Non-Paraneoplastic Autoimmune Retinopathy: The First Case Report in Korea

Autoimmune retinopathy (AIR) is an immune-mediated retinopathy, resulting from an immunologic process caused by the aberrant recognition of retinal antigens as autoantigens. The diagnosis of AIR involves the detection of antiretinal antibodies with concurrent clinical and electrophysiological evidence of retinopathy. A 40-year-old patient presented with progressive loss of bilateral vision over several months. A fundus examination was unremarkable. Spectral domain optical coherence tomography revealed a blurred photoreceptor ellipsoid zone at the subfoveal region in both eyes with more prominent disruption in the left eye. Full-field electroretinography (ERG) showed relatively normal rod and cone responses in the right eye, and decreased photopic bwaves with minimal attenuation of a-waves in the left eye. Multifocal ERG demonstrated slightly reduced amplitude of the inner segment ring in the right eye and decreased amplitudes and delayed latencies of all modalities in the left eye. The patient was suspected to have AIR and it was supported by positive Western blots for 23-kDa protein, enolase (46-kDa), aldolase (40-kDa), 62-kDa and 78-kDa proteins and by immunohistochemical staining of human retinal bipolar and ganglion cells. Despite the immunosuppressive treatment, the destruction of the retinal photoreceptors progressed, and immunosuppressive interventions produced very little visual improvement. We report on what is, to the best of our knowledge, the very first case of serologically confirmed nonparaneoplastic AIR in Korea.

Construction of CD19-CAR retroviral vector and modification of its transduction of human T-lymphocytes / 中华血液学杂志

<p><b>OBJECTIVE</b>To improve the MigR1-CD19-CAR (chimeric antigen receptor) that contains a single chain variable region (scFv) which targeted to CD19 through a retroviral vector transduction efficiency of T-lymphocytes.</p><p><b>METHODS</b>Insert the CD19-CAR fragment into the retroviral vector (MigR1) through recombinant DNA technology, after transfecting plat-A packaging cell lines, viral supernatant was collected to transduce K562 cell line and activated human T-lymphocytes. We used flow cytometry to determine the transduction efficiency and RT-PCR to confirm the transcription of CD19-CAR gene. The ability of the transduced T cells to produce IFN-γ and TNF-α in a CD19-specific manner was measured in an enzyme-linked immunosorbent (ELISA) assay.</p><p><b>RESULTS</b>(1)Using MigR1-CD19-CAR retroviral vector to produce the high titer retrovirus. (2)MigR1-CD19-CAR transduction efficiency of K562 cell line was significantly higher than human T-lymphocytes (P<0.01). (3)120 min centrifugation could significantly improve transduction efficiency of T-lymphocytes to (54.5±14.6)%. (4)Transduction efficiency could be improved by deciding transduce time according to T-lymphocytes proliferation fold in vitro individually, and the highest transduction efficiency in the study was 69.3%. The CD19-CAR gene sequence was transcripted specificly with high efficiency. (5) IFN-γ and TNF-α released by CD19-CAR transduced T-lymphocytes significantly increased to (13 230±1 543) pg/ml and (4 217±211) pg/ml when coculture with CD19-K562 cells.</p><p><b>CONCLUSION</b>We have successfully constructed a second generation CAR which targeted to CD19 through a retroviral vector called MigR1 (MigR1-CD19-CAR). Deciding transduce time according to T-lymphocytes proliferation fold in vitro individually and 120 min centrifugation could improve the CAR transduction efficiency of T-lymphocytes. RT-PCR confirmed that the CD19-CAR gene was specificly transcripted with high efficiency. IFN-γ and TNF-α released by CD19-CAR transduced T-lymphocytes significantly increased when activated by target cells.</p>

Cytotoxity of pomalidomide combined CAR-T cell for multiple myeloma cell RPMI8226 and U266 / 中华血液学杂志

<p><b>OBJECTIVE</b>To observe the cytotoxity of CD138-CAR-T cells on human multiple myeloma cell RPMI8226 and U266 cells and explore the impact of pomalidomide on the cytotoxity of CD138-CAR-T on RPMI8226 and U266 cells.</p><p><b>METHODS</b>The cytotoxity of CD138-CAR-T and CD138-CAR-T combined pomalidomide on RPMI8226 and U266 was detected by CFSE/7AAD. The effctor cells were co-cultured with target cells at 5:1 for 18 h, and then the supernatant were collected and used for ELISA assays.</p><p><b>RESULTS</b>After 18 h co-culture, the cytotoxity of CD138-CAR-T on RPMI8226 and U266 was significantiy higher than control (P<0.01). There was no significant change on the cytotoxity of pomalidoide combined with CD138-CAR-T on RPMI8226 and U266. The results showed that co-cultured system contribted to a markedly increased production of IFN-γ, after adding pomalidomide to the co-cultured system. It can significantly enhance the production of IFN-γ, compared with CD138-CAR-T alone.</p><p><b>CONCLUSION</b>CD138-CAR-T had significantly cytotoxity on U266 and RPMI8226. Pomalidomide could promote CD138-CAR-T cells IFN-γ production.</p>

Intravenously Administered Anti-recoverin Antibody Alone Does Not Pass through the Blood-Retinal Barrier

PURPOSE: Cancer-associated retinopathy is a paraneoplastic retinal degeneration which may primarily result from auto-immune mediated apoptosis. It has been hypothesized that high titer of auto-antibodies are able to cross the blood-retinal barrier (BRB) and to enter retinal cells to activate apoptotic pathway which has been already well-established. However, it still remains to be elucidated whether auto-antibodies could cross BRB in the retina. Herein, we demonstrated that intravenously administrated anti-recoverin antibodies could not pass through BRB and not lead to retinal cell death. METHODS: Anti-recoverin antibody was intravenously injected to C57BL/6 mice, which were sacrificed 1 and 7 days to obtain eye. Vascular endothelial growth factor was intravitreally injected to induce BRB breakdown, which was confirmed by fluorescein angiography and western blotting for zonula occludens (ZO)-1, ZO-2 and occludin. To investigate the location of anti-recoverin antibody in the retina, immunofluorescein was performed. The retinal toxicity of intravenous anti-recoverin antibody was evaluated by histological examination and transferase-mediated dUTP nick-end labeling. Immunofluorescein staining for glial fibrillary acidic protein was done to address glial activation as well. RESULTS: Intravenously administrated anti-recoverin antibodies were exclusively distributed on retinal vessels which were co-localized with CD31, and led to neither increase of glial fibrillary acidic protein expression, as an indicator of retinal stress, nor apoptotic retinal cell death. Moreover, even in the condition of vascular endothelial growth factor-induced BRB breakdown, anti-recoverin antibodies could not migrate across BRB and still remained on retinal vessels without retinal cytotoxicity. CONCLUSIONS: Our results suggest that high titer of intravascular anti-recoverin antibodies could not penetrate into the retina by themselves, and BRB breakdown mediated by dysregulation of tight junction might not be sufficient to allow anti-recoverin antibodies to pass through BRB.

Comparison of two specific immunoglobulin E test systems in the diagnosis of allergic rhinitis / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To evaluate the diagnostic performance of the AllergyScreen system from Mediwiss An alytic GmbH and the Pharmacia CAP system from Pharmacia Diagnostics for the detection of four inhalant allergens in the diagnosis of allergic rhinitis.@*METHOD@#All 35 serum samples were collected from patients who were referred to the allergist for a suspected allergic rhinitis between January to March in 2007. Patients were classified as study diagnosis-positive for inhalant allergy if they both had a positive clinical examination/history and a related positive skin test for the suspected inhalant allergen.@*RESULT@#Comparing with the reference standard, the diagnostic indexes (sensitivity, specificity, accuracy, predictive value of positive, and predictive value of negative) of the CAP system and the AllergyScreen system were 0.96 vs 0.89, 0.84 vs 0.75, 0.89 vs 0.80, 0.78 vs 0.65 and 0.93 vs 0.93 respectively. The CAP system method had higher sensitivity, specificity and accuracy than AllergyScreen system method, but there have no statistical difference between two systems (P>0.05).@*CONCLUSION@#This data support the use of ImmunoCAP system and AllergyScreen system to identify potentially significant individual allergens in the diagnosis of allergic rhinitis. The diagnostic indexes between the two systems have no statistical difference. As a simple, rapid turnaround time and low-cost system, AllergyScreen system can test multi-allergens in one time, so it can be used as a complementary with the ImmunoCAP system.

Altered Expression of Parvalbumin in Neural Retina of the Insulin-dependent Diabetic Rat / 대한해부학회지

Calcium-binding proteins in the nervous system are functioned in Ca2+ buffering and Ca2+ transport and regulation of various enzyme systems. They potentially have a number of different effects on cells includingaltering the duration of action potentials, promoting neuronal bursting activity and protecting cells against the damaging effects of excessive calcium influx. The present study has been designed to clarify the differential responding patterns of parvalbumin immunoreactive neurons in the rat retina following diabetic injury, for better understandings of role of parvalbumin in the retinal circuitry and in calcium homeostasis. Experimental diabetes was induced by a single intravenous injection of streptozotocin in a dose of 60 mg/kg body weight. Diabetic rats showing high blood glucose levels (above 300 mg/dL) were cared for 1, 4, 8, 12 and 24 weeks, respectively. The retinas at each time point were processed for immunohistochemistry and Western blotting using antiparvalbumin antibody. In the rat retina at normal, parvalbumin immunoreactivity appeared in AII amacrine cells, amacrine cells of a widefield type and displaced amacrine cells. A few bipolar cells are also showed the reactivity. During diabetes, the intensity of parvalbumin immunoreactivity is decreased especially in the AII amacrine cells. The cell number of parvalbumin immunoreactive neurons has showed no large changes throughout the diabetes, except that of bipolar cells. That population of parv immunoreactive of bipolar cells has increased remarkably at later diabetic periods. The protein levels of parvalbumin have showed transiently a slight increase at earlier diabetic periods, and then decreased to lower than that of normal. Parvalbumin immunoreactive bipolar cells at diabetes are co-localized not with PKC-alpha or recoverin, but with glutamate transporter Glt-1b. AII amacrine cell processes were joined with each other and with axon terminals of presumed cone bipolar cells by gap junction. These results suggest that the calcium buffering activity of parvalbumin is shifted from AII amacrine cells to a certain type of cone bipolar cells, in response to diabetes. This event may be occurred through electrically coupled gap junction in between these cell processes.

Differentiation of Recoverin Immunoreactive Cone Bipolar Cells and Their Timing of Synaptic Formation with GABAergic Amacrine Cells in the Rat Retina / 대한해부학회지

Recoverin is a member of the large family of EF-hand calcium binding proteins (Baimbridge et al., 1992), and it is thought to be involved in the regulation of phosphodiesterase in photoreceptors and in the phosphorylation of activated rhodopsin (Polans et al., 1996). Although the functional significance of recoverin in cone bipolar cells is not fully understood, the antiserum against recoverin has been widely used to identify a certain population of cone bipolar cells (Milam et al., 1993; Sasso's Pognetto et al., 1994; Euler & W sle, 1995). GABA is well known to act as major neurotransmitters in the mammalian central nervous system including retina. This study was conducted to identify the development process of recoverin-labeled cone bipolar cells, and the timing points of synaptic formation of the labeled bipolar cells and GABAergic amacrine cells in the rat retina. The results were as follows; In the adult rat retina, recoverin-labeled cone bipolar cells were subdivided into twotypes; type 2 cells with axon terminal stratified in sublamina a of the inner plexiform layer (IPL), and type 8 cells with axon terminals stratified in sublamina b of the IPL. Recoverin-labeled cone bipolar cells began to appear from postnatal day 5. The axon terminals of recoverin-labeled type 2 cone bipolar cells stratified at postnatal day 10, while those of type 8 cone bipolar cells stratified at postnatal day 13. The axon terminals of type 2 cone bipolar cells made ribbon synapses onto GABAergic amacrine cells in the IPL at postnatal day 10. These results demonstrate that recoverin-labeled type 2 cone bipolar cells differentiate earlier than recoverin-labeled type 8 cone bipolar cells, and suggest that GABAergic amacrine cells may play important roles in visual processing of recoverin-labeled type 2 cone bipolar cells by making synapse onto these cells at early stage. Synapses between type 2 cone bipolar cells and GABAergic amacrine cells are formed about the time of postnatal day 10 for visual processing.