapse of the transposed necrotic lesion after ARO remain largely unknown. Therefore, we performed a retrospective study of ARO to determine: (1) if preoperative collapse influences collapse of the transposed necrotic area, (2) if any other factor may influence collapse of the transposed necrotic area. HYPOTHESIS: We hypothesized the degree of preoperative femoral head collapse influences progressive collapse of the transposed necrotic lesion after ARO. MATERIALS AND METHODS: We reviewed 47 hips in 42 patients with ONFH treated with ARO between 2000 and 2005 with a mean follow-up of 11.4 years (10-14 years). The occurrence of progressive collapse of the transposed necrotic lesion after ARO was examined using lateral radiographs taken at least once every year after ARO. The following factors were statistically analyzed: age, sex, body mass index, Harris Hip Score (HHS), preoperative level of collapse, extent of the necrotic lesion and postoperative intact ratio (ratio of the transposed intact articular surface of the femoral head). RESULTS: Progressive collapse of the transposed necrotic lesion (progressive collapse group) was seen in 17 hips (36%) during a mean period of 1.8 years (0.5-3.7 years) after ARO, which has developed within 4 years in all cases. Preoperative level of collapse in the progressive collapse group (4.4±1.4mm) was significantly larger than that in the non-progressive collapse group (2.1±1.0mm), which was independently associated with progressive collapse of the transposed necrotic lesion in multivariate analysis (P<0.0001) with cut off point of 2.98mm. In univariate analysis, lower preoperative HHS, severe extent of the necrotic lesion and the lower postoperative intact ratio were also associated with progressive collapse of the transposed necrotic lesion, but were not associated as independent factors in multivariate analysis. DISCUSSION: The current study suggests that progressive collapse of the transposed necrotic lesion after ARO depends mainly on the preoperative level of collapse (cut-off point=2.98mm). LEVEL OF EVIDENCE: IV; retrospective case series.

Effects of intertrochanteric osteotomy plane and preoperative femoral anteversion on the postoperative morphology of the proximal femur in transtrochanteric anterior rotational osteotomy: 3D CT-based simulation study.

BACKGROUND: Transtrochanteric anterior rotational osteotomy (ARO) is joint-preserving surgery for patients with osteonecrosis of the femoral head (ONFH). During ARO, femoral neck-shaft varus angulation by changing intertrochanteric osteotomy plane is often designed to obtain a sufficient postoperative intact ratio. However, the effect of intertrochanteric osteotomy plane on postoperative femoral anteversion has not been well examined. Therefore, we performed a simulation study of ARO to determine how intertrochanteric osteotomy plane and preoperative femoral anteversion affect both femoral neck-shaft varus angle and postoperative femoral anteversion. HYPOTHESIS: Both femoral neck-shaft varus angle and postoperative femoral anteversion are predicted by intertrochanteric osteotomy plane and preoperative femoral anteversion in ARO. MATERIALS AND METHODS: Using CT-data obtained from 10 hips in 10 patients with ONFH, ARO was simulated. On anteroposterior view, basic intertrochanteric osteotomy line (AP-view line) was defined as the perpendicular line to the femoral neck axis. On lateral view, basic intertrochanteric osteotomy line (lateral-view line) made through the cut surface of greater trochanter was defined as the perpendicular line to the lateral axis of the femur. By changing either AP-view or lateral-view line, 49 ARO models/hip were produced, in which femoral neck-shaft varus angle and postoperative femoral anteversion were assessed. RESULTS: With increase in the vertically-inclined degree of AP-view line, both neck-shaft varus angle and postoperative femoral anteversion increased. With increase in the posteriorly-tilted degree of lateral-view line, neck-shaft varus angle increased, whereas postoperative femoral anteversion decreased. The approximation equations based on the multiple regression analyses were as follows: neck-shaft varus angle≈vertically-inclined degree of AP-view line×0.9+posteriorly-tilted degree of lateral-view line×0.8+preoperative femoral anteversion×0.7; postoperative femoral anteversion≈vertically-inclined degree of AP-view line×1.1-posteriorly-tilted degree of lateral-view line×0.8. DISCUSSION: The postoperative morphology of proximal femur was nearly defined by intertrochanteric osteotomy plane with preoperative femoral anteversion, which is useful for preoperative planning in terms of both achieving a sufficient postoperative intact ratio and maintaining femoral anteversion. LEVEL OF EVIDENCE: Level IV case series without control group.

Analysis of corneal inflammation induced by cauterisation in CCR2 and MCP-1 knockout mice.

AIM: To elucidate the role of CCR2/MCP-1 in corneal inflammation. METHODS: A cauterisation induced corneal inflammation model was used. The corneas were cauterised with silver nitrate in CCR2 knockout (KO) mice, MCP-1 KO mice, and control mice. Clinical signs such as corneal oedema and opacity were examined 96 hours after cauterisation and the phenotypes of the cells infiltrating the cornea were analysed by flow cytometry. Corneal inflammation in neutrophil depleted mice was also analysed. RESULTS: After cauterisation both CCR2 KO and MCP-1 KO mice showed the same levels of corneal oedema and opacity as control mice. Flow cytometry revealed that in control mice most of the infiltrating cells were neutrophils and macrophages, whereas in both CCR2 KO mice and MCP-1 KO mice, the number of macrophages infiltrating the cornea were markedly reduced. However, prominent infiltrates of neutrophils were still observed in the cornea in CCR2 KO mice and MCP-1 KO mice. The depletion of neutrophils significantly reduced the oedema and opacity induced in the cornea by cauterisation. CONCLUSION: The CCR2 and MCP-1 molecules are not essential for cauterisation induced corneal inflammation. Neutrophils, rather than migrated macrophages, are the final effector cells involved in inducing inflammation in this model.

GEP oncogene promotes cell proliferation through YAP activation in ovarian cancer.

G-protein-coupled receptors (GPCRs) and their ligands function in the progression of human malignancies. Gα12 and Gα13, encoded by GNA12 and GNA13, respectively, are referred to as the GEP oncogene and are implicated in tumor progression. However, the molecular mechanisms by which Gα12/13 activation promotes cancer progression are not fully elucidated. Here, we demonstrate elevated expression of Gα12/13 in human ovarian cancer tissues. Gα12/13 activation did not promote cellular migration in the ovarian cancer cell lines examined. Rather, Gα12/13 activation promoted cell growth. We used a synthetic biology approach using chimeric G proteins and GPCRs activated solely by artificial ligands to selectively trigger signaling pathways downstream of specific G proteins. We found that Gα12/13 promotes proliferation of ovarian cancer cells by activating the transcriptional coactivator YAP, a critical component of the Hippo signaling pathway. Furthermore, we reveal that inhibition of YAP by short hairpin RNA or a specific inhibitor prevented the growth of ovarian cancer cells. Therefore, YAP may be a suitable therapeutic target in ovarian cancer.

The characterisation of hyalocytes: the origin, phenotype, and turnover.

AIM: To determine the characterisation of hyalocytes: the origin, phenotype, and turnover in the rodent. METHODS: To characterise the ultrastructure and distribution of hyalocytes, transmission and scanning electron microscopy was performed in rat eyes. Immunophenotypical analysis was performed by either anti-ED1 or ED2 antibodies. To examine the origin of the hyalocytes, the chimeric mice were created and were used to transplant the bone marrow (BM) cells from enhanced green fluorescent protein (EGFP) transgenic mice. The turnover of hyalocytes was examined at 0, 4, 6, 7, and 12 months after BM transplantation. RESULTS: Hyalocytes were distributed especially in the vitreous cortex and had an irregular shape with a spherical granule. Immunophenotypical studies demonstrated that most of the hyalocytes in rat eyes expressed ED2 but not ED1. In the chimeric mice, the hyalocytes were GFP negative right after BM transplantation. Interestingly, more than 60% of hyalocytes were replaced within 4 months and approximately 90% within 7 months after BM transplantation. CONCLUSIONS: The rodent hyalocytes were shown to express tissue macrophage marker, were derived from BM, and totally replaced within 7 months. These data provide the characterisation of hyalocytes in physiological conditions, especially their origin, distribution, and turnover, and may contribute to the better understanding of the pathogenesis of vitreoretinal disease.

Tumor-associated antigen 22-1-1 expression in the uterine cervical squamous neoplasias.

We have reported that a novel tumor-associated antigen (Ag), 22-1-1, was expressed in cancer cells derived mainly from the uterus and ovary [K. Sonoda et al., Cancer (Phila.), 77: 1501-1509, 1996]. The 22-1-1 Ag existed not only in adenocarcinomas but also in squamous cell carcinomas in the uterine cervix. Here, a relationship between tumor progression and invasion and 22-1-1 Ag expression was investigated in squamous cell neoplasms of the uterine cervix using immunohistochemical staining. The 22-1-1 Ag was not detected in normal uterine cervix (0 of 10 total cases) and dysplasias (0 of 47 total cases). However, 20% of carcinoma in situ (4 of 20 total cases) and 16.7% of microinvasive carcinomas (2 of 12 total cases) stained positively for 22-1-1 Ag. Moreover, areas depicting microinvasion on histology in uterine cancers (stage Ia) were more strongly stained than carcinoma in situ lesions. 22-1-1 Ag expression was found to be more frequent in invasive squamous cell carcinomas (82.6%; 57 of 69 total cases). The 22-1-1 Ag existed both in the cytoplasm and on the membrane of cancer cells. These findings suggest that 22-1-1 Ag expression might be related to tumor cell progression and invasion in the uterine cervical squamous cell epithelium.

The prognostic significance of tumor-associated antigen 22-1-1 expression in adenocarcinoma of the uterine cervix.

We previously established (K. Sonoda et al., Int. J. Oncol., 6: 1099-1104, 1995) a novel monoclonal antibody, 22-1-1, generated from adenocarcinoma of the uterine cervix, and 22-1-1 antigen (Ag) was expressed in cancer cells derived mainly from the uterus and ovary. In this report, a relationship between 22-1-1 Ag expression and clinicopathological variables and the prognostic significance of 22-1-1 Ag were immunohistochemically investigated in adenocarcinoma of the cervix. Of 56 cases, the 22-1-1 Ag was negative in 7, 1+ in 14, 2+ in 26 and 3+ in 9 instances. The 22-1-1 Ag existed both in the cytoplasm and on the membrane of cancer cells. There was no correlation between 22-1-1 Ag expression and age, stage, grade, myometrial invasion, lymph-vascular space invasion, lymph node metastasis, and parametrial invasion. The estimated 5-year overall survival (OS) of patients with low 22-1-1 Ag expression (-/+) and high 22-1-1 Ag expression (++/ ) were 90.5 and 71.4%, respectively. Patients with high 22-1-1 Ag expression had significantly worse OS than those with low 22-1-1 Ag expression (log-rank test, P = 0.0193). In addition, lymph-node metastasis, age, and clinical stage were significantly related to OS in univariate analysis. Multivariate analysis for OS revealed a prognostic significance in 22-1-1 Ag expression, stage, age, and grade. These data suggest that 22-1-1 Ag expression may be related to prognosis in adenocarcinoma of the cervix.

Regulation of adaptive immune responses by innate cells expressing NK markers and antigen-transporting macrophages.

A continuing theme of work done in our laboratory involves regulation of adaptive immune response by innate cells, in general, and immuneregulation by natural killer (NK) and NKT cells, in particular. Studies include work with the lung and the eye. In addition to immune surveillance of tumor cells, the NK cell is often associated with secreting cytokines that contribute to the creation of microenvironments conducive to Th1 responses and with defense mechanisms that lessen the initial infecting viral load. Reported studies show that the NKT cells support both T helper cell responses (type 1 and 2), as well as their being absolutely central to the development of antigen-specific T-regulatory cells involved in peripheral tolerance. Because of the multifunctional capabilities of the NKT cell, we propose that yet another cell, such as the antigen-presenting cell (APC), may influence the effector pathway of the NKT cell. We postulate that the APC that transports the antigen from the entry environment provides both trafficking and activation signals for innate cells in the secondary lymphoid organs. Evidence is presented that macrophage-derived signals selectively recruit NKT cells and bias their cytokine synthesis. Data imply that, just as occurs in immune inflammation, a collection of innate and adaptive immune cells interact within the secondary lymphoid tissue to generate antigen-specific tolerance in the periphery.

Necrotic enlargement of cone photoreceptor cells and the release of high-mobility group box-1 in retinitis pigmentosa.

Retinitis pigmentosa (RP) refers to a group of inherited retinal degenerations resulting form rod and cone photoreceptor cell death. The rod cell death due to deleterious genetic mutations has been shown to occur mainly through apoptosis, whereas the mechanisms and features of the secondary cone cell death have not been fully elucidated. Our previous study showed that the cone cell death in rd10 mice, an animal model of RP, involves necrotic features and is partly mediated by the receptor interacting protein kinase. However, the relevancy of necrotic cone cell death in human RP patients remains unknown. In the present study, we showed that dying cone cells in rd10 mice exhibited cellular enlargement, along with necrotic changes such as cellular swelling and mitochondrial rupture. In human eyes, live imaging of cone cells by adaptive optics scanning laser ophthalmoscopy revealed significantly increased percentages of enlarged cone cells in the RP patients compared with the control subjects. The vitreous of the RP patients contained significantly higher levels of high-mobility group box-1, which is released extracellularly associated with necrotic cell death. These findings suggest that necrotic enlargement of cone cells is involved in the process of cone degeneration, and that necrosis may be a novel target to prevent or delay the loss of cone-mediated central vision in RP.

Acquired anhaptoglobinemia in panhypopituitarism.

This report describes the first case of acquired anhaptoglobinemia observed in panhypopituitarism (Sheehan's syndrome). Anhaptoglobinemia was completely reversed by the administration of hydrocortisone. These findings suggest that haptoglobin synthesis and/or secretion are modulated by hydrocortisone.

Protective effects of trifluoperazine on the microcirculation of cold-stored livers.

Previous studies have shown a protective effect of trifluoperazine (TFP), a calmodulin inhibitor, upon the microcirculation of cold-stored kidneys. The present study points to similar beneficial effects of TFP on the microcirculation of cold-stored livers; 25 canine livers were preserved for 24 hr with Euro-Collins' solution (EC) (n = 8), University of Wisconsin solution (UW) (n = 7), or UW + TFP (n = 10). The stored livers underwent heterotopic transplantation (HLTX); hepatic-artery and portal-vein pressure and flow were monitored; oxygen consumption and extraction were measured before HLTX and at 15-min intervals after reperfusion, for 1 hr. Mean hepatic-artery and portal-vein flow (HAF & PVF) prior to donor hepatectomy were 172 and 530 cc/min, respectively. Poor HAF and PVF occurred in EC-HLTX (mean 35, 175 cc/min, respectively). The damaged EC-flushed livers could not compensate to the decreased hepatic blood flow by increased oxygen extraction (oxygen consumption and extraction, 8.7 vol.% and 48%, respectively). Light and electron microscopy showed severe liver necrosis and periportal hemorrhages. Improved hepatic-artery and portal-vein flows were seen in UW HLTX (105 and 254 cc/min), and oxygen consumption and extraction were 16.4 vol.% and 66%, respectively. Liver biopsy taken just before reperfusion revealed well-preserved liver architecture. Liver biopsy obtained 1 hr after reperfusion revealed marked edema of the portal triad, sinusoid congestion, and hemorrhage. Electron-microscopy biopsies obtained during reperfusion at 15-min intervals revealed severe vasospasm of the terminal hepatic arterioles and progressive damage to the liver microcirculation. The addition of TFP to the UW-flush solution resulted in excellent protection of the liver microcirculation. Marked increase in hepatic-artery and portal-vein blood flow was noted after reperfusion (mean 167 and 421 cc/min, respectively (P 0.02 vs. UW: P 0.001 vs. EC). The recovery of metabolic activity was evident by the high oxygen consumption and extraction (25.8 vol.% and 80%, respectively). And serial liver biopsies obtained after reperfusion have shown excellent protection of liver architecture and the absence of hepatic arteriolar vasospasm. Taken together, these data suggest that the addition of TFP to the UW solution protects the liver microcirculation by rendering the hepatic microcirculation insensitive to vasospastic stimuli during reperfusion, thus permitting better metabolic recovery after transplantation.

Acute rejection of orthotopic corneal xenografts in mice depends on CD4(+) T cells and self-antigen-presenting cells.

PURPOSE: Guinea pig corneal xenografts have been reported to be rejected acutely in eyes of normal adult mice. Rejection of this type is independent of xenoreactive antibodies, and mice deficient in CD8(+) and NK T cells are unable to reject guinea pig corneal grafts acutely. Therefore, a study was conducted to determine the extent and manner by which CD4(+) T cells are responsible for rejection of orthotopic corneal xenografts. METHODS: Xenogeneic corneas were prepared from eyes of normal guinea pigs and grafted orthotopically into normal eyes of C57BL/6 mice, class II major histocompatibility complex (MHC) knockout (KO) mice, and class II MHC KO mice reconstituted with syngeneic (C57BL/6) CD4(+) T cells and/or bone marrow cells. Graft survival was assessed clinically, and success of cellular reconstitution was assayed using flow cytometric analysis of peripheral blood leukocytes. T cells from rejector mice were analyzed for proliferative responses to guinea pig xenoantigens in vitro. RESULTS: Median survival times (MST) of corneal xenografts in MHC class II KO mice was significantly delayed (31 days) compared with grafts in wild-type C57BL/6 eyes (9 days). Acute rejection was restored almost completely when MHC class II KO mice were reconstituted simultaneously with C57BL/6 bone marrow and CD4(+) T cells, but not when the KO mice were reconstituted with either CD4(+) T cells or bone marrow cells alone. Mice that rejected guinea pig corneas possessed only CD4(+) T cells capable of responding to guinea pig xenoantigens in vitro. CONCLUSIONS: Acute rejection of orthotopic corneal xenografts in mice is mediated by CD4(+) T cells that detect guinea pig xenoantigens that are presented on MHC class II(+) syngeneic antigen-presenting cells. These results strongly suggest that rejection occurs exclusively through the indirect pathway of T-cell activation.

Inhibition of corneal inflammation by the topical use of Ras farnesyltransferase inhibitors: selective inhibition of macrophage localization.

PURPOSE: Ras farnesyltransferase inhibitors are known to block the membrane translocalization of oncogenic Ras protein. They inhibit the cytoplasmic mitogen-activated protein kinase signaling cascade related to Ras protein. Thus far, Ras farnesyltransferase inhibitors have been exclusively regarded with the anticancer drugs. The object of this study was to elucidate the role of Ras farnesyltransferase inhibitors on the corneal opacity induced by an inflammatory stimulus. METHODS: We used a cauterization-induced corneal inflammation model. The central corneas of BALB/c mice were cauterized with silver nitrate (1 mm in diameter). Ras farnesyltransferase inhibitors, either manumycin or gliotoxin eye drops (each drug dissolved in balanced salt solution [BSS] at concentrations of 1 mM), were topically delivered to the cauterized cornea every 8 hours; BSS eye drops were used as a control. Clinical signs such as corneal edema, opacity, and corneal neovascularization, which are major causes of visual disturbance, were then examined 96 hours after the cauterization. The corneal edema and opacity were clinically scored under a stereoscopic microscope. The corneal neovascularization was evaluated by the length of the blood vessels from the limbus and the sum of extension central angle of vascularized limbus. Furthermore, the corneas were examined histologically, and the phenotypes of the cornea-infiltrating cells were analyzed by flow cytometry. RESULTS: The control corneas showed prominent edema, neovascularization, and opacity. Histologic analysis revealed corneal epithelial and endothelial cell loss and a large amount of inflammatory cell infiltration into the corneal stroma. Flow cytometric analysis revealed that most of the infiltrating cells were neutrophils and macrophages. In contrast, the degree of corneal edema, neovascularization, and opacity was significantly less in the manumycin- or gliotoxin-treated corneas than in the control corneas. Histologically, the manumycin- and gliotoxin-treated corneas showed minimum edema and good epithelialization. Flow cytometric analysis showed corneal infiltration of macrophages to be selectively and clearly inhibited. Neither manumycin nor gliotoxin produced any side effects in the noncauterized normal cornea either clinically or histologically. CONCLUSIONS: Ras proteins play an important role in cauterization-induced corneal inflammation and the opacity it induces. Ras farnesyltransferase inhibitors thus have a great potential for improving the treatment of corneal opacity induced by a corneal inflammatory stimulus.

The promotion of invasion through the basement membrane of cervical carcinoma cells by fibronectin as a chemoattractant.

An in vitro migration and invasion assay was used as the model system to study the effect of 3T3 fibroblast conditioned medium (FCM) and purified human fibronectin on the invasion of cervical carcinoma cells. The 3T3 FCM significantly enhanced both the migration and the invasion of a cervical carcinoma cell line, HeLa. This enhancement of migration and invasion was inhibited by anti-fibronectin antibody. Purified fibronectin alone enhanced the invasion in a dose-dependent manner for all cervical carcinoma cell lines, HeLa, CAC-1 and TMCC. The pretreatment of cells with cell binding aminosequences, GRGDSP and/or YIGSR blocked the enhancement of cell invasion. The implication of these findings for the invasion of cervical carcinoma is discussed.

Correlation between reduced p21(WAF1/CIP1) expression and abnormal p53 expression in esophageal carcinomas.

p21(WAF1/CIP1) is a potent inhibitor of various cyclin-dependent kinases, the expression of which is transcriptionally regulated by tumor suppressor gene product p53. We immunohistochemically examined the expression of p53 and p21(WAF1/CIP1) in 61 esophageal squamous cell carcinomas. p53 protein was expressed in 37 (61%) of 61 carcinomas. p21(WAF1/CIP1) was consistently expressed in the normal stratified esophageal mucosa. In the carcinomas, the expression of p21(WAF1/CIP1) protein was markedly reduced or not expressed in 33 (54%) cases. Clinicopathologic analyses revealed that no significant correlation exists either between p53-positive and -negative cases or between p21(WAF1)/(CIP1)-positive and -negative cases. Twenty-four cases were p53-positive/p21(WAF1/CIP1) negative, 15 were p53-negative/p21(WAF1/CIP1)-positive, 13 were positive for both and 9 were negative for both, and these findings thus showed an inverse correlation of the positivity between p53 and p21(WAF1/CIP1) (p<0.05). Furthermore, of the 13 cases with positive staining for both, the distribution of the expression was mutually exclusive in 6 cases and coincidental in the remaining 7 cases. These findings showed the p53-dependent expression of p21(WAF1/CIP1) was observed in esophageal squamous cell carcinomas, while the lack of an absolute correlation between abnormal p53 expression and p21(WAF1/CIP1) expression suggested that the p53-independent expression of p21(WAF1/CIP1) might also occur in some portions of the esophageal squamous carcinomas, suggesting that the molecular mechanisms of esophageal carcinogenesis appear to be complicated.