Alfredo De Ioannes - Academia.edu (original) (raw)

Papers by Alfredo De Ioannes

Research paper thumbnail of Preliminary Structural Characterization of Pacific Hagfish Immunoglobulin

Advances in Experimental Medicine and Biology, 1975

The immune capacities of primitive fishes of the class Agnatha, the lamprey and the hagfish, have... more The immune capacities of primitive fishes of the class Agnatha, the lamprey and the hagfish, have been under study for nearly a decade. Early results from Good and his collaborators suggested that lampreys, but not hagfish, possessed an adaptive immune response (1, 2). However, subsequent work of Linthicum and Hildemann (3) and Hildemann and Thoenes (4, 5) showed that hagfish (Eptatretus stoutii) could also respond specifically to certain immunogens and could reject allografts under appropriate conditions.

Research paper thumbnail of Síntesis y secreción del antígeno de superficie del virus de hepatitis B en cultivo de células animales

Research paper thumbnail of Allergens of the urushiol family promote mitochondrial dysfunction by inhibiting the electron transport at the level of cytochromes b and chemically modify cytochrome c1

Biological Research, 2021

BackgroundUrushiols are pro-electrophilic haptens that cause severe contact dermatitis mediated b... more BackgroundUrushiols are pro-electrophilic haptens that cause severe contact dermatitis mediated by CD8+effector T-cells and downregulated by CD4+T-cells. However, the molecular mechanism by which urushiols stimulate innate immunity in the initial stages of this allergic reaction is poorly understood. Here we explore the sub-cellular mechanisms by which urushiols initiate the allergic response.ResultsElectron microscopy observations of mouse ears exposed to litreol (3-n-pentadecyl-10-enyl-catechol]) showed keratinocytes containing swollen mitochondria with round electron-dense inclusion bodies in the matrix. Biochemical analyses of sub-mitochondrial fractions revealed an inhibitory effect of urushiols on electron flow through the mitochondrial respiratory chain, which requires both the aliphatic and catecholic moieties of these allergens. Moreover, urushiols extracted from poison ivy/oak (mixtures of 3-n-pentadecyl-8,11,13 enyl/3-n-heptadecyl-8,11 enyl catechol) exerted a higher inhi...

Research paper thumbnail of Immunodetection of acrosin during the acrosome reaction of hamster, guinea-pig and human spermatozoa

Biological research, 1992

Mammalian sperm acrosomes contain a trypsin-like protease called acrosin which causes limited and... more Mammalian sperm acrosomes contain a trypsin-like protease called acrosin which causes limited and specific hydrolysis of the extracellular matrix of the mammalian egg, the zona pellucida. Acrosin was localized on hamster, guinea-pig and human sperm using monoclonal and polyclonal antibodies to human acrosin labelled with colloidal gold. This was visualized directly with transmission electron microscopy, and with light and scanning microscopy after silver enhancement of the colloidal gold probe. Four distinct labelling patterns were found during capacitation and the acrosome reaction in hamster and guinea-pig spermatozoa, and three patterns were found in human spermatozoa. In the hamster, acrosin was not detected on the inner acrosomal surface after the completion of the acrosome reaction, thus correlating with the observation that hamster spermatozoa lose the ability to penetrate the zona after the acrosome reaction. With guinea-pig and human spermatozoa, acrosin was still detected ...

Research paper thumbnail of Electron Microscopy of the Natural IGM-like Hemagglutinin of the Ratfish (Callorhynchus callorhynchus)

Developmental & Comparative Immunology, 1981

Research paper thumbnail of Visible Light Anaerobic Photoconversion of Tyrosine Sensitized by Riboflavin. Cytotoxicity on Mouse Tumoral Cells

Photochemistry and Photobiology, 2008

The anaerobic phototransformation of tyrosine under visible light sensitized by riboflavin is rep... more The anaerobic phototransformation of tyrosine under visible light sensitized by riboflavin is reported. The cytotoxicity of the anaerobic photoproducts on in vitro-cultured myeloid mouse tumoral cells was demonstrated. A radical mechanism is proposed. Dityrosine was identified as one of the main anaerobic photoproducts by using absorption, emission and 'H-NMR spectra.

Research paper thumbnail of Apoptosis Induction in Nonirradiated Human HL-60 and Murine NSO/2 Tumor Cells by Photoproducts of lndole-3-acetic Acid and Riboflavin

Photochemistry and Photobiology, 1999

The effect of the photoproducts of indole-3-acetic acid sensitized by riboflavin on nonirradiated... more The effect of the photoproducts of indole-3-acetic acid sensitized by riboflavin on nonirradiated human HL-60 and murine NS0/2 tumor cells was studied. Severe damage with a dose-response effect was observed on both cell types. The effect was greater than that previously described for the tryptophan riboflavin photoproducts. Electron microscopy studies and flow cytometry analysis of DNA fragmentation allowed us to conclude that the photoproducts studied in this work induce cell death by an apoptotic mechanism.

Research paper thumbnail of Females of the communally breeding rodent, Octodon degus, transfer antibodies to their offspring during pregnancy and lactation

Journal of Reproductive Immunology, 2007

Females in numerous rodent species engage in communal nesting and breeding, meaning that they sha... more Females in numerous rodent species engage in communal nesting and breeding, meaning that they share a nest to rear their young together. One potential benefit to communally nesting mothers is that infants improve their immunocompetence. Thus, suckling from two or more females might provide newborns with a more diverse array of antibodies and defensive cells. As a first step toward testing the immunocompetence hypothesis, we assessed whether female degus (Octodon degus), a communally nesting and breeding caviomorph rodent, transfer immunoglobulins to their young through the yolk sac or placenta while in the uterus and, during lactation, through milk. With this aim, adult degu females were immunized with four antigens, including two mollusk hemocyanins from Concholepas and Megathura (CCH and KLH, respectively), porcine thyroglobulin and tetanus toxoid. Specific antibodies against the experimental antigens were used to track the origin of antibodies in the young. To establish the presence of specific antibodies of IgG and IgA isotypes in sera and milk of animals, an indirect enzyme-linked immunosorbent assay (ELISA) was developed. Degu females produced specific antibodies against antigens not found in their natural environment, and mothers were able to transfer the induced antibodies to their litters during pregnancy (IgG) and during lactation (IgA). However, we recorded only limited evidence of degu offspring acquiring antibodies from lactating mothers other than their own, giving little support to the increased immunocompetence hypothesis.

Research paper thumbnail of Procedure for Radiolabeling Gizzerosine and Basis for a Radioimmunoassay

Journal of Agricultural and Food Chemistry, 1999

A method for the labeling of gizzerosine (GZ), a biogenic amine found in fish meal, is described.... more A method for the labeling of gizzerosine (GZ), a biogenic amine found in fish meal, is described. The labeling procedure with 125 I using a water-soluble Bolton-Hunter reagent and a mild water-insoluble oxidant (Iodogen) reagent is rapid and reproducible. The 125 I-GZ hapten was demonstrated to be immunologically active in a radioimmunoassay developed with polyclonal antibodies to GZ absorbed with a histamine-Sepharose column. The curves were linear in the range of 0.0001 and 0.1 µg/mL. Samples of fish meal previously extracted of histamine with methanol and submitted to acid hydrolysis were contaminated with known amounts of GZ and submitted to the assay. The fish meal samples contaminated with GZ show a dose-response effect similar to the standard curve, and apparently the other component present in the sample did not interfere with the binding of the antibodies to 125 I-GZ. These data indicate the suitability of the radioimmunoassay to determine specifically GZ in fish meal.

Research paper thumbnail of The role of acrosin and antibodies to acrosin in gamete interactions

Journal of Reproductive Immunology, 1989

Research paper thumbnail of Sperm antigens relevant to human fertility : final report

Research paper thumbnail of Hemocyanin of the Molluscan Concholepas concholepas Exhibits an Unusual Heterodecameric Array of Subunits

Journal of Biological Chemistry, 2004

Research paper thumbnail of The oxygen-binding properties of hemocyanin from the mollusk Concholepas concholepas

Biochimica et biophysica acta, Dec 24, 2017

Hemocyanins have highly conserved copper-containing active sites that bind oxygen. However, struc... more Hemocyanins have highly conserved copper-containing active sites that bind oxygen. However, structural differences among the hemocyanins of various mollusks may affect their physicochemical properties. Here, we studied the oxygen-binding cooperativity and affinity of Concholepas concholepas hemocyanin (CCH) and its two isolated subunits over a wide range of temperatures and pH values. Considering the differences in the quaternary structures of CCH and keyhole limpet hemocyanin (KLH), we hypothesized that the heterodidecameric CCH has different oxygen-binding parameters than the homodidecameric KLH. A novel modification of the polarographic method was applied in which rat liver submitochondrial particles containing cytochrome c oxidase were introduced to totally deplete oxygen of the test solution using ascorbate as the electron donor. This method was both sensitive and reproducible. The results showed that CCH, like other hemocyanins, exhibits cooperativity, showing an inverse relat...

Research paper thumbnail of Product and Composition Containing a Concholepas Concholepas Hemocyanin (CCH) Subunit A, and a Method of Use Thereof

Research paper thumbnail of CD8p T Cells Are the Effectors of the Contact Dermatitis Induced by Urushiol in Mice and Are Regulated by CD4p T Cells

Int Arch Allergy Immunol, 1998

Research paper thumbnail of Novel hemocyanin from the Fissurella latimarginata exhibits an outstanding immunogenicity and non-specific immunomodulatory effects in a melanoma model

The Journal of Immunology, May 1, 2012

Research paper thumbnail of Product and composition containing a Concholepas concholepas hemocyanin (CCH) subunit, and a method of use thereof

Research paper thumbnail of Development of anti-human B blood group monoclonal antibodies suitable as a blood typing reagent

Hybridoma, 1994

An improved procedure for the generation of high-avidity anti-human B blood group monoclonal anti... more An improved procedure for the generation of high-avidity anti-human B blood group monoclonal antibodies (MAbs) was developed. One of them, termed 7A1-2, showed excellent qualities of titer, avidity, and intensity required for use as human B blood typing reagent. Hemagglutination inhibition studies with monosaccharides and oligosaccharides were carried out to determine the specificity of the MAb 7A1-2. These studies indicate that the antibody reacts with the immunodominant region of the antigen which is known to confer the serologic specificity of this blood group.

Research paper thumbnail of A Light-Induced Tryptophan-Riboflavin Binding: Biological Implications

Advances in Experimental Medicine and Biology, 1991

We review here the covalent photo-binding induced by visible light between the essential amino ac... more We review here the covalent photo-binding induced by visible light between the essential amino acid tryptophan and the vitamin riboflavin. We discuss the biological implications of this photoadduct in relation to the hepatotoxic and cytotoxic effect associated to parenteral nutrients and to culture media exposed to the action of light, respectively. We also analyze the formation of a photo-binding between riboflavin and the residues of tryptophan present in the proteins of the eye lens, a tissue which is permanently exposed to visible light.

Research paper thumbnail of Inhibitory effect of nordihydroguaiaretic acid and its tetra-acetylated derivative on respiration and growth of adenocarcinoma TA3 and its multiresistant variant TA3MTX-R

In vivo (Athens, Greece)

The effects of nordihydroguaiaretic acid (NDGA) and its tetraacetylated derivative (NDGATA) on th... more The effects of nordihydroguaiaretic acid (NDGA) and its tetraacetylated derivative (NDGATA) on the growth, oxygen consumption, adenosine 5'-triphosphate (ATP) level and viability of mouse mammary adenocarcinoma TA3 and its multiresistant variant TA3-MTX-R cell lines were determined. NDGA inhibited mitochondrial carbonyl cyanide m-chlorophenylhydrazone (CCCP)-stimulated oxygen consumption in mouse liver and tumor cells when glutamate plus malate or succinate was added as substrate. The effects were considerably weaker when respiration was supported by duroquinol, indicating that NDGA inhibited primarily mitochondrial electron flow located at some point before ubiquinone. Although NDGATA only inhibited the electron flow through complex I, it was more efficient and selective than NDGA because mouse liver mitochondria were significantly less sensitive to it than both tumor cell lines tested. NDGA and NDGATA inhibited mitochondrial ATP synthesis and, consequently, cell viability and ...

Research paper thumbnail of Preliminary Structural Characterization of Pacific Hagfish Immunoglobulin

Advances in Experimental Medicine and Biology, 1975

The immune capacities of primitive fishes of the class Agnatha, the lamprey and the hagfish, have... more The immune capacities of primitive fishes of the class Agnatha, the lamprey and the hagfish, have been under study for nearly a decade. Early results from Good and his collaborators suggested that lampreys, but not hagfish, possessed an adaptive immune response (1, 2). However, subsequent work of Linthicum and Hildemann (3) and Hildemann and Thoenes (4, 5) showed that hagfish (Eptatretus stoutii) could also respond specifically to certain immunogens and could reject allografts under appropriate conditions.

Research paper thumbnail of Síntesis y secreción del antígeno de superficie del virus de hepatitis B en cultivo de células animales

Research paper thumbnail of Allergens of the urushiol family promote mitochondrial dysfunction by inhibiting the electron transport at the level of cytochromes b and chemically modify cytochrome c1

Biological Research, 2021

BackgroundUrushiols are pro-electrophilic haptens that cause severe contact dermatitis mediated b... more BackgroundUrushiols are pro-electrophilic haptens that cause severe contact dermatitis mediated by CD8+effector T-cells and downregulated by CD4+T-cells. However, the molecular mechanism by which urushiols stimulate innate immunity in the initial stages of this allergic reaction is poorly understood. Here we explore the sub-cellular mechanisms by which urushiols initiate the allergic response.ResultsElectron microscopy observations of mouse ears exposed to litreol (3-n-pentadecyl-10-enyl-catechol]) showed keratinocytes containing swollen mitochondria with round electron-dense inclusion bodies in the matrix. Biochemical analyses of sub-mitochondrial fractions revealed an inhibitory effect of urushiols on electron flow through the mitochondrial respiratory chain, which requires both the aliphatic and catecholic moieties of these allergens. Moreover, urushiols extracted from poison ivy/oak (mixtures of 3-n-pentadecyl-8,11,13 enyl/3-n-heptadecyl-8,11 enyl catechol) exerted a higher inhi...

Research paper thumbnail of Immunodetection of acrosin during the acrosome reaction of hamster, guinea-pig and human spermatozoa

Biological research, 1992

Mammalian sperm acrosomes contain a trypsin-like protease called acrosin which causes limited and... more Mammalian sperm acrosomes contain a trypsin-like protease called acrosin which causes limited and specific hydrolysis of the extracellular matrix of the mammalian egg, the zona pellucida. Acrosin was localized on hamster, guinea-pig and human sperm using monoclonal and polyclonal antibodies to human acrosin labelled with colloidal gold. This was visualized directly with transmission electron microscopy, and with light and scanning microscopy after silver enhancement of the colloidal gold probe. Four distinct labelling patterns were found during capacitation and the acrosome reaction in hamster and guinea-pig spermatozoa, and three patterns were found in human spermatozoa. In the hamster, acrosin was not detected on the inner acrosomal surface after the completion of the acrosome reaction, thus correlating with the observation that hamster spermatozoa lose the ability to penetrate the zona after the acrosome reaction. With guinea-pig and human spermatozoa, acrosin was still detected ...

Research paper thumbnail of Electron Microscopy of the Natural IGM-like Hemagglutinin of the Ratfish (Callorhynchus callorhynchus)

Developmental & Comparative Immunology, 1981

Research paper thumbnail of Visible Light Anaerobic Photoconversion of Tyrosine Sensitized by Riboflavin. Cytotoxicity on Mouse Tumoral Cells

Photochemistry and Photobiology, 2008

The anaerobic phototransformation of tyrosine under visible light sensitized by riboflavin is rep... more The anaerobic phototransformation of tyrosine under visible light sensitized by riboflavin is reported. The cytotoxicity of the anaerobic photoproducts on in vitro-cultured myeloid mouse tumoral cells was demonstrated. A radical mechanism is proposed. Dityrosine was identified as one of the main anaerobic photoproducts by using absorption, emission and 'H-NMR spectra.

Research paper thumbnail of Apoptosis Induction in Nonirradiated Human HL-60 and Murine NSO/2 Tumor Cells by Photoproducts of lndole-3-acetic Acid and Riboflavin

Photochemistry and Photobiology, 1999

The effect of the photoproducts of indole-3-acetic acid sensitized by riboflavin on nonirradiated... more The effect of the photoproducts of indole-3-acetic acid sensitized by riboflavin on nonirradiated human HL-60 and murine NS0/2 tumor cells was studied. Severe damage with a dose-response effect was observed on both cell types. The effect was greater than that previously described for the tryptophan riboflavin photoproducts. Electron microscopy studies and flow cytometry analysis of DNA fragmentation allowed us to conclude that the photoproducts studied in this work induce cell death by an apoptotic mechanism.

Research paper thumbnail of Females of the communally breeding rodent, Octodon degus, transfer antibodies to their offspring during pregnancy and lactation

Journal of Reproductive Immunology, 2007

Females in numerous rodent species engage in communal nesting and breeding, meaning that they sha... more Females in numerous rodent species engage in communal nesting and breeding, meaning that they share a nest to rear their young together. One potential benefit to communally nesting mothers is that infants improve their immunocompetence. Thus, suckling from two or more females might provide newborns with a more diverse array of antibodies and defensive cells. As a first step toward testing the immunocompetence hypothesis, we assessed whether female degus (Octodon degus), a communally nesting and breeding caviomorph rodent, transfer immunoglobulins to their young through the yolk sac or placenta while in the uterus and, during lactation, through milk. With this aim, adult degu females were immunized with four antigens, including two mollusk hemocyanins from Concholepas and Megathura (CCH and KLH, respectively), porcine thyroglobulin and tetanus toxoid. Specific antibodies against the experimental antigens were used to track the origin of antibodies in the young. To establish the presence of specific antibodies of IgG and IgA isotypes in sera and milk of animals, an indirect enzyme-linked immunosorbent assay (ELISA) was developed. Degu females produced specific antibodies against antigens not found in their natural environment, and mothers were able to transfer the induced antibodies to their litters during pregnancy (IgG) and during lactation (IgA). However, we recorded only limited evidence of degu offspring acquiring antibodies from lactating mothers other than their own, giving little support to the increased immunocompetence hypothesis.

Research paper thumbnail of Procedure for Radiolabeling Gizzerosine and Basis for a Radioimmunoassay

Journal of Agricultural and Food Chemistry, 1999

A method for the labeling of gizzerosine (GZ), a biogenic amine found in fish meal, is described.... more A method for the labeling of gizzerosine (GZ), a biogenic amine found in fish meal, is described. The labeling procedure with 125 I using a water-soluble Bolton-Hunter reagent and a mild water-insoluble oxidant (Iodogen) reagent is rapid and reproducible. The 125 I-GZ hapten was demonstrated to be immunologically active in a radioimmunoassay developed with polyclonal antibodies to GZ absorbed with a histamine-Sepharose column. The curves were linear in the range of 0.0001 and 0.1 µg/mL. Samples of fish meal previously extracted of histamine with methanol and submitted to acid hydrolysis were contaminated with known amounts of GZ and submitted to the assay. The fish meal samples contaminated with GZ show a dose-response effect similar to the standard curve, and apparently the other component present in the sample did not interfere with the binding of the antibodies to 125 I-GZ. These data indicate the suitability of the radioimmunoassay to determine specifically GZ in fish meal.

Research paper thumbnail of The role of acrosin and antibodies to acrosin in gamete interactions

Journal of Reproductive Immunology, 1989

Research paper thumbnail of Sperm antigens relevant to human fertility : final report

Research paper thumbnail of Hemocyanin of the Molluscan Concholepas concholepas Exhibits an Unusual Heterodecameric Array of Subunits

Journal of Biological Chemistry, 2004

Research paper thumbnail of The oxygen-binding properties of hemocyanin from the mollusk Concholepas concholepas

Biochimica et biophysica acta, Dec 24, 2017

Hemocyanins have highly conserved copper-containing active sites that bind oxygen. However, struc... more Hemocyanins have highly conserved copper-containing active sites that bind oxygen. However, structural differences among the hemocyanins of various mollusks may affect their physicochemical properties. Here, we studied the oxygen-binding cooperativity and affinity of Concholepas concholepas hemocyanin (CCH) and its two isolated subunits over a wide range of temperatures and pH values. Considering the differences in the quaternary structures of CCH and keyhole limpet hemocyanin (KLH), we hypothesized that the heterodidecameric CCH has different oxygen-binding parameters than the homodidecameric KLH. A novel modification of the polarographic method was applied in which rat liver submitochondrial particles containing cytochrome c oxidase were introduced to totally deplete oxygen of the test solution using ascorbate as the electron donor. This method was both sensitive and reproducible. The results showed that CCH, like other hemocyanins, exhibits cooperativity, showing an inverse relat...

Research paper thumbnail of Product and Composition Containing a Concholepas Concholepas Hemocyanin (CCH) Subunit A, and a Method of Use Thereof

Research paper thumbnail of CD8p T Cells Are the Effectors of the Contact Dermatitis Induced by Urushiol in Mice and Are Regulated by CD4p T Cells

Int Arch Allergy Immunol, 1998

Research paper thumbnail of Novel hemocyanin from the Fissurella latimarginata exhibits an outstanding immunogenicity and non-specific immunomodulatory effects in a melanoma model

The Journal of Immunology, May 1, 2012

Research paper thumbnail of Product and composition containing a Concholepas concholepas hemocyanin (CCH) subunit, and a method of use thereof

Research paper thumbnail of Development of anti-human B blood group monoclonal antibodies suitable as a blood typing reagent

Hybridoma, 1994

An improved procedure for the generation of high-avidity anti-human B blood group monoclonal anti... more An improved procedure for the generation of high-avidity anti-human B blood group monoclonal antibodies (MAbs) was developed. One of them, termed 7A1-2, showed excellent qualities of titer, avidity, and intensity required for use as human B blood typing reagent. Hemagglutination inhibition studies with monosaccharides and oligosaccharides were carried out to determine the specificity of the MAb 7A1-2. These studies indicate that the antibody reacts with the immunodominant region of the antigen which is known to confer the serologic specificity of this blood group.

Research paper thumbnail of A Light-Induced Tryptophan-Riboflavin Binding: Biological Implications

Advances in Experimental Medicine and Biology, 1991

We review here the covalent photo-binding induced by visible light between the essential amino ac... more We review here the covalent photo-binding induced by visible light between the essential amino acid tryptophan and the vitamin riboflavin. We discuss the biological implications of this photoadduct in relation to the hepatotoxic and cytotoxic effect associated to parenteral nutrients and to culture media exposed to the action of light, respectively. We also analyze the formation of a photo-binding between riboflavin and the residues of tryptophan present in the proteins of the eye lens, a tissue which is permanently exposed to visible light.

Research paper thumbnail of Inhibitory effect of nordihydroguaiaretic acid and its tetra-acetylated derivative on respiration and growth of adenocarcinoma TA3 and its multiresistant variant TA3MTX-R

In vivo (Athens, Greece)

The effects of nordihydroguaiaretic acid (NDGA) and its tetraacetylated derivative (NDGATA) on th... more The effects of nordihydroguaiaretic acid (NDGA) and its tetraacetylated derivative (NDGATA) on the growth, oxygen consumption, adenosine 5'-triphosphate (ATP) level and viability of mouse mammary adenocarcinoma TA3 and its multiresistant variant TA3-MTX-R cell lines were determined. NDGA inhibited mitochondrial carbonyl cyanide m-chlorophenylhydrazone (CCCP)-stimulated oxygen consumption in mouse liver and tumor cells when glutamate plus malate or succinate was added as substrate. The effects were considerably weaker when respiration was supported by duroquinol, indicating that NDGA inhibited primarily mitochondrial electron flow located at some point before ubiquinone. Although NDGATA only inhibited the electron flow through complex I, it was more efficient and selective than NDGA because mouse liver mitochondria were significantly less sensitive to it than both tumor cell lines tested. NDGA and NDGATA inhibited mitochondrial ATP synthesis and, consequently, cell viability and ...