Granulocyte‐macrophage colony‐stimulating factor,a potent adjuvant for polarization to Th‐17 pattern: an experience on HIV‐1 vaccine model

Cytokines are mediators for polarization of immune response in vaccines. Studies show that co‐immunization of DNA vaccines with granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) can increase immune responses. Here, experimental mice were immunized with HIV‐1_{tat/pol/gag/env} DNA vaccine with GM‐CSF and boosted with recombinant vaccine. Lymphocyte proliferation with Brdu and CTL activity, IL‐4, IFN‐γ, IL‐17 cytokines, total antibody, and IgG1 and IgG2a isotypes were assessed with ELISA. Results show that GM‐CSF as adjuvant in DNA immunization significantly increased lymphocyte proliferation and IFN‐γ cytokines, but CTL response was tiny increased. Also GM‐CSF as adjuvant decreased IL‐4 cytokine vs mere vaccine group. IL‐17 in the group that immunized with mixture of DNA vaccine/GM‐CSF was significantly increased vs DNA vaccine group. Result of total antibody shows that GM‐CSF increased antibody response in which both IgG1 and IgG2a increased. Overall, results confirmed the beneficial effect of GM‐CSF as adjuvant to increase vaccine immunogenicity. The hallmark result of this study was to increase IL‐17 cytokine with DNA vaccine/GM‐CSF immunized group. This study for the first time provides the evidence of the potency of GM‐CSF in the induction of IL‐17 in response to a vaccine, which is important for control of infection such as HIV‐1.     

Etiologic and Epidemiologic Pattern of Urolithiasis in North Iran;Review of 10-Year Findings

Objective: To determine epidemiologic and metabolic characteristics of renal stone in the northern Iran. Methods: We prospectively analyzed demographic, clinical and metabolic findings in children less than 16 years old with renal stone revealed by ultrasonography from September 2003 to May 2012. Evaluations included serum and urine measurement of main elements predisposing patients to stone formation. Findings : 271 children (160 males) aged 2 months to 16-years (mean 30 months) were evaluated. 91 (33.6%) had a positive family history, abdominal discomfort (18.8%), UTI (11.8%) and hematuria (11.4%) were main presenting features. 45 children were diagnosed accidentally without any specific compliant. Nearly all (99%) stones lay in kidney., 35.1% had metabolic, 10% infective and 4.1% obstructive trends, 110 children had no definable etiology. Hypercalciuria (25.5%) hyperoxaluria (18.4%) and hypocitraturia (18.1%) were more frequent than uricosuria (8.5%) and cystinuria (3.1%) Conclusion: Metabolic derangement plays significant role in stone formation in our area. Patients should be carefully evaluated considering this point of view.Key Words: Nephrolithiasis, Kidney Stone, Hypercalciuria, Hyperoxaluria, Cystinuria, Hypocitraturia     

Cell surface engineering using glycosylphosphatidylinositol anchored tissue inhibitor of matrix metalloproteinase‐1 stimulates cutaneous wound healing

The balance between matrix metalloproteinases and their endogenous tissue inhibitors (TIMPs) is an important component in effective wound healing. The biologic action of these proteins is linked in part to the stoichiometry of TIMP/matrix metalloproteinases/surface protein interactions. We recently described the effect of a glycosylphosphatidylinositol (GPI) anchored version of TIMP‐1 on dermal fibroblast biology. Here, cell proliferation assays, in vitro wound healing, electrical wound, and impedance measurements were used to characterize effects of TIMP‐1‐GPI treatment on primary human epidermal keratinocytes. TIMP‐1‐GPI stimulated keratinocyte proliferation, as well as mobilization and migration. In parallel, it suppressed the migration and matrix secretion of dermal myofibroblasts, and reduced their secretion of active TGF‐β1. Topical application of TIMP‐1‐GPI in an in vivo excisional wound model increased the rate of wound healing. The agent positively influenced different aspects of wound healing depending on the cell type studied. TIMP‐1‐GPI counters potential negative effects of overactive myofibroblasts and enhances the mobilization and proliferation of keratinocytes essential for effective wound healing. The application of TIMP‐1‐GPI represents a novel and practical clinical solution for facilitating healing of difficult wounds.     

Optimal parameters for the clinical test of dynamic visual acuity in patients with a unilateral vestibular deficit

OBJECTIVE: To determine the influence of frequency and direction of head movement and type of vision chart on the score of a clinical test of dynamic visual acuity (DVA). METHODS: The subjects were 31 healthy individuals (22 to 79 years old) and 10 patients (19 to 70 years old) with a unilateral vestibular deficit owing to surgical resection of an acoustic neuroma. They read a Snellen or an E-chart while their head was passively moved +/- 20 degrees back and forth in the horizontal or vertical direction at one of four frequencies (0.5, 1.0, 1.5, and 2.0 Hz). The DVA score was the difference in the number of lines on the vision chart that could be read with the head passively moved versus with the head immobile. RESULTS: Four healthy subjects had a low DVA score during horizontal head movements at the fastest frequency (2.0 Hz) with the Snellen chart. In patients, DVA scores significantly decreased as head movement frequency increased from 0.5 to 1.0 Hz and from 1.0 to 1.5 Hz, during horizontal and vertical movements, and with both vision charts (p < .001). The DVA scores of healthy subjects were more consistent across three trials with the E-chart than with the Snellen chart at 1.0 and 0.5 Hz (horizontal movements, p < .01) and at 1.5 and 1.0 Hz (vertical movements, p < .01). CONCLUSIONS: This study provides new indications on the optimal parameters for the clinical test of DVA. From the results, it is recommended that DVA be tested during horizontal and vertical head movements at a frequency of 1.5 Hz with the E-chart.     

Identification and quantification of antigens associated with HLA class I molecules on a bladder tumour cell line. Implications for vaccine design

The aim of this study was to isolate and identify antigenic peptides associated with HLA class I molecules on a bladder tumour cell line. HLA-A1 molecules were purified using an immunobead-purification technique and following elution of nonapeptides associated with the complex, their HPLC profile was determined by Tandem mass spectrometry (MS/MS). Three peptides were identified namely: (1) P991 (VTDPGNLLY); (2) P1041.5 (LTDLGFLVY), and (3) P1057.7 (LTDPHLLSY); these sequences matched elements of hepatitis B, MAGE1-A1 and herpes simplex viruses. These antigens had half-lives of approximately 120 min which is within the theoretical range of such short peptides. These results indicate that identification of MHC-associated peptides is possible without using an assay for cytotoxic T cells. Although this approach was applied on a relatively small scale, broader applications can be foreseen.     

Selective binding and presentation of CCL5 by discrete tissue microenvironments during renal inflammation

T cells are differentially recruited to the tubulointerstitium during renal inflammation. The selective presentation of chemokines by surface structures may in part underlie this phenomenon. In an attempt to better characterize the presentation of chemokines by tissue environments an exemplary chemokine with a well-defined structure was selected, and a binding assay for the protein on fixed archival tissue sections was developed. This article describes the selective binding of the chemokine CCL5 to renal structures. CCL5 was shown to bind to endothelial regions, interstitial extracellular matrix, tubular epithelial cells, and tubular basement membranes but rarely to glomerular structures in well-preserved kidneys. In contrast, binding of CCL5 to glomerular components was seen in renal biopsies with acute allograft glomerulitis (in which T cells accumulate in glomeruli). The N terminus mediates receptor binding, whereas two clusters of basic amino acid residues ((44)RKNR(47) and (55)KKWVR(59)) are involved in the presentation of CCL5 by extracellular structures. Mutation of either loop abrogated CCL5 binding to tissue sections. Variations of the N terminus and a mutation that prevents higher order oligomerization did not change the binding pattern. The data suggest that renal compartments differ in their capacity to present chemokines, which may help explain the differential recruitment of leukocytes during allograft injury. Both clusters of basic residues in CCL5 are necessary for sufficient binding of CCL5 to tissue sections.     

Behcet's disease in Iran: analysis of 6500 cases

Objective: To identify the clinical picture of Behcet’s disease in a large cohort of patients (6500) in Iran, over a period of 35 years, and compare them with other large series from around the world. Methods: Patients with Behcet’s disease from all over Iran were seen in the Behcet’s Disease Research Unit by a multidisciplinary team (rheumatologists, dermatologists, and ophthalmologists). Diagnosis was based on ‘expert opinion’. Data were collected on a standardized data sheet (105 items), and stored in an electronic database. Data were updated at each follow‐up. Results: Male to female ratio was 1.22 : 1.00. The mean age at onset was 26 years ± 11.3. The frequency of symptoms were: oral aphthosis 97.3%, genital aphthosis 64.6%, skin manifestations 64.9% (pseudofolliculitis 54.5%, erythema nodosum 22.5%, other lesions 7%), pathergy phenomenon 52.5%, ophthalmologic manifes‐tations 56.8% (anterior uveitis 41.2%, posterior uveitis 44.9%, retinal vasculitis 32.1%), joint manifestations 37.4% (arthralgia 17.2%, monoarticular arthritis 7.6%, oligoarthritis 16.8%, ankylosing spondylitis 2%), neurological manifestations 3.8% (central manifestations 3.5%, mononeuritis multiplex 0.3%), gastrointestinal manifestations 7.4%, vascular involvement 8.3% (phlebitis 5.7%, superficial phlebitis 2.2%, large vein thrombosis 1.1%, arterial thrombosis 0.154%, aneurysm 0.5%), epididymitis 4.7%, cardiac involvement 0.6%, and pulmonary involvement 0.9%. Sedimentation rate was normal in 46.5% of patients. Abnormal urine sediment was detected in 12.2%. HLA‐B5 was present in 53.3% and HLA‐B51 in 47.9% of patients. Conclusion: Behcet’s disease is mainly seen in young people. The most frequent symptoms are mucocutaneous, ocular and joint manifestations. Comparison with large series did not show major differences.     

Impact of the positive pathergy test on the performance of classification/diagnosis criteria for Behcet’s disease

Background

The only diagnostic test that currently exists for Behcet’s disease (BD) is the pathergy test. A positive pathergy test (PPT) is an important component of many of the 16 sets of classification/diagnosis criteria used to diagnose BD. The aim of this study was to determine the importance of a PTT in the performance of the diagnosis/classification criteria for BD.

Patients and methods

All patients listed in the BD registry of the Rheumatology Research Center, Tehran (6,727) and 4,648 BD controls were enrolled in the study. The diagnosis was clinical when no other diagnosis could explain the patient’s manifestations. The criteria were tested with and without PPT results. Sensitivity, specificity, and accuracy were calculated.

Results

Without PPT, all sets of criteria lost sensitivity, gained specificity, and lost accuracy, with the exception of the Cheng–Zhang criteria. The largest loss in sensitivity was for the Hubault–Hamza (35 %) and Dilsen (17.3 %) criteria; the least was for the Curth (1.9 %) and ICBD (6.5 %) criteria. The largest gain on specificity was for the Dilsen (4.7 %) and Curth (3.1 %) criteria; the least was for the Japan (0.1 %) and Japan revised (0.1 %) criteria. The greatest loss in accuracy was for the Hubault–Hamza (20.4 %) and Dilsen revised (9.3 %) criteria; the least was for the ICBD (3.6 %), while Curth gained 0.3 %.

Conclusion

Without PPT as a criterion for the diagnosis of BD, the sensitivity and accuracy of the sets of classification/diagnosis criteria decrease, while the specificity improves.