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Objective:Generation and identification of hybridoma to produce monoclonal antibody against MICA/B. Methods:SP2/0 cells were fused with murine splenocyte immunized with recombinant protein rMICA?012 to get hybridoma cell lines. The titer of the monoclonal antibody produced by 9B10 cell line was determined by ELISA and its specificity was tested by Western blot,Luminex mutiplex microsphere-based immunoassay and immunofluorescence assay. Results:Six hybridoma cell lines were selected by ELISA screening test. The minimum reaction concentration of mAb 9B10 was 0. 02 ng/μl,and the specificity of mAb 9B10 was determinated by Western blot,Luminex mutiplex microsphere-based immunoassay and immunofluorescence. Conclusion:The monoclonal antibody 9B10 was available to apply for the detection of MICA and MICB expression.
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Objective:Generation and identification of hybridoma to produce monoclonal antibody against MICA/B. Methods:SP2/0 cells were fused with murine splenocyte immunized with recombinant protein rMICA?012 to get hybridoma cell lines. The titer of the monoclonal antibody produced by 9B10 cell line was determined by ELISA and its specificity was tested by Western blot,Luminex mutiplex microsphere-based immunoassay and immunofluorescence assay. Results:Six hybridoma cell lines were selected by ELISA screening test. The minimum reaction concentration of mAb 9B10 was 0. 02 ng/μl,and the specificity of mAb 9B10 was determinated by Western blot,Luminex mutiplex microsphere-based immunoassay and immunofluorescence. Conclusion:The monoclonal antibody 9B10 was available to apply for the detection of MICA and MICB expression.
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Background The pathological change of the anterior uveitis is the disruption of blood-aqueous barrier.Slit lamp examination appears to be limited for the evaluation of inflammatory response,and fluorescine angiography is an objective approach.However,there are few relative studies up to now in China.Objective Aim of this study was to observe the characteristics and assess the clinical applications of iris fluorescein angiography (IFA)in Chinese uveitis with brown iris.Methods Forty eyes of 40 normal subjects and 21 eyes of 13 patients with the anterior uveitis were collected in this study.IFA,slit-lamp examination and iris photograph were performed on the subjects.All individuals were informed consented at the initiation of this study.Results In normal eyes,fluorescence in iris vessels was blocked by the melanin pigment,but peripupillary weak fluorescent leakage was seen in the normal eyes with the age of ＞60 years old.The multiple patterns of fluorescence leakage were found in the patients suffered from uveitis of various etiologies although the negative slit-lamp finding,including the leakage of fluorescein around the pupillary margin and radial iris vessels in the eyes with mild diseases,transmitted fluorescence of regular iris vessels in the eyes with diffuse and local iris atrophy,and vascular tufts of the pupillary margin with coiled interwind tight clusters of thin vessels at the early phase in the eyes with dilated capillaries,microvascular anomalies and new vessel formation.The hyperfluorescence remained throughout the IFA duration.Conclusions IFA findings in uveitis vary depending on the topography,type and severity of inflammation.IFA has a good clinical applying value because of its objective assessment ability of the degree of the blood-aqueous barrier breakdown and iris neovascularization breakdown.It can exhibit the unvisible lesion under the slit-lamp and monitor the efficacy of medical theraphy in patients with active or quiescent uveitis.
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Background Researches showed that multifocal electroretinogram (mfERG) is able to assess the retinal function in the eyes with acute Vogt-Koyanagi-Harada ( VKH ) syndrome. But the mfERG characteristics of convalescence stage of VKH are still below clear. Objective Present study was to compare and follow up the variation process of visual acuity and mfERG in acute and recovery stages of VKH syndrome. Methods This was a clinic-based retrospective study. Visual acuity, mfERG and fundus fluorescence angiography ( FFA ) were recorded from 35 eyes of 18 acute VKH cases. The period of follow-up in recovery stage lasted about 18 months with the repetitive recording results for 4 times. Results In this study, the visual acuity range in acute stage VKH was 0. 01 to 1.0, and 91.4% (32/35 eyes) was below 0.6. Compared with normal control group, the visual acuity was significantly decreased (P＜0.01). The response densities (amplitudes) of N1 ,P1 waves of the first-order kernel were significantly lowed in all the 6 rings,and the implicit times of 1-4 rings of both waves were significantly prolonged in acute VKH eyes(P＜0. 05). The abnormalities of retinal function showed a regional difference at the posterior pole retina with the dominant change in the first ring,showing a cutting off78% in the P1 amplitude. The abnormal degree of mfERG was more serious as the the increase of retinal eccentricity. In 2 months of convalescence after glucocorticosteroids therapy,the range of visual acuity were 0. 1-1.2 ,and the amplitudes of N1, P1 of 1-2 rings were greatly elevated in comparison with acute on-set (P＜0. 05 ). However, there was still a remarkable difference in the amplitudes of from 1 through 6 rings,comparing with normal. The response density of P1 wave from whole recording region was only 44% of normal. Though the visual acuity was stable during the follow-up duration, a decreasing tendency in N1 and P1 amplitudes were seen. The implicit times of both wave shortened only in 1-3 rings in recovery stages of VKH (P＜0.05). Conclusion VKH syndrome cause serious damage of posterior retinal function.Macular region is the site with greater retinal functional lesion and restore before and after medication. This hardly recovery of retinal function can last over one and half year,even satisfied visual acuity is stable after proper treatment.
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<p><b>BACKGROUND</b>There is no detailed report about the angiographic leakage of polypoidal choroidal vasculopathy (PCV) lesions on indocyanine green (ICG) angiography. This study aimed to investigate the angiographic leakage of polypoidal lesions in PCV on ICG angiography.</p><p><b>METHODS</b>One hundred and forty-four eyes of 137 patients diagnosed as PCV were prospectively observed. Fundus examination, fluorescein angiography, and ICG angiography were performed. Leakage of polypoidal lesions and clinical features were recorded according to the angiograms.</p><p><b>RESULTS</b>In all 144 eyes, 110 eyes showed angiographic leakage (leakage group) on ICG angiography and three subtypes of leakage group were noted, which were polypoidal dilations leakage (47 eyes, 42.7%), branching vascular networks leakage (14 eyes, 12.7%) and leakage of both (49 eyes, 44.5%). The other 34 eyes showed regression of polypoidal lesions (regression group). In leakage group, the rates of pigment epithelial detachment (PED), best corrected visual acuity (BCVA) < 0.1 and old subretinal hemorrhages were 56.4% (62 eyes), 19.1% (21 eyes), and 4.6% (5 eyes) respectively, compared with 8.8% (3 eyes), 50% (17 eyes) and 38.2% (13 eyes) of regression group (P < 0.001). The history of regression group was significantly longer (P < 0.001).</p><p><b>CONCLUSIONS</b>Angiographic leakage and regression can be observed in PCV lesions. Leakage of both polypoidal dilations and branching vascular networks is the most common subtype in leakage group. PCV in leakage group is more likely to be related to PED, better BCVA and shorter history, while PCV regression group tends to relevant to old subretinal hemorrhage, worse BCVA and longer history. This may reflect that the former is active or in the early course while the later is resting or in the late phase of PCV.</p>