Search PubMed⌕ Search

Biomedical subjects

R Vilella

Publications and source records attributed to R Vilella.

43 records · Page 3Linked to original sources
PubMed1987

Hybridoma process: ultrastructural cytology of different stages.

In order to define the ultrastructure of the hybridoma cell and to learn more about the plasmocytic differentiation process, a scanning (SEM) and transmission (TEM) electron microscopy study of several cell types involved in the production of monoclonal antibodies was performed. Cells of the three different stages in hybridoma process were studied. These cells included NS/1 murine myeloma cells, 40-3A4 in vitro cultured hybridoma and 33-1D2 ascitic tumor hybridoma cells. A stereological analysis of the Sv parameter (surface of RER per volume unit of cytoplasm) was performed in the murine myeloma line, the in vitro cultured hybridoma and the ascitic tumor hybridoma. In order to comparatively evaluate the plasmocytic differentiation of these cells the same methodology was applied to splenic lymphocytes from immunized mouse and to mature human myelomatous plasma cells. As expected, during the hybridoma process, a progressive increase in the amount of RER was detected. This was in contrast with the surface characteristics of the cells which become progressively smooth when the hybridoma was cloned, either in vitro or in vivo. From these results it can be inferred that the amount of RER is a more reliable parameter than surface blebs as a morphological element indicative of plasmocytic differentiation. On the other hand, numerous viral particles were present not only in murine myeloma line but also in hybridoma clones secreting monoclonal antibodies.

Animals↗
PubMed1984

An antiplatelet monoclonal antibody that inhibits ADP and epinephrine-induced aggregation.

A monoclonal antibody (Mab) named EDU-3, was produced by fusing splenocytes from one Balb/c mouse, immunized with a mixture of platelets and non-T cells from heparinized human peripheral blood, with the HAT-sensitive myeloma line P3-NS1/1.Ag4.1. By indirect immunofluorescence (IF) it was seen that this Mab reacted with all normal human platelets and bone marrow megakaryocytes, but did not react with lymphoid cells from normal donors, or platelets from Glanzmann's thrombasthenia (GT) patients. Immunoprecipitation and SDS-PAGE experiments demonstrated that this Mab recognized an epitope on the IIb-IIIa glycoprotein complex (GPC). EDU-3 inhibited platelet aggregation and release of ATP induced by ADP and epinephrine. Aggregation induced by arachidonic acid, ristocetin and bovine factor VIII were not inhibited by EDU-3. The difference between EDU-3 and other Mab directed against the IIb-IIIa GPC is discussed.

Adenosine Diphosphate↗
PubMed1983

Monoclonal antibody against HLA-Aw32 + A25. Is HLA-Aw32 an allele with no unique antigenic determinant?

In the present paper we describe the production and characterization of a monoclonal antibody (Mab) recognizing HLA-Aw32 + A25 antigens. NS1 murine myeloma cells were fused with splenocytes from a BALB/c mouse immunized with normal human peripheral blood (PB) lymphocytes of phenotype A1, Aw32; B7,B37,Cw-,Cw-;DR2,DRw10. Supernatants were first screened against Cr51-labeled immunizing cells by complement dependent cytotoxicity 51Cr-CDC). Cultures identified as producing cytotoxic antibodies were subcultured and the supernatants tested against a selected panel of HLA typed cells by the NIH microcytotoxicity method. One culture producing antibody reacting with an HLA polymorphism was detected. This hybrid, designated CATA 1, was cloned twice by limiting dilution and obtained in ascitic form. Specificity of CATA 1 Mab was evaluated against a panel of 120 PB T cells from normal donors. CATA 1 reacted with cells bearing HLA-A25 or HLA-Aw32 antigens. In addition, a reaction was observed with a cell of phenotype A2,Aw31; B17,Bw49. Isoelectric focusing revealed the monoclonal nature of CATA 1, with immunofixation identifying it as an IgG molecule. Absorption studies have demonstrated that CATA 1 recognizes a common determinant on HLA-A25 and HLA-Aw32. The finding that this Mab recognizes the same CREG as alloantisera against HLA-Aw32 suggests that this antigen has no unique epitopes.

Alleles↗
PubMedDate not supplied

Specific immune response to Phleum pratense plant profilin in atopic patients and control subjects.

BACKGROUND: Phleum pratense (Phl p) pollen is a known cause of allergic disease worldwide. Profilins have been identified as functional plant pan-allergens. The role of Phl p profilin in the specific immune response in sensitized Phl p patients is unknown. METHODS: Skin prick test and specific serum IgE levels were performed in 26 patients allergic to Phl p and in 18 nonallergic control donors. Peripheral blood mononuclear cells were isolated from both groups and stimulated with crude extract or highly purified Phl p profilin, and the production of type I and type II cytokines was determined in patients and controls stimulated with specific and polyclonal stimulus. T-cell lines specific to Phl p profilin were established from PBMCs and cross-reactivity with another highly purified profilin from Parietaria judaica (Pj) was evaluated. RESULTS: Patients allergic to Phl p profilin showed increased T-cell-proliferative responses to this profilin compared with control subjects. The production of IL-4 and IFN-gamma in response to the specific stimulus was undetectable. However, the production of IL-4 and IFN-gamma in response to a polyclonal stimulus (PHA) was measurable and different for atopic patients and control subjects: IL-4 was higher (p < 0.001) in allergic patients and IFN-gamma lower (although not significant) in controls. Neither the T-cell responses nor the production of IL-4 in response to a polyclonal stimulus (PHA) correlated with the individual degree of cutaneous response to Phl p profilin or to the levels of specific Phl p IgE. The T-cell lines tested did not show any cross-reactivity with Pj profilin. CONCLUSIONS: Phl p profilin is in part responsible for the T-cell mediated immunological response in patients allergic to Phl p. The response is very specific since Phl p profilin specific T-cell lines did not show cross-reactivity with a highly homologous profilin from Parietaria judaica (Pj). The lack of correlation between the proliferative T-cell response and polyclonal IL-4 production with allergen-specific serum IgE and SPT probably indicates that some of the responding T-cells may be involved in immune reactions other than the support of IgE production.

Adult↗
PubMedDate not supplied

Specific immune response to Parietaria judaica plant profilin: a low T cell proliferative response supports high IgE and skin prick test.

BACKGROUND: allergic disease caused by Parietaria judaica (Pj) has been widely documented in Mediterranean area. Profilins have been identified as widely distributed allergenic proteins. The role of Pj profilin in specific immune response in Pj-sensitized patients is unknown. METHODS: skin prick test and determination of specific and total IgE levels in serum were performed in all patients (n = 28) and non-allergic controls (n = 18). Peripheral blood mononuclear cells (PBMC) were isolated from both groups and stimulated with crude extract or highly purified Pj profilin. The production of type I and type II cytokines was determined by specific and polyclonal stimuli in patients and controls. T-cell lines specific to Pj profilin were established and cross-reactivity with another highly purified profilin from Phleum pratense (Phl p) was evaluated. RESULTS: Pj profilin-sensitized patients showed a small but significantly increased in T-cell proliferative response to this profilin compared with non-atopic controls. The production of interleukin (IL)-4 and interferon (IFN)-γ in response to the specific stimulus was undetectable. However, the production of IL-4 in response to a polyclonal stimulus [phytohemagglutinin (PHA)] was significantly higher in atopic patients than in controls. The T-cell response did not correlate with the magnitude of response to skin prick tests with Pj profilin or with Pj-specific serum IgE levels. In addition, the production of IL-4 in response to a polyclonal stimulus (PHA) did not correlate with the individual skin prick tests to Pj profilin or with Pj-specific IgE levels in serum. The T-cell lines tested showed no cross-reactivity with Phl p profilin. CONCLUSIONS: our results suggest that Pj profilin is partly responsible for the T-cell-mediated response in patients allergic to Pj. The high skin reactivity to Pj profilin is these patients was accompanied by a small increase in the T-cell response to this profilin. The response was highly specific since Pj profilin specific T-cell lines showed no cross-reactivity with a highly homologous profilin from Phl p. The lack of correlation between the proliferative T-cell response and polyclonal IL-4 production with allergen-specific serum IgE and skin reactivity probably indicates that some of the responding T-cells may be involved in immune reactions other than those supporting IgE production.

Adult↗