Selective elimination of alloreactivity in vitro and in vivo while sparing other T-cell-mediated immune responses.

Selective elimination of alloreactive cells was carried out in the set-up of T-cell-mediated immunotherapy in an effort to gain the benefits of hematopoietic allogeneic transplantation while reducing the risk of GVHD. Low MW chemical compounds were screened for their effect on T-cell-mediated immune responses of murine- and human-derived lymphocytes. Selected compounds were further tested in secondary MLR assays in which sensitization to alloantigens was carried out in vitro, in the presence or absence of a given compound, followed by exposure to related and unrelated alloantigens or T-cell mitogenic stimulation. At a low concentration of <1 µM, a quinazoline derivative named AO#349 [2-(3,4,5-trimethoxyphenyl)-N-p-tolylquinazolin-4-amine], was able to induce 78-90% inhibition of a selective allogeneic response while retaining >92% immune reactivity to unrelated alloantigens and mitogenic stimuli in vitro. Following allogeneic sensitization in the presence of AO#349, elimination of alloreactivity to the priming alloantigens was also proved in a murine model of GVHD: 10 out of 15 sub-lethally irradiated mice inoculated with these sensitized cells were GVHD-free for >200 days. AO#349 was efficient in induction of a selective elimination of alloreactivity and should be considered for clinical application in allogeneic cell-mediated immunotherapy.

Putative non-uniting double central retinal artery system.

PURPOSE: To report a case of probable non-uniting double retinal arterial circulation. METHODS: Case report. A 66-year-old man suffered central retinal artery occlusion. RESULTS: The medical findings and angiographic workup supported the existence of double central retinal circulation not uniting before entering the globe and independently branching in the retina. CONCLUSIONS: In previously recognized unusual anatomic structures of the central retinal artery two parallel arteries were recognized, both of which united before entering the globe. We have described a rare anatomic variant in which those arteries remained separated after entering the globe, including while branching in the retina.

Immune reconstitution following allogeneic stem cell transplantation in recipients conditioned by low intensity vs myeloablative regimen.

We have investigated the immune status of patients with hematologic malignancies treated with a low intensity conditioning in preparation for allogeneic stem cell transplantation. Conditioning consisted of fludarabine, anti-T lymphocyte globulin and low-dose busulfan, followed by infusion of allogeneic blood stem cells. This protocol resulted in rapid engraftment and complete replacement of host with donor hematopoietic cells. Immunological parameters of these patients were compared to those patients who were conditioned by an aggressive myeloablative regimen. Distribution of cell surface markers of lymphocyte subsets from both groups of patients was similar, but different from that of normal control cells. Reduced intensity or non-myeloablative conditioning prior to allogeneic stem cell transplantation (NST), hardly lowered the normal T cell-dependent mitogenic response even during the early period following transplant, while the myeloablative treatments resulted in a suppressed mitogenic reaction and in slow immune recovery. Reactivity of non-MHC restricted cytotoxic T cells was also at a normal level in patients who were treated with NST. We conclude that stem cell engraftment following reduced conditioning may result in early reconstitution of immune responses assessed in vitro. We hypothesize that clinical application of NST may lead to faster development of effective immune responses against residual host-type malignant and abnormal non-malignant hematopoietic cells, although the role of fludarabine on post-transplant infections remains to be investigated in a larger cohort of patients.

Cell therapy with preimmunized effector cells mismatched for minor histocompatible antigens in the treatment of a murine mammary carcinoma.

Cell therapy with allogeneic donor cells mismatched for minor histocompatible (MiHC) antigens was applied to a murine mammary carcinoma (4T1) model to test the feasibility of graft versus tumor (GVT) effect against metastatic epithelial tumor cells. BALB/c mice bearing a 4T1 tumor of BALB/c origin were given syngeneic or MiHC-mismatched splenocytes. GVT effects were determined in secondary recipients of adoptively transferred lung cells derived from primary hosts who had previously been inoculated intravenously with 4T1 cells, and injected with one of the following: 1) naive BALB/c splenocytes, 2) naive DBA/2 splenocytes, 3) 4T1-immune DBA/2 splenocytes, or 4) DBA/2 splenocytes immunized with host-derived BABL/c spleen cells. Naive DBA/2 splenocytes inhibited tumor growth only slightly and only slightly prolonged the survival of secondary recipients, in comparison with fully matched tumor/host BALB/c spleen cells. An efficient GVT reaction was demonstrated in vitro and in vivo with MiHC-mismatched DBA/2 splenocytes from mice presensitized by multiple injections of irradiated tumor or BALB/c-derived spleen cells. All 30 mice adoptively inoculated with lung cells from primary hosts that had previously been treated with these presensitized effector cells were tumor free for >250 days. Secondary recipients inoculated with lung cells from mice given naive BALB/c or DBA/2 spleen cells died of metastatic tumors within 33 to 46 days. These results suggest that preimmunized donor cells represent an effective tool against metastatic disease; hence, the next goal should be to control graft-versus-host disease while exploiting the GVT potential.

Immunotherapy of relapsed resistant chronic myelogenous leukemia post allogeneic bone marrow transplantation with alloantigen pulsed donor lymphocytes.

Allogeneic cell-mediated immunotherapy with donor lymphocyte infusion (DLI) can successfully reverse chemoradiotherapy-resistant relapse in patients with chronic myeloid leukemia treated by allogeneic bone marrow transplantation (BMT). We describe the first successful attempt in 1992 to treat DLI-resistant relapse in a patient with CML in full hematologic relapse, using immunized donor lymphocytes. Donor lymphocytes were pulsed in vitro with a mixture of irradiated peripheral blood lymphocytes (PBL) obtained from both parents, in order to trigger alloactivation of donor lymphocytes against host alloantigens presented by parental cells, using as stimulating cells maternal PBL expressing the shared maternal haplotype and paternal PBL expressing the shared paternal haplotype of the patient. Full hematologic, cytogenetic and molecular remission was induced for the first time, independently of GVH, and has persisted for more than 9 years. To the best of our knowledge, this report represents the first successful immunotherapy with donor lymphocytes activated against host-type antigens. We suggest that immune donor PBL may be superior to DLI, possibly effective even when all other modalities fail, perhaps even independently of GVHD.

Paediatric chiasmal neuritis--typical of post-Epstein-Barr virus infection?

A case of chiasmal neuritis with bilateral, asymmetric optic nerve involvement is presented. Association with recent Epstein-Barr virus infection is suggested. The clinical picture, possible aetiological factors, and causative relation to Epstein-Barr virus are presented.

Multiple retinal arteriolar occlusions associated with coexisting primary antiphospholipid syndrome and factor V Leiden mutation.

PURPOSE: To investigate a case of a young woman with both primary antiphospholipid syndrome and factor V Leiden mutation who developed multiple retinal arteriolar occlusions. METHOD: Case report of a 25-year-old woman with history and laboratory tests confirming the diagnosis of both primary antiphospholipid syndrome and factor V Leiden mutation who presented with blurred vision in both eyes. RESULTS: Multiple retinal arteriolar occlusions were observed in both of her eyes. The patient was treated first with heparin and then with warfarin. CONCLUSIONS: Primary antiphospholipid syndrome and factor V Leiden mutation, as well as other forms of thrombophilia, should be considered in the differential diagnosis of unexplained retinal vascular occlusions. The coexistence of several thrombophilic disorders may carry a particularly high risk for thrombotic manifestations.

Induction of antitumor immunity by indomethacin.

Irradiated tumor cells given, together with indomethacin, to syngeneic mice induced an antitumor response and conferred protection against a challenge of a lethal dose of murine mammary (4T1) and lung (3LL) carcinoma cells. Continuous administration of indomethacin was crucial throughout the entire period of immunization and challenge, as no protection was achieved when the drug was given during only one of these procedures. Antitumor immunity was long-lasting and, when tested in the 4T1 model, 48% of mice were resistant to a second challenge of lethal tumor cells. Tumor-free immune mice that were given indomethacin for more than 300 days remained healthy with normal white blood cell counts and normal spleen size. Cells isolated from immune mice were able to kill tumor cells in culture after in vitro activation by interleukin-2, in a manner similar to cells from naive normal control mice. In addition, the mitogenic response of their T cells was as high as that of the control naive mice. While indomethacin was able to induce antitumor immunity to 4T1 and 3LL murine carcinoma cells, both of which contain a high concentration of endogenic prostaglandin E(2) (PGE2), no such immunity was achieved to murine tumor cells with a low concentration of endogenic PGE2. These results suggest a correlation between PGE2 concentration and the ability of indomethacin to induce antitumor immunity. We therefore suggest that an immunotherapy protocol with long-term dispensation of a tolerable dose of an immunomodulator, given together with irradiated autologous tumor cells, may stimulate antitumor responses to tumors containing high concentrations of endogenic PGE2.

Bacteriological profile of ophthalmic infections in an Israeli hospital.

AIM: To find the optimal antibiotic treatment for ophthalmic infections in an Israeli hospital. METHODS: In a retrospective study from our laboratory, which serves as both a primary and secondary referral center, we analyzed the bacteriological profile and the antibiotic sensitivity of ophthalmic infections using the computerized laboratory reports of 331 consecutive ophthlamic bacteriological cultures from patients with various ophthalmic infections. RESULTS: Microbiological growth was obtained in 113 samples (34.1%). The most commonly isolated organism was coagulase-negative staphylococcus (19.5%), followed by coagulase-positive staphylococcus (16.8%), Enterobacteriaceae (14.2%), Pseudomonas aeruginosa (13.3%), and streptococcal species (8.9%). Pseudomonas species were the most common isolates from the lacrimal pathways (20.0%). Streptococci were the most common isolates cultured from the conjunctiva (27.3%). Coagulase-positive staphylococcus was the most common isolate from corneal ulcers (33.3%), and coagulase-negative staphylococcus from the vitreous (30.8%). The overall antibiotic sensitivity of common ophthalmic pathogens was similar to that reported from other parts of the world. CONCLUSIONS: Although essentially similar to previous series, this report from the Middle East differs as follows. Firstly, Pseudomonas species were the most common isolates from the lacrimal pathways. Secondly, the overall rate of streptococcal isolates was lower than in previous reports. Thirdly, streptococcal species were rarely isolated from corneal samples. Although other studies from the region have described the causative organisms of ocular infections in specific ocular sites, this is the first study from the Middle East to summarize the full bacteriological profile of ocular infections in one medical center.

Tc-99m labeled red blood cells scintigraphy: a diagnostic method for orbital cavernous hemangioma.

PURPOSE: To evaluate Tc-99m labeled red blood cells scintigraphy (Tc-99m RBC scintigraphy) as a diagnostic method for orbital cavernous hemangioma. PATIENTS AND METHODS: Retrospective analysis of the medical records of all patients who were diagnosed as suffering from orbital cavernous hemangioma over the last 16 years. RESULTS: Medical records of 12 patients with orbital cavernous hemangioma were identified. In all twelve patients, the tentative diagnosis of orbital cavernous hemangioma was made by CT scan. In all of them, the diagnosis was established by Tc-99m RBC scintigraphy, demonstrating a typical picture of "perfusion blood- pool mismatch" (This is normal radionuclide angiography, followed by a variable radionuclide uptake during the early blood-pool study and an intensive radionuclide uptake during the delayed blood-pool study). The tumor was surgically removed in all 12 patients. This diagnostic method was found to be reliable as no single case was recorded in which the preoperative diagnosis was not confirmed pathologically after surgical removal of the tumor. CONCLUSIONS: Tc-99m RBC scintigraphy is a reliable method for diagnosing orbital cavernous hemangioma. It should be included in our arsenal of diagnostic techniques, particularly for cases which are otherwise difficult to diagnose.

Visual disturbances and pathologic ocular findings in primary antiphospholipid syndrome.

OBJECTIVE: To delineate the features and prevalence of the systemic and ophthalmic involvement of primary antiphospholipid syndrome (PAS). DESIGN: Prospective case series. PARTICIPANTS: A total of 39 consecutive patients with PAS. METHODS: During an 18-month period, the medical records of all patients with PAS who attended a clinic for thrombosis and hemostasis were reviewed on arrival. The patients were then referred to a retinal clinic and examined prospectively. The first 20 consecutive patients underwent retinal fluoroangiography. MAIN OUTCOME MEASURES: The prevalence of the various organs affected by the disease was noted in the group, as well as the prevalence and type of ophthalmic involvement. RESULTS: The most common forms of systemic involvement were fetal loss in 11 female patients (46%), central nervous system involvement in 17 patients (44%), and venous thrombosis in 16 patients (41%). Ophthalmic findings were as follows: 13 patients (33%) were symptomatic, and 12 of them complained of visual disturbances. In 10 (83%) of these 12 patients, the visual symptoms were transient, and 7 of them were nonocular in nature. Pathologic signs on ophthalmic examination were found in 5 of the 39 patients (13%), two of whom had intraocular pathologic findings related to PAS, and these were in the form of mild retinopathy. Of the 36 patients who were either asymptomatic or had transient visual disturbances, pathologic ocular findings were detected in only 2 (6%). Routine retinal fluoroangiography did not reveal any additional information that could not otherwise have been detected by funduscopy. CONCLUSIONS: Ocular involvement in PAS is uncommon. Transient visual disturbances are common, although most of them are related to central nervous system rather than ocular ischemia. Pathologic ophthalmic findings are unlikely to be found in asymptomatic patients or in patients with transient visual disturbances. Routine retinal fluoroangiography performed on patients with PAS is unproductive.

Surgical outcome of transconjunctival cryosurgical extraction of orbital cavernous hemangioma.

BACKGROUND AND OBJECTIVE: Transconjunctival cryosurgical extraction (TCE) is not an accepted method for the removal of most intraorbital tumors, and thus is not routinely performed. This study evaluates TCE for orbital cavernous hemangioma (OCH), which, due to its specific vascular structure, is suitable for this technique. PATIENTS AND METHODS: In this retrospective case series, the medical charts of 12 consecutive patients with OCH who underwent TCE were reviewed. The operative complications and the surgical outcome were analyzed. RESULTS: Eleven tumors were intraconal and 1 was extraconal. No tumor involved the orbital apex. In 10 patients (83%), the operation was uneventful. One patient lost vision due to intraoperative cilioretinal arterial occlusion. In another patient, the tumor could not be identified and was subsequently removed via lateral orbitotomy. CONCLUSION: TCE, a relatively minor operation, is as safe as more extensive surgical procedures for the removal of OCH not located in the posterior third of the orbital space.

Nd:YAG laser treatment for premacular hemorrhage.

The authors describe two cases of premacular hemorrhage secondary to retinal macroaneurysm, with significant loss of vision. A few low-energy bursts of ND:YAG laser were applied to perforate the anterior surface of the hemorrhage, subsequently draining the blood into the vitreous cavity. The blood gradually cleared from the vitreous, with restoration of good vision. Nd:YAG laser seems to be a safe and efficient method for treating premacular hemorrhage and should be considered as a therapeutic option.

Lack of effect of prophylactic gentamicin treatment on intraocular and extraocular fluid cultures after pars plana vitrectomy.

BACKGROUND AND OBJECTIVE: To analyze the effect of preoperative prophylatic 0.3% gentamicin sulfate eyedrops on intraocular and extraocular fluid cultures after pars plana vitrectomy. PATIENTS AND METHODS: Forty consecutive patients undergoing pars plana vitrectomy were randomly assigned to receive either 0.3% gentamicin eyedrops or placebo preoperatively. Fluids from the vitreous cavity and from the conjunctival sac were collected for bacteriologic studies. RESULTS: Positive cultures from extraocular fluids were obtained in 30.4% of the gentamicin group and 35.3% of the placebo group (P = .75). Furthermore, 1 (3%) of 40 cultures of intraocular fluid and 13 (33%) of 40 cultures of extraocular fluid were positive. Culture growth positivity was not influenced by the patients' age, by diabetes, or by the duration of the operation. CONCLUSIONS: Prophylactic gentamicin treatment by the method applied in this study is not effective on reducing growth positivity. Contaminated extraocular fluid may play an important role in the pathogenesis of bacterial endophthalmitis following pars plana vitrectomy.

Cryoextraction of orbital capillary hemangioma diagnosed by technetium-99m-labeled red blood cell scintigraphy.

The authors report a case of an infant in whom orbital capillary hemangioma was diagnosed using technetium-99m-labeled red blood cell scintigraphy. The tumor was subsequently removed by transconjunctival cryoextraction.

The application of the macular photocoagulation study eligibility criteria for laser treatment in age-related macular degeneration.

BACKGROUND AND OBJECTIVE: To analyze the application of the Macular Photocoagulation Study eligibility criteria for laser photocoagulation of choroidal neovascularization, in view of the expansion of these criteria in recent years. PATIENTS AND METHODS: The authors prospectively analyzed 50 eyes of 47 consecutive patients with exudative age-related macular degeneration (AMD) to determine their suitability for treatment. RESULTS: Fifteen eyes (30%) were found to be suitable for laser photocoagulation. Patients eligible for treatment were more likely to have experienced visual symptoms for a months or less (P = .006), to have a visual acuity of 20/200 or better (P = .009), and to be younger in age (P = .02). Visual symptoms experienced for a month or less were more prevalent in extrafoveal exudative lesions compared with the subfoveal type (P = .01) CONCLUSIONS: Despite recent advances, laser photocoagulation still can be applied only to a minority of the patients with neovascular AMD. Prompt ocular examination following the onset of visual symptoms is essential.

Activated long-term peripheral blood cultures as preparation for adoptive alloreactive cell therapy in cancer patients.

Different modes of in vitro activation of peripheral blood mononuclear cells (PBMC) were compared for their effect on long-term propagation. PBMC cultures were activated by short exposures to the mitogen phytohemagglutinin (PHA) and the CD3 complex, with or without secondary signals provided by ligands of CD28 costimulatory molecules. Activation and long-term cultures were carried out in the presence of recombinant interleukin-2 (rIL-2). Addition of supernatant derived from IL-2-activated PBMC improved culture cell yield. Cumulative fold expansions ranged between 10(3) and 10(5) within 21 days. The highest cell yield was found after PHA activation. Fewer cells were obtained after activation with a combination of CD3 and CD28, and even fewer were obtained after CD3 activation alone. An increase in CD8+ and CD56+ cells, without change in CD4+ cells, was found in activated cultures when compared with fresh PBMC. Non-MHC-restricted cytotoxic activity was documented in all activated cultures. Cytotoxic activity per culture was highest in PHA-activated PBMC because of the high cell yield on the day of harvest. Successful in vitro expansion of PBMC might be helpful for gene transfer into T lymphocytes, as well as for the induction of an antitumor response, particularly for prevention and treatment of relapse of hematologic malignancies following allogeneic or autologous bone marrow transplantation.

[Prognostic factors and surgical results in traumatic cataract].

The visual outcome in 23 men and 2 women with traumatic cataracts was analyzed retrospectively. Their average age was 33 and they ranged from 10 to 69 years. Surgical results were either very good or very poor. Associated retinal injuries significantly decreased final visual acuity (p = 0.001). Those with initial visual acuity restricted to finger counting had better visual results than those with initial visual acuity restricted to light perception (p = 0.01) and hand motions (p = 0.02). Usually the lens was removed via the pars plana; the most common mode of optical correction was contact lenses.

[Motor vehicle accidents and eye injuries].

The medical records of 24,632 patients treated in our surgical emergency service over a 3-year period were reviewed to determine the frequency and characteristics of ocular trauma caused by motor vehicle accidents (MVA). MVA-related injuries accounted for 13.9% of all visits to the service and involved 1106 of the patients (33%), of whom 77% were young males. At least 1 pathological finding was found in 858 (77.6%) and 169 (15.2%) were admitted. 16 patients sustained very severe ocular injuries which resulted in poor vision.