Optimization of Large-Scale Expansion and Cryopreservation of Human Natural Killer Cells for Anti-Tumor Therapy

Allogeneic natural killer (NK) cell therapy is a potential therapeutic approach for a variety of solid tumors. We established an expansion method for large-scale production of highly purified and functionally active NK cells, as well as a freezing medium for the expanded NK cells. In the present study, we assessed the effect of cryopreservation on the expanded NK cells in regards to viability, phenotype, and anti-tumor activity. NK cells were enormously expanded (about 15,000-fold expansion) with high viability and purity by stimulating CD³⁺ T cell-depleted peripheral blood mononuclear cells (PBMCs) with irradiated autologous PBMCs in the presence of IL-2 and OKT3 for 3 weeks. Cell viability was slightly reduced after freezing and thawing, but cytotoxicity and cytokine secretion were not significantly different. In a xenograft mouse model of hepatocellular carcinoma cells, cryopreserved NK cells had slightly lower anti-tumor efficacy than freshly expanded NK cells, but this was overcome by a 2-fold increased dose of cryopreserved NK cells. In vivo antibody-dependent cell cytotoxicity (ADCC) activity of cryopreserved NK cells was also demonstrated in a SCID mouse model injected with Raji cells with rituximab co-administration. Therefore, we demonstrated that expanded/frozen NK cells maintain viability, phenotype, and anti-tumor activity immediately after thawing, indicating that expanded/frozen NK cells can provide ‘ready-to-use’ cell therapy for cancer patients.

Novel T cells with improved in vivo anti-tumor activity generated by RNA electroporation

The generation of T cells with maximal anti-tumor activities will significantly impact the field of T-cell-based adoptive immunotherapy. In this report, we found that OKT3/IL-2-stimulated T cells were phenotypically more heterogeneous, with enhanced anti-tumor activity in vitro and when locally administered in a solid tumor mouse model. To further improve the OKT3/IL-2-based T cell manufacturing procedure, we developed a novel T cell stimulation and expansion method in which peripheral blood mononuclear cells were electroporated with mRNA encoding a chimeric membrane protein consisting of a single-chain variable fragment against CD3 and the intracellular domains of CD28 and 4-1BB (OKT3-28BB). The expanded T cells were phenotypically and functionally similar to T cells expanded by OKT3/IL-2. Moreover, co-electroporation of CD86 and 4-1BBL could further change the phenotype and enhance the in vivo anti-tumor activity. Although T cells expanded by the co-electroporation of OKT3-28BB with CD86 and 4-1BBL showed an increased central memory phenotype, the T cells still maintained tumor lytic activities as potent as those of OKT3/IL-2 or OKT3-28BB-stimulated T cells. In different tumor mouse models, T cells expanded by OKT3-28BB RNA electroporation showed anti-tumor activities superior to those of OKT3/IL-2 T cells. Hence, T cells with both a less differentiated phenotype and potent tumor killing ability can be generated by RNA electroporation, and this T cell manufacturing procedure can be further optimized by simply co-delivering other splices of RNA, thus providing a simple and cost-effective method for generating high-quality T cells for adoptive immunotherapy.

The Impact of ABCB1 Gene Polymorphism on Steroid Responsiveness in Acute Rejection in Kidney Transplantation / 대한이식학회지

BACKGROUND: Steroid pulse therapy has been used for patients with acute rejection after kidney transplantation. The ABCB1 gene codes for P-glycoprotein, a transporter that is involved in the metabolism of steroids. However, the role of ABCB1 polymorphisms has not been investigated in patients with acute rejection after kidney transplantation. METHODS: Among 763 patients that received kidney or simultaneous pancreas-kidney transplantation at Seoul National University Hospital between May 1996 and July 2009, 684 patients agreed to genetic sampling for polymorphisms. Acute rejection was defined as biopsy-proven, acute cellular rejection with increased serum creatinine, or in the context of delayed or slow graft function. Steroid-resistance was defined as no improvement in serum creatinine, need for additional OKT3 or ATG treatment, or repeated acute rejection within 30 days. Three polymorphisms of ABCB1 gene (C1236T, C3435T, G2677T/A) were assessed. RESULTS: C allele frequency of C3435T was 59.3% and of C1236T 40.1%. Patients who were steroid-resistant (n=37) had higher serum creatinine at kidney biopsy compared to those who were steroid-sensitive (n=49, P<0.001). The frequency of ABCB1 gene polymorphisms (C1236T and C3435T) did not differ significantly between patients who were steroid-sensitive and those who were resistant. An association with G2677T/A could not be analyzed due to a high failure rate of genotyping. CONCLUSIONS: ABCB1 gene polymorphisms (C1236T and C3435T) were not associated with steroid resistance in patients with acute cellular rejection after kidney transplantation.

Ex vivo expansion of tumor-infiltrating lymphocytes from nasopharyngeal carcinoma patients for adoptive immunotherapy / 癌症

Establishing Epstein-Barr virus(EBV)-specific cytolytic T lymphocytes(EBV-CTLs) from peripheral blood mononuclear cells(PBMCs) for adoptive immunotherapy has been reported in EBV-associated malignancies including Hodgkin's lymphoma and nasopharyngeal carcinoma(NPC). In the current study, we performed ex vivo expansion of tumor-infiltrating lymphocytes(TILs) obtained from NPC biopsy specimens with a rapid expansion protocol using anti-CD3 monoclonal antibody(OKT3), recombinant human interleukin(IL)-2, and irradiated PBMCs from healthy donors to initiate the growth of TILs. Young TIL cultures comprised of more than 90% of CD3+ T cells, a variable percentage of CD3+CD8+ and CD3+CD4+ T cells, and less than 10% of CD3-CD16+ natural killer cells, a similar phenotype of EBV-CTL cultures from PBMCs. Interestingly, TIL cultures secreted high levels of the Th1 cytokines, interferon gamma (IFNγ) and tumor necrosis factor-alpha (TNF-α), and low levels of the Th2 cytokines, IL-4 and IL-10. Moreover, young TILs could recognize autologous EBV-transformed B lymphoblast cell lines, but not autologous EBV-negative blast cells or allogeneic EBV-negative tumor cells. Taken together, these data suggest that ex vivo expansion of TILs from NPC biopsy tissue is an appealing alternative method to establish T cell-based immunotherapy for NPC.

CD25 Monoclonal Antibody (Basiliximab) Used in High Risk Kidney Recipient (Single Center Experience) / 대한이식학회지

PURPOSE: CD25 monoclonal antibody (basiliximab, BA) has been reported an excellent effect on induction immunosuppression than ALG, ATG, OKT3 in renal transplantation. Since we can use BA only in the group of high risk kidney recipient, we want to evaluate the effect of BA treated group by comparing with conventional 3 drugs combination group (calcineurin inhibitor/cyclosporine or tacrolimus, mycophenolate mofetil and prednisolon). METHODS: Total 103 recipients were treated by BA and conventional 3 drugs from March 2000 through February 2006, and these cases were compared with 122 recipients without BA. Government guidelines for using BA were in cadaveric transplantation, more than 4 HLA mismatching, zero DR antigen matching, retransplantation and more than 80% positive PRA. The episode of acute rejection (AR), steriod resistant acute rejection, change of serum creatinine, maintaining dosage of calcineurin inhibitor (CNI) and other side effects were compared between groups. RESULTS: The rate of AR within 12 months after transplantation was 11.7% in BA group while 9.0% in control group (P=0.451). The steroid resistant acute rejection was 16.6% in BA group and 27.3% in control group (P=0.119). Rejection free graft survival adjusted various clinical risk factors by Cox-regression test were no significance between groups. Serum creatinine level at one year after transplantation was 1.61+/-.1 and 1.46+/-.7 mg/dL in BA and control group, and recipients number whose SCr over 1.5 mg/dL were 39.0% and 32.7% (P=0.326). Cyclosporin dosage at one year in BA and control group were 4.21 and 3.62 mg/kg (P=0.202) and 0.11 and 0.17 mg/kg in tacrolimus group (P=0.291). Incidence of PTDM and viral infection were all no difference statistically between groups. CONCLUSION: In conclusion, BA induction immunosuppression with CNI, mycophenolate mofetil and prednisolon used in high risk kidney recipient effectively control the acute rejection and steroid resistant acute rejection for one year at least the same as control group. But since there was no more beneficial effect in BA added to Tac group than conventional Tac based immunosuppression, this 4 drug combination is better avoided if possible.

Detection, enrichment and expansion of T lymphocytes mediating alloresponse based on cytokine / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To detect, enrich and expand the cytokine secreting T lymphocytes after allogeneic PBMNCs stimulation.</p><p><b>METHODS</b>The novel cytokine secretion assay (CKSA) was applied to detect T lymphocytes secreting IFN-gamma at single cell level in human mixed lymphocytes reaction. IFN-gamma secreting T cells were enriched by means of magnetic sorting system and expanded with OKT(3), anti-CD(3)mAb and IL-2 combination. Antigen specificity of the expanded cells was confirmed using enzyme linked immunospot assay.</p><p><b>RESULTS</b>A sizable proportion of IFN-gamma secreting T lymphocytes could be detected [(1.12 +/-0.13)% compared with (0.23 +/-0.07)%] and be further enriched to (67.3 +/-10.5)%, or (93.8 +/-22.1) fold. T lymphocytes could be expanded up to 600-fold within 21-28 days and the specific IFN-gamma response of expanded cells was confirmed with stimulation of the relevant allogeneic PBMNC, which was significantly higher than the irrelevant PBMNC control.</p><p><b>CONCLUSION</b>It is feasible to detect significantly increased IFN-gamma secreting T lymphocytes after allogeneic PBMNCs stimulation based on the CKSA technique at single cell level and these cells can be efficiently enriched and expanded for further research.</p>

Desorden linfoproliferativo en el paciente con transplante de órgeno sólido: manifestaciones cutáneas / Lymphoproliferative disorders in a solid organ transplant patient: cutaneous manifestations

El desorden linfoproliferativo en el paciente transplantado es una entidad caracterizada por una proliferación del linfocito B infectado por el virus de Epstein Barr (EBV), que provoca manifestaciones extracutáneas y cutáneas. La forma cutánea de presentación es infrecuente. Presentamos dos pacientes transplantados, renal y cardiopulmonar, que presentaron nódulos en la pierna derecha y en el tronco. El estudio histopatológico reveló la presencia de un infiltrado de blastos linfoides de estirpe B. El tratamiento se basó en el descenso de la inmunosupresión, además de radioterapia y quimioterapia. La evolución del cuadro fue favorable en ambos pacientes

OKT3-Induced Cytokine Release Attenuation by Interleukin-10

Purpose: OKT3 is a very powerful immunosuppressive drug in acute allograft rejection treatment but its side-effects such as fever, nausea, vomiting, and pulmonary edema are strongly linked with its inducing cytokines such as tumor necrosis factor alphaTNF-alpha, interleukin-6(IL-6), and interferon-gammaIFN-gamma. Interleukin-10(IL-10) inhibits proinflammatory cytokines which are produced by activated monocyte/ macrophages and prevents production of cytokines in acute inflammatory states. The purpose of this study is to determine the effect of exogenous administration of the anti- inflammatory cytokine, IL-10 on TNF-alpha IL-6, and IFN-gammaproduction and pulmonary injury after OKT3 injection. METHODS: For experiment, Sprague-Dawley rat weighed 300~400 gm was injected either OKT3(0.6mg/kg i.v.) only or recombinant IL-10(0.5microgram/rat i.p. one hour before the injection of OKT3(IL-10/OKT3). The rats were divided into three groups; control group(n=5); normal saline injected group(1.0ml/rat i.v.), OKT3 group(n=5); OKT3 only injected group, and IL-10/OKT3 group; IL-10 plus OKT3 injected group. After two hours of injection, all animals were sacrificed and submitted for a study of serologic and histologic changes. Student t-test was used for statistical analysis. To evaluate the cytokine production the serum levels of TNF-alpha, IL-6, and IFN-gamma level were measured. The serum levels of TNF-alpha, IL-6, and IFN-gamma were also significantly decreased in IL-10/ OKT3 group(109.6+/-38.0, 65.2+/-14.1, 96.2+/-17.3pg/ml) compared with OKT3 group(399.8+/-71.4, 155.4+/-45.1, 297+/-87.0pg/ml)(p<0.05). To determine the pulmonary injury, wet/dry ratio and microscopic findings for the lung tissue were analyzed. RESULTS: The wet/dry ratio of the lung tissue was significantly decreased in IL-10 /OKT3 group (3.52+/-0.31) compared with OKT3 group(4.16+/-0.48)(p<0.05). Microscopic findings of lung tissue revealed severe neutrophilic infiltration and microvascular congestion in the OKT3 group, but in IL-10/ OKT3 group, neutrophilic infiltration and microvascular congestion were decreased. CONCLUSION: In this study the inhibitory effect of IL-10 on the production of proinflammatory cytokines by OKT3 treatment was significant. This results suggest that the exogenous IL-10 injection may decrease the complications associated with OKT3 treatment of allograft rejection in organ transplantation.

Prevention and treatment of rejection after simultaneous pancreas-kidney transplantation / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To explore methods of preventing and reversing rejection after simultaneous pancreas-kidney (SPK) transplantation.</p><p><b>METHODS</b>Seventeen patients underwent SPK transplantation from September 1999 to September 2003 were reviewed retrospectively. Immunosuppression was achieved by a triple drug regimen consisting of cyclosporine, mycophenolate mofteil (MMF), and steroids. Three patients were treated with anti-CD3 monoclone antibody (OKT3, 5 mg x d(-1)) for induction therapy for a mean period of 5-7 days. One patients received IL-2 receptor antibodies (daclizumab) in a dose of 1 mg x kg(-1) on the day of transplant and the 5th day posttransplant. One patient was treated with both OKT3 and daclizumab for induction.</p><p><b>RESULTS</b>No primary non-functionality of either kidney or pancreas occurred in this series of transplantations. Function of all the kidney grafts recovered within 2 to 4 days after transplantation. The level of serum creatinine was 94 +/- 11 micromol/L on the 7th day posttransplant. One patient experienced the accelerated rejection, resulting in the resection of the pancreas and kidney grafts because of the failure of conservative therapy. The incidence of the first rejection episodes at 3 months was 47.1% (8/17). Only the kidney was involved in 35.3% (6/17); and both the pancreas and kidney were involved in 11.8% (2/17). All these patients received a high-dose pulse of methylprednisone (0.5 g x d(-1)) for 3 days. OKT3 (0.5 mg x d(-1)) was administered for 7-10 days in two patients with both renal and pancreas rejection. All the grafts were successfully rescued.</p><p><b>CONCLUSION</b>Rejection, particularly acute rejection, is the major cause influencing graft function in SPK transplantation. Monitoring renal function and pancreas exocrine secretion, and reasonable application of immunosuppressants play important roles in the diagnosis and treatment of rejection.</p>

Post-Transplant Lymphoproliferative Disorder / 대한이식학회지

Post-transplant lymphoproliferative disorder (PTLD) is a heterogeneous group of conditions characterized by Epstein-Barr virus (EBV)-driven lymphoid proliferation in the face of impaired T-cell immune surveillance. Most of PTLD are of B- cell origin. Sero-negative recipients of sero-positive organs, and recipients receiving OKT3 are at increased risk. The clinical features are multiple and varied and can range from a benign mononucleosis-like illness to a fulminant non- Hodgkin's lymphoma. Frequently, an extranodal presentation is found with a predilection for brain and allograft. The presentation of forms localized to the renal graft may be incidentally discovered during imaging of the kidney; because the masses may also produce a pressure effect on the collecting system, the patient may present with renal obstruction. To establish diagnosis of PTLD, tissue biopsy is necessary. The mainstay of treatment is immunosuppressive dose reduction. For patients failing to respond to a reduction in immunousppression, a second step of treatment can be used such as interferon alpha, chemotherapy, anti-CD20 antibody (rituximab), and EBV-specific cytotoxic T-cells. Chemotherapy is currently the most commonly used therapeutic alternative. Anti-viral drug therapy has been shown to be of very limited benefit and there are theoretical reasons why it may be ineffective. There appears to be a correlation between PTLD and EBV viral load measured by polymerase chain reaction (PCR) of the peripheral blood, and quantitative PCR may be a useful guide in the management of PTLD. Multicenter randomized trials and standardized diagnostic and therapeutic strategies are required to improve our understanding of PTLD.

Rituximab Rescue for Refractory Antibody Mediated Rejection after Kidney Transplantation / 대한이식학회지

PURPOSE: Antibody mediated rejection (AMR), although less common than acute cellular rejection (ACR), may be recalcitrant to conventional rescue therapy. AMR is caused by de novo B-cell mediated production of immunoglobulin G antibody (IgG) targeted against specific allograft antigen in a presensitized recipient. Rituximab is a chimeric murine- human anti-CD20 monoclonal antibody which targets CD-20 positive B-cells for elimination. Rituximab has been described to improve allograft salvage for refractory AMR. METHODS: From January 2002 to May 2004, 11 patients were diagnosed with AMR. The first 5 patients (non-rituximab group: NRG) were treated with high dose steroids, plasmapheresis followed by IVIG (500 mg/kg/dose) in addition to OKT3 and/or rabbit antithymocyte globulin. The latter 6 patients (rituximab group: RG) were given Rituximab (375 mg/m2) with IVIG following plasmapheresis. All patients had biopsy proven AMR. RESULTS: Four patients received allografts from living donors and one patient from cadaveric donor in NRG. Each three patients received allografts from living or cadaveric donors in RG. One patient of RG had a positive anti-HLA B-cell crossmatch by CDC (complement dependent cytotoxicity). The anti-donor antibody was reduced to zero with negative CDC and flowcytometry through a desensitization protocol prior to transplantation. The time to diagnosis of AMR in both groups were 17.8+/-18.17 days (NRG); 11+/-2.5 days (RG). ACR was identified in conjunction with AMR in 2 (40%: NRG), 4 patients (66.7%: RG), respectively. All patients had biopsies with classic features of AMR on light microscopy, including C4d staining. Three (50%) patients of RG had positive post-transplantation CDC and donor-specific antibody (DSA) identified. Mean serum creatinine (SCr) upon diagnosis of AMR were 4.3+/-1.71 mg/dL (NRG); 5.77+/-2.65 mg/dL (RG). The rescue rate of RG was superior than NRG (83% vs. 40%, P>0.05). The time to rescue from AMR in both groups were 40.5 +/-28.99 days (NRG); 48+/-54.67 days (RG). Mean SCr of the rescued patients were 1.65+/-0.07 mg/dL (NRG); 2.2+/-1.4 (RG) with median follow up of 120 days (range 33~319 days). Allograft nephrectomies were performed in 3 patients of NRG. CONCLUSION: Rescue therapy with Rituximab improves allograft salvage after AMR and should be considered early in the treatment of biopsy proven AMR.

Tacrolimus in Delayed Graft Function in Cadaveric Renal Transplantation / 대한신장학회잡지

BACKGROUND: In the presence of anticipated or established acute tubular necrosis (ATN) immediately after cadaveric kidney transplantation, induction with monoclonal or polyclonal antibody is recommended in preparation of increased risk of acute rejection caused by ATN. Tacrolimus is a potent immunosuppressive agent than cyclosporine. In this study, we analyzed retrospectively the clinical outcome of patients who had taken tacrolimus as a replacement of cyclosporine in the period of delayed graft function(DGF) to determine the eligibility of tacrolimus instead of antilymphocyte antibody in this situation. METHODS: Between March 1, 1991 and August 31, 2000, DGF developed in eighteen first cadaveric renal transplant recipients in our center. During DGF period, twelve patients received tacrolimus based immunosuppression without OKT3. We reviewed the complete clinical course of the 12 patients. RESULTS: Among the 12 patients, 1 patient underwent graft nephrectomy at postoperative 27 days, because of poor renal function and concomitant aspergillosis infection. In the remaining 11 patients, however, for whom tacrolimus was maintained continuously without OKT3 therapy, renal function was recovered successfully. One acute rejection developed at postoperative 15 months. One patient died at postoperative 5 months with functioning graft. One-year graft survival rate was 83%. CONCLUSION: Tacrolimus could be used in replacement of cyclosporine for the prevention of acute rejection in DGF. This could provide a graft survival comparable to that by the monoclonal or polyclonal antibodies without the potential risk of life- threatening side effects in this situation.

Clinical Experiences of 52 Cases Cadaveric Donor Kidney Transplantation / 대한이식학회지

PURPOSE: The organ procurement organization (OPO) and transplant coordinator were established in Chonnam National University Hospital in 1993 to promote organ donation and to manage organ procurement. At the same time, the first protocol and planning transplant was performed simultaneously. We performed this study to know the predicting factors and survival rate of cadaveric kidney transplantation. METHODS: First cadaveric donor kidney transplantation was performed at May 1993 in Chonnam National University Hospital. Thereafter 52 cases of cadaveric kidney transplantation were performed using 28 cadaver donors till December 1999. The most frequent cause of brain death was head injury by traffic accident. Male to female donor ratio was 1.8 : 1. 52 recipients (29 males, 23 females), aged 20 to 65 years (median age 36 years) were the subjects of this study. The immunosuppressive regimens consisted of cyclosporin, mycophenolate mofetil, and prednisone. Acute rejection was treated with three consecutive bolus of 1.0 gram methylprednisolone or 5 mg/day for 10 days of OKT3. Three HLA mismatchs were 7 cases (13.5%) and six mismatches were 6 cases (11.5%). There were 2 cases of multiple renal arteries. RESULTS: There was no primary non functioning graft. In the first 7 post operative days, urine amount less than 4,000 mL per day was noted in 29 patients (55.8%) and serum creatinine over 1.5mg/dL was noted in 13 patients (25%). No significant proteinuria and hematuria was observed. Postoperative medical complications were occured in 12 patients (23.1%) and minor surgical complications in 3 patients (5.8 %). One patient was performed reoperation because of urinary leakage. CMV infections were noted in 15 patients (28.8%). Acute rejection episodes were 17 cases (32.7%). We lost 4 grafts within 1 year. The major cause of graft loss was patient death. Recipient age was significant risk variable for graft and patient survival in multivariate analysis (P=0.012). one and five year graft survival rates were 92.15%. CONCLUSION: To achive better results, continued attention should be paid to the cadaveric donor organ procurement.

Comparison of Prevention for Acute Rejection in Renal Transplantation between Mycophenolate Mofetil(MMF) and Azathioprine / 대한신장학회잡지

BACKGROUND: Acute renal allograft rejection is not only risk factor of chronic rejection but is also a significant cause of graft loss and patient death. MMF has been shown to reduce the incidence and severity of acute rejection. METHODS: To compare the risk of acute rejection and side effects of MMF with azathioprine(AZA), a total of 108 patients, who received living transplants, were divided in two groups : MMF(n=48) and AZA group(n=60). Cyclosporin microemulsion(Neoral) and steroid were administered concomitantly to all patients. RESULTS: The MMF group was significantly lower rate of acute rejection compared with AZA group during the first 3 months after renal transplantation(14.6% vs 30.0%, p=0.005). 54.5% of patients in the MMF group and 44% in the AZA group were treated only with steroid pulsing for acue rejection. 45.5% in the MMF group, compared to 56% in the AZA group, required OKT3 or Atgam for treatment of severe acute rejection, the difference is not significant. Treatment failure occurred among 31.3% of the MMF group compared with 55% in the AZA group(p=0.013). Serum creatinine of 6 months after transplantation was significantly lower in the MMF group than in the others(1.31+/-.27 vs 1.50+/-.28 mg/dL, p=0.017). The incidence of opportunistic infection was similar in both groups. Gastrointestinal side effects were more common in the MMF group 14.6% than in the AZA group 3.3%(p=0.035), while leukopenia was more common in the AZA group 21.7% than in the MMF group 4.3%(p=0.017). CONCLUSION: MMF reduced the incidence of acute rejection without notable side effects. Long-term follow up will be needed to establish the protective effect of MMF against immunological attack.

Effect of addition of proton carriers in culture medium on growth and secretion of hybridoma cell line OKT3.

Monoclonal antibodies (MAb) constitute the centre of all in-vitro diagnostic measures and almost all in-vivo therapeutic manoeuvres now. Production emphasis for these antibodies is having a current shift from animal-based large-scale culture to in-vitro bioreactor-based high-density culture. One of the major difficulties in high-density culture is end-metabolite accumulation in batch and fed-batch cultures in the forms of H+, NH4+ etc.. thereby reducing cellular growth and secretions. In the present study, effects of added proton carries--NAD and NADP--over and above the metabolic pools of the molecules, were examined on the cellular growth and secretion kinetics. Although NADP fortification showed a remarkable improvement in cellular growth (time dependent 200-300% improvements compared to controls) and size, cumulative MAb titre was better with NAD fortification. Combined additional loads of the proton carriers would be interesting to study in high density culture conditions.

O destino dos rins transplantados tratados com OKT3 para rejeiçäo córtico-resistente / The fate of trenal allgrafts treated with OKT3 for steroid-resistan rejection

Objetivo: Avaliar o efeito do anticorpo monoclonal anti-CD3 (OKT3), utilizado para tratamento de rejeição aguda córtico-resistente em pacientes transplantados renais, em relação à função do rim transplantado e à sobrevida do enxerto e do paciente a longo prazo...

Postrenal Transplantation Cyclosporine Associated Hemolytic Uremic Syndrome in End Stage Renal Failure Patient Caused by Systemic Lupus Erythematosus / 대한이식학회지

Although the outcome and the possibility of renal recurrence of disease in systemic lupus erythematosus is still a matter of controversy, kidney transplantation is generally regarded as a proper indication for the treatment of patients with end stage renal failure caused by systemic lupus erythematosus. Hemolytic uremic syndrome is characterized by the symptoms of sudden onset of hemolytic anemia, thrombocytopenia, and deteriorating renal function. Many patients with postrenal transplantation hemolytic uremic syndrome have lost their grafts because of no known established treatment modality. Although the substitution of cyclosporine to FK506 is reported as a successful strategy for the treatment of cyclosporine associated hemolytic uremic syndrome in many reported cases, we cannot find the constant reports because FK506 or even OKT3 is also known as the cause of postrenal transplantation hemolytic uremic syndrome. But cyclosporine associated hemolytic uremic syndrome can be treated by the proper choice of immunosuppressant and conservative treatment. In this report, a patient with end stage renal failure caused by systemic lupus erythematosus experienced cyclosporine associated postrenal transplantation hemolytic uremic syndrome. He has recovered from the symptoms by withdrawal of cyclosporine and reduced dose of FK506 and at the same time, conservative treatment. We report this case with literature review.

A Case of Polyoma Virus(PV) Infection in a Renal Allograft Recipient / 대한신장학회잡지

We report one case of renal PV infection after renal allograft transplantation leading to graft dysfunction. According to prior reports, PV induced interstitial nephritis might be a cause of graft loss. Pathologic findings show varying degrees of interstitial infiltration and tubular degenerative changes, which resemble acute cellular rejection. Therapeutic strategies have not yet been developed. Case ; A 23 years old male underwent renal transplantation from his HLA haploidentical 25 year old sister. His renal function had been good with cyclosporin, steroid and azathioprine until 9 months after transplantation, when his serum creatinine level rose to 2.2mg/dl. The renal biopsy revealed diffuse lymphocyte infiltration in the interstitium and feature of the tubulitis. Also, giant tubular epithelial cells with large, hyperchromic nuclei were present. Despite steroid pulsing and OKT3, renal function progressively de- teriorated. After 10 days of OKT3 therapy, the patient suffered from high fever, dyspnea and general aches. A chest X-ray revealed interstitial infiltration in both lung fields and the cytomegalovirus PCR (polymerase chain reaction) test of serum and blood was positive. Intravenous ganciclorvir was administered and immunosuppressants were tapered. 4 months after admission, he lost his graft function and underwent hemodialysis. The aforementioned renal biopsy was retested immunohistochemically. Nuclear inclusions in renal tubular epithelial cells were shown and these inclusions were reacted positively with PV monoclonal antibodies.

Effectiveness of Cipol-N(R) in Primary Living Donor Kidney Transplant Patients: Open-Label, Multi-Center Study / 대한이식학회지

Cyclosporine (CsA) has been one of the main immunosuppressants after kidney transplantation since its introduction in Korea. There was remarkable improvement of graft survival in kidney transplantation with CsA, compared with azathioprine. Cipol-N(R)(Chong Kun Dang, Korea), microemulsion gelatin capsule formulation of CsA, is a new generic drug. This pure domestic brand of CsA was tested for bioequivalence in healthy adults compared with the reference drug of the same formulation, Sandimmun-Neoral(R)(Novartis, Switzerland) in 1997. This open-label, multi-center study is designed to evaluate the efficacy of Cipol-N(R) in primary kidney transplant recipients for 6 months after transplantation. A total of 59 patients from 4 medical centers were enrolled in the study. Maintenance immunosuppressive protocol was based on CsA and steroid dual therapy, which was induced 2 days prior to the operation. Acute rejection was diagnosed with clinical or pathological clue. Clinical criteria for the diagnosis of acute rejection were oliguria, graft swelling and tenderness, rising serum creatinine, fever, and papillary swelling and increased vascular resistant index on Doppler ultrasonography. Steroid pulse therapy was used as primary treatment. Steroid resistant acute rejection was treated with anti-lymphocyte agents such as OKT3, ATG, or ALG. The primary efficacy endpoint was onset of acute rejection or treatment failure, defined as graft loss, death, or premature termination from the study for any reason. Incidence and severity of acute rejection, actual survival rate of patient and graft, function of the graft, pharmacokinetics of the Cipol-N(R), and the primary efficacy variables were evaluated 6 months after transplantation. All enrolled patients were included in the primary analyses of efficacy on the basis of intent to treat. Mean age of the patients was 37.1 10.4 years old. Male and female ratio was 42:17. There were 38 related pairs, which included 5 HLA identical and 33 HLA haplo-identical matches, and 21 unrelated pairs. A total of 10 patients were withdrawn from the study before post- transplant 6 months. The causes for premature withdrawal were patient's request without specific reason (6), partially rescued acute rejection (3), and patient's death (1). There were 27 episodes of acute rejection in 25 patients, which were diagnosed clinically (11) and pathologically (16). Steroid pulse therapy and anti-lymphocyte agent were used in 24 and 3 cases respectively. There were 4 patients, who showed partial rescue but no graft loss due to acute rejection. Patient and graft survival was 98.3% at post-transplant 6 months. Serum creatinine concentration showed 1.3-1.7 mg/dl all through the study period, which meant relatively stable graft function. Mean daily doses of Cipol-N(R) at post-transplant 1 and 6 months were 325 and 300 mg respectively. With this short term study, we can report that Cipol-N(R) showed relatively good efficacy in primary living donor kidney transplantation. Further study is needed for the evaluation of long term efficacy and safety.