RESUMEN
OBJECTIVES: To review the scientific rationale for the clinical use of recombinant bactericidal permeability-increasing protein (rBPI21) and to discuss the results, implications, and lessons learned during the clinical development of rBPI21 for adjunctive treatment of children with severe meningococcemia. DATA SOURCES: The published medical literature. STUDY SELECTION: Of the phase I/II and phase III trials in humans, preclinical experimental studies were selected. Data from these sources are presented in the context of the authors' experiences as principal investigators in the phase I/II and/or phase III clinical trials. DATA EXTRACTION AND DATA SYNTHESIS: Bactericidal permeability-increasing protein and N-terminal fragments of bactericidal permeability-increasing protein, such as rBPI21, bind and neutralize endotoxin and are potently bactericidal against both smooth and rough forms of Gram-negative bacteria, including Neisseria meningitidis. Based on these properties and compelling preclinical data indicating that administration of rBPI21 reduced mortality in several models of sepsis, we initiated clinical trials by using rBPI21 as adjunctive therapy for children with severe meningococcemia. Data from the phase III, randomized, placebo-controlled trial indicate that rBPI21 reduces clinically significant morbidities and improves the functional outcome of children with severe meningococcemia. No statistically significant benefit in mortality was demonstrated; however, because of the rare incidence of disease and the rapidity of death in this study, the trial was substantially underpowered to detect a statistically significant mortality advantage. Before the completion of the trial, the probability that the study might have been underpowered to detect a significant reduction in mortality was recognized. An attempt at selecting a previously unvalidated composite end point to increase the meaningful event rate for the primary end point proved unsuccessful. Significant improvements were seen in other prospectively defined outcome variables that suggest an overall substantial benefit of therapy with rBPI21 in children with severe meningococcemia. CONCLUSIONS: As the largest therapeutic trial conducted in pediatric critical care, the phase III trial of rBPI21 demonstrates important principles that can influence the design of future trials targeting rare, life-threatening diseases.
Asunto(s)
Bacteriemia/tratamiento farmacológico , Proteínas de la Membrana/uso terapéutico , Infecciones Meningocócicas/tratamiento farmacológico , Actividades Cotidianas , Animales , Bacteriemia/clasificación , Bacteriemia/complicaciones , Bacteriemia/mortalidad , Niño , Ensayos Clínicos Fase III como Asunto , Personas con Discapacidad , Modelos Animales de Enfermedad , Evaluación Preclínica de Medicamentos , Humanos , Proteínas de la Membrana/farmacología , Infecciones Meningocócicas/clasificación , Infecciones Meningocócicas/complicaciones , Infecciones Meningocócicas/mortalidad , Ensayos Clínicos Controlados Aleatorios como Asunto , Índice de Severidad de la Enfermedad , Análisis de Supervivencia , Resultado del TratamientoRESUMEN
BACKGROUND: Endotoxin is a primary trigger of the inflammatory processes that lead to shock, multiorgan failure, and purpura fulminans in meningococcal sepsis. Bactericidal/permeability-increasing protein (BPI) is a natural protein, stored within the neutrophil granules, that binds to and neutralises the effects of endotoxin in vitro, in laboratory animals, and in humans. To establish whether a recombinant 21-kDa modified fragment of human BPI (rBPI21), containing the active antimicrobial and endotoxin-neutralising moiety, would decrease death and long-term disability from meningococcal sepsis, we did a randomised, double-blind, placebo-controlled trial of rBPI21 in children with severe meningococcal sepsis. METHODS: We enrolled children (2 weeks to 18 years of age) presenting to 22 centres in the UK and the USA with a clinical picture suggestive of meningococcal sepsis, and with evidence of severe disease. Children were randomly assigned rBPI21 (2 mg/kg over 30 min followed by 2 mg/kg over 24 h) or placebo (0.2 mg/mL human albumin solution) in addition to conventional medical therapy. Primary outcome variables were mortality, amputations, and change in paediatric overall performance category (POPC) from before illness to day 60. Analysis was by intention to treat. FINDINGS: Of 1287 patients screened, 892 were excluded, including 57 patients who died or who met criteria for imminent death before receiving the study drug. 190 patients received rBPI21, and 203 placebo. 34 (8.7%) of 393 patients died during the study: 14 (7.4%) in the rBPI21 group and 20 (9.9%) in the placebo group (odds ratio 1.31 [95% CI 0.62-2.74], p=0.48). Compared with patients randomised to placebo, fewer patients treated with rBPI21 had multiple severe amputations (six of 190 [3.2%] vs 15 of 203 [7.4%], odds ratio 2.47 [0.94-6.51], p=0.067), and more had a functional outcome similar to that before illness (as measured by the POPC scale) at day 60 (136 of 176 [77.3%] vs 126 of 190 [66.3%], p=0.019). INTERPRETATION: Because most deaths occurred in the interval between identification of patients and study drug administration, the mortality rate in the placebo group was substantially lower than predicted. The trial was therefore underpowered to detect significant differences in mortality. However, patients receiving rBPI21 had a trend towards improved outcome in all primary outcome variables. Given the excellent severity match between placebo and rBPI21 groups at study entry, the results overall indicate that rBPI21 is beneficial in decreasing complications of meningococcal disease.
Asunto(s)
Bacteriemia/tratamiento farmacológico , Proteínas de la Membrana/uso terapéutico , Infecciones Meningocócicas/tratamiento farmacológico , Adolescente , Amputación Quirúrgica/estadística & datos numéricos , Bacteriemia/clasificación , Bacteriemia/mortalidad , Quimioterapia Adyuvante , Niño , Preescolar , Método Doble Ciego , Endotoxinas , Femenino , Escala de Coma de Glasgow , Humanos , Lactante , Masculino , Infecciones Meningocócicas/clasificación , Infecciones Meningocócicas/mortalidad , Resultado del Tratamiento , Reino Unido , Estados UnidosRESUMEN
STUDY OBJECTIVES: Recent data indicate that increases in inflammatory cytokines are seen in patients with diverse cardiac diseases. However, the primary stimulus for cytokine secretion during cardiac illness remains unknown. Since bacterial endotoxin is a potent inducer of cytokines, we determined the incidence, magnitude, and clinical relevance of endotoxemia in children with congenital heart disease before and after surgical repair. DESIGN: A prospective, observational study. SETTING: A large, urban, university-affiliated, tertiary-care children's hospital. PATIENTS: Thirty children with a variety of congenital heart defects (median age, 59 days; median weight, 4.0 kg) were sequentially enrolled. INTERVENTIONS: Blood was sampled prior to surgery, and at 1, 8, 24, 48, and 72 h following cardiopulmonary bypass. Assays included plasma endotoxin, lipopolysaccharide-binding protein (LBP), and interleukin-6 (IL-6). MEASUREMENTS AND RESULTS: Twenty-nine of 30 patients (96%) had evidence of endotoxemia during the study period. Twelve of the 30 patients (40%) were significantly endotoxemic prior to surgery. LBP, a plasma marker that responds to bacteria and endotoxin, rose significantly following cardiopulmonary bypass, as did the plasma levels of IL-6. Fifteen of 30 patients met prospectively defined criteria for experiencing a severe hemodynamic disturbance in their postoperative course. These patients had significantly higher preoperative plasma LBP (p < 0.02) and plasma endotoxin levels (p < 0.05), compared to patients with less-severely disturbed hemodynamics. Mortality was 25% in patients with preoperative endotoxemia, compared with no mortality in patients who were not endotoxemic before surgery (p = 0.05). CONCLUSIONS: These data demonstrate that endotoxemia in children with congenital heart disease is more common than previously suspected, and is associated with clinical outcomes. We conclude that clinical trials targeting endotoxin will be necessary to determine if endotoxin is a causal, etiologic agent in the disease process.
Asunto(s)
Proteínas de Fase Aguda , Endotoxemia/diagnóstico , Cardiopatías Congénitas/cirugía , Glicoproteínas de Membrana , Complicaciones Posoperatorias/diagnóstico , Puente Cardiopulmonar , Proteínas Portadoras/sangre , Citocinas/sangre , Endotoxemia/inmunología , Endotoxemia/mortalidad , Endotoxinas/sangre , Femenino , Cardiopatías Congénitas/inmunología , Cardiopatías Congénitas/mortalidad , Hemodinámica/fisiología , Humanos , Lactante , Interleucina-6/sangre , Masculino , Complicaciones Posoperatorias/inmunología , Complicaciones Posoperatorias/mortalidad , Pronóstico , Estudios Prospectivos , Tasa de SupervivenciaRESUMEN
CONTEXT: Knowledge and understanding of gram-negative sepsis have grown over the past 20 years, but the ability to treat severe sepsis successfully has not. OBJECTIVE: To assess the efficacy and safety of E5 in the treatment of patients with severe gram-negative sepsis. DESIGN: A multicenter, double-blind, randomized, placebo-controlled trial conducted at 136 US medical centers from April 1993 to April 1997, designed with 90% power to detect a 25% relative risk reduction, incorporating 2 planned interim analyses. SETTING: Intensive care units at university medical centers, Veterans Affairs medical centers, and community hospitals. PATIENTS: Adults aged 18 years or older, with signs and symptoms consistent with severe sepsis and documented or probable gram-negative infection. INTERVENTION: Patients were assigned to receive 2 doses of either E5, a murine monoclonal antibody directed against endotoxin (n = 550; 2 mg/kg per day by intravenous infusion 24 hours apart) or placebo (n = 552). MAIN OUTCOME MEASURES: The primary end point was mortality at day 14; secondary end points were mortality at day 28, adverse event rates, and 14-day and 28-day mortality in the subgroup without shock at presentation. RESULTS: The trial was stopped after the second interim analysis. A total of 1090 patients received study medication and 915 had gram-negative infection confirmed by culture. There were no statistically significant differences in mortality between the E5 and placebo groups at either day 14 (29.7% vs 31.1%; P = .67) or day 28 (38.5% vs 40.3%; P = .56). Patients presenting without shock had a slightly lower mortality when treated with E5 but the difference was not significant (28.9% vs 33.0% for the E5 and placebo groups, respectively, at day 28; P = .32). There was a similar profile of adverse event rates between E5 and placebo. CONCLUSIONS: Despite adequate sample size and high enrollment of patients with confirmed gram-negative sepsis, E5 did not improve short-term survival. Current study rationale and designs should be carefully reviewed before further large-scale studies of patients with sepsis are conducted.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Infecciones por Bacterias Gramnegativas/tratamiento farmacológico , Sepsis/tratamiento farmacológico , Método Doble Ciego , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Análisis de SupervivenciaRESUMEN
Plasma endotoxin and lipopolysaccharide-binding protein (LBP) levels were measured in a group of 253 patients at the onset of severe sepsis and/or septic shock. Endotoxin levels were significantly greater than control levels (n=33; mean +/- SD, 5.1+/-7.3 pg/mL) in 78.3% of patients. Median endotoxin levels in patients with sepsis were 300 pg/mL (25%-75% interquartile range, 110-726 pg/mL). LBP levels were elevated in 97% of patients compared with normal control values of 4.1+/-1.65 microgram/mL. Median LBP levels in patients with sepsis were 31.2 microgram/mL (interquartile range, 22.5-47.7 microgram/mL). Median endotoxin levels at study entry were more highly elevated (515 vs. 230 pg/mL; P<.01), and LBP levels were less highly elevated (28.0 vs. 33.2 microgram/mL; P<.05) in nonsurvivors than survivors over the 28-day study period. No correlation was found between endotoxin and LBP levels. The quantitative level of both endotoxin and LBP may have prognostic significance in patients with severe sepsis.
Asunto(s)
Proteínas de Fase Aguda , Bacteriemia/sangre , Proteínas Portadoras/sangre , Endotoxinas/sangre , Fungemia/sangre , Lipopolisacáridos/sangre , Glicoproteínas de Membrana , Choque Séptico/sangre , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , PronósticoRESUMEN
BACKGROUND: Infection and organ failure are the most common causes of death or serious complication in trauma patients surviving initial resuscitation and operation. Of the many possible causes of these complications, bacterial translocation and release of harmful cytokines and oxygen free radicals may play an important role in the pathogenesis of the complications associated with traumatic hemorrhage. Recombinant human bactericidal/permeability-increasing protein (rBPI21) has antibacterial and antiendotoxin properties, reduces cytokine levels, and increases survival in animal models of hemorrhagic shock. The primary objective of this study was to evaluate the safety and efficacy of prophylactic rBPI21 infusion in patients with hemorrhage due to trauma. METHODS: This was a phase II, multicenter, randomized, double-blind, placebo-controlled trial. Patients who required at least 2 U of blood were randomized to receive rBPI21 (4 mg x kg(-1) x d(-1) for 2 consecutive days) or an equivalent volume of placebo by continuous infusion within 12 hours of injury. The primary efficacy end point was mortality or serious complication occurring during the first 15 days of the study. Safety was monitored clinically and by laboratory panels during the study period. RESULTS: A total of 401 patients were treated (202 in the rBPI21 group and 199 in the placebo group). The composite end point rate of mortality or serious complication through day 15 was 46% in the placebo group and 39% in the rBPI21 group (hazard ratio = 0.79; p = 0.13). Secondary analysis, which adjusted for age, mechanism of injury, Injury Severity Score (1990 version), and units of blood received before study drug infusion showed similar results (hazard ratio = 0.79; p = 0.14). The proportion of patients who developed at least one serious organ dysfunction was 22% in the placebo group and 16% in the rBPI21 group (hazard ratio = 0.71; p = 0.14). The proportion of patients who developed either pneumonia or acute respiratory distress syndrome was 32% in the placebo group and 22% in the rBPI21 group (hazard ratio = 0.66; post hocp = 0.03). The beneficial trends of rBPI21 were observed in both blunt and penetrating trauma and were generally observed across different age groups, Injury Severity Scores, and units of blood transfused. No treatment difference was observed in mortality or resource utilization in this phase II study. CONCLUSION: rBPI21 was well-tolerated and demonstrated a favorable trend in reducing the composite primary end point of mortality or serious complication through day 15, especially respiratory complications, in patients with hemorrhage due to trauma. A phase III study is currently in progress.
Asunto(s)
Hemorragia/tratamiento farmacológico , Proteínas de la Membrana/uso terapéutico , Heridas y Lesiones/complicaciones , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Esquema de Medicación , Femenino , Escala de Coma de Glasgow , Hemorragia/clasificación , Hemorragia/etiología , Hemorragia/mortalidad , Humanos , Infusiones Intravenosas , Masculino , Proteínas de la Membrana/administración & dosificación , Proteínas de la Membrana/efectos adversos , Persona de Mediana Edad , Síndrome de Dificultad Respiratoria/etiología , Índice de Severidad de la Enfermedad , Estados UnidosRESUMEN
OBJECTIVE AND SUMMARY BACKGROUND DATA: The recombinant fragment of bactericidal/permeability-increasing protein, rBPI21, has potent bactericidal activity against gram-negative bacteria as well as antiendotoxin (lipopolysaccharide [LPS]) action. On the basis of these activities, the authors sought to discover whether rBPI21 would be protective in baboons with live Escherichia coli-induced sepsis and whether the potential protective effects of rBPI21 (together with antibiotics) would be more closely related to its antibacterial or LPS-neutralizing effects. METHODS: In a prospective, randomized, placebo-controlled subchronic laboratory study, the efficacy of rBPI21 or placebo was studied over 72 hours in chronically instrumented male baboons infused with live E. coli under antibiotic therapy. RESULTS: Intravenous rBPI21 attenuated sepsis-related organ failure and increased survival significantly. Bacteremia was significantly reduced in the rBPI21 group at 2 hours after the start of the E. coli infusion, whereas circulating LPS was less affected. The in vivo formation of tumor necrosis factor was significantly suppressed by the rBPI21 treatment regimen. Microcirculation and organ function were improved. CONCLUSIONS: In baboon live E. coli sepsis, the salutary effect of rBPI21 results from a more prevalent antibacterial than antiendotoxin activity.
Asunto(s)
Infecciones por Escherichia coli/prevención & control , Proteínas de la Membrana/farmacología , Sepsis/prevención & control , Animales , Hemodinámica/efectos de los fármacos , Pulmón/efectos de los fármacos , Pulmón/fisiología , Masculino , Proteínas de la Membrana/uso terapéutico , Papio , Distribución Aleatoria , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
BACKGROUND: Meningococcal sepsis remains an important cause of morbidity and mortality. We hypothesised that children with severe meningococcaemia might benefit from inhibition of the inflammatory processes thought responsible for fulminant disease. rBPI21 is a recombinant, N-terminal fragment of human bactericidal/permeability-increasing protein, which kills meningococci and binds to and clears bacterial endotoxin, these being the primary inducers of the systemic inflammation. The aim of this study was to determine the safety and kinetics of rBPI21 in children with severe meningococcaemia and to make a preliminary assessment of clinical outcome. METHODS: In this open-label, dose-escalation, phase I/II trial in severe meningococcaemia (Glasgow meningococcal prognostic septicaemia score [GMSPS] > or = 8), 26 patients aged 1-18 years, who had received their first dose of antibiotics no more than 8 hours earlier were given rBPI21 by infusion at total doses of 1.0, 2.0, and 4.0 mg/kg. FINDINGS: The patients had significantly raised plasma concentrations of bacterial endotoxin and cytokines. Peak and steady state BPI concentrations were comparable with pharmacokinetic data in healthy adults. All complications were compatible with the expected pattern for severe meningococcal sepsis. Only one patient died. This outcome was found to compare favourably with a predicted mortality of > or = 30% by GMSPS, > or = 15% by plasma endotoxin values, > or = 28% by plasma interleukin-6 concentrations, 29-49% by severity of coagulopathy, and 20% (11/54) by comparison with recent historical patients consecutively treated in participating centres before this study. INTERPRETATION: This, the first clinical trial or rBPI21, shows that rBPI21 can be safely administered to children with severe meningococcaemia and that the pharmacokinetics are consistent with patterns seen in healthy adults. Predicted mortality, on the basis of GMSPS, laboratory indices of inflammation and coagulopathy, and historical controls, was for between four and eight deaths. These findings have prompted a phase III randomised trial.
Asunto(s)
Antiinfecciosos/uso terapéutico , Bacteriemia/tratamiento farmacológico , Proteínas Sanguíneas/uso terapéutico , Proteínas de la Membrana , Infecciones Meningocócicas/tratamiento farmacológico , Adulto , Antiinfecciosos/efectos adversos , Antiinfecciosos/farmacocinética , Péptidos Catiónicos Antimicrobianos , Bacteriemia/microbiología , Bacteriemia/mortalidad , Actividad Bactericida de la Sangre , Proteínas Sanguíneas/efectos adversos , Proteínas Sanguíneas/farmacocinética , Niño , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Masculino , Infecciones Meningocócicas/mortalidad , Proteínas Recombinantes/efectos adversos , Proteínas Recombinantes/farmacocinética , Proteínas Recombinantes/uso terapéutico , Resultado del TratamientoRESUMEN
Acute graft-versus-host disease (GVHD) is most often treated with high dose glucocorticoids, but less than half of patients have durable overall improvement. Previous phase I and phase II studies suggested that treatment with a CD5-specific immunotoxin (XomaZyme-CD5 Plus) could ameliorate symptoms of GVHD. In a randomized, double-blind trial, we compared XomaZyme-CD5 Plus and glucocorticoids versus placebo and glucocorticoids as initial therapy for 243 patients who developed acute GVHD after allogeneic marrow transplantation. The study drug (XomaZyme. CD5-Plus or an identical appearing placebo) was administered at a dose of 0.1 mg/kg body weight on each of 14 consecutive days. All patients were treated concomitantly with a standard regimen of methylprednisolone. At the time of entry on study, 94% of patients had a rash, 56% had hyperbilirubinemia, 61% had diarrhea, and 84% had nausea and vomiting. At 3, 4, and 5 weeks after starting treatment, symptom severity was less in the CD5 group than in the placebo group. At 4 weeks, 40% of patients assigned to the CD5 group had complete response compared with 25% of those assigned to the control group (P = .019). At 6 weeks, 44% of patients assigned to the CD5 group had complete response as compared with 38% in the placebo group (P = .36). Clinical extensive chronic GVHD developed in 65% of patients in the CD5 group compared with 72% in the control group (P = .35). Survival at 1 year after treatment was 49% in the CD5 group and 45% in the control group (P = .68). Side effects required close monitoring and appropriate adjustment of treatment. The combined administration of a CD5-specific immunotoxin and glucocorticoids controls GVHD manifestations more effectively than treatment with glucocorticoids alone during the first 5 weeks after starting treatment. Use of this immunotoxin does not result in any long-term clinical benefit for patients with acute GVHD.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Trasplante de Médula Ósea/efectos adversos , Antígenos CD5/inmunología , Enfermedad Injerto contra Huésped/terapia , Inmunotoxinas/uso terapéutico , Depleción Linfocítica , Ricina/uso terapéutico , Enfermedad Aguda , Adolescente , Adulto , Anticuerpos Monoclonales/efectos adversos , Niño , Preescolar , Terapia Combinada , Método Doble Ciego , Femenino , Enfermedad Injerto contra Huésped/tratamiento farmacológico , Enfermedad Injerto contra Huésped/etiología , Enfermedad Injerto contra Huésped/mortalidad , Humanos , Inmunosupresores/uso terapéutico , Inmunotoxinas/efectos adversos , Lactante , Enfermedades Renales/inducido químicamente , Masculino , Metilprednisolona/uso terapéutico , Persona de Mediana Edad , Ricina/efectos adversos , Linfocitos T Citotóxicos/efectos de los fármacos , Resultado del TratamientoRESUMEN
H65-RTA is an immunoconjugate that consists of the A chain of ricin (RTA), a ribosomal-inhibiting protein, coupled to a murine monoclonal antibody (H65) directed against the pan-T-cell antigen CD5. The CD5 antigen is heterogeneously expressed on cutaneous T-cell lymphoma tumor cells, but is not expressed on normal cells except lymphocytes. A phase I trial was therefore conducted in which 14 patients with cutaneous T-cell lymphoma progressive on other therapies were treated with up to three cycles of H65-RTA. The maximal tolerated dose (MTD) of H65-RTA was 0.33 mg/kg/d administered intravenously for 10 days as defined by dyspnea at rest at higher doses. Other reversible side effects included myalgia, mild hypoalbuminemia with weight gain, pedal edema, fatigue, fevers, and chills. Six patients received more than one cycle of H65-RTA without increased side effects compared with the first cycle. Pharmacokinetic analysis showed that peak serum drug levels were dose-dependent, and ranged from 1.13 to 5.56 micrograms/mL, with a terminal half-life ranging from 1.0 to 2.9 hours. The development of antibodies against the immunoconjugate was associated with a lower peak drug level, but not with enhanced side effects. Partial responses lasting from 3 to 8 months were documented in four patients. Three of the responding patients received more than one cycle of H65-RTA in the presence of anti-immunoconjugate antibodies. The results from this phase I trial suggest that H65-RTA is an active drug in the treatment of cutaneous T-cell lymphoma. The immunoconjugate may be safely administered repeatedly, even in the presence of anti-immunoconjugate antibodies, with responses noted. Additional studies at the MTD are needed to define the response rate in this disease.
Asunto(s)
Inmunotoxinas/uso terapéutico , Leucemia de Células T/terapia , Ricina/uso terapéutico , Neoplasias Cutáneas/terapia , Adulto , Anciano , Anticuerpos Monoclonales/efectos adversos , Anticuerpos Monoclonales/farmacocinética , Anticuerpos Monoclonales/uso terapéutico , Evaluación de Medicamentos , Femenino , Humanos , Inmunofenotipificación , Inmunotoxinas/efectos adversos , Inmunotoxinas/farmacocinética , Infusiones Intravenosas , Leucemia de Células T/inmunología , Masculino , Persona de Mediana Edad , Ricina/efectos adversos , Ricina/farmacocinéticaRESUMEN
Human antibody responses to immunotoxin components were evaluated in 21 melanoma patients who were treated with XomaZyme-MEL, a murine monoclonal antimelanoma antibody-ricin A chain conjugate. Twenty of the 21 melanoma patients produced antibodies against ricin A chain, while 15 of 21 produced antibodies reactive with the murine monoclonal antibody component. Both IgM and IgG antibody responses were produced. Immunoglobulin responses were usually detected 1 to 2 weeks following initiation of therapy, with peak levels generally attained 2 to 4 weeks posttherapy. Titers of the anti-ricin A chain antibodies were generally higher than those of the antimurine monoclonal antibodies for the dose range tested. There was no clear correlation between the dose of immunotoxin administered and the antibody titer. By use of a competitive flow cytometry assay, antiidiotype responses were demonstrated in eight of 10 melanoma patients who had antimurine antibodies. Both the kinetics of appearance and the relative titers of the antiidiotype responses generally corresponded to the antimurine responses. The development of antimmunotoxin antibodies can reduce the therapeutic potential of immunotoxins through several mechanisms. The development of antibodies in a significant number of patients suggests that optimally effective, repeated courses of therapy will require some procedure for suppressing or abrogating the response against the immunotoxin.
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Anticuerpos Monoclonales/inmunología , Inmunotoxinas/inmunología , Melanoma/tratamiento farmacológico , Ricina/inmunología , Anticuerpos Antiidiotipos/biosíntesis , Anticuerpos Monoclonales/uso terapéutico , Humanos , Inmunoglobulina G/biosíntesis , Inmunoglobulina M/biosíntesis , Inmunotoxinas/uso terapéutico , Cinética , Melanoma/inmunología , Ricina/uso terapéuticoRESUMEN
Monoclonal antibody 791T/36, recognizing a Mr 72,000 antigen on the surface of colon carcinoma cells, has been used to construct an immunotoxin by conjugating to it the ribosomal inhibitor protein, ricin toxin A chain. The antibody 791T/36 has been shown to bind to membranes of freshly disaggregated tumor cells from human colon tumors, and to localize in tumors in vivo. Subacute toxicology testing in rats receiving immunotoxin i.v. showed, at highest doses, weight loss, decreased serum albumin, and hepatocyte vacuolization without elevation in liver function tests. A Phase I dose escalation study was carried out in which 17 patients with metastatic colorectal cancer were treated with doses of immunotoxin ranging from 0.02 to 0.2 mg/kg/day in 1-h i.v. infusions for a 5-day course. Side-effects included a composite of signs and symptoms thought to be generic to ricin A chain immunotoxins, including decreased serum albumin, mild fever, and flu-like symptoms, all being reversible. Two additional findings, reversible proteinuria and mental status changes, were also noted which may be characteristic of this immunotoxin. By 10-20 days after therapy, most patients developed IgM and IgG antibodies against both the ricin toxin A chain and the immunoglobulin portion of the immunotoxin, which were asymptomatic. A strong anticombining site antibody response was seen. Biological activity manifest as mixed tumor regression was seen in five patients.
Asunto(s)
Anticuerpos Monoclonales/efectos adversos , Neoplasias del Colon/terapia , Inmunotoxinas/efectos adversos , Neoplasias Hepáticas/secundario , Neoplasias Pulmonares/secundario , Ricina/efectos adversos , Adulto , Anciano , Animales , Anticuerpos Monoclonales/uso terapéutico , Anticuerpos Monoclonales/toxicidad , Formación de Anticuerpos , Antígeno Carcinoembrionario/análisis , Neoplasias del Colon/inmunología , Evaluación de Medicamentos , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunoglobulina G/análisis , Inmunoglobulina M/análisis , Inmunotoxinas/uso terapéutico , Inmunotoxinas/toxicidad , Dosificación Letal Mediana , Pruebas de Función Hepática , Neoplasias Hepáticas/inmunología , Neoplasias Hepáticas/terapia , Neoplasias Pulmonares/inmunología , Neoplasias Pulmonares/terapia , Masculino , Ratones , Ratones Endogámicos BALB C , Persona de Mediana Edad , Metástasis de la Neoplasia , Ratas , Ratas Endogámicas , Ricina/uso terapéutico , Ricina/toxicidad , Albúmina Sérica/análisisRESUMEN
Ricin A chain immunotoxin (IT) 45-2D9-RTA mediates regression of spontaneous pulmonary metastases and lung colonies from K-ras transformed rat fibroblasts (TRF cells). However, residual metastases are frequently noted after IT therapy, and therefore, possible mechanisms mediating tumor cell escape were investigated. Individual lung colonies were dissected from lungs of BALB/c mice, and single-cell suspensions of fresh cells from short-term cultures (eight passages) were tested. Immunoperoxidase staining with 45-2D9 monoclonal antibody showed that stable loss of surface antigen by cells cultured from IT-treated mice did not occur after four injections of specific IT. Sensitivity to specific IT in vitro was equal for metastatic tumor cells from mice treated with either two or four doses of specific IT compared to cells from nonspecific IT-treated mice and to parental cells. Clones derived from metastases of IT-treated mice were not resistant to IT. Clones derived from metastases of specific IT-treated mice internalized bound antibody or IT at the same rate as untreated cells. Freshly disaggregated cells from specific IT-treated mice were as sensitive to specific IT as were cells from nonspecific IT-treated or untreated mice. Specific IT successfully mediated reduction of lung colonies derived from fresh suspensions of lung colony TRF cells from IT-treated mice. This reduction was equivalent to that seen for cells not previously exposed to specific IT. Immunoperoxidase stains of lung sections with 45-2D9 showed that colonies consisting entirely of unstained TRF cells were present in both specific IT and phosphate buffered saline-treated mice. There was a trend toward a higher percentage of antigen-negative colonies in mice treated with IT, although 9 days following specific IT therapy, greater than 80% of lung colonies expressed gp74 antigen. When TRF cells were grown on agar plugs, which promoted three-dimensional growth, groups of cells showing absence of immunoperoxidase staining with antibody to gp74 were identified during 2 weeks of growth. Thus, stability of antigen-negative variants is favored by three-dimensional growth conditions and the selective pressure of IT administration. Our results also suggest that impaired trafficking of IT to antigen-positive cells may also contribute to escape from IT therapy.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Antígenos de Neoplasias/inmunología , Glicoproteínas/inmunología , Inmunotoxinas/inmunología , Neoplasias Experimentales/terapia , Animales , Anticuerpos Antineoplásicos/uso terapéutico , Transformación Celular Neoplásica , Técnicas para Inmunoenzimas , Inmunoterapia , Neoplasias Pulmonares/inmunología , Neoplasias Pulmonares/secundario , Ratones , Peso Molecular , Neoplasias Experimentales/inmunología , Ricina/administración & dosificación , Factores de TiempoRESUMEN
Monoclonal antibody 791 (XMMCO-791) recognizes a colorectal tumour-associated antigen. Antibody 791-ricin A chain immunotoxin (XMMCO-791-RTA) inhibits growth of human tumour xenografts and it is therefore being evaluated for the treatment of colorectal cancer. One of the problems with therapy with mouse monoclonal antibodies is they stimulate humoral responses in patients. However antigens linked to ricin are cytotoxic for B cells and therefore XMMCO-791-RTA may not be immunogenic. The humoral antibody response to murine monoclonal antibody XMMCO-791 (IgG2b) conjugated to the plant toxin, ricin A chain (RTA), was measured in colorectal cancer patients in a phase I clinical trial. All patients produced strong responses to the XMMCO-791 immunoglobulin and to RTA. The predominant response to the antibody was against the idiotypic determinant although anti-subclass and anti-mouse antibodies were also detected. A component of the anti-idiotypic immunoglobulin response in the colorectal cancer patients was directed against the combining site of XMMCO-791. These antibodies inhibited in-vitro binding of XMMCO-791 to target 791 cells and so may be inhibitors of repeated immunotoxin therapy. Immunotoxins do not abrogate the immune response to mouse immunoglobulin in vivo but instead are highly immunogenic.
Asunto(s)
Neoplasias Colorrectales/inmunología , Inmunotoxinas/inmunología , Ricina/inmunología , Adulto , Anciano , Animales , Anticuerpos Antiidiotipos/biosíntesis , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/uso terapéutico , Neoplasias Colorrectales/terapia , Evaluación de Medicamentos , Humanos , Inmunoglobulina G/biosíntesis , Inmunoglobulina M/biosíntesis , Inmunotoxinas/uso terapéutico , Persona de Mediana EdadAsunto(s)
Anticuerpos Monoclonales/uso terapéutico , Trasplante de Médula Ósea , Enfermedad Injerto contra Huésped/prevención & control , Inmunotoxinas/uso terapéutico , Depleción Linfocítica , Ricina/uso terapéutico , Linfocitos T , Adulto , Trasplante de Médula Ósea/inmunología , Evaluación de Medicamentos , Enfermedad Injerto contra Huésped/inmunología , Prueba de Histocompatibilidad , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva/inmunología , Leucemia Mielógena Crónica BCR-ABL Positiva/cirugía , Leucemia Mielógena Crónica BCR-ABL Positiva/terapia , Masculino , Complicaciones Posoperatorias/prevención & control , Ricina/inmunologíaRESUMEN
The A chain of the toxin ricin has been conjugated by a disulfide bond to a murine monoclonal antibody that recognizes the CD5 (T,p67) antigen present on 95% of peripheral blood T lymphocytes. This immunotoxin was used to treat a patient with severe grade III-IV, steroid-resistant, acute graft-vs-host disease (GvHD) after an allogeneic, human leukocyte antigen-identical bone marrow transplant for acute myelogenous leukemia. Immunotoxin therapy produced a complete clinical response in the skin and gastrointestinal tract. The patient tolerated a 14-day course without symptoms or signs of toxic effects. After two days of therapy, circulating T cells could not be demonstrated by indirect immunofluorescence. After therapy, acute GvHD did not recur. However, seven months after therapy the patient demonstrated mild signs of chronic GvHD that were easily controlled with low-dose immunosuppressive therapy. These findings indicate that an anti-T-cell ricin A chain immunotoxin can be given safely for treatment of acute GvHD and may be an effective therapy for this significant posttransplant complication.
Asunto(s)
Enfermedad Injerto contra Huésped/terapia , Inmunotoxinas/uso terapéutico , Ricina/uso terapéutico , Enfermedad Aguda , Anticuerpos Monoclonales/uso terapéutico , Reacciones Antígeno-Anticuerpo , Trasplante de Médula Ósea , Niño , Resistencia a Medicamentos , Femenino , Técnica del Anticuerpo Fluorescente , Enfermedad Injerto contra Huésped/etiología , Humanos , Leucemia Mieloide Aguda/terapia , Metilprednisolona/uso terapéutico , Linfocitos T/inmunologíaRESUMEN
We developed a model to assess the therapeutic effects of the 45-2D9-ricin A-chain immunotoxin (RTA) on pulmonary metastases. The 45-2D9 mouse monoclonal antibody recognizes a Mr 74,000 glycoprotein highly expressed by rat fibroblasts transformed with the Kirsten sarcoma virus (transformed rat fibroblasts). These cells metastasize spontaneously and form lung colonies in nu/nu and irradiated BALB/c mice. Injection i.v. of 45-2D9-RTA specifically reduced formation of spontaneous pulmonary metastases and lung colonies originating from freshly disaggregated tumor cells or cultured cells. Antibody alone or mixed with unconjugated ricin A chain and an immunotoxin that recognizes a melanoma-associated antigen were ineffective. Unconjugated 45-2D9 antibody specifically blocked the 45-2D9-RTA activity in vivo. Administration of the lysosomotrophic agents ammonium chloride and chloroquine in vivo did not potentiate immunotoxin-mediated reduction in lung colonies although they were effective in vitro. Monensin potentiated 45-2D9-RTA activity in vitro and in vivo.
Asunto(s)
Inmunotoxinas/uso terapéutico , Neoplasias Pulmonares/secundario , Monensina/uso terapéutico , Ricina/uso terapéutico , Cloruro de Amonio/farmacología , Animales , Antígenos de Neoplasias/análisis , Cloroquina/farmacología , Dimetilsulfóxido/farmacología , Sinergismo Farmacológico , Femenino , Neoplasias Pulmonares/terapia , Ratones , Ratones Endogámicos BALB CRESUMEN
Nine patients with suspected gram-negative bacterial sepsis were studied to determine the safety, pharmacokinetics, and immunogenicity of XMMEN-0E5, a murine immunoglobulin M monoclonal antibody directed against the core lipid A region of bacterial endotoxin. Antibody was administered by single intravenous infusion of 1 to 4 h duration at doses ranging from 0.1 to 15 mg/kg. Five patients had positive blood cultures for gram-negative bacteria, one patient had Torulopsis septicemia, one patient had gram-negative bacterial meningitis, and two patients were culture negative. No evidence of antibody-mediated toxicity was observed at any dose level. The serum half-life of the antibody was approximately 10 h at doses of 0.1 to 7.5 mg/kg and approximately 18 h at a dose of 15 mg/kg. No apparent difference in clearance of antibody was observed between bacteremic and nonbacteremic patients. Human anti-mouse antibodies were detected in the sera of three evaluable patients that received doses equal to or greater than 2.0 mg/kg but not in patients that received lower doses of antibody. This study demonstrates that XMMEN-0E5 is well tolerated at doses from 0.1 to 15 mg/kg and may be immunogenic at doses of 2.0 mg/kg and above. Controlled trials to establish the efficacy of this antibody in the treatment of gram-negative bacteremia are indicated.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Lípido A/inmunología , Sepsis/terapia , Adulto , Anciano , Anticuerpos Monoclonales/efectos adversos , Anticuerpos Monoclonales/inmunología , Evaluación de Medicamentos , Femenino , Bacterias Gramnegativas , Semivida , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Immunotoxin constructed by conjugating ricin A chain to monoclonal antibody 791T/36 specifically inhibits growth of human tumor xenografts which express the gp72 antigen recognized by the antibody component. Dose schedule tests showed that the major response was obtained during the first 5 days of treatment and further prolonged treatment did not improve therapy. Expression of gp72 antigen on tumor cells derived from xenografts in immunotoxin-treated mice was not markedly altered indicating that treatment did not lead to the expansion of tumor antigen deficient tumor cells. The experiments indicate that treatment for short duration with immunotoxin may be the most effective protocol.