RESUMEN
This manuscript reports the results of preclinical studies in the rat with robenacoxib, a novel selective cyclooxygenase (COX)-2 inhibitor. Robenacoxib selectively inhibited COX-2 in vitro as evidenced from COX-1:COX-2 IC50 ratios of 27:1 in purified enzyme preparations and >967:1 in isolated cell assays. Binding to COX-1 was rapid and readily reversible (dissociation t(1/2) << 1 min), whilst COX-2 binding was slowly reversible (t(1/2) = 25 min). In vivo, robenacoxib inhibited PGE2 production (an index of COX-2 inhibition) in lipopolysaccharide (LPS)-stimulated air pouches (ID50 0.3 mg/kg) and for at least 24 h in zymosan-induced inflammatory exudate (at 2 mg/kg). Robenacoxib was COX-1 sparing, as it inhibited serum TxB2 synthesis ex vivo (an index of COX-1 inhibition) only at very high doses (100 mg/kg but not at 2-30 mg/kg). Robenacoxib inhibited carrageenan-induced paw oedema (ID50 0.40-0.48 mg/kg), LPS-induced fever (ID50 1.1 mg/kg) and Randall-Selitto pain (10 mg/kg). Robenacoxib was highly bound to plasma protein (99.9% at 50 ng/mL in vitro). After intravenous dosing, clearance was 2.4 mL/min/kg and volume of distribution at steady-state was 306 mL/kg. Robenacoxib was preferentially distributed into inflammatory exudate; the AUC for exudate was 2.9 times higher than for blood and the MRT in exudate (15.9 h) was three times longer than in blood (5.3 h). Robenacoxib produced significantly less gastric ulceration and intestinal permeability as compared with the reference nonsteroidal anti-inflammatory drug (NSAID), diclofenac, and did not inhibit PGE2 or 6-keto PGF(1alpha) concentrations in the stomach and ileum at 30 mg/kg. Robenacoxib also had no relevant effects on kidney function at 30 mg/kg. In summary, results of preclinical studies in rats studies suggest that robenacoxib has an attractive pharmacological profile for potential use in the intended target species, cats and dogs.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Inhibidores de la Ciclooxigenasa 2/farmacología , Ciclooxigenasa 2/efectos de los fármacos , Difenilamina/análogos & derivados , Fenilacetatos/farmacología , Animales , Antiinflamatorios no Esteroideos/efectos adversos , Antiinflamatorios no Esteroideos/farmacocinética , Área Bajo la Curva , Línea Celular , Ciclooxigenasa 1/efectos de los fármacos , Ciclooxigenasa 1/metabolismo , Ciclooxigenasa 2/metabolismo , Inhibidores de la Ciclooxigenasa 2/efectos adversos , Inhibidores de la Ciclooxigenasa 2/farmacocinética , Inhibidores de la Ciclooxigenasa/efectos adversos , Inhibidores de la Ciclooxigenasa/farmacocinética , Inhibidores de la Ciclooxigenasa/farmacología , Difenilamina/efectos adversos , Difenilamina/farmacocinética , Difenilamina/farmacología , Modelos Animales de Enfermedad , Edema/inducido químicamente , Edema/patología , Fiebre/inducido químicamente , Fiebre/patología , Humanos , Isoenzimas , Masculino , Dolor/inducido químicamente , Dolor/patología , Fenilacetatos/efectos adversos , Fenilacetatos/farmacocinética , Unión Proteica , Distribución Aleatoria , Ratas , Ratas Endogámicas Lew , Ratas Sprague-Dawley , Ratas WistarRESUMEN
OBJECTIVE: To apply quantitative analytical methods to the evaluation of radiographic images in experimental arthritis. METHODS: Adjuvant was used to induce arthritis in rats. Arthritis progression was followed by conventional methods. In addition, digitized images of radiographs of the calcaneus were examined for changes in the mean and in the distribution pattern of gray values. Periosteal new bone formation was measured as an increase in image area of the calcaneus. RESULTS: Significant changes in the gray value profile and increases in periosteal bone formation occurred in arthritic rats. More extensive changes occurred in Lewis rats than in Sprague-Dawley rats. Analysis of serial radiographs revealed an initial decrease in the density of juxtaarticular bone, followed by progressive increases in gray value variation due to concurrent bone loss and bone formation. Eventually, bone formation in arthritic rats resulted in increased gray values above those in nonarthritic rats. CONCLUSION: Image analysis represents a sensitive, quantitative method for detecting radiographic changes in experimental arthritis.
Asunto(s)
Artritis Experimental/diagnóstico por imagen , Animales , Artritis Experimental/patología , Calcáneo/diagnóstico por imagen , Enfermedad Crónica , Progresión de la Enfermedad , Femenino , Procesamiento de Imagen Asistido por Computador , Intensificación de Imagen Radiográfica , Ratas , Ratas Endogámicas Lew , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: To determine the effects of peptidyl fluoromethyl ketones on the in vitro activity of purified cathepsins B and L, on tissue cysteine proteinase activity, and on cartilage and bone destruction in experimental arthritis. METHODS: The effects of the fluoroketones on cathepsins B and L in vitro and the effects of oral administration of fluoroketones on ex vivo cysteine proteinase activity in tissue homogenates were determined by measuring the inhibition of fluorogenic substrate cleavage. To determine the effects on arthritis, animals were injected with adjuvant or type II collagen, treated orally with the fluoroketones, and the severity of arthritis was assessed by clinical, histologic, and radiologic methods. RESULTS: All of the fluoroketones tested were potent inhibitors of purified cathepsins B and L activity. Oral administration of the fluoroketones reduced tissue cysteine proteinase activity by up to 77%. In addition, fluoroketone treatment significantly reduced the severity of clinical joint disease and decreased the destruction of articular cartilage and bone. Quantitative analysis of radiographic images indicated that treatment significantly reduced soft tissue changes, periosteal proliferation, and bone erosion, but only partially reduced juxtaarticular osteoporosis. CONCLUSION: These studies suggest that cysteine proteinase inhibitors may limit tissue destruction in diseases such as rheumatoid arthritis.
Asunto(s)
Artritis/patología , Huesos/efectos de los fármacos , Huesos/patología , Cartílago Articular/efectos de los fármacos , Cartílago Articular/patología , Inhibidores de Cisteína Proteinasa/farmacología , Morfolinas , Animales , Artritis/inducido químicamente , Artritis Experimental/diagnóstico por imagen , Artritis Experimental/fisiopatología , Catepsinas/antagonistas & inhibidores , Enfermedad Crónica , Colágeno , Dipéptidos/farmacología , Femenino , Cetonas/farmacología , Ratones , Ratones Endogámicos DBA , Radiografía , Ratas , Ratas Sprague-DawleyRESUMEN
Peptidyl fluoromethyl ketones with the structure carbobenzoxy (Z)-L-phenylalanine-L-alanine-CH2F (MDL 201,053), and its diastereomer Z-L-phenylalanine-D-alanine-CH2F (MDL 201,117), were synthesized and evaluated in vitro for inhibition of purified human cathepsin B. MDL 201,053 was shown to be a potent inhibitor of cathepsin B activity, whereas MDL 201,117 was more than 100-fold less active. In rats with adjuvant induced arthritis, oral administration of MDL 201,053 (13 or 34 mg/kg/day), but not MDL, 201,117 (28 mg/kg/day), significantly decreased the severity of gross clinical arthritis and reduced histologically graded articular cartilage and bone destruction by 76 to 100%. Quantitative image analysis of radiographs indicated that MDL 201,053 treatment significantly reduced bone density changes and inhibited focal bone erosion that normally occur during the course of adjuvant disease. MDL 201,117 had no significant effect on cartilage or bone destruction by any of the evaluation methods used. The effects of MDL 201,053 treatment were dose dependent and treatment was at least partially effective when initiated after the onset of disease. Our studies suggest that inhibitors of cathepsin B may be useful for the treatment of chronic inflammatory joint disease.
Asunto(s)
Artritis Experimental/tratamiento farmacológico , Catepsina B/antagonistas & inhibidores , Dipéptidos/farmacología , Cetonas/farmacología , Animales , Artritis Experimental/patología , Artritis Experimental/fisiopatología , Huesos/efectos de los fármacos , Huesos/patología , Cartílago Articular/efectos de los fármacos , Cartílago Articular/patología , Dipéptidos/uso terapéutico , Relación Dosis-Respuesta a Droga , Femenino , Cetonas/uso terapéutico , Ratas , Ratas Sprague-Dawley , Índice de Severidad de la EnfermedadRESUMEN
The potent irreversible inhibitor of S-adenosyl-L-homocysteine hydrolase (Z)-5'-fluoro-4',5'-didehydro-5'-deoxyadenosine (MDL 28,842) was examined for its effect on the development and treatment of collagen-induced arthritis in mice. We have previously shown that MDL 28,842 inhibits T cell activation without affecting B cell activation. Animals were dosed with MDL 28,842 at 5, 2.5, or 1 mg/kg/day p.o. in water beginning 1 day before immunization with chick type II collagen (CII) and continuing through day 51 postimmunization. None of the animals treated with MDL 28,842 at 5 or 2.5 mg/kg/day developed arthritis compared with 87.5% of the controls. Animals treated with 1 mg/kg MDL 28,842 had a delay in the development of the disease and a decreased incidence of arthritis (55%) during the course of treatment. After the treatment was discontinued, 40% of the mice in the 5-mg/kg treatment group, 60% of the mice who had previously received 2.5 mg/kg MDL 28,842, and 27% of the mice in the 1-mg/kg treatment group remained free of any signs of arthritis. Treatment with MDL 28,842 also lowered serum anti-CII IgG levels. In addition, T cells taken from animals immunized with CII and treated with 2.5-mg/kg/day MDL 28,842 had a lower proliferative response to denatured CII in vitro than controls. Therapeutically, MDL 28,842 was administered to animals at 2.5 mg/kg/day p.o., beginning at the first clinical signs of arthritis and continuing for 4 wk. Over the course of treatment, there was significantly less clinical disease in animals given MDL 28,842. In addition, at the end of treatment, hind paws were removed from the animals and examined radiographically and histologically for joint pathology. Animals treated with MDL 28,842 had significantly fewer bone lesions than control animals. These results suggest that inhibitors of S-adenosyl-L-homocysteine hydrolase may be effective anti-arthritic agents.
Asunto(s)
Adenosina/análogos & derivados , Artritis/tratamiento farmacológico , Artritis/prevención & control , Enfermedades Autoinmunes/tratamiento farmacológico , Enfermedades Autoinmunes/prevención & control , Colágeno/inmunología , Hidrolasas/antagonistas & inhibidores , Terapia de Inmunosupresión/métodos , Inmunosupresores , Adenosina/farmacología , Adenosilhomocisteinasa , Animales , Autoanticuerpos/biosíntesis , Activación de Linfocitos/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos DBA , Linfocitos T/inmunologíaRESUMEN
Peptidyl fluoromethyl ketones (FMKs), with the amino acid sequence Phe-Ala held constant but with variable N-terminal groups, were synthesized and tested for inhibition of the cysteine proteinase cathepsin B. The FMKs were effective in inhibiting cathepsin B activity in vitro. The inhibition was time dependent and was not reversed by dialysis, suggesting covalent modification of the enzyme. Cathepsin B activity present in livers and kidneys of rats treated with FMKs was reduced by 22-91% 4 hr after a single oral dose of 25 mg/kg. The FMKs inhibited the severity of inflammation and the extent of cartilage and bone damage in adjuvant-induced arthritis. These effects were seen during the late-stage of the disease with no effect on onset or incidence of disease. This is consistent with inhibition of protease-mediated damage. These FMKs or derivatives may be of clinical value in the treatment of arthritis.
Asunto(s)
Catepsina B/antagonistas & inhibidores , Dipéptidos/farmacología , Cetonas/farmacología , Riñón/efectos de los fármacos , Hígado/efectos de los fármacos , Animales , Artritis Reumatoide/tratamiento farmacológico , Peso Corporal/efectos de los fármacos , Dipéptidos/síntesis química , Dipéptidos/uso terapéutico , Evaluación Preclínica de Medicamentos , Femenino , Cetonas/síntesis química , Cetonas/uso terapéutico , Riñón/enzimología , Cinética , Hígado/enzimología , Ratas , Ratas Sprague-Dawley , Relación Estructura-ActividadRESUMEN
Protein antigens of Mycobacterium leprae have been identified by screening the lambda gt11, pYA626 and pHC79::M. leprae genomic libraries with pooled sera from leprosy patients and with antiserum to M. leprae cell wall protein (CWP) aggregate. Immunological screening of the lambda gt11 library with pooled sera from 21 lepromatous (LL) leprosy patients resulted in the identification of 19 antigens that are apparently different from previously identified M. leprae antigens. Five additional antigens were identified by screening the lambda gt11 library with pooled sera from 30 borderline tuberculoid or tuberculoid patients. Four other antigens were identified by screening the lambda gt11 library with anti-CWP. Two groups of recombinant cosmids were identified by screening the pHC79 library with LL patients' sera: one group specified proteins that reacted with monoclonal antibodies (mAb) against the 65-kDa protein and against the 18-kDa protein; the other group specified a 15-kDa protein that did not react with any of the mAb that were tested. One pYA626 clone also specified a 15-kDa protein that reacted with LL patients' sera, but did not react with any mAb. Genes specifying several of these antigens have been subcloned into the Asd+ plasmid vector pYA292 and have been introduced into a delta cya delta crp delta asd Salmonella typhimurium strain to evaluate the ability of individual M. leprae proteins to elicit immune responses against M. leprae infection.
Asunto(s)
Antígenos Bacterianos/aislamiento & purificación , Proteínas Bacterianas/inmunología , Mycobacterium leprae/inmunología , Animales , Anticuerpos Antibacterianos , Antígenos Bacterianos/genética , Proteínas Bacterianas/genética , Proteínas Bacterianas/aislamiento & purificación , Vacunas Bacterianas/genética , Vacunas Bacterianas/aislamiento & purificación , Humanos , Lepra/inmunología , Ratones , Mycobacterium leprae/genética , Salmonella typhimurium/genética , Salmonella typhimurium/inmunología , Vacunas Sintéticas/genética , Vacunas Sintéticas/aislamiento & purificaciónRESUMEN
Antigenic determinants of Mycobacterium leprae were identified by screening a lambda gt11::M. leprae genomic library with two separate pools of sera from leprosy patients. A total of 45 recombinant clones were detected with pooled sera from 21 lepromatous (LL) leprosy patients and 5 additional clones specified polypeptides that reacted with antibodies in pooled sera from 30 borderline tuberculoid or tuberculoid leprosy patients. The recombinant clones that specified antigenic determinants that reacted with sera from LL patients were condensed into eight groups on the basis of DNA hybridization experiments among the M. leprae DNA insert fragments. In addition, 11 of the 45 recombinant clones did not hybridize to members of the eight groups nor to one another; these represent unique recombinant clones. None of the recombinant clones identified by screening with sera from tuberculoid leprosy patients hybridized to each other or to any of the 45 LL recombinant clones. The polypeptides specified by the recombinant clones were usually fusion proteins with beta-galactosidase, ranging in size from 117 to 175 kilodaltons (kDa). Members of hybridization group III specified nonfusion proteins of 45 kDa. Only members of hybridization group I reacted with any of 30 monoclonal antibodies prepared against M. leprae proteins; recombinant proteins from these clones reacted with a single monoclonal antibody directed against the M. leprae 65-kDa protein. Thus, at least 22 new antigenic determinants of M. leprae have been identified on the basis of their reactivity to antibodies in sera from LL patients or sera from tuberculoid leprosy patients or both.
Asunto(s)
Anticuerpos Antibacterianos/inmunología , Antígenos Bacterianos/genética , Lepra/inmunología , Mycobacterium leprae/inmunología , Antígenos Bacterianos/inmunología , Proteínas Bacterianas/inmunología , Western Blotting , Clonación Molecular , Epítopos , Biblioteca de Genes , HumanosRESUMEN
We have utilized the 6-day air-pouch model in rats to study the local tissue response to interleukin-1 exposure. Injection of either recombinant human interleukin 1 alpha (rIL-1 alpha) or interleukin 1 beta (rIL-1 beta) directly into preformed air pouches caused a 10- to 100-fold increase in the number of white blood cells present within the pouch. On a weight basis, rIL-1 beta was more active than rIL-1 alpha. Polymorphonuclear neutrophils (PMN) represented the majority of cells entering the pouch following either a single injection or repeated daily injections of rIL-1 alpha or rIL-1 beta. Significant increases in the number of mononuclear cells present were observed only following repeated injections. Repeated injections of rIL-1 beta, but not rIL-1 alpha, also caused the accumulation of large amounts of fluid within preformed pouches and a grossly apparent thickening of the connective-tissue lining of the pouch. Microscopic examination of stained sections of pouch lining tissue indicated a proliferation of the connective-tissue elements of the lining and deposition of large quantities of extracellular collagen within the pouch wall. These findings are entirely consistent with a role for interleukin 1 in the development and perpetuation of inflammatory reactions.
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Inflamación/inducido químicamente , Interleucina-1/toxicidad , Aire , Animales , Modelos Animales de Enfermedad , Femenino , Humanos , Inyecciones , Fenilglioxal , Ratas , Ratas Endogámicas , Proteínas Recombinantes/toxicidadRESUMEN
Rat ankle joints injected intraarticularly with 5 micrograms of group A streptococcal peptidoglycan-polysaccharide (PG-APS) developed an acute course of arthritis. Recurrence of arthritis was induced in 100% of these joints by intravenous injection of as little as 10 micrograms of Salmonella typhimurium lipopolysaccharide (LPS) 3 wk after intraarticular injection. This reaction was similar in athymic and euthymic rats. Buffalo rats were less susceptible than Lewis or Sprague-Dawley rats. Neisseria gonorrhoeae, Yersinia enterocolitica, and Escherichia coli LPS, and S. typhimurium Re mutant LPS, were also active. Re mutant LPS activity was greatly reduced by mixing with polymyxin B. E. coli lipid A was weakly active. An acute synovitis of much less incidence, severity, and duration was seen in contralateral joints injected initially with saline, and in ankle joints of naive, previously uninjected rats after intravenous LPS injection. The intravenous injection of the muramidase mutanolysin on day 0 or 7 after intraarticular PG-APS injection prevented LPS-induced recurrence of arthritis. These studies suggest that the phlogistic activities of lipid A and peptidoglycan might interact in an inflammatory disease process, and that LPS may play a role in recurrent episodes of rheumatoid arthritis or reactive arthritis.
Asunto(s)
Artritis/inducido químicamente , Lipopolisacáridos/toxicidad , Peptidoglicano/toxicidad , Polisacáridos Bacterianos/toxicidad , Animales , Endopeptidasas/farmacología , Femenino , Lípido A/toxicidad , Ratas , Ratas Endogámicas BUF , Ratas Endogámicas Lew , Recurrencia , Especificidad de la Especie , Sinovitis/inducido químicamente , Linfocitos T/fisiologíaRESUMEN
The in vivo degradation and persistence of 125I-labeled peptidoglycan-polysaccharide (PG-PS) fragments from the cell walls of group A and D streptococci were compared by group after intraperitoneal injection into rats. The quantity of PG-PS in the livers and spleens of group D PG-PS-injected rats was less than the quantity in rats injected with group A PG-PS throughout the course of the experiment. Gel filtration analyses of liver and spleen homogenates indicated that group A PG-PS was relatively resistant to degradation, whereas group D PG-PS was extensively degraded to yield a heterogeneous mixture of fragments of lower molecular weight. There was no significant difference in the content of group A PG-PS versus that of group D in joints or blood samples. Analysis of fragment sizes in these tissues also indicated more extensive degradation of group D PG-PS. However, the majority of group A PG-PS in blood samples and joints was a lower molecular weight than that found in the livers or spleens. We conclude that group A PG-PS undergoes a significant but low level of degradation and that group D PG-PS is much less persistent and more extensively degraded than group A PG-PS is in vivo. These differences in PG-PS catabolism may account, in part, for the capacity of group A PG-PS to induce chronic, recurrent arthritis of longer duration than that induced by group D PG-PS.
Asunto(s)
Peptidoglicano/metabolismo , Polisacáridos Bacterianos/metabolismo , Streptococcus pyogenes/metabolismo , Streptococcus/metabolismo , Animales , Pared Celular/metabolismo , Femenino , Articulaciones/metabolismo , Hígado/metabolismo , Tasa de Depuración Metabólica , Peso Molecular , Ratas , Especificidad de la Especie , Bazo/metabolismo , Distribución TisularRESUMEN
The muralytic enzyme mutanolysin can act in vivo to eliminate chronic erosive arthritis induced in rats by polymers of peptidoglycan-polysaccharide isolated from group A streptococci (PG-APS). The amounts of PG-APS in the livers and spleens of rats treated with mutanolysin were significantly reduced compared with the amounts in control rats treated with phosphate-buffered saline. However, the amounts of PG-APS in the limbs of mutanolysin- and phosphate-buffered saline-treated rats were comparable. PG-APS polymers extracted from the livers, spleens, and limbs of mutanolysin-treated rats were extensively degraded, whereas PG-APS extracted from phosphate-buffered saline-treated rats had a high molecular weight. We propose that mutanolysin abrogates arthritis in rats by degrading PG-APS polymers to a size which is no longer able to induce chronic erosive arthritis, even though the polymers are still present in the limbs.
Asunto(s)
Artritis Experimental/prevención & control , Artritis/prevención & control , Pared Celular/metabolismo , Endopeptidasas/metabolismo , Peptidoglicano/metabolismo , Polisacáridos Bacterianos/metabolismo , Animales , Artritis Experimental/microbiología , Edema/microbiología , Extremidades/metabolismo , Femenino , Hígado/metabolismo , Peso Molecular , Ratas , Ratas Endogámicas Lew , Bazo/metabolismoRESUMEN
Cell wall polymers isolated from group A streptococci, as well as lipopolysaccharide from Salmonella typhimurium and synthetic muramyl dipeptide, were injected into the ankle joints of rats. The inflammatory responses were assessed by gross and histologic examination, and edema was measured by accumulation of radiolabeled albumin in the limbs. The isolated group-specific polysaccharide induced extensive edema of the articular and periarticular tissue immediately after injection, and this resolved in 24 hours. The peptidoglycan moiety did not produce early edema, but induced an acute exudative reaction followed by a proliferative synovitis which resolved after 5 days. Reactions induced by covalently bound complexes of peptidoglycan and the group-specific polysaccharide (PG-APS) varied, depending on the size of the complex. Small fragments, derived from mutanolysin digestion, caused both an acute edematous reaction and transient arthritis. Larger fragments did not cause the immediate edematous reaction, but induced an acute arthritis that appeared within 24 hours and evolved into a chronic process. Episodes of recurrent inflammation, a distinctive feature of joint inflammation induced by systemic injection of PG-APS polymers, were not observed following intraarticular injection of any of the cell wall polymers. The relative susceptibility of different rat strains to arthritis induced by intraarticular injection paralleled the responses to systemic injection of PG-APS. These results demonstrate that variations in arthropathogenicity are due, in part, to inherent differences in the phlogistic activities of different cell wall polymers, and that the genetic control of susceptibility involves regulation of the inflammatory responses rather than the quantity of cell wall distributed to the joint.
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Artritis/inducido químicamente , Peptidoglicano , Polisacáridos Bacterianos , Acetilmuramil-Alanil-Isoglutamina , Animales , Artritis/patología , Pared Celular , Edema/inducido químicamente , Femenino , Hiperplasia , Lipopolisacáridos , Peso Molecular , Neutrófilos/patología , Fragmentos de Péptidos , Ratas , Ratas Endogámicas , Salmonella typhimurium , Streptococcus pyogenesRESUMEN
Joint inflammation initially induced by intraarticular injection of an aqueous suspension of peptidoglycan-polysaccharide (PG-PS) fragments isolated from Streptococcus pyogenes was reactivated by systemic injection of a normally subarthropathic dose of homologous or heterologous cell wall polymers, including muramyl dipeptide and lipopolysaccharide. Reactivation was not correlated with the severity of the initial inflammatory reaction. Results of studies utilizing 125I-labeled PG-PS fragments suggested that reactivation was associated with increased localization of PG-PS fragments in the joint following reinjection. These results indicate that the initial injury of the joint by S pyogenes PG-PS fragments increases the susceptibility of the joint to subsequent injury. Furthermore, once the inflammatory reaction is initiated, it can be perpetuated by a variety of ubiquitous cell wall polymers derived from normal flora as well as from pathogenic bacteria.
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Artritis/etiología , Membrana Celular/análisis , Peptidoglicano , Polímeros , Streptococcus pyogenes , Acetilmuramil-Alanil-Isoglutamina , Animales , Artritis/patología , Fraccionamiento Celular , Susceptibilidad a Enfermedades , Femenino , Miembro Posterior , Inyecciones Intraarteriales , Inyecciones Intravenosas , Lipopolisacáridos , Ratas , Ratas Endogámicas , Factores de TiempoRESUMEN
The rosette inhibition assay for immunosuppressive activity of antilymphocyte globulin or plasma has been studied in an effort to improve its reliability. Important changes include the elimination of calcium and magnesium ions from salt solutions used in the assay, the use of deoxyribonuclease to prevent lymphocyte clumping, and pretreatment of plasma samples (heating at 63 C for 10 min followed by acrinol precipitation) to prevent nonspecific inhibition of rosette formation. The use of a graded dose response potency assay against a house standard is discussed. A significant correlation was established between the in vitro activity of several series of antilymphocyte globulin or antithymocyte globulin preparations and their ability to prolong skin graft survival in primates. The best correlation was achieved using a potency estimate relative to a house standard, rather than the conventional titer estimate.
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Reacción de Inmunoadherencia/métodos , Trasplante de Piel , Inmunología del Trasplante , Animales , Anticoagulantes/normas , Suero Antilinfocítico/normas , Calcio , Separación Celular , Cromatografía DEAE-Celulosa , Haplorrinos , Caballos/inmunología , Calor , Inmunoglobulinas , Terapia de Inmunosupresión/normas , Linfocitos/inmunología , Macaca , Magnesio , Timo/citología , Trasplante HomólogoRESUMEN
Tuberculosis lesions were found in necropsied monkeys used in studies involving immunosuppression. Correlations were made between the demonstration of reduced numbers of circulating T cells, the duration of allograft survival and the appearance of T cell-dependent tuberculin sensitivity. The immunosuppressed state of these monkeys may have resulted in false tuberculin reactions. The report confirms the necessity for sequential tuberculin tests on rhesus monkeys prior to their being used in experiments.