Anatomic and histologic analysis of paca larynx (Cuniculus paca, Linnaeus 1766) / [Análise anatômica e histológica da laringe da paca (Cuniculus paca, Linnaeus 1766)]

Paca (Cuniculus paca) has encouraged research as an experimental model both in the human medicine and veterinary, as well as the economic exploitation of its meat cuts, which favored its zootechnical use. There are no anatomical, microscopic descriptions and measurements of the larynx in this rodent. Eight pacas were dissected from the wild animal's sector of the Faculty of Agricultural and Veterinary Sciences of the University of the State of São Paulo. The larynx was observed located in the ventral region of the neck, ventral to the esophagus, connecting the pharynx to the trachea, with cylindrical and irregular shape. Laryngeal cartilages (epiglottic, thyroid, cricoid and arytenoid) are interconnected and have different shapes. Thyroid showed greater length and width, compared to the others. Laryngeal cartilages were submitted to histological processing and stained with hematoxylin-eosin and Masson's trichrome. The epiglottic cartilage was stained with toluidine blue. Laryngeal cartilages thyroid, cricoid and the lower portion of the arytenoids are of hyaline origin and, in contrast, the epiglottis and the upper portion of the arytenoids are elastic. This latter cartilage demonstrated taste buds. The results will be able to auxiliate in veterinary care and as well as the conservation programs for this rodent.(AU)

A paca (Cuniculus paca) tem encorajado inúmeras pesquisas, tornando-a modelo experimental tanto em humanos como na veterinária, além da exploração econômica de seus cortes cárneos, que favoreceu diretamente sua importância zootécnica. Não há descrições anatômicas, microscópicas e mensurações da laringe desse roedor. Foram dissecadas oito laringes de pacas, provenientes do setor de Animais Silvestres da Faculdade de Ciências Agrárias e Veterinárias - Universidade Estadual Paulista. Observou-se que a laringe localiza-se na região ventral do pescoço, ventral ao esôfago, conectando a faringe à traqueia, possuindo formato cilíndrico e irregular. As cartilagens laríngeas (epiglote, tireóide, cricóide e aritenóidea) são interligadas e possuem formatos variados. A cartilagem tireóide demonstrou-se maior em comprimento e largura, comparativamente às demais. As cartilagens laríngeas foram submetidas a processamento histológico e coradas em hematoxilina-eosina e tricrômio de Masson. A cartilagem epiglote foi corada em azul de toluidina. As cartilagens laríngeas tireóide, cricóide e a porção inferior das aritenóides são de origem hialina; em contrapartida, a epiglote e a porção superior das aritenóides, de origem elástica. Esta última cartilagem demonstrou corpúsculos gustativos. Os resultados poderão auxiliar tanto nos atendimentos veterinários quanto nos programas de conservação desse roedor.(AU)

Morfologia do pâncreas da paca (Cuniculus paca, Linnaeus, 1766) / Morphology of the pancreas of pacas (Cuniculus paca, Linnaeus, 1766)

Este trabalho se baseou na descrição morfológica da paca (Cuniculus paca), espécie selvagem pertencente à ordem dos roedores, típica de regiões tropicais, sendo o seu conhecimento anatômico detalhado muito importante para sua conservação, experimentação e comercialização, pois ainda são escassos na literatura os trabalhos sobre morfologia de espécies selvagens. Macroscopicamente, o pâncreas da paca é formado por lóbulos, organizados em ramificações difusas, e, microscopicamente, tal órgão possui uma camada de tecido conjuntivo frouxo, a partir da qual partem septos, que dividem a glândula em lóbulos, compostos de ácinos e ilhotas pancreáticas.(AU)

The present study aimed to describe the morphology of pacas (Cuniculus paca), a wild species belonging to the order Rodentia, found in tropical areas. The detailed anatomical knowledge of pacas is very important for conservation, experimental studies and commercialization because studies in literature about wild species are scarce. The pancreas of paca has lobes organized in diffuse branches at the macroscopic level and microscopically this organ has a loose connective tissue layer, originating septa that divide the gland into lobes composed of acini and pancreatic islets.(AU)

Cost comparison of pressure ulcer preventive dressings: hydrocolloid dressing versus transparent polyurethane film.

OBJECTIVE: To evaluate and compare the costs of using a transparent polyurethane film (PF) and hydrocolloid dressing (HD) in the prevention of pressure ulcers (PUs). METHOD: This descriptive, observational, longitudinal, comparative study was conducted in the intensive care units, coronary care unit and medical clinic of a charity hospital in Brazil. Data were collected during a 30-day study period, consisting of physical examination, assessment of risk factors for PU development and application of the Braden scale, which were performed at inclusion in the study and once daily during hospitalisation. Either PF or HD was applied bilaterally in the sacral and trochanteric regions for prevention of PUs in patients at a moderate to high risk of PUs according to the Braden scale, and costs of using PU preventive dressings were estimated. RESULTS: The mean total costs per dressing change per patient when using the HD and PF to prevent PUs were 413.60 BRL and 74.04 BRL, respectively. There were significant between-group differences in mean costs for all variables, except for saline solution and nurse-technician services. CONCLUSION: Results showed that the mean cost per dressing change per patient was lower when using the transparent PF than when using the HD.

The Hindlimb Arterial Vessels in Lowland paca (Cuniculus paca, Linnaeus 1766).

This study aims to describe the origin and distribution of the hindlimb arterial vessels. Five adult lowland pacas (Cuniculus paca) were used. Stained and diluted latex was injected, caudally to the aorta. After fixation in 10% paraformaldehyde for 72 h, we dissected to visualize and identify the vessels. It was found out that the vascularization of the hindlimb in lowland paca derives from the terminal branch of the abdominal aorta. The common iliac artery divides into external iliac and internal iliac. The external iliac artery emits the deep iliac circumflex artery, the pudendal epigastric trunk, the deep femoral artery; the femoral artery originates the saphenous artery, it bifurcates into cranial and caudal saphenous arteries. Immediately after the knee joint, the femoral artery is called popliteal artery, which divides into tibial cranial and tibial caudal arteries at the level of the crural inter-osseous space. The origin and distribution of arteries in the hindlimb of lowland paca resembles that in other wild rodents, as well as in the domestic mammals.

Morfologia do estômago e do duodeno da paca (Cuniculus paca Linnaeus, 1766) / Morphology of the stomach and duodenum of paca (Cuniculus paca Linnaeus, 1766)

Considerando a importância da paca (Cuniculus paca, Linnaeus, 1766), tanto como fonte proteica alternativa para a população quanto pela possibilidade de vir a se tornar um animal de experimentação e pela falta de informações sobre a sua anatomia, objetivou-se descrever a morfologia macroscópica e histológica do estômago e do duodeno desse roedor, reconhecendo as relações, forma e posição que esses órgãos estabelecem entre si e com outros órgãos. O estômago da paca é unicavitário, está no plano médio mais voltado para o antímero esquerdo, transversalmente, na região hipocôndrica em posição ventrocaudal, interposto entre o esôfago e o duodeno. O duodeno da paca se inicia em sequência ao estômago, segue caudalmente até o nível da quinta ou sexta vértebra lombar, na altura das quais se curva e toma direção cranial, dirigindo-se até o nível da transição entre a última vértebra torácica e a primeira vértebra lombar, onde se continua como jejuno. Histologicamente, o estômago e o duodeno da paca possuem o padrão característico da arquitetura dos órgãos ocos, apresentando as seguintes túnicas: mucosa, submucosa, muscular e serosa. Da forma que se conduziu este estudo, conclui-se que o estômago e duodeno da paca, de forma geral, possuem características morfológicas macroscópicas e histológicas semelhantes às dos animais domésticos e de outros roedores selvagens.

Considering the importance of paca (Cuniculus paca, Linnaeus, 1766) as an alternative protein source for the population as well as the possibility to become an experimental model and the lack of information regarding to the anatomy of this species, the present study aims to describe the gross anatomy and the histology of the stomach and duodenum of this rodent, recognizing the relations, shape and position that these organs establish between themselves and among other organs. The paca stomach is monocavitary; it is located at the median plane, more toward the left antimere, transversally, at the hypochondriac region in a ventrocaudal position, interposed between the esophagus and duodenum. The duodenum of paca begins in sequence of the stomach, following caudally until the fifth or sixth lumbar vertebra, at this level it curves and takes the cranial direction, going until the transition level between the last thoracic vertebra and the first lumbar vertebra where it continues as jejune. Histologically, the stomach and duodenum of paca have the characteristic pattern of the hollow organs architecture, presenting the following layers: mucosa, submucosa, muscular and serous. As this study was conducted, it is possible to conclude that the stomach and duodenum of paca, in general, present gross anatomical and histological characteristics similar to the domestic animals and to the other wild rodents.

Morfologia do estômago do cervo-do-pantanal (Blastocerus dichotomus, Illiger 1815) / Morphology of the marsh deer's stomach (Blastocerus dichotomus, Illiger 1815)

Em razão da falta de informações sobre a morfologia do cervo-do-pantanal, objetivou-se apresentar a morfologia das câmaras gástricas desse cervídeo. Macroscopicamente, o estômago do cervo-do-pantanal é formado pelo rúmen, retículo, omaso e abomaso, assemelhando-se aos ruminantes domésticos. Microscopicamente, o rúmen e o abomaso são semelhantes aos animais domésticos, já o retículo e o omaso apresentam características específicas, como acentuada queratinização no ápice das pequenas projeções epiteliais do retículo e pregas omasais revestidas por discretas papilas.(AU)

Based on the lack of information regarding the morphology of marsh deer, this work aims to describe some morphological aspects of the gastric chamber in this species, collaborating with future investigations, mainly related to rational handling in this cervid. This work aimed to describe the morphology of the gastric chamber of the marsh deer, characterizing the external and internal macroscopical details and the microscopical architecture of these structures by light microscopy. Macroscopically, the marsh deer stomach is formed by the rumen, reticulum, omasum and abomasum similar to the domestic ruminants. Microscopically, rumen and abomasum are similar to the domestic ruminants. The reticulum and the omasum, however, present specific characteristics such as keratin on the top of the reticulum, small epithelial projections and omasum folds covered with discrete papillae.(AU)

O uso do peritônio de paca conservado em solução supersaturada de açúcar a 300 por cento ou glicerina a 98 por cento implantados na parede abdominal de ratos / The use of paca peritoneum preserved in supersaturated sugar solution 300 percent or glycerine 98 percent implanted in the abdominal wall of rats

Na busca de material biológico alternativo para a realização de implantes, objetivou-se com o presente estudo avaliar comparativamente a implantação do peritônio de paca, uma nova opção de biomaterial, conservado em solução supersaturada de açúcar a 300 por cento e conservado em glicerina a 98 por cento na parede abdominal de ratos Wistar. Foram utilizados 60 ratos, machos, da linhagem Wistar, pesando entre 150 e 200 gramas, organizados nos seguintes grupos experimentais: grupo controle (GI), grupo peritônio conservado em solução supersaturada de açúcar a 300 por cento (GII) e grupo peritônio conservado em glicerina a 98 por cento (GIII), cada um com 20 animais. Os grupos GII e GIII receberam o enxerto de peritônio da paca conservado em solução de açúcar 300 por cento e glicerina 98 por cento, respectivamente, e o grupo GI não recebeu a membrana. Cinco ratos de cada grupo foram submetidos à eutanásia em quatro momentos distintos: sete, 15, 30 e 60 dias de pós-operatório para avaliações macroscópicas e microscópicas da interface implante-tecido nativo. Apesar de reações adversas observadas em 57,5 por cento dos animais do grupo GII e GIII, em 95 por cento dos animais desses grupos houve boa cicatrização da membrana. Na análise histológica, verificou-se a presença de grande infiltrado inflamatório nos períodos iniciais (sete e 15 dias) e grande presença de tecido conjuntivo nos momentos finais (30 e 60 dias). Concluiu-se que o peritônio da paca como membrana biológica conservado nos meios estudados pode ser utilizado com segurança na parede abdominal de ratos; ainda, que sua conservação em solução supersaturada de açúcar a 300 por cento permitiu melhor maleabilidade no ato cirúrgico...

In the search for alternative biological material to perform implants, this study aimed to compare the implantation of paca peritoneum, a new biomaterial option, preserved in supersaturated sugar solution 300 percent and preserved in glycerin 98 percent in the abdominal wall of Wistar rats. A total of 60 male rats from the Wistar strain weighing between 150 and 200 grams were housed into three different experimental groups: control group (GI), peritoneum preserved in supersaturated sugar solution 300 percent group (GII) and peritoneum preserved in glycerin 98 percent group (GIII), with 20 animals each. The GII and GIII received the paca peritoneum graft preserved in sugar solution 300 percent and glycerin 98 percent, respectively and group GI did not receive any membrane. Five rats from each group were euthanized at four different times: seven, 15, 30 and 60 days post-surgery for macroscopic and microscopic evaluations in graft-native tissue interface. Despite the adverse reactions observed in 57.5 percent of GII and GIII, there was good healing of the membrane in 95 percent of the animals in these groups. Upon histological examination there was a large presence of inflammatory infiltrates in the initial periods (seven and 15 days) and a large presence of connective tissue in the final stages (30 and 60). It was concluded that the paca peritoneum as biological membrane preserved as presented in this study can be used safely in the abdominal wall of rats, the preservation in supersaturated sugar solution 300 percent allowed better flexibility during surgery...

Avaliação macroscópica e microscópica de implante de Biomembrana em estômago de coelhos (Oryctolagus cuniculus) / Macroscopic and microscopic evaluation biomembrane implant in the stomach of rabbits (Oryctolagus cuniculus)

Neste estudo foi implantado um retalho de biomembrana de látex natural em substituição a um fragmento da parede de estômago de coelhos albinos, raça Nova Zelândia, adultos, machos não castrados (n=12), com o intuito de avaliar o processo de reparação tecidual no que se refere à biocompatibilidade, à capacidade de reparação tecidual e a possíveis complicações. Aos 15, 30 e 60 dias de pós-operatório, os animais foram sacrificados, mediante o emprego de tiopental sódico (200mg kg-1), para posterior avaliação macroscópica e análise histopatológica da interface do implante com o tecido nativo. Macroscopicamente, aos 15, 30 e 60 dias de pós-operatório observou-se presença de aderências na face serosa. Aos 60 dias de pós-operatório, em todos os animais avaliados, a biomembrana não foi encontrada. Sob microscopia de luz, aos 15 e 30 dias, verificaram-se descontinuidade das camadas muscular e mucosa, presença de infiltrado inflamatório polimorfonuclear. Foram visualizados vasos sanguíneos e fibras musculares. Aos 60 dias, as camadas mucosa, muscular estavam completamente reconstituídas. O implante foi biocompatível e forneceu arcabouço para orientação e desenvolvimento das camadas teciduais do estômago, mediante processos de reparação, restabelecendo a estrutura do órgão.

In this study, biomembrane of natural latex was utilized to replace a section of the stomach wall of New Zealand rabbits, adult and non-castrated males (n=12), in order to evaluate the tissue repair process in regards to its biocompatility, scar formation ability and possible complications. The animals were euthanized at fifteen, 30 and 60 days post operation, by use of sodium thiopental (200mg kg-1), followed by macroscopic and histopathological analysis of the implant interface with the native tissue. Macroscopically, at fifteen, 30, and 60 days post operation adherence was observed in the serosal wall. At 60 days post operation, the biomembrane is not in the stomach. Under light microscopy, at fifteen and 30 days, discontinuity of muscle layer and mucosa layer, and presence of polimorfonuclear population of inflammatory cells was observed. New vessels and muscle fibers were observed. At 60 days, the mucosa and muscle layers were complete reconstituted. The implants were biocompatible and had provided the mainframe for orientation and development of the tissue layers through repairing processes, thus reestablishing the organ structure.

Toll-like receptors regulate B cell cytokine production in patients with diabetes.

AIMS/HYPOTHESIS: Understanding cellular and molecular events in diabetes mellitus will identify new approaches for therapy. Immune system cells are important modulators of chronic inflammation in diabetes mellitus, but the role of B cells is not adequately studied. The aim of this work was to define the function of B cells in diabetes mellitus patients through focus on B cell responses to pattern recognition receptors. METHODS: We measured expression and function of Toll-like receptors (TLRs) on peripheral blood B cells from diabetes mellitus patients by flow cytometry and multiplexed cytokine analysis. We similarly analysed B cells from non-diabetic donors and periodontal disease patients as comparative cohorts. RESULTS: B cells from diabetes mellitus patients secrete multiple pro-inflammatory cytokines, and IL-8 production is significantly elevated in B cells from diabetic patients compared with those from non-diabetic individuals. These data, plus modest elevation of TLR surface expression, suggest B cell IL-8 hyperproduction is a cytokine-specific outcome of altered TLR function in B cells from diabetes mellitus patients. Altered TLR function is further evidenced by demonstration of an unexpected, albeit modest 'anti-inflammatory' function for TLR4. Importantly, B cells from diabetes mellitus patients fail to secrete IL-10, an anti-inflammatory cytokine implicated in inflammatory disease resolution, under a variety of TLR-stimulating conditions. Comparative analyses of B cells from patients with a second chronic inflammatory disease, periodontal disease, indicated that some alterations in B cell TLR function associate specifically with diabetes mellitus. CONCLUSIONS/INTERPRETATION: Altered TLR function in B cells from diabetes mellitus patients increases inflammation by two mechanisms: elevation of pro-inflammatory IL-8 and lack of anti-inflammatory/protective IL-10 production.

Comparison of Schistosoma mansoni irradiated cercariae and Sm23 DNA vaccines.

Immunization with defined antigens is generally less effective at inducing host protection against experimental infection with Schistosoma mansoni than vaccination with attenuated infective cercariae. We predicted that quantitative and/or qualitative differences existed between the immune responses generated to attenuated cercariae and those induced by defined antigens. Thus, we compared immune responses typically associated with protection in the murine model between animals vaccinated with attenuated cercariae and mice immunized with DNA encoding Sm23, a schistosome integral membrane protein that has previously been shown to confer protection. Mice vaccinated three times with attenuated cercariae demonstrated higher levels of protection than Sm23-vaccinated animals but spleen cells from Sm23 DNA vaccinated mice produced significantly higher levels of schistosome antigen-specific IFN-gamma. Both vaccines induced similar levels of Sm23-specific antibody and post-challenge dermal inflammation. However, the pulmonary inflammatory responses following challenge were much less pronounced in DNA immunized animals compared to those receiving irradiated cercariae. Thus, although Sm23 DNA vaccination effectively induced parasite-specific IFN-gamma and antibody responses, it failed to evoke other critical responses needed for optimal vaccine efficacy.

Differential induction of nitric oxide synthase in various organs of the mouse during endotoxaemia: role of TNF-alpha and IL-1-beta.

BALB/c mice injected intraperitoneally with bacterial lipopolysaccharide (LPS) developed lethal septic shock. This was accompanied by significantly elevated concentrations of nitrite and nitrate in the plasma and expression of high levels of nitric oxide (NO) synthase activity in the lungs, heart, spleen and peritoneal macrophages. Mice pretreated with anti-tumour necrosis factor-alpha (TNF-alpha) monoclonal antibody or anti-interleukin-1 beta (IL-1 beta) polyclonal antibody were protected, in a dose-dependent manner, from endotoxin-induced mortality. This effect was accompanied by a significant reduction in plasma levels of nitrite and nitrate. Antibody treatment also reduced the level of NO synthase activity in peritoneal macrophages, spleen and heart but had no effect on enzyme expression in the lung. These results demonstrate that TNF-alpha and IL-1 beta play an important role in the induction of NO following administration of LPS and in the development of endotoxin-induced shock. In addition, NO synthase activity is differentially expressed in various organs and this may not always require TNF-alpha and IL-1 beta.

Pre-exposure of murine macrophages to lipopolysaccharide inhibits the induction of nitric oxide synthase and reduces leishmanicidal activity.

Murine macrophages produce nitric oxide (NO) from L-arginine on stimulation with lipopolysaccharide (LPS), alone or with interferon-gamma (IFN-gamma). The effect of incubation of macrophages with low concentrations of LPS on NO synthesis on subsequent stimulation was investigated, using a murine macrophage cell line, J774, and peritoneal macrophages from CBA mice. Cells which had been incubated with LPS produced significantly lower amounts of NO, and expressed lower levels of NO synthase activity, following stimulation with IFN-gamma and LPS, or with a high concentration of LPS. This effect was not reversed by tumor necrosis factor-alpha. The ability of CBA macrophages to kill the intracellular parasite Leishmania major was markedly reduced by pre-incubation with LPS. Reduced NO production by macrophages previously exposed to LPS is a manifestation of endotoxin tolerance, and may represent an important means of regulation of NO synthesis and thus a survival mechanism for intracellular parasites.

Induction of macrophage parasiticidal activity by Staphylococcus aureus and exotoxins through the nitric oxide synthesis pathway.

Murine peritoneal macrophages stimulated in vitro with killed Gram-positive bacteria Staphylococcus aureus or its membrane components in the presence of interferon-gamma (IFN-gamma) expressed high levels of nitric oxide (NO) synthase and produced large amounts of NO in a dose-dependent manner. This is not due to the contamination by Gram-negative endotoxin because the stimulatory activity was not affected by the addition of polymyxin B. The expression of the NO synthase and the synthesis of NO by macrophages stimulated with toxic shock syndrome toxin-1 (TSST), lipoteichoic acid (LTA) or killed whole S. aureus together with IFN-gamma was inhibited by the glucocorticoid, dexamethasone or by the specific inhibitor of NO synthesis, L-N-iminoethyl-ornithine (L-NIO). The exotoxins together with IFN-gamma also activated macrophages to kill the intracellular parasite Leishmania major. The leishmanicidal activity was completely inhibited by L-NIO.

Interleukin-4 transgenic mice of resistant background are susceptible to Leishmania major infection.

The outcome of cutaneous leishmaniasis is dependent on the balance of Th1 and Th2 cells. In the murine model, Th1 cells are host-protective whereas the Th2 cells are disease-promoting. However, the in vivo role of interleukin-4 (IL-4), a signature product of Th2 cells, is uncertain. We compared the course of Leishmania major infection in the genetically resistant 129/Sv mice and the mutant 129/Sv mice transgenic for the murine IL-4 gene under the control of the immunoglobulin heavy chain enhancer and promoter. We report here that in contrast to their wild-type parents, the IL-4 transgenic mice are susceptible to L. major infection. This is associated with the development of inexorably progressive lesions and parasite loads. Spleen cells from infected transgenic mice produced significantly higher levels of IL-4 but lower amounts of interferon-gamma when stimulated in vitro with leishmanial antigens compared to those from infected normal 129/Sv mice. Furthermore, sera from the infected transgenic mice contained higher levels of IL-4 and IgE than the sera of infected normal 129/Sv mice. These results, therefore, establish in a new animal model that IL-4 promotes disease development in murine cutaneous leishmaniasis.

Reconstitution of C.B-17 scid mice with BALB/c T cells initiates a T helper type-1 response and renders them capable of healing Leishmania major infection.

C.B-17 scid mice, which were found to be very susceptible to infection with Leishmania major, were reconstituted with various doses of T cells, T plus B cells or unfractionated spleen cells from nonhealer BALB/c mice. All reconstitution protocols, except for the transfer of very high numbers of BALB/c spleen cells, led to a spontaneously healing infection and resistance to reinfection, rather than the lethal, nonhealing infection typical of BALB/c mice. These healing responses were associated with a strong T helper 1 (Th1)-like response characterized by delayed-type hypersensitivity (DTH) responsiveness, but no elevation of serum IgE, and by the production of high levels of interferon-gamma (IFN-gamma), but no interleukin-4 (IL-4) by lymph node and spleen cells after restimulation with antigen in vitro. The development of this Th1 response from BALB/c Th cells requires IFN-gamma during the initial infection period. Treatment of scid mice with a single injection of neutralizing anti-IFN-gamma antibody prior to infection and reconstitution prevented healing and permitted the development of a Th-2 like response as indicated by elevated serum IgE, but no DTH, and by the production of IL-4, but very little IFN-gamma, after antigen stimulation in vitro. As few as 10(4) transferred T cells led to a Th1-like response, suggesting that the IFN-gamma is of host rather than donor origin. The transfer of very high numbers (7.5 x 10(7)) of BALB/c spleen cells overcame the effects of the IFN-gamma and led to the nonhealing infection and cytokine pattern characteristic of BALB/c mice. The enrichment or depletion of B cells from the transferred T cells had no measurable effect upon the development of a healing response in reconstituted scid mice.

Interactions between CD3 and Thy1 T cell activation pathways: blockade of CD3-mediated T lymphocyte activation induced by immobilized anti-Thy1 antibodies.

Resting murine T cell activation induced by either CD3 complexes or Thy1 molecules was investigated in vitro, using surface-bound anti-CD3 mAb as the stimulus. One mitogenic anti-Thy 1 mAb (G7) lost mitogenicity when presented to T cells immobilized on a plastic surface, even in the presence of phorbol ester. Moreover, T cell activation induced by immobilized anti-CD3 was potently blocked by coimmobilized anti-Thy 1 mAb. Nonmitogenic anti-Thy 1 mAb also blocked CD3-induced activation when coimmobilized with anti-CD3. Control experiments showed that anti-Thy 1 specifically blocked T cell activation, even in the presence of measurable and functional concentrations of plastic-bound anti-CD3. Coimmobilized anti-Thy 1 potently blocked IL2 secretion stimulated by anti-CD3. Addition of exogenous rIL2 completely prevented anti-Thy 1-mediated blockade. On the other hand, while completely blocking T cell proliferation, immobilized anti-Thy 1 only partially blocked secretion of IL3-like activity by the T cells. One IgM anti-Thy 1 mAb (2A3) induced secretion of IL3-like activity by T cells when immobilized in the absence of bound anti-CD3. These results indicate that extensive aggregation of Thy 1 molecules delivers a potent negative signal which antagonizes CD3-mediated T cell activation and growth.

Independent and cooperative behavior of Thy1 and CD3:TcR T cell signalling pathways.

Normal resting spleen T lymphocytes from mice were stimulated in vitro by monoclonal antibodies (mAbs) against either Thy 1 or CD3:TcR surface protein molecules. Although both mAbs were mitogenic, anti-Thy 1 activation generated 5 times more IL2 secretion than anti-CD3 activation under similar conditions. Production of IL3-like activity was comparable for both Thy1 and CD3-mediated activation. In addition, non-mitogenic doses of anti-CD3 and anti-Thy1 (0.16 micrograms/ml and 0.0125% ascites, respectively) mAbs induced T cell activation when provided together. These results indicate that Thy1 signalling cooperates with the CD3:TcR pathway to activate T cells. However, the Thy1 pathway is also regulated independently since IL2 production is larger when stimulated by anti-Thy1 than anti-CD3 mAbs.

Independent and cooperative bheravior of THY1 and CD3:TcR T cell signalling pathways

Normal resting spleen T lymphocytes form mice were stimulated in vitro by monoclonal antibodies (mAbs) against either Thy1 or CD3:TcR surface protein molecules. Although both mAbs were mitogenic, anti-Thy1 activation generated 5 times more IL2 secretiom than anti-CD3 activation under similar conditions. Priduction of IL-like activity was comparable for both Thy1 and CD3-mediated activation. In addition, non-mitogenic doses of anti-CD3 and anti-Thy1 (0.16microng/ml and 0.0125% ascites, repectively) mAbs induced T cell activation when provided together. These reults indicate that Thy1 signalling cooperates with the CD3:TcR pathway to activate T cells. However, the pathway is also regulated independently since IL2 production is larger when stimulated by anti-Thy1 than anti-CD3mAbs