Anti-4-1BB×PD-L1 Bispecific Antibody Reinvigorates Tumor-Specific Exhausted CD8+ T Cells and Enhances the Efficacy of Anti-PD-1 Blockade.

PURPOSE: To overcome the limited efficacy of immune checkpoint blockade, there is a need to find novel cancer immunotherapeutic strategies for the optimal treatment of cancer. The novel anti-4-1BB×PD-L1 bispecific antibody-ABL503 (also known as TJ-L14B)-was designed to simultaneously target PD-L1 and 4-1BB, and demonstrated strong antitumor T-cell responses without considerable toxicity. Here, we investigated how the combination of ABL503 and anti-PD-1 blockade affected the reinvigoration of exhausted tumor-infiltrating CD8+ T cells (CD8+ TILs) and anti-tumor efficacy. EXPERIMENTAL DESIGN: Single cell suspensions of hepatocellular carcinoma and ovarian cancer from treatment-naive patients were used for immunophenotyping of CD8+ TILs and in vitro functional assays. Humanized hPD-1/hPD-L1/h4-1BB triple knock-in mice were used to evaluate the effects of ABL503 and anti-PD-1 blockade in vivo. RESULTS: We observed that ABL503 successfully restored the functions of 4-1BB+ exhausted CD8+ TILs, which were enriched for tumor-specific T cells but unresponsive to anti-PD-1 blockade. Importantly, compared to anti-PD-1 blockade alone, the combination of ABL503 and anti-PD-1 blockade further enhanced the functional restoration of human CD8+ TILs in vitro. Consistently, the combination of ABL503 with anti-PD-1 in vivo significantly alleviated tumor growth, and induced enhanced infiltration and activation of CD8+ TILs. CONCLUSIONS: ABL503-a PD-L1 and 4-1BB dual-targeting bispecific antibody-elicits pronounced additive tumor growth inhibition, with increased infiltration and functionality of exhausted CD8+ T cells, which in turn enhances the anti-cancer effects of anti-PD-1 blockade. These promising findings suggest that ABL503 (TJ-L14B) in combination with PD-1 inhibitors will likely further enhance therapeutic benefit in clinical trials.

CLDN18.2 and 4-1BB bispecific antibody givastomig exerts antitumor activity through CLDN18.2-expressing tumor-directed T-cell activation.

BACKGROUND: Claudin18.2 (CLDN18.2) is a tight junction protein that has been identified as a clinically proven target in gastric cancer. Stimulation of 4-1BB with agonistic antibodies is also a promising strategy for immunotherapy and 4-1BB+ T cells were reported to be present within the tumor microenvironment of patients with gastric cancer. However, hepatotoxicity-mediated by 4-1BB activation was observed in clinical trials of agonistic anti-4-1BB monoclonal antibodies. METHODS: To specifically activate the 4-1BB+ T cells in tumor and avoid the on-target liver toxicity, we developed a novel CLDN18.2×4-1BB bispecific antibody (termed 'givastomig' or 'ABL111'; also known as TJ-CD4B or TJ033721) that was designed to activate 4-1BB signaling in a CLDN18.2 engagement-dependent manner. RESULTS: 4-1BB+ T cells were observed to be coexisted with CLDN18.2+ tumor cells in proximity by multiplex immunohistochemical staining of tumor tissues from patients with gastric cancer (n=60). Givastomig/ABL111 could bind to cell lines expressing various levels of CLDN18.2 with a high affinity and induce 4-1BB activation in vitro only in the context of CLDN18.2 binding. The magnitude of T-cell activation by givastomig/ABL111 treatment was closely correlated with the CLDN18.2 expression level of tumor cells from gastric cancer patient-derived xenograft model. Mechanistically, givastomig/ABL111 treatment could upregulate the expression of a panel of pro-inflammatory and interferon-γ-responsive genes in human peripheral blood mononuclear cells when co-cultured with CLDN18.2+ tumor cells. Furthermore, in humanized 4-1BB transgenic mice inoculated with human CLDN18.2-expressing tumor cells, givastomig/ABL111 induced a localized immune activation in tumor as evident by the increased ratio of CD8+/regulatory T cell, leading to the superior antitumor activity and long-lasting memory response against tumor rechallenge. Givastomig/ABL111 was well tolerated, with no systemic immune response and hepatotoxicity in monkeys. CONCLUSIONS: Givastomig/ABL111 is a novel CLDN18.2×4-1BB bispecific antibody which has the potential to treat patients with gastric cancer with a wide range of CLDN18.2 expression level through the restricted activation of 4-1BB+ T cells in tumor microenvironment to avoid the risk of liver toxicity and systemic immune response.

Polymorphism, gigantism, and cannibalism, one stylonychid ciliate (Ciliophora, Hypotricha) to rule them all.

The morphology, ontogenesis, and molecular phylogeny of the polymorphic and cannibalistic giant forming Tetmemena polymorpha n. sp., found in a brackish water sample in South Korea, were investigated. The present species has long been misidentified as "Oxytricha bifaria." The new investigation shows that the species produces three morphologically different morphs. The small morph is bacterivorous and characterized by its small body size and slim body and it is found only in the stationary and decline phases of the culture. The large morph has a wide body, larger oral apparatus, and feeds on small eukaryotes such as yeast cells and small ciliates. It divides very quickly and produces the other two morphs and found in the exponential phase of the cultures. The giant morph is characterized by its huge body and oral apparatus. It feeds on the small morph cells of the same species and other ciliates, and occurs together with the small morph. Phylogenetic analyses based on the 18S rRNA gene sequences show that the new species is placed in a sister subclade to that containing other Tetmemena sequences. Moreover, Tetmemena indica Bharti et al., 2019 nov. stat. is raised to species level based on the differences in the cyst morphology and the dorsal ciliature to the authoritative Tetmemena pustulata population.

LAG-3xPD-L1 bispecific antibody potentiates antitumor responses of T cells through dendritic cell activation.

Several preclinical studies demonstrate that antitumor efficacy of programmed cell death-1 (PD-1)/programmed death-ligand 1 (PD-L1) blockade can be improved by combination with other checkpoint inhibitors. Lymphocyte-activation gene 3 (LAG-3) is an inhibitory checkpoint receptor involved in T cell exhaustion and tumor immune escape. Here, we describe ABL501, a bispecific antibody targeting LAG-3 and PD-L1 in modulating immune cell responses against tumors. ABL501 that efficiently inhibits both LAG-3 and PD-L1 pathways enhances the activation of effector CD4+ and CD8+ T cells with a higher degree than a combination of single anti-LAG-3 and anti-PD-L1. The augmented effector T cell responses by ABL501 resulted in mitigating regulatory-T-cell-mediated immunosuppression. Mechanistically, the simultaneous binding of ABL501 to LAG-3 and PD-L1 promotes dendritic cell (DC) activation and tumor cell conjugation with T cells that subsequently mounts effective CD8+ T cell responses. ABL501 demonstrates its potent in vivo antitumor efficacy in a humanized xenograft model and with knockin mice expressing human orthologs. The immune profiling analysis of peripheral blood reveals an increased abundance of LAG-3hiPD-1hi memory CD4+ T cell subset in relapsed cholangiocarcinoma patients after gemcitabine plus cisplatin therapy, which are more responsive to ABL501. This study supports the clinical evaluation of ABL501 as a novel cancer immunotherapeutic, and a first-in-human trial has started (NCT05101109).

A new "flagship" ciliate, Pseudostylonychia obliquocaudata n. gen., n. sp. (Ciliophora, Hypotricha), from South Korea.

The morphology, ontogenesis, and molecular phylogeny of a new ciliate, Pseudostylonychia obliquocaudata n. gen., n. sp., discovered in a semiterrestrial habitat in South Korea, were investigated. The new species is characterized by an obovate or triangular body with pointed posterior end. The caudal area is distinctly obliquely truncated rightwards. Pseudostylonychia n. gen. is established and characterized by the obovate or triangular body, Stylonychia-like infraciliature and macronuclear pattern, and undulating membranes in Laurentiella pattern. It is also characterized by the involvement of cirri III/2 and IV/3 in primordia formation. SEM investigations show that the resting cyst has high, spirally arranged ridges. Phylogenetic analyses based on SSU rRNA gene sequences show that the new species is placed as a sister to the clade containing sequences of Stylonychia mytilus complex, Stylonychia (Metastylonychia), and Coniculostomum. Furthermore, Stylonychia koreana Kumar et al., 2016 nov. stat. is raised to species level based on the differences in the cyst morphology between the two subspecies of S. ammermanni. Also, Metastylonychia Kumar and Foissner, 2017 nov. stat. is raised to genus rank based on the new molecular data and the morphological and ontogenetic differences to Coniculostomum and Stylonychia.

Factors associated with the effectiveness of part-time patching for intermittent exotropia in children.

PURPOSE: To evaluate the factors associated with the efficacy of low-dose part-time patching in children with intermittent exotropia (IXT). METHODS: In this prospective observational study, we enrolled 186 patients diagnosed with IXT. Outcome measures included office based control scales, magnitude of exo-deviation, and stereoacuity at near and distance after daily patching for 2 h. We analyzed the clinical data and demographic factors association with improvement of IXT. RESULTS: The study was completed by 152 subjects of total enrolled patients on a consecutive basis followed up for 1 year. Decrease in the magnitude of exo-deviation, improvement of control, and or gain of stereoacuity were observed in 31.6% patients of the recruited subjects after part-time patching. Multivariate analyses showed that prognostic factors determining improvement to part-time patching included convergence insufficiency (CI) type IXT (p = 0.016), poor distance stereopsis (p = 0.044), and large exotropic deviation at distance (p = 0.025). CONCLUSIONS: CI-type exotropia, large distance magnitude of exo-deviation, or poor distance stereopsis appear to be associated with a better response to part-time patching. Therefore low dose part-time patching may be a useful non-surgical treatment alternative to delay surgery in these cases.

Novel anti-4-1BB×PD-L1 bispecific antibody augments anti-tumor immunity through tumor-directed T-cell activation and checkpoint blockade.

BACKGROUND: Stimulation of 4-1BB with agonistic antibodies is a promising strategy for improving the therapeutic efficacy of immune checkpoint inhibitors (ICIs) or for overcoming resistance to ICIs. However, dose-dependent hepatotoxicity was observed in clinical trials with monoclonal anti-4-1BB agonistic antibodies due to the activation of 4-1BB signaling in liver resident Kupffer cells. METHODS: To avoid this on-target liver toxicity, we developed a novel bispecific antibody (4-1BB×PD-L1 bispecific antibody, termed "ABL503") uniquely designed to activate 4-1BB signaling only in the context of PD-L1, while also blocking PD-1/PD-L1 signaling. RESULTS: Functional evaluation using effector cells expressing both 4-1BB and PD-1 revealed superior biological activity of ABL503 compared with the combination of each monoclonal antibody. ABL503 also augmented T-cell activation in in vitro assays and further enhanced the anti-PD-L1-mediated reinvigoration of tumor-infiltrating CD8+ T cells from patients with cancer. Furthermore, in humanized PD-L1/4-1BB transgenic mice challenged with huPD-L1-expressing tumor cells, ABL503 induced superior anti-tumor activity and maintained an anti-tumor response against tumor rechallenge. ABL503 was well tolerated, with normal liver function in monkeys. CONCLUSION: The novel anti-4-1BB×PD-L1 bispecific antibody may exert a strong anti-tumor therapeutic efficacy with a low risk of liver toxicity through the restriction of 4-1BB stimulation in tumors.

B7-H3×4-1BB bispecific antibody augments antitumor immunity by enhancing terminally differentiated CD8+ tumor-infiltrating lymphocytes.

Cancer immunotherapy with 4-1BB agonists has limited further clinical development because of dose-limiting toxicity. Here, we developed a bispecific antibody (bsAb; B7-H3×4-1BB), targeting human B7-H3 (hB7-H3) and mouse or human 4-1BB, to restrict the 4-1BB stimulation in tumors. B7-H3×m4-1BB elicited a 4-1BB-dependent antitumor response in hB7-H3-overexpressing tumor models without systemic toxicity. BsAb primarily targets CD8 T cells in the tumor and increases their proliferation and cytokine production. Among the CD8 T cell population in the tumor, 4-1BB is solely expressed on PD-1+Tim-3+ "terminally differentiated" subset, and bsAb potentiates these cells for eliminating the tumor. Furthermore, the combination of bsAb and PD-1 blockade synergistically inhibits tumor growth accompanied by further increasing terminally differentiated CD8 T cells. B7-H3×h4-1BB also shows antitumor activity in h4-1BB-expressing mice. Our data suggest that B7-H3×4-1BB is an effective and safe therapeutic agent against B7-H3-positive cancers as monotherapy and combination therapy with PD-1 blockade.

Budget Impact of Switching to Biosimilar Trastuzumab (CT-P6) for the Treatment of Breast Cancer and Gastric Cancer in 28 European Countries.

BACKGROUND: As the economic burden of treating cancer patients has been soaring in European countries, performing a budget impact analysis is becoming one of the requirements for payers' application dossiers. OBJECTIVE: The objective of this study was to estimate the budgetary impact of introducing the biosimilar trastuzumab (CT-P6) from the payer's perspective and to determine the number of additional patients who could be treated with resulting savings in 28 European countries. METHODS: A budget impact model was developed to analyze the financial impact of switching from originator trastuzumab to biosimilar CT-P6 in the treatment of early and metastatic breast cancer and metastatic gastric cancer with a time horizon of 1-5 years. Budgetary savings and the number of patients potentially affected were measured based on epidemiological and sales volume data. The base-case analysis assumed that the price of CT-P6 is 70% of the originator price, the switching rate of originator to CT-P6 in the first year is 20%, and the annual growth in the switching rate for each subsequent year is 5%. RESULTS: For analyses using the base-case scenario following CT-P6 introduction, the total estimated budgetary savings over a 5-year period (depending on the scenario) ranged from €1.13 billion to €2.27 billion based on epidemiological data, or from €0.91 billion to €1.82 billion based on sales volume data. In the first year only, the projected budgetary savings ranged from €58 million to €136 million, and the number of additional patients who could be treated using the savings ranged from 3503 to 7078 by sensitivity analysis. CONCLUSIONS: The conducted budget impact analysis assessing a switch from originator trastuzumab to biosimilar CT-P6 in 28 European countries indicates that budget savings could be between €0.91 billion and €2.27 billion over the next 5 years. These savings could be used to help improve patient access to local biologics in their respective countries while simultaneously strengthening the overall public health landscape across the European Union.

Monographic treatment of Paraholosticha muscicola (Ciliophora, Keronopsidae), including morphological and molecular biological characterization of a brackish water population from Korea.

Paraholosticha muscicola, type species of Paraholosticha Wenzel, inhabits mainly terrestrial habitats, but also freshwater. A brackish water population from Korea is described, the first record from such a habitat. Principal component analysis shows that this population is more similar to a terrestrial population from Denmark than to a population from Antarctic soil. Keronopsids have two strong morphological/ontogenetic apomorphies (frontal corona formed from anlagen I-III; division in cysts). However, the SSU rRNA sequence of the Korean population does not cluster with that of the Antarctic population in the phylogenetic tree, but both branch off consecutively and immediately before a mixture of other non-dorsomarginalian hypotrichs, including two further keronopsids. Furthermore, the keronopsids cluster in the phylogenetic network, providing phylogenetic conflicts, which cannot be exemplified in the conventional gene tree. To complete the picture of P. muscicola, we provide a detailed overview about nomenclature, history, taxonomy, and its geographic distribution. From the four synonyms proposed so far, we tentatively accept only P. lichenicola and P. ovata. Paraholosticha algivora is likewise very similar. Thus we propose to include these three taxa as members of the P. muscicola complex. Stylonethes sterkii and P. algivora are transferred to Paraholosticha Wenzel. A key to the Paraholosticha species is provided.

Genotype and Phenotype Spectrum of FRMD7-Associated Infantile Nystagmus Syndrome.

Purpose: We investigate the genotype and phenotype spectrum of FRMD7-associated infantile nystagmus syndrome in Korean probands. Methods: A total of 37 patients with infantile nystagmus syndrome were recruited prospectively for genetic analysis. We performed polymerase chain reaction (PCR)-based direct sequencing and haplotype analysis for FRMD7. Detailed ophthalmic examinations and eye movement recordings were compared between FRMD7 and non-FRMD7 groups. Results: In 13 (35%) of 37 patients, five different mutations of FRMD7 were detected: start codon mutation c.1A>G, splice site mutation c.162+6T>C, and three missense mutations (c.575A>C, c.722A>G, and c.875T>C). The latter mutation was identified in seven unrelated patients, and always was accompanied with two single nucleotide polymorphisms of exon 12 (rs6637934, rs5977623). Compared to non-FRMD7 groups, a cup-to-disc ratio was significantly decreased in FRMD7 groups (P < 0.001), and a disc-macula distance to disc diameter ratio markedly increased in the FRMD7 group (P = 0.015). Most patients in the FRMD7 group had at least two types of the nystagmus waveforms, and the most common type was unidirectional jerk nystagmus (75%), such as pure jerk and jerk with extended foveation, followed by pendular (25%), bidirectional jerk (19%), and dual jerk (6%) nystagmus. No significant differences were observed between FRMD7 and non-FRMD7 groups in terms of the nystagmus waveform, presence of periodic alternating nystagmus, and mean foveation time. Conclusions: We identified five FRMD7 mutations in 35% of our infantile nystagmus syndrome cohort, expanding its mutational spectrum. The missense mutation c.875T>C may be a common mutation arisen from the founder effect in Korea. Optic nerve dysplasia associated with FRMD7 mutations suggests that the abnormal development of afferent visual systems may affect neural circuitry within the oculomotor system.

Long-Term Surgical Outcomes of Early Surgery for Intermittent Exotropia in Children Less than 4 Years of Age.

PURPOSE: To investigate the long-term outcomes of intermittent exotropia surgery for children less than 4 years of age. MATERIALS AND METHODS: Consecutive patients who underwent surgery for intermittent exotropia and had follow-up durations longer than 2 years were recruited. The patients were classified according to age at surgery-the patients of group 1 had undergone surgery before 4 years of age and those of group 2 at or after 4 years of age. Motor success was defined by exodeviation < 10 prism diopters (PD) and esodeviation < 5 PD at distance at 2 years postoperatively. Stereoacuity was considered as success at a value ≤ 60 arc seconds. The motor and sensory success rates as well as the surgical complications were compared. RESULTS: Of the 73 patients, 36 were allocated to group 1 and 37 to group 2. At 2 years after surgery, 13 of the 36 (36.1%) patients in group 1 and 12 of the 37 (32.4%) in group 2 had achieved successful alignment; 32 (88.9%) patients in group 1 and 35 (94.6%) in group 2 achieved normal stereoacuity. No significant differences in the motor or sensory success rates were observed between the two groups (p = 0.46 and 0.32, respectively). CONCLUSIONS: The surgical success rates for intermittent exotropia were comparable between the patients operated upon before 4 years of age and those operated upon after 4 years of age. The incidence of postsurgical complications was low and not significantly different between the two study groups.

Strabismus in children with white matter damage of immaturity: MRI correlation.

BACKGROUND/AIMS: To investigate the correlation between strabismus and the severity of white matter damage of immaturity (WMDI), based on MRI findings. Although strabismus is commonly associated with WMDI, its clinical features are not well established. METHODS: This cross-sectional study involved 73 consecutive patients who visited the department of ophthalmology and were diagnosed with WMDI. The severity of WMDI was graded based on the MRI findings of the patients. All of the patients underwent complete ophthalmic examination, and strabismus was characterised in terms of direction, constancy, and angle of deviation. The prevalence and the characteristics of strabismus and their correlation with the grade of WMDI were investigated. RESULTS: The perinatal characteristics, age at MRI, and the number of MRIs per child did not differ between different grades of WMDI. Refractive errors, found in 56 (76.7%) patients, did not differ between the grades of WMDI either. Strabismus was observed in 38 (52.1%) patients, and its prevalence increased with the grade of the disorder; 20 patients had exotropia and 18 had esotropia. Constant strabismus was found more frequently in patients with higher grade WMDI. However, the direction and angle of deviation did not differ depending on the grade of WMDI. CONCLUSIONS: The prevalence of strabismus increased with the severity of WMDI and was higher among patients with WMDI than among healthy individuals. The severity of WMDI might be related to the presence and constancy of strabismus.

Pseudonotohymena antarctica n. g., n. sp. (Ciliophora, Hypotricha), a New Species from Antarctic Soil.

A new soil ciliate, Pseudonotohymena antarctica n. g., n. sp., from King George Island, Antarctica, is described based on live observation, protargol impregnation, and its 18S rRNA gene. The new genus Pseudonotohymena is morphologically similar to the genus Notohymena Blatterer and Foissner in the following characteristics: 18 fronto-ventral-transverse cirri, a flexible body, undulating membranes, dorsomarginal kineties, and the number of cirri in the marginal rows. However, Pseudonotohymena differs from Notohymena particularly in the dorsal ciliature, that is, in possessing a nonfragmented dorsal kinety (vs. fragmented). In addition, the molecular phylogenetic relationship of the new species differs from that of Notohymena species. On the basis of the morphological features, the genetic data, and morphogenesis, we establish P. antarctica n. g., n. sp. In addition, the cyst morphology of this species is described.

The effects of vascular endothelial growth factor (VEGF) on human orbital preadipocyte.

PURPOSE: To investigate the presence of the Vascular Endothelial Growth Factor (VEGF) and its receptor (VEGFR) in human orbital preadipocytes, and to evaluate the effect of VEGF on human orbital preadipocyte differentiation and adipogenesis in vitro. RESULTS: Four isoforms of VEGF (VEGF121, 155, 189, and 206), VEGFR-1, VEGF-2, and neuropilin-1 were expressed in human orbital preadipocytes. Treatment with 100 ng/ml VEGF induced higher expressions of C/EBPα and LPL than the non-treated control (p = 0.03 and p = 0.01) or treatment with 50ng/ml (p = 0.04 for both). At both concentrations VEGF enhanced the accumulation of intra-cytoplasmic lipid versus the control, and treatment with 100 ng/ml VEGF induced more lipid accumulation than treatment with 50 ng/ml VEGF (p = 0.03). CONCLUSIONS: VEGF and VEGFR were observed in human orbital preadipocytes, and exogenous VEGF enhanced adipogenesis in these cells. These results suggest that VEGF plays a role as an autocrine or paracrine growth factor during human orbital preadipocyte differentiation.

Intracranial fibromuscular dysplasia presenting as various ocular manifestations.

Fibromuscular dysplasia (FMD) is a nonatherosclerotic, noninflammatory vascular disease that mainly affects renal, extracranial carotid, and vertebral arteries. Intracranial FMD is uncommon unlike extracranial or renal FMD, and the primary manifestation of intracranial FMD is intracranial aneurysm. We report an unusual case of intracranial FMD showing various ocular manifestations, including central retinal artery occlusion, transient monocular blindness, and oculomotor nerve palsy without renal involvement.

Morphology, morphogenesis, and molecular phylogeny of Anteholosticha multicirrata n. sp. (Ciliophora, Spirotrichea) with a note on morphogenesis of A. pulchra (Kahl, 1932) Berger, 2003.

Two marine urostylid ciliates, Anteholosticha multicirrata n. sp. and Anteholosticha pulchra (Kahl, 1932) Berger, 2003, were collected from South Korea. These species were identified based on morphology, morphogenesis, and SSU rRNA gene sequence comparison. Anteholosticha multicirrata n. sp. is characterized by the following features: body size 90-125 × 30-45 µm in vivo, shape slender to ellipsoidal in outline, with yellow-greenish cortical granules distributed along and between dorsal kineties and cirri; single contractile vacuole positioned on left at mid-body; three frontal, five to seven frontoterminal, one buccal, one to two pretransverse and four to six transverse cirri; three complete dorsal kineties; one left and one right marginal cirral row; about 117 macronuclear nodules; and three to four micronuclei observed during morphogenesis. In addition, based on the observations of morphogenesis, we found that A. pulchra has pretransverse cirri, which were not described in detail in previous studies. Nuclear small subunit ribosomal RNA (SSU rRNA) gene was used to analyse their phylogenetic relationship, and the gene tree supports that the genus Anteholosticha is a highly polyphyletic group.

A case of fungal arthritis caused by Hansenula anomala.

Hansenula anomala (H. anomaly) is part of the normal flora in the alimentary tract and throat. It has been reported to be an organism causing opportunistic infections in immunocompromised patients. However, cases of fungal arthritis caused by H. anomala are rare. We encountered a case of H. anomala arthritis in a 70-year-old man who was treated with an empirical antibiotic treatment and surgery under the impression of septic arthritis. However, the patient did not improve after antibiotic therapy and surgery. Consequently, knee joint aspiration was performed again, which identified fungal arthritis caused by H. anomala. It was treated successfully with amphotericin B and fluconazole. When treating arthritis patients with diabetes, it is important to consider the possibility of septic arthritis by H. anomala and provide the appropriate treatment.

Tumor targeting of humanized fragment antibody secreted from transgenic rice cell suspension culture.

The tumor-associated glycoprotein 72 (TAG 72) has been shown to be expressed in the majority of human adenocarcinomas. In an effort to develop a technique for the safe and inexpensive production of large quantities of anti-TAG 72 humanized antibody fragments (hzAb) as a future source of clinical-grade proteins, we developed a transgenic rice cell suspension culture system. The in vivo assembly and secretion of hzAb were achieved in a transgenic rice cell culture under the control of the rice alpha amylase 3D (RAmy 3D) expression system, and the biological activities of plant-derived hzAb were determined to be quite similar to those of animal-derived antibody. Purified hzAb was shown to bind to the recombinant antigen, TAG 72, and to bind specifically to human LS 174T colon adenocarcinoma cells expressing the TAG 72 antigen, and this binding occurred to the same extent as was seen with animal-derived antibody. Plant-derived hzAb proved as effective as animal-derived antibody in targeting tumors of xenotransplanted LS 174T cells in nude mice. The results of this study indicate that the hzAb derived from plant cell suspension cultures may have great potential for pharmaceutical applications in the development of future cancer therapeutic and diagnostic protocols.

Decompression retinopathy after trabeculectomy.

PURPOSE: To present a case of a unilateral diffuse retinal hemorrhage in a 15-year-old girl, who underwent bilateral trabeculectomy for steroid induced glaucoma. METHODS: Despite the maximally tolerable medical treatment, IOP in the right eye remained above 50 mmHg for four months, and was simultaneously elevated in the left eye. So we performed bilateral trabeculectomy. RESULTS: On the first postoperative day, diffuse retinal hemorrhages were observed in the right eye; however, no retinal hemorrhage was found in the left eye. The hemorrhages resolved completely without consequences two months later. CONCLUSIONS: In the case of high IOP for a long period, sudden lowering of IOP may acutely increase the blood flow and consequently rupture multiple retinal capillaries because of altered autoregulatory function. Special care is therefore needed to prevent an abrupt fall in IOP before, during, and after surgery, especially when IOP has been highly elevated for an extended period.