Long-term immunogenicity of a quadrivalent human papillomavirus vaccine in systemic lupus erythematosus.

OBJECTIVES: To evaluate the 5-year immunogenicity of a quadrivalent human papillomavirus (HPV) vaccine (GARDASIL) in patients with systemic lupus erythematosus (SLE). METHODS: Female SLE patients and controls, aged 18-35 years, who received GARDASIL in 2011 and sero-converted 12 months post-vaccination were followed for persistence of immunogenicity. Antibody measurement to HPV serotypes 6, 11, 16, 18 was repeated at 5 years. The rate of sero-reversion was compared between patients and controls, and factors associated with sero-reversion of the anti-HPV antibodies were studied. RESULTS: 50 SLE patients and 50 controls were vaccinated with GARDASIL. Among subjects who sero-converted at 1 year and consented for this study, antibodies to HPV serotypes 6, 11, 16 and 18 at 5 years were persistent in 24/27 (89%), 26/31 (84%), 32/34 (94%) and 24/25 (96%) of the SLE patients; and 32/33 (97%), 32/33 (97%), 32/32 (100%) and 23/24 (96%) of the controls, respectively. Antibody titers to HPV-6 and 16 were significantly lower in patients than controls. Seven (21%) SLE patients had sero-reversion of ≥1 anti-HPV antibodies. Sero-reverted patients experienced significantly more SLE flares, particularly renal, and had received significantly higher cumulative doses of prednisolone, mycophenolate mofetil and tacrolimus than those with persistent immunogenicity. The cumulative doses of prednisolone correlated inversely and significantly with the anti-HPV 6, 11, and 16 titers at 5 years. CONCLUSIONS: Immunogenicity of the quadrivalent HPV vaccine was retained in a high proportion of SLE patients at 5 year. Patients with more SLE renal flares and had received more immunosuppression were more likely to have sero-reversion of the anti-HPV antibodies. CLINICAL TRIAL REGISTRATION NUMBER: US ClinicalTrials.gov (NCT00911521 & NCT02477254).

Integrin α11ß1 regulates cancer stromal stiffness and promotes tumorigenicity and metastasis in non-small cell lung cancer.

Integrin α11ß1 is a stromal cell-specific receptor for fibrillar collagens and is overexpressed in carcinoma-associated fibroblasts (CAFs). We have investigated its direct role in cancer progression by generating severe combined immune deficient (SCID) mice deficient in integrin α11 (α11) expression. The growth of A549 lung adenocarcinoma cells and two patient-derived non-small cell lung carcinoma (NSCLC) xenografts in these α11 knockout (α11(-/-)) mice was significantly impeded, as compared with wild-type (α11(+/+)) SCID mice. Orthotopic implantation of a spontaneously metastatic NCI-H460SM cell line into the lungs of α11(-/-) and α11(+/+) mice showed significant reduction in the metastatic potential of these cells in the α11(-/-) mice. We identified that collagen cross-linking is associated with stromal α11 expression, and the loss of tumor stromal α11 expression was correlated with decreased collagen reorganization and stiffness. This study shows the role of integrin α11ß1, a receptor for fibrillar collagen in differentiation of fibroblasts into CAFs. Furthermore, our data support an important role for α11 signaling pathway in CAFs, promoting tumor growth and metastatic potential of NSCLC cells and being closely associated with collagen cross-linking and the organization and stiffness of fibrillar collagen matrices.

Referral strategy for early recognition of axial spondyloarthritis: consensus recommendations from the Hong Kong Society of Rheumatology.

Low back pain is one of commonest problems prompting a visit to the family physician. Up to 5% of patients with chronic low back pain in the primary care setting are diagnosed as having spondyloarthritis, which includes the prototype disease ankylosing spondylitis. Making a diagnosis of ankylosing spondylitis is often delayed for years, leading to significant pain, impairment of quality of life, disability and productivity loss. A recent breakthrough in the treatment of spondyloarthritis is the anti-tumor necrosis factor-alpha biologics, which lead to rapid relief of pain and inflammation, and improvement in all clinical parameters of the disease. Patients with early spondyloarthritis often respond better than those with late established disease. With proper recognition of inflammatory back pain, and the use of magnetic resonance imaging, spondyloarthritis can now be diagnosed much earlier before features are evident on plain radiographs. Referral to the rheumatologist based on onset of back pain (> 3 months) before the age of 45 years, and an inflammatory nature of the pain, or the presence of human leukocyte antigen-B27, or sacroiliitis by imaging, have been confirmed in multi-center international studies to be a pragmatic approach to enable early diagnosis of spondyloarthritis. This referral strategy has recently been adopted by the Hong Kong Society of Rheumatology for primary care physicians and non-rheumatology specialists.

Combined low-dose mycophenolate mofetil and tacrolimus for lupus nephritis with suboptimal response to standard therapy: a 12-month prospective study.

OBJECTIVE: The objective of this paper is to evaluate the efficacy of combined mycophenolate mofetil (MMF) and tacrolimus (TAC) for lupus nephritis with suboptimal response to standard therapy. METHODS: Inclusion criteria for patients: (1) biopsy-confirmed active lupus nephritis; and (2) inadequate response to ≥ 2 immunosuppressive regimens. While prednisolone (≤ 10 mg/day) and angiotensin-converting enzyme inhibitors were continued, immunosuppressive agents were replaced by combined MMF (1 g/day) and TAC (4 mg/day). Patients were followed every 2 months for the clinical response and adverse events at 12 months. RESULTS: Twenty-one patients were recruited (20 women; age 35.8 ± 9.2 years; systemic lupus erythematosus (SLE) duration 111 ± 51 months). The histological classes of lupus nephritis were: IV/III (33%), V+III/IV (33%) and pure V (33%). The creatinine clearance (CrCl), urine protein-to-creatinine ratio (uP/Cr) and serum albumin was 82.4 ± 33 ml/min (<90 ml/min in 57%), 3.27 ± 1.5 and 30.1 ± 5.9 g/l, respectively. Thirteen (62%) patients had active urinary sediments and 17 (81%) patients had active lupus serology. At 12 months, eight (38%) patients had very good response, one (5%) patient had good response and five (24%) patients had partial response. Significant improvement in uP/Cr, albumin, complement C3 and anti-dsDNA titer, and stabilization of CrCl was observed in the responders. Thirty-three adverse events were reported in 18 patients: major infection requiring hospitalization (6%), infection not requiring hospitalization (27%), herpes infection (9%), diarrhea (12%), cramps (9%), dyspepsia (6%), transient increase in serum Cr (6%), alopecia (4%), facial twitching (3%), tremor (3%) and diabetes mellitus (3%). None of these had led to protocol withdrawal. CONCLUSIONS: Combined low-dose MMF and TAC is an option for lupus nephritis that fails to respond adequately to standard regimens, with two-thirds of patients improving after 12 months. Longer-term observation is needed to confirm its efficacy and safety.

Changes in body composition after glucocorticoid therapy in patients with systemic lupus erythematosus.

The aim of this study was to evaluate the changes in body composition after glucocorticoid treatment in patients with systemic lupus erythematosus (SLE). Consecutive SLE patients were recruited for serial measurements (baseline, months 2 and 6) of bone mineral density (BMD) and body composition [bone mineral content (BMC), fat and lean mass] by dual energy X-ray absorptiometry scan after high-dose oral glucocorticoid therapy. Factors correlated with changes in body composition were evaluated. 29 SLE patients were studied (age 39.7 +/- 11.5 years; 83% women with 29% postmenopausal; SLE duration 80.1 +/- 80 months). Fourteen patients (48%) were glucocorticoid-naive. The mean maximum daily dosage of prednisolone was 32.9 +/- 6.5 mg and the cumulative prednisolone dosage in 6 months was 2.7 +/- 0.7 g. At 6 months, a significant drop in BMC of the trunk (-5.0 +/- 2.2%; P = 0.04) and whole body (-1.2 +/- 0.4%; P = 0.002) compared with baseline was observed, and so was the BMD of the hip (-1.7 +/- 0.6%; P = 0.006) and whole body (-0.7 +/- 0.3%; P = 0.01). A significant increase in the fat mass of the trunk (+14.5 +/- 4.1%; P = 0.001) and limbs (+10.0 +/- 3.2%; P = 0.004), but a non-significant drop in lean mass of the trunk (-3.3 +/- 1.8%; P = 0.08) and limbs (-0.8 +/- 2.4%; P = 0.75) also occurred. The changes in whole body BMC correlated significantly with age (rho = -0.51; P = 0.02) and changes in total fat mass (rho = 0.44; P = 0.02) but not with lean mass (rho = -0.21; P = 0.27), gender, body mass index, smoking, prednisolone dosages or changes in BMD. In SLE patients, high-dose glucocorticoids lead to an early and rapid drop in bone mass, which is more serious in older patients and correlates with an increase in body fat.

A new strategy to replace the natural vitreous by a novel capsular artificial vitreous body with pressure-control valve.

PURPOSE: The current vitreous substitutes such as silicone oil, heavy silicone oil, and polymeric gels that are directly injected into vitreous cavity frequently cause severe intraocular complications. There is a very urgent need to find a more suitable artificial vitreous substitute for pars plana vitrectomy (PPV) surgery. METHODS: We have devised a novel capsular artificial vitreous using tailor-made silicone rubber elastomer. The novel device was implanted into the vitreous cavity of rabbit after PPV and the eye was examined by ophthalmoscopy, fundus photography, and tonometry during an 8-week treatment period. B-scan ultrasonography, electroretinogram (ERG), and histological studies by light microscopy were also performed at the end of 8 weeks. RESULTS: The novel artificial vitreous body consists of a thin vitreous-like capsule with a silicone tube-valve system. The capsule can be folded and implanted into vitreous cavity through 1.5 mm incision on sclera. Physiological balanced solution (PBS) was then injected into the capsule and inflated to support retina and control intraocular pressure (IOP) through the tube-valve system subsequently fixed under the conjunctiva. Experiments using rabbits showed that the novel vitreous body could effectively support the retina and apparently induced no significant pathological changes in the eye over 8 weeks. CONCLUSION: This approach may provide a new research strategy in the vitreous replacement technology. The novel artificial vitreous body device can effectively support retina, control IOP, and has good biocompatibility. It may be a good alternative to injecting artificial vitreous although its tamponade properties and usefulness still have to be proven in complex vitreoretinal diseases.

Is antibody clustering predictive of clinical subsets and damage in systemic lupus erythematosus?

OBJECTIVE: To examine autoantibody clusters and their associations with clinical features and organ damage accrual in patients with systemic lupus erythematosus (SLE). METHODS: The study group comprised 1,357 consecutive patients with SLE who were recruited to participate in a prospective longitudinal cohort study. In the cohort, 92.6% of the patients were women, the mean +/- SD age of the patients was 41.3 +/- 12.7 years, 55.9% were Caucasian, 39.1% were African American, and 5% were Asian. Seven autoantibodies (anti-double-stranded DNA [anti-dsDNA], anti-Sm, anti-Ro, anti-La, anti-RNP, lupus anticoagulant (LAC), and anticardiolipin antibody [aCL]) were selected for cluster analysis using the K-means cluster analysis procedure. RESULTS: Three distinct autoantibody clusters were identified: cluster 1 (anti-Sm and anti-RNP), cluster 2 (anti-dsDNA, anti-Ro, and anti-La), and cluster 3 (anti-dsDNA, LAC, and aCL). Patients in cluster 1 (n = 451), when compared with patients in clusters 2 (n = 470) and 3 (n = 436), had the lowest incidence of proteinuria (39.7%), anemia (52.8%), lymphopenia (33.9%), and thrombocytopenia (13.7%). The incidence of nephrotic syndrome and leukopenia was also lower in cluster 1 than in cluster 2. Cluster 2 had the highest female-to-male ratio (22:1) and the greatest proportion of Asian patients. Among the 3 clusters, cluster 2 had significantly more patients presenting with secondary Sjögren's syndrome (15.7%). Cluster 3, when compared with the other 2 clusters, consisted of more Caucasian and fewer African American patients and was characterized by the highest incidence of arterial thrombosis (17.4%), venous thrombosis (25.7%), and livedo reticularis (31.4%). By using the Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index, the greatest frequency of nephrotic syndrome (8.9%) was observed in patients in cluster 2, whereas cluster 3 patients had the highest percentage of damage due to cerebrovascular accident (12.8%) and venous thrombosis (7.8%). Osteoporotic fracture (11.9%) was also more common in cluster 3 than in cluster 2. CONCLUSION: Autoantibody clustering is a valuable tool to differentiate between various subsets of SLE, allowing prediction of subsequent clinical course and organ damage.

High-throughput array production using precision glass syringes.

The advantages of using 1, 96, or 384 precision glass syringes in automated high-throughput microdispensers in creating highly uniform and reproducible DNA, protein, and organic compound array filters and slides are described. Using the Hydra Microdispenser and Tango Liquid Handling system, 0.1-5 ng (in 50-300 nL) PCR-amplified, human cancer-related genes and housekeeping genes were spotted onto nylon membranes and coated slides. Protein solutions of 50 microg/mL to 1 mg/mL were spotted onto coated slides or onto MaxiSorp 96-well plates. Up to 6144 spots/membrane and up to 1000 spots/slide were printed. The size of the spots created by glass syringes was uniform and reproducible (precision variation of less than 5%) from spot to spot and membrane to membrane. Using a Tango 384 system, a total of ten 6144-spot filters can be produced in approximately 25 min, translating into a spotting speed of 2.5 min/membrane.

A YRBS survey of youth risk behaviors at alternative high schools and mainstream high schools in Hong Kong.

In Hong Kong, prevocational schools serve as an alternative to mainstream schools to provide education with more emphasis on practical and technical subjects. In this paper, health-risk behaviors of prevocational school (PVS) students were identified, and comparisons of health-risk behaviors with or without adjusting the demographic factors from prevocational schools and mainstream schools were made. The PVS students were at higher risk for most categories of health-risk behaviors such as unintentional and intentional injuries, smoking, alcohol drinking, glue sniffing, inadequate physical activity, insufficient consumption of fresh fruits and vegetables, and early sexual activity with multiple partners. Female students of PVS reported higher prevalence of emotional problems and substance abuse. Findings suggest that the school environment is an influential factor on the lifestyle behavior of students. Comprehensive health education and intervention programs are needed for youth in Hong Kong prevocational schools.

Electrophysiology and glucose transport of human peritoneal mesothelial cells: implications for peritoneal dialysis.

OBJECTIVE: To elucidate ionic and glucose transport across human peritoneal mesothelium, we utilized an Ussing chamber setup and studied the electrophysiological characteristics and tissue permeabilities of human peritoneal mesothelial cells (HPMC) to L- and D-glucose. METHODS: Human mesothelial cells were grown on polyester filters (snapwell; Costar, Cambridge, MA, U.S.A.) that, upon confluence, were fitted into Ussing chambers. Transmesothelial resistance and resting potential were determined using electrophysiological techniques. Radiolabeled glucose was added to one side of the chamber and the permeabilities determined by serial sampling in the receptive compartment. RESULTS: The transmesothelial potential and resistance were 0.54 +/- 0.07 mV (apical positive) and 20.4 +/- 3.2 ohms x cm2 respectively (mean +/- SEM, n = 36). The course of overall transfer of D- and L-glucose was examined using L-glucose as a positive diffusion-plus-leak marker. The permeabilities of HPMC to D-glucose were 3.00 +/- 0.26 cm/sec (apical-to-basolateral) and 3.25 +/- 0.27 cm/sec (basolateral-to-apical) [n = 6 experiments, p = not significant (NS)], which were not different from those of L-glucose: 3.00 +/- 0.30 cm/sec (apical-to-basolateral) and 2.71 +/- 0.24 (basolateral-to-apical) (n = 6 experiments, p = NS). CONCLUSIONS: The transepithelial resistance of HPMC is low and the ionic gradient, although it exists, is small and inconsequential. Passive paracellular flow accounts for the majority of transmesothelial glucose transport. The existence of a large paracellular shunt precludes the mesothelial membrane as a clinically relevant osmotic barrier.

Model of ionic transport for bovine ciliary epithelium: effects of acetazolamide and HCO.

The possible existence of transepithelial bicarbonate transport across the isolated bovine ciliary body was investigated by employing a chamber that allows for the measurement of unidirectional, radiolabeled fluxes of CO2 + HCO. No net flux of HCO was detected. However, acetazolamide (0.1 mM) reduced the simultaneously measured short-circuit current (I(sc)). In other experiments in which (36)Cl- was used, a net Cl- flux of 1.12 microeq. h(-1). cm(-2) (30 microA/cm(2)) in the blood-to-aqueous direction was detected. Acetazolamide, as well as removal of HCO from the aqueous bathing solution, inhibited the net Cl- flux and I(sc). Because such removal should increase HCO diffusion toward the aqueous compartment and increase the I(sc), this paradoxical effect could result from cell acidification and partial closure of Cl- channels. The acetazolamide effect on Cl- fluxes can be explained by a reduction of cellular H+ and HCO (generated from metabolic CO2 production), which exchange with Na+ and Cl- via Na+/H+ and Cl-/HCO exchangers, contributing to the net Cl- transport. The fact that the net Cl- flux is about three times larger than the I(sc) is explained with a vectorial model in which there is a secretion of Na+ and K+ into the aqueous humor that partially subtracts from the net Cl- flux. These transport characteristics of the bovine ciliary epithelium suggest how acetazolamide reduces intraocular pressure in the absence of HCO transport as a driving force for fluid secretion.

A rapid method of preparing the plasma membrane enriched fraction of the bovine retinal pigment epithelial cells.

PURPOSE: To devise a rapid method of isolating the plasma membrane enriched fraction (PMEF) of the bovine retinal pigment epithelial (RPE) cells with Percoll centrifugation medium. METHODS: The bovine RPE was homogenised with a tight fit Dounce homogeniser and centrifuged in a 16.7% Percoll gradient for 20 minutes. The RPE particulate fractions were characterised in terms of their protein concentrations, Na/K-ATPase and bicarbonate stimulated ATPase activities. RESULTS: The total protein recovery was 88.7% of the RPE homogenate. The nucleus layer was identified at the first band. The mitochondrial fraction was at the second layer according to its bicarbonate stimulated ATPase activity. The 3rd and 4th bands were enriched with plasma membranes and their Na/K-ATPase activities were 31.5 and 34.6 mumol/mg/h respectively. The Na/K-ATPase activities were about six times that of the RPE homogenate. CONCLUSIONS: A rapid method of isolating the bovine RPE PMEF has been devised which involved a single centrifugation procedure in a Percoll gradient.

Differential expression of Met/hepatocyte growth factor receptor in subtypes of non-small cell lung cancers.

Hepatocyte growth factor (HGF)/scatter factor (SF) is a multifunctional factor that stimulates epithelial cell motility, invasion and morphogenesis. Its receptor is a transmembrane tyrosine kinase encoded by the Met proto-oncogene. Several studies have suggested a possible role for HGF/Met in tumor development and progression. To investigate the potential roles of Met in human lung cancer biology, we have studied the mRNA and protein expression of Met in normal lung tissue, primary non-small cell lung carcinoma (NSCLC), and NSCLC cell lines. The results indicated a differential pattern of Met expression among various subtypes of NSCLC. The majority of squamous cell carcinoma (SQCC), either in vivo or in vitro, expressed Met mRNA and its protein product at levels much lower than or similar to normal lung tissue or bronchial epithelium. Moreover, SQCC characteristically over-expressed a variant Met mRNA which corresponds to a 5' partially deleted transcript produced by alternative splicing. In contrast, the expression of Met mRNA and its protein product in adenocarcinoma (ADC) and large cell undifferentiated carcinoma were more heterogeneous. Overexpression was demonstrated in approximately 35% and 20% of these subtypes of NSCLC, respectively. Among ADC, intermediate to high levels of Met immunoreactivity correlated with greater degree of tumor differentiation. Furthermore, an accentuation of Met immunoreactivity was often noted in cancer cells at the advancing edge of tumors. These findings support a role for Met in lung cancer cell invasion and differentiation in vivo, but its expression and functions may be modified by the differentiation phenotype of the tumor cells.

The roles of hepatocyte growth factor/scatter factor and met receptor in human cancers (Review).

The protein product of c-met proto-oncogene, Met, is a tyrosine kinase receptor for the hepatocyte growth factor (HGF), also known as the scatter factor (SF). HGF/SF-Met signaling has multifunctional effects on mammalian cells. These include stimulation or inhibition of cellular proliferation, promotion of cell movement, invasion into extracellular matrix, and induction of glandular/tubular morphogenesis by epithelial cells. There is a substantial body of experimental evidence that supports the oncogenic role of HGF/SF-Met signaling pathways. This is putatively mediated by autocrine or paracrine mechanisms that promote tumor cell growth, invasion and angiogenesis. We review the evidence that HGF/SF and Met receptor play significant roles in the pathogenesis and biology of human cancers.