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F Santoro

Publications and source records attributed to F Santoro.

At least 73 records · Page 4Linked to original sources
PubMed1988

Protective role of IgE in immunocompromised rat toxoplasmosis.

In contrast to euthymic adult Fischer rats, immunocompromised Nu/Nu animals develop a lethal infection when inoculated with the RH strain of the protozoan Toxoplasma gondii. However, a significant period of survival is obtained when Nu/Nu rats are passively transferred with sera from 28-day infected Fischer +/+ (euthymic) animals. Specific IgE are involved since IgE-depleted sera are unable to afford such a protection. Only excreted/secreted Ag or living tachyzoites are able to induce a significant protective IgE response in intact animals. In addition, platelets or, to a lesser extent, eosinophil-rich populations from Toxoplasma infected or excreted-secreted Ag-immunized euthymic animals bear surface IgE and are cytotoxic for the parasite in vitro. Also, adoptive transfer of immune platelets confers a significant degree of protection to Toxoplasma-infected Nu/Nu animals. Our results clearly show the key role of Ag present in both living parasites and excreted-secreted Ag to induce, in this model, a protective IgE response. In addition, as in other parasitic infections, platelets and probably eosinophils are the effector cells involved in controlling parasitic dissemination during Toxoplasma infection in immunocompromised rats.

Animals↗
PubMed1988

The major surface protein of Leishmania promastigotes is a fibronectin-like molecule.

The major surface glycoprotein of Leishmania chagasi promastigotes showed crossreactivity with fibronectin (Fn), a large glycoprotein that is a major constituent of the extracellular matrix of most mononuclear cells. Polyclonal and monoclonal antibodies against Fn precipitated two molecules of 63-58 kDa from the lysates of both 125I and [35S]methionine-labeled promastigotes. In addition, a monoclonal antibody against a 15-kDa fragment of Fn containing the Arg-Gly-Asp-Ser (RGDS) sequence and several polyclonal monospecific mouse antibodies against a synthetic RGDS peptide also recognized the above two molecules. The attachment of Leishmania promastigotes to mouse peritoneal macrophages in vitro was partially inhibited when promastigotes were treated with F(ab')2 fragment of an anti-Fn IgG. Identical results were obtained by saturating the Fn receptors on macrophages using different peptides containing the RGDS sequence. Moreover, antigen preparations rich in glycoprotein 63 could efficiently promote the attachment and spreading of 3T3 mouse fibroblasts to surfaces coated with the antigen. These results clearly suggest that the gp63 of L. chagasi promastigotes is an Fn-like molecule that shares certain biological and molecular characteristics with Fn.

Animals↗
PubMed1988

Killer cells in human cutaneous leishmaniasis.

In French Guiana, American cutaneous leishmaniasis is localized in the skin. The host response appears to be effective since few extra- or intracellular organisms can be found in tissue lesions, and we never observed any cutaneous dissemination or visceral involvement. However, this response is not fully effective since lesions may last for months. By using immunoperoxidase techniques and monoclonal antibodies directed against various cell populations, we examined the local immune response in skin biopsies. We found a high percentage of cells with the K/NK phenotype, a variable but usually high percentage of cells with the T cell phenotype bearing TAC receptors, and moderate numbers of monocytes and B cells. These results suggest that K/NK cells could play a role in the local control of parasite dissemination.

Adult↗
PubMed1988

Induction of a protective antibody-dependent response against toxoplasmosis by in vitro excreted/secreted antigens from tachyzoites of Toxoplasma gondii.

Toxoplasma gondii is a worldwide protozoan parasite which causes severe disease in congenitally infected children and in immunocompromised patients. Besides the well-defined cytoplasmic and membrane antigens of tachyzoites, we felt that excreted/secreted antigens could play a major role in the immune response. We first report the development of a well-controlled procedure for obtaining tachyzoite excreted/secreted antigens (E/SA) in cell-free incubation media. The E/SA immunogenic in human, rat and mouse toxoplasmosis were then characterized. The major E/SA recognized by human sera from the chronic phase of toxoplasmosis had molecular weights of 108, 97, 86, 69, 60, 57, 42, 39, 28.5, 27 and 26 kD. When injected into +/+ Fischer rats, E/SA elicited high antibody titres. In addition, passive transfer of these sera to highly susceptible nu/nu littermates induced a significant degree of protection towards the virulent RH strain of T. gondii. This work, which demonstrates the key role played by E/SA in the protective immune response, suggests that these antigens should be of value both for diagnostic purposes and for the development of new strategies for immunization against toxoplasmosis.

Animals↗
PubMed1988

Requirements of defined cultivation conditions for standard growth of Leishmania promastigotes in vitro.

The growth characteristics of L. chagasi (MHOM/BR/79/LI01) and L. braziliensis (MHOM/BR/72/1670), the causative agents of visceral and muco-cutaneous leishmaniases, respectively, were compared. Inoculum size clearly influences the growth course of both Leishmania species, whatever the culture medium used (serum-supplemented media: GLSH or RPMI, and a chemically defined medium: LITR9). Cultures initiated with low concentrations failed to promote cell growth, while typical growth curves were obtained when higher promastigote inocula were used. For all the species tested, the higher the initial density of flagellates in the medium, the shorter were the periods covered by the latent and particularly by the logarithmic growth phases. In contrast, using constant inocula, variations in the volume of the incubation medium did not change the time-course of the different culture phases of either Leishmania species, provided that the ratio of incubation medium to total flask volume was comparable. Only cell division time significantly increased with the culture volume. We also determined whether or not the growth characteristics of the promastigotes of L. chagasi or L. braziliensis could be generalized to other members of the genus. Our results show that, whatever the culture medium used, L. infantum behaves in the same way as does L. chagasi, whereas L. panamensis, L. guyanensis, L. mexicana and L. amazonensis display growth patterns similar to that of L. braziliensis.

Animals↗
PubMed1987

Infectivity of Leishmania braziliensis promastigotes is dependent on the increasing expression of a 65,000-dalton surface antigen.

Sequential development of Leishmania braziliensis promastigotes from a noninfective to an infective stage was demonstrated. The generation of infective forms was related to their growth cycle and restricted to stationary stage organisms. Using immunofluorescence techniques, we have noticed that the binding of a monoclonal antibody (mAb) against L. braziliensis (VD5/25) increased progressively as the promastigotes developed in culture and was maximal with the infective forms. This antigenic differentiation was not detected with an anti-L. braziliensis polyclonal rabbit antiserum, suggesting that only a few epitopes, including that recognized by VD5/25, have their expression effectively increased on the surface of infective promastigotes. Immunoprecipitation of lysates of surface-iodinated L. braziliensis promastigotes with this mAb revealed two proteins of apparent 65,000 and 50,000 Mr, the 50,000 Mr protein probably representing the unreduced form of the major surface glycoprotein described in several species of Leishmania (GP65). The increasing expression of this epitope was not found with L. chagasi promastigotes, but seems to occur with the parasites from the L. mexicana complex. Intracellular survival of L. braziliensis was completely inhibited when the infective promastigotes were treated with VD5/25. It appears, therefore, that the increasing expression of GP65 on the promastigote surface represents an essential mechanism of leishmania survival in the macrophage.

Animals↗
PubMed1987

Leishmania chagasi: in vitro differentiation of promastigotes monitored by flow cytometry.

A sequential development from a less infective to an infective stage of Leishmania promastigotes growing in culture has been previously reported. The aim of this work was to investigate whether freeze-fracture electron microscopy and flow cytometry would be able to provide some reliable morphological markers of in vitro differentiation of Leishmania chagasi promastigotes. The flow cytometry technique discriminates between the L. chagasi promastigotes from the different stages of their in vitro differentiation. The "forward scatter" intensity of the parasite, very high 15 hr after seeding when the parasites were very condensed and with a high DNA content per particle, strongly decreased during the culture course. Parallel experiments have shown a striking correlation between forward scatter intensity, growth curves, and infectivity of promastigote populations. By contrast, freeze-fracture techniques showed that in either less infective or infective promastigote plasma membranes, the intramembrane particles density in protoplasmic fracture faces (about 2800/micron 2) and in exoplasmic fracture faces (about 1000/micron 2) was independent of the time of cultivation. The amount of filipin lesions, which reflects the cholesterol content within the plasma membrane, was also constant throughout the culture course. Both data suggest that the architecture of the plasma membrane is an intrinsic characteristic of the promastigote stage. This study shows that whereas freeze-fracture electron microscopy does not provide markers for the differentiation of Leishmania promastigotes, flow cytometry may on the other hand be of value as a screening test for promastigote populations allowing the characterization of their developmental stages in in vitro cultures.

Animals↗
PubMed1987

Identification of a major 72 kilodalton surface antigen in twelve isolates of Leishmania braziliensis braziliensis.

The study of the surface antigens of Leishmania braziliensis braziliensis revealed a great homogeneity among ten strains isolated from Bolivia and two reference strains from Brazil and Belize. A 72 kDa major protein, present in all L. b. braziliensis strains, was recognized by both cutaneous and mucocutaneous human sera, but was not recognized by Kala-azar and chagasic sera. No cross-reactive antigens were found among strains of Leishmania braziliensis guyanensis, Leishmania braziliensis panamensis, Leishmania mexicana amazonensis and Leishmania donovani chagasi testing these strains with hamster and human anti-L. b. braziliensis sera. Moreover, these strains possessed major antigens with molecular weights different from those of L. b. braziliensis strains. A microheterogeneity of L. b. braziliensis surface antigens was detected for the high molecular weight antigens and seemed to be related to the isoenzymic microheterogeneity.

Animals↗
PubMed1987

[Infection of the athymic rat by Toxoplasma gondii].

In contrast to normal rats which are resistant to T. gondii infection (10(7) tachyzoïtes), athymic rats did not survive an intraperitoneal infection with 10(3) toxoplasma. When nude rats were injected intravenously with lymph node cells from hirsute littermates, they became resistant in a dose-dependent manner to the infection. In addition, reconstituted athymic rats having survived for more than 4 months the first infection were also protected against a second challenge with 10(5) tachyzoïtes. Anti-T. gondii antibody levels detected in reconstituted athymic rats were related to protection. These preliminary findings suggest that T-dependent immunity is essential in the development of effector mechanisms involving antibodies in resistance to toxoplasmosis.

Animals↗
PubMed1986

[A new ELISA method for the diagnosis of toxoplasmosis. Assay of serum IgM by immunocapture with an anti-Toxoplasma gondii monoclonal antibody].

The diagnosis of acute toxoplasmosis is based exclusively on the detection of IgM anti-Toxoplasma antibodies. The principal of the immunoenzymatic test reported here is the capture of serum IgM antibodies which are detected indirectly by the sequential addition of antigen and a monoclonal antibody directed against the immunodominant epitope at the surface of the tachyzoite. This test combines the sensitivity of enzyme-based assays and the specificity of monoclonal reagents and represents an important contribution to the diagnosis of Toxoplasma gondii infection.

Acute Disease↗
PubMed1986

The immunodominant epitope of the major membrane tachyzoite protein (P30) of Toxoplasma gondii.

Several physicochemical characteristics of the repeated epitope of the major surface protein (P30) of Toxoplasma gondii were investigated with an anti-P30 mAb by two different methods: a one-site/inhibition assay that detects molecules containing single or multiple epitopes and a two-site/one antibody radiometric assay that is only effective with antigenic molecules containing two or more identical epitopes. Using both techniques, the repeated epitope within purified P30 was stable after 1 h at 63 degrees C, but labile at 100 degrees C. It was also resistant to successive freezing and thawing, and not affected after one year at -70 degrees C. Lyophilization and acidic or basic treatment had no effect. This epitope was also resistant to 20% trichloroacetic acid precipitation (activity recovered in the pellet) and to precipitation with cold acetone. To investigate the immunodominance of this repeated epitope during the humoral immune response against T. gondii, competition binding assays between anti-P30 mAb and polyclonal antibodies, from rabbits immunized with either purified P30 or total Toxoplasma extract and from patients with toxoplasmosis, have been used. We found that the mAb inhibited 50-95% of the binding of the IgG antibodies from both rabbits to purified P30. In addition, the binding of both human IgG and IgM antibodies to P30 was significantly inhibited by the mAb. It appears, therefore, that a single region of P30 contains most of the immunogenic activity.

Animals↗